Comparison of 3 DNA extraction methods for extracting DNA from an adult Culex quinquefasciatus (Diptera: Culicidae).

The aim of this study was to compare 3 DNA extraction methods: the PureLink Genomic DNA kit, DNAzol Direct reagent, and a microwave-based method, for extracting DNA from an adult Culex quinquefasciatus by focusing on the quantity and purity of DNA, cost, and time required. Ten mosquitoes were individually used for DNA extraction by each method. Based on the results obtained, DNA was extracted from each method using specific primers, resulting in a polymerase chain reaction (PCR) product with a length of 274 bp. The DNA quantity extracted using the DNAzol Direct (179.08 ±â€…3.77 ng/µl) differs significantly from that of the commercial kit (115.98 ±â€…4.57 ng/µl) and a microwave-based method (119.26 ±â€…3.06 ng/µl). The absorbance ratio of DNA extracted using the PureLink Genomic DNA kit, the DNAzol Direct, and the microwave-based methods was 1.92 ±â€…0.02, 1.79 ±â€…0.01, and 1.87 ±â€…0.01, respectively. Among the 3 methods evaluated, the microwave-based method is simpler, less expensive, and more time efficient. This is the first evaluation of the microwave-based method for extracting DNA from an adult mosquito. This study provides a useful guide for alternative DNA extraction methods for PCR-based assays, especially in low-resource settings.

The Prevalence of Intestinal Protozoan Infections among Children in Southwest Sumba Based on the Type of Water Sources.

BACKGROUND: Intestinal protozoan infection is a public health problem in Indonesia, particularly in regions lacking clean water sources such as Perokonda and Perobatang villages, Southwest Sumba Regency, East Nusa Tenggara Province. The residents of Perokonda and Perobatang villages use wells and unprotected open springs respectively. Therefore, this study aims to determine the prevalence of intestinal protozoan infections in these two villages. MATERIALS AND METHODS: This was a cross-sectional study, carried out in Perokonda and Perobatang from August 2016 to August 2017. A total of 291 children aged 6 months to 18 years were used as subjects. Furthermore, fecal examinations were carried out by preparing smears, which were stained with lugol solution and examined under the microscopes by four experienced technicians. The data were processed using SPSS version 20 and analyzed by chi-square test. RESULTS: Among the 291 samples, the prevalence of intestinal protozoan infections was 20%, with 23.4% in Perokonda and 17.2% in Perobatang. The predominant species found were Blastocystis hominis (34.5%), Giardia lamblia (19.0%), and Entamoeba coli (15.5%). Moreover, approximately 69.0% of single infections were found while 31.0% were mixed and associated with 2 to 3 protozoan species. There were no significant differences between the prevalence of intestinal protozoan infections and the children in Perokonda and Perobatang villages (confidence interval 0.83 - 2.62, P = 0.19). CONCLUSION: The type of water sources has no relationship with the species of intestinal protozoan infecting children in Perokonda and Perobatang.

Wound Healing in Diabetic Foot Ulcer Patients Using Combined Use of Platelet Rich Fibrin and Hyaluronic Acid, Platelet Rich Fibrin and Placebo: An Open Label, Randomized Controlled Trial.

BACKGROUND: Autologous platelet-rich fibrin (A-PRF) is an adjunctive method for diabetic foot ulcer (DFU) in addition to glycaemic control and debridement. This study aimed to evaluate the role of A-PRF + hyaluronic acid (HA), A-PRF and sodium chloride 0.9% (control) in DFU wound healing. Nowaday, the use of  PRF autologous consider as adjuvant therapy in DFU treatment. METHODS: This open-label randomized controlled trial was conducted at Koja District Hospital and Gatot Soebroto Hospital from July 2019 to April 2020. DFU patients with wound duration of three months, Wagner-2, and ulcer size < 40 cm2 were recruited and randomly assigned into A-PRF + AH, A-PRF and control group. On day-0, day-3 and day -7, samples and photographs were taken. Samples were analysed with ELISA and photographs were analysed with ImageJ to calculate granulation index (GI). Statistical analysis was performed using SPSS version 20. RESULTS: Topical therapy with A-PRF + AH was associated with a significant increase in VEGF from day 0 (232.8 pg/mg) vs day 7 (544.5 pg/mg) compared to A-PRF on day 0 (185.7 pg/mg) vs day 7 (272.8 pg/mg), and the controls on day 0 (183.7 pg/mg) vs day 7 (167.4 pg/mg). On evaluation  of VEGF swab, there is increasing significantly in A-PRF+HA group compare others group in day -3 ( p=0.022) and day -7 (p= 0.001).In the A-PRF + AH group, there was a significant decrease in IL-6 from day 0 (106.4 pg/mg) vs day 7 (88.7 pg/mg) compared with PRF on day 0 (91.9 pg/mg) vs day 7 (48,8 pg/mg). IL-6 was increased in the control group from day 0 (125.3 pg/mg) vs day 7 (167.9 pg/mg). On evaluation  of IL-6 swab, there is decreasing significantly in A-PRF+HA group compare others group in day -7 (p= 0.041). CONCLUSION: The PRF + HA combination increased angiogenesis and reduced inflammation in DFUs and may represent a new DFU therapy.

The role of VEGF, PDGF and IL-6 on diabetic foot ulcer after Platelet Rich Fibrin + hyaluronic therapy.

BACKGROUND: Current standard management of diabetic foot ulcers (DFUs) consists of surgical debridement followed by soak NaCl 0.9% gauzes tight infection and glycaemic control. Nowadays the use of advanced platelet-rich fibrin (A-PRF) has emerged as an adjunctive method for treating DFUs. This study was conducted to demonstrate the ability of combine A-PRF + HA as a complementary therapy in DFUs healing related with angiogenesis,inflammation and granulation index process. METHODS: This open label randomized controlled trial was conducted in Koja District Hospital and Gatot Soebroto Hospital Jakarta, Indonesia on July 2019-April 2020. DFUs patients with wound duration of three months, Wagner-2, with size of ulcer less than 40 cm2 were included in the study. The number of subjects was calculated based on the rule of thumb and allocated randomly into three groups, namely topical A-PRF + HA, A-PRF and Sodium Chloride 0.9% as a control, for each of 10 subjects. A-PRF made by 10 mL venous blood, centrifuge 200 G in 10 min, meanwhile A-PRF + HA though mix both them with vertex machine around 5 min. Biomarker such as VEGF, PDGF and IL-6 examined from DFU taken by cotton swab and analysis using ELISA. Granulation Index was measured using ImageJ. Biomarkers and granulation index were evaluated on day 0, 3, 7 and 14. Data were analysed using SPSS version 20 with Anova and Kruskal Wallis test to compare the angiogenesis and inflammation effect between the three groups. RESULT: In topical dressing A-PRF + HA, there is an increase in delta VEGF on day-3 (43.1 pg/mg protein) and day-7 (275,8 pg/mg protein) compared to A-PRF on day-3 (1.8 pg/mg protein) and day-7 (104.7 pg/mg protein), also NaCl (control) on day-3 (-4.9 pg/mg protein) and day-7 (28.3 pg/mg protein). So that the delta VEGF of A-PRF + HA group increase significantly compared with others on day-3 (p = 0.003) and day- 7 (p < 0.001). Meanwhile A-PRF + AH group, there is also a decrease in delta IL-6 after therapy on day-3 (-10.9 pg/mg protein) and day-7 (-18.3 pg/mg protein) compared to A-PRF in delta IL-6 on day- 3 (-3.7 pg/mg protein) and on day-7 (-7.8 pg/mg protein). In NaCl (control) group there is a increase delta IL-6 on day-3 (4.3 pg/mg protein) and on day-7 (35.5 pg/mg protein). So that the delta IL-6 of A-PRF + HA group decrease significantly compared with others only on day- 7 (p = 0.015). In PDGF le level analysis, A-PRF + HA group increase significantly (p = 0.012) only in day -7 compare with other group (5.5 pg/mg protein). CONCLUSION: The study shows the superior role of combined A-PRF + HA in the treatment DFU though increase angiogenesis and decrease inflammation pathway. The advantage of using A-PRF + HA is that it accelerates wound healing by increasing granulation tissue compared to A-PRF alone.

Molecular typing and antifungal susceptibility study of Aspergillus spp. in intensive care unit (ICU) patients in Indonesia.

INTRODUCTION: Aspergillus exhibits a wide variation of susceptibility against antifungals according to genetic and environmental factors. Identification to the species level is necessary for appropriate treatment. Our objective was to determine the Aspergillus species involved in invasive pulmonary aspergillosis (IPA) among ICU patients in Jakarta, Indonesia. METHODOLOGY: The incidence of IPA in ICU patients at six hospitals in Jakarta from October 2012 - January 2015 was investigated. It involved a collection of endotracheal aspirates (ETA), nasal swabs and environmental samples around the hospitals, phenotypic screening, molecular characterization, and antifungal susceptibility testing. RESULTS: Of the 405 patients investigated, 31 patients (7.7%) were diagnosed with putative IPA, from whom 45 Aspergillus isolates were collected. Aspergillus isolates were identified from pulmonary secretions in 24 patients, from nasal swabs in 7 patients and from both pulmonary secretions and nasal swabs in 7 patients. The phenotypic method showed 33 isolates of Aspergillus flavus (73.4%), nine Aspergillus fumigatus (20%), two Aspergillus niger (4.4%), and one Aspergillus nidulans (2.2%) isolate. Molecular identification showed 27 isolates of A. flavus (60.0%), eight isolates of A. fumigatus (17.8%), two isolates of A. niger (4.4%) and one isolate of A. nidulans (2.2%), while seven isolates (15.6%) were cryptic species or mixed isolates. CONCLUSIONS: Susceptibility testing showed all isolates were susceptible to amphotericin B, azoles and micafungin. Aspergillus flavus was the main causative organism in IPA cases in Jakarta, followed by A. fumigatus.

Protein intake and number of children associated with nutritional status.

OBJECTIVE: This study aimed to evaluate the association between protein intake and number of children in a family, based on nutritional status of children aged 2 to 12 years of age. METHODS: This was a cross-sectional study with consecutive sampling. Weight and height were measured, and the 24-hour food recall was gathered with a questionnaire. Nutritional status was assessed using weight-for-age curve of Centers for Disease Control and Prevention (CDC) growth chart. Protein intake was assessed using NutriSurvey. Data were analysed with chi-square test. RESULTS: There were 99 children, 52 girls and 47 boys. From these, 57 (57.6%) were undernourished and 42 (42.4%) subjects had a good nutritional status. Sixty-five (65.6%) children had poor protein intake, while 34 (34.3%) had adequate protein intake. There was no association between protein intake and nutritional status (p = 0.805) or number of children in the family and nutritional status (p = 0.414). The principal protein source was squid, but squid is often sold for additional income. CONCLUSIONS: The nutritional status of children in Pero Konda Village could be considered undernourished. There was no association between protein intake and number of children based on nutritional status. The villagers had poor dietary habits, so good dietary habits should be promoted.

Effect of melatonin supplementation in combination with neoadjuvant chemotherapy to miR-210 and CD44 expression and clinical response improvement in locally advanced oral squamous cell carcinoma: a randomized controlled trial.

BACKGROUND: Squamous cell carcinoma of the oral cavity (OSCC) is the sixth most common malignancy. Surgery is mainstay treatment for oral cancers. Surgery in locally advanced OSCC presents many challenges primarily because the head and neck have critical structures that can be damaged by tumor or treatment. It is thought that neoadjuvant chemotherapy (NC) in locally advanced OSCC is able to shrink tumor size. Chemoresistancy is a problem due to hypoxic microenvironment characterized by increased expression of HIF-1α. It is also regulated by miR-210 as well as increased expression of CD44 and CD133. Melatonin has a powerful antioxidant and oncostatic effects that are expected to improve tumor hypoxia and clinical response. Fifty patients with OSCC were included and randomized. miR-210 and CD44 expression were measured before and after intervention using qRT-PCR absolute quantification, and clinical response was evaluated according to RECIST 1.1 criteria. This study aims to determine the effect of melatonin in improving the clinical response of patients with locally advanced oral squamous cell carcinoma (OSCC) after neoadjuvant chemotherapy to miR-210 and CD44 expression. RESULTS: Melatonin administration reduced miR-210 levels but not significant (p = 0.767). CD44 expression also decreased in the melatonin group compared with placebo yet was not significant (p = 0.103). There was a decrease in the expression of miR-210 and CD44 followed by a decrease in the percentage of residual tumor but not significant (p = 0.114). CONCLUSION: In OSCC, the addition of 20-mg melatonin to neoadjuvant chemotherapy (NC) reduced the expression of miR-210 and CD44 and decreased the percentage of tumor residue; however, no statistically significant result was observed. TRIAL REGISTRATION: This study is registered to ClinicalTrials.gov under trial registration number: NCT04137627 with date of registration on October 22, 2019-retrospectively registered, accessible from: https://clinicaltrials.gov/ct2/show/NCT04137627.

Characterization and Synergistic Antimicrobial Evaluation of Lipopeptides from Bacillus amyloliquefaciens Isolated from Oil-Contaminated Soil.

Lipopeptides show great potential for biomedical application. Several lipopeptides exhibit narrow and broad-spectrum inhibition activities. The aim of the study is to characterize the lipopeptides produced by B. amyloliquefaciens strain MD4-12 and evaluate the synergistic antimicrobial activity in combination with a conventional antibiotic against Gram-negative bacteria. B. amyloliquefaciens strain MD4-12 was isolated from oil-contaminated soil. The isolate was cultivated in McKeen medium, and the lipopeptides were isolated by precipitation and extraction with methanol. Characterization of the lipopeptides by ESI-MS gave nine mass ion peaks with m/z 994-1072, resulted from protonating of the main ions in [M + H]+ and [M + Na]+ ion form. These mass ion peaks attributed to surfactin homologs. By tandem mass spectrometry, five variants of surfactin with the same amino acid sequence in peptide moiety could be revealed. The peptide moiety contains seven amino acids identified as Glu-Leu/Ile-Leu-Val-Asp-Leu-Leu/Ile while the fatty acid moiety comprises a different length of chain from C12 to C16. Surfactin showed antibacterial activity against various Gram-positive and Gram-negative bacteria. Combination surfactin with ampicillin showed a synergistic effect against P. aeruginosa ATCC 27853.

The Effectiveness of Triple Dose Albendazole in Treating Soil Transmitted Helminths Infection.

Soil transmitted helminths (STH) infection is a major health problem in tropical countries such as Indonesia. Albendazole is an effective and widely used anthelmintic agent to treat STH; however, it is not effective towards T. trichiura and its effectiveness varies between populations. Hence, we conducted a study to determine the effectiveness of triple dose albendazole in children of Perobatang village, Southwest Sumba, Indonesia. A pre-post study was carried out in Perobatang village on July 2016. Children aged 1-15 years old were enrolled in the study and asked to collect stool samples which were then examined using Kato-Katz method. The children infected with STH were given albendazole 400 mg for three consecutive days. From 246 subjects examined, 192 (78%) were positive for any STH consisting of T. trichiura (64%), A. lumbricoides (60%), and hookworms (10%). After treatment, the prevalence of STH decreased significantly (McNemar test, p<0.001) to 27%, T. trichiura 25%, A. lumbricoides 2%, and hookworm 0%. Cure rate for T. trichiura, A. lumbricoides, and hookworms was 61%, 97%, and 100%, respectively. Significant decrease of eggs per gram of feces was found in all STH (Wilcoxon test, p value <0.001 for A. lumbricoides and T. trichiura, p value = 0.027 for hookworms); egg reduction rate for T. trichiura was 91%, A. lumbricoides was 100%, and hookworms was 100%. In conclusion, triple dose albendazole is effective in controlling STH in children of Perobatang village, Southwest Sumba, Indonesia.

A novel diagnosis scoring model to predict invasive pulmonary aspergillosis in the intensive care unit.

OBJECTIVES: To improve the quality of invasive pulmonary aspergillosis (IPA) management for intensive care unit (ICU) patients using a practical diagnostic scoring model. METHODS: This nested case-control study aimed to determine the incidence of IPA in 405 ICU patients, between July 2012 and June 2014, at 6 hospitals in Jakarta, Indonesia. Phenotypic identifications and galactomannan (GM) tests of sera and lung excreta were performed in mycology laboratory, Parasitology Department, Faculty of Medicine, Universitas Indonesia in Jakarta, Indonesia. RESULTS: The incidence of IPA in the ICUs was 7.7% (31 of 405 patients). A scoring model used for IPA diagnosis showed 4 variables as the most potential risk factors: lung excreta GM index (score 2), solid organ malignancy (score 2), pulmonary tuberculosis (score 2), and systemic corticosteroids (score 1). Patients were included in a high-risk group if their score was greater than 2, and in a low-risk group if their score was less than 2. CONCLUSION: This study provides a novel diagnosis scoring model to predict IPA in ICU patients. Using this model, a more rapid diagnosis and treatment of IPA may be possible. The application of the diagnosis scoring should be preceded by specified pre-requisites.

Universal health coverage in Indonesia: concept, progress, and challenges.

Indonesia is a rapidly growing middle-income country with 262 million inhabitants from more than 300 ethnic and 730 language groups spread over 17 744 islands, and presents unique challenges for health systems and universal health coverage (UHC). From 1960 to 2001, the centralised health system of Indonesia made gains as medical care infrastructure grew from virtually no primary health centres to 20 900 centres. Life expectancy improved from 48 to 69 years, infant mortality decreased from 76 deaths per 1000 livebirths to 23 per 1000, and the total fertility rate decreased from 5·61 to 2·11. However, gains across the country were starkly uneven with major health gaps, such as the stagnant maternal mortality of around 300 deaths per 100 000 livebirths, and minimal change in neonatal mortality. The centralised one size fits all approach did not address the complexity and diversity in population density and dispersion across islands, diets, diseases, local living styles, health beliefs, human development, and community participation. Decentralisation of governance to 354 districts in 2001, and currently 514 districts, further increased health system heterogeneity and exacerbated equity gaps. The novel UHC system introduced in 2014 focused on accommodating diversity with flexible and adaptive implementation features and quick evidence-driven decisions based on changing needs. The UHC system grew rapidly and covers 203 million people, the largest single-payer scheme in the world, and has improved health equity and service access. With early success, challenges have emerged, such as the so-called missing-middle group, a term used to designate the smaller number of people who have enrolled in UHC in wealth quintiles Q2-Q3 than in other quintiles, and the low UHC coverage of children from birth to age 4 years. Moreover, high costs for non-communicable diseases warrant new features for prevention and promotion of healthy lifestyles, and investment in a robust integrated digital health-information system for front-line health workers is crucial for impact and sustainability. This Review describes the innovative UHC initiative of Indonesia along with the future roadmap required to meet sustainable development goals by 2030.

TOAST Subtypes of Ischemic Stroke and Its Risk Factors: A Hospital-Based Study at Cipto Mangunkusumo Hospital, Indonesia.

BACKGROUND AND PURPOSE: Stroke is a leading cause of death and disability, with ischemic stroke as the highest prevalent cases in Indonesia. Ischemic stroke can be classified further into five subtypes according to TOAST classification. Numerous studies have revealed that stroke risk factor has variable correlation with different stroke subtype. Currently, there is no data regarding this phenomenon in Indonesia. The aim of study is to identify characteristic of ischemic stroke subtypes and the risk factors in TOAST classification. METHODS: A retrospective, cross-sectional study of patients diagnosed with ischemic stroke at Cipto Mangunkusumo Hospital from January till December 2016. Demographic data, ischemic stroke subtypes, risk factors, and other relevant data were documented. Bivariate and multivariate analysis was done using SPSS 23. RESULTS: 235 recorded data patients were included. Large artery atherosclerosis (LAA) was the most prevalent stroke subtypes at 59,6%, followed with small vessel disease (SVD) at 26,7%, undetermined etiology at 9,8%, cardioembolism (CE) at 2,1%, and other determined etiology at 0,9%. Hypertension was the most common vascular risk factor. However, it was only significant in SVD (p=0,023) and undetermined etiology subtypes (p<0,001). Significant risk factor in LAA was diabetes (55%; p=0,016) while in CE subtypes was atrial fibrillation (60%;p<0,001). In multivariate analyses, hypertension (OR 3; 95% CI 1,12-8,05) was the only variable that was related to SVD while in CE it was atrial fibrillation (OR 113,5; 95% CI 13,6-946,5). CONCLUSION: LAA was the most common stroke ischemic subtypes. Associated risk factor in LAA was diabetes while in SVD and undetermined etiology subtypes it was hypertension. Atrial fibrillation was associated with cardioembolism.

Post-Ocular Trauma Corneal Staphyloma in a Child Living in an Underdeveloped Region of Eastern Indonesia.

Ocular trauma occurring in children often leads to visual impairment or blindness when it is not properly managed. This often occurs in underdeveloped regions with difficult access to professional care at local health facilities. We report a case of post-trauma corneal staphyloma in an underdeveloped region of eastern Indonesia to illustrate the importance of proper management of ocular trauma in areas lacking such expertise and where patients have difficulty accessing even basic health care. During a community health outreach in Southwest Sumba, eastern Indonesia in May 2017, a 7-year-old boy presented with white protrusion of the left eye of 3 years' duration following an event of sharp trauma. The patient lived in an inaccessible and impoverished area. Upon contemporary examination, visual acuity of the left eye was 1/300 while that of the right eye was 6/6. Anterior examination revealed corneal staphyloma of the left eye, and its posterior segment could not be evaluated. The patient was transported to an eye care center in Jakarta, receiving a stock prosthesis implant while awaiting a corneal evisceration procedure with dermofat graft. Ocular trauma is one of the most common causes of visual impairment in children. This case illustrates the consequences of inadequate post-trauma management and the importance of prevention of infection.

The Effect of Deworming Using Triple-Dose Albendazole on Nutritional Status of Children in Perobatang Village, Southwest Sumba, Indonesia.

High prevalence of STH leads to malnutrition, anemia, cognitive impairment, and growth disorders. Triple-dose albendazole 400 mg is a broad-spectrum anthelminthic; however, its effectiveness varies in every region. This study aims to determine the benefits of deworming using triple-dose albendazole on children's nutritional status in Perobatang Village, Southwest Sumba, Indonesia. This pre-post study was conducted in July 2016 and January 2017. Children aged 1-15 years were asked to collect stool for diagnosis of STH infection (Kato-Katz method), were measured for anthropometry status to obtain the nutritional status, and took albendazole 400 mg for three consecutive days. Data was analyzed with SPSS version 20. Prevalence of STH prior to the treatment was 95.4%: T. trichiura 85.2%, A. lumbricoides 71.6%, and hookworm 18.2%. After treatment, prevalence of STH decreased significantly (McNemar test, p < 0.001) to 53.4%, (T. trichiura 39.8%, A. lumbricoides 22.7%, and hookworm 1.1%). Before treatment, 33% participants were in normal nutritional status, 47.7% underweight, and 19.3% severely underweight. After treatment, children in normal nutritional status increased to 75%, underweight children decreased to 25%, and there were no severely underweight children. In conclusion, deworming with triple-dose albendazole 400 mg is effective in improving the nutritional status of children in Perobatang Village.

Heavy burden of intestinal parasite infections in Kalena Rongo village, a rural area in South West Sumba, eastern part of Indonesia: a cross sectional study.

BACKGROUND: Intestinal parasitic infections (IPIs) are one of the major public health problems, especially in the rural area of developing countries with low socio-economic status and poor sanitation. The study was aimed to determine the prevalence of IPIs among the inhabitants of a rural area in South West Sumba, eastern part of Indonesia. METHODS: A cross-sectional study was done in Kalena Rongo village, South West Sumba in April 2014. Stool samples were collected and examined for IPIs using direct smear method. RESULTS: Faecal samples were collected from 424 of 473 inhabitants of the village, age 2 months to 80 years. About 95.5 % (405/424) of the participants had any IPIs. The most prevalent parasites found were Ascaris lumbricoides 65.8 % (279/424), Trichuris trichiura 60.4 % (256/424), hookworms 53.5 % (227/424), Blastocystis hominis 34.4 % (146/424), Entamoeba histolytica 17.9 % (76/424), and Giardia lamblia 4.5 % (19/424). The villagers used no latrine and defecated in their backyard. Clean water sources were scarce and far from the village. CONCLUSIONS: In Kalena Rongo village, the rural area in eastern part of Indonesia, the finding of IPIs was conspicuous and therefore expressed the poor hygiene and absence of proper sanitation in the area. Integrated efforts, such as improving infrastructure to provide clean water source and educating the inhabitants for appropriate hygienic lifestyle are needed.