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Detection of Knockdown-Resistance Mutations (V1016G and F1534C) in Dengue Vector from Urban Park, Surabaya, Indonesia Shifa Fauziyah; Sri Subekti; Budi Utomo; Teguh Hari Sucipto; Hebert Adrianto; Aryati Aryati; Puspa Wardhani; Soegeng Soegijanto
Journal of Tropical Biodiversity and Biotechnology Vol 6, No 3 (2021): December
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jtbb.65357

Abstract

An urban park is potentially a source of vector-borne disease transmission due to it being a natural and artificial mosquito breeding habitats combined with people's continuous presence. Thus, this study aims to screen the occurrence of knockdown-resistance (kdr) mutant alleles (V1016G and F1534C) in mosquito populations collected from urban parks in Surabaya, Indonesia. Cross sectional study was conducted in July 2019. A total of 28 ovitraps were installed in seven urban parks, having four ovitraps installed in each park. In total, 1,662 eggs were collected, and only 187 emerged into adult mosquitoes, consisting of 97 Aedes (Stegomyia) aegypti and 90 Aedes (Stegomyia) albopictus. All-female adult mosquitoes (n=55) were tested using allele-specific polymerase chain reaction assay (AS-PCR) to detect voltage gated sodium channel (VGSC) gene mutations. This study found no mutations in Valine to Glysine mutation in point 1016 (V1016G) and Phenylalanine to Cysteine in point 1534 (F1534C) alleles in both two species. All of mosquito samples have wild type genotype of kdr alleles (V1016V and F1534F). Data were analysed using R Studio 1.4 Version by Genetics package. Results showed that the frequency of resistant alleles (G1016 and C1534) was zero, and the frequency of susceptible allele was 1 (V1016 and F1534). Insecticide bioassay could not be established due to the limited number of adult mosquitoes, so insecticide resistance status could not be determined. However, this study can be used as preliminary monitoring for the vector control program.
RNA Isolation of Dengue Virus Type 2 with Different Precipitation Solvents : Methanol, Chloroform, and 2-Isopropanol. Yovilianda Maulitiva Untoro; Teguh Hari Sucipto; Harsasi Setyawati; Siti Churrotin; Ilham Harlan Amarullah; Puspa Wardhani; Aryati Aryati; Shuhai Ueda; Soegeng Soegijanto
Jurnal Kimia Riset Vol. 3 No. 1 (2018): Juni
Publisher : Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (416.739 KB) | DOI: 10.20473/jkr.v3i1.7455

Abstract

Dengue virus distributed in tropical and subtropical regions in the world. DENV viruses are transmitted between humans primarily by Aedes aegypti and Aedes albopictus mosquitoes and are endemic in most areas in which the vectors occur. Four serotypes of dengue virus are DENV-1, DENV-2, DENV-3 and DENV-4. DENV-2 is comprised of six genotypes. Ethanol precipitation is a commonly used technique for concentrating and de-salting nucleic acids (DNA or RNA) preparations in aqueous solution. RNA isolation by combining Guanidinium thiocyanate and phenol reported has been reported. In this report, we investigated RNA isolation from DENV-2 using QIAamp Mini Kit with 2-Isopropanol, Methanol, Chloroform precipitation solvent. Electrophoregram showed DNA band as  the result of RNA isolation with methanol and 2-isopropanol are produced quite well. Dna band of the of RNA isolation with chloroform solvent has the lowest intensity than methanol and 2-isopropanol. This study showed that methanol and 2-isopropanol  can used as precipitation solvent for isolating RNA.
Precipitation Solvents for RNA Extraction of Dengue Virus Type 3: Dimethylformamide, Ethylenediamintetraacetic Acid, and Ultrapure H2O Rizqidhana Juliana Putri; Teguh Hari Sucipto; Harsasi Setyawati; Siti Churrotin; Ilham Harlan Amarullah; Puspa Wardhani; Aryati Aryati; Soegeng Soegijanto
Jurnal Kimia Riset Vol. 3 No. 2 (2018): Desember
Publisher : Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (477.838 KB) | DOI: 10.20473/jkr.v3i2.9353

Abstract

Dengue is a disease caused by a virus from the family Flaviviradae, carried by a female mosquito of Aedes aegypti species. Dengue fever is widespread in the tropic areas. It caused by rainfall, temperature and unplanned urbanization. According to the ministry of health , almost all provinces in Indonesia are endemic areas of dengue fever. In 2014, up to mid-December Dengue Hemorrhagic Fever (DHF) patients in 34 provinces in Indonesia are 71,668 people and 641. This figure is lower than the previous year, 2013 with 112,511 people and 871 deaths . This disease consists of four types of serotypes, namely DENV-1, DENV-2, DENV-3, and DENV-4. This disease can be identified using a variety of methods, one of the method is Reverse Transcription - Polymerase Chain Reaction (RT-PCR) method. This study aims to determine the ability of Dimethylformamide (DMF), Ethylenediamintetraacetic Acid (EDTA), and Ultrapure H2O as the substitute of  Ethanol for precipitation in RNA extraction process. The sample used in this research obtained from Surabaya. RNA extraction itself can be done by using a special kit for RNA extraction. In Reverse Transcription - Polymerase Chain Reaction method, first RNA is extracted and then transcribed back (Reverse Transcription) which then form cDNA that later will be amplified by using PCR method. In this study used specific primers for dengue virus type 3 (DENV-3). The results of this study show that DMF, EDTA, and Ultrapure H2O can be used as the substitute of Ethanol for precipitation on RNA extraction. The result is evidenced by the formation of viral DNA bands on gel electrophoresis results.
Comparison of pregnancy rates on day 3 and day 5 embryo transfer in In Vitro Fertilization (IVF) Audia Mumtaz Rifasky; Puspa Wardhani; Ashon Sa’adi; Ninik Darsini; Hamdani Lunardhi; Zakiyatul Faizah
Majalah Obstetri dan Ginekologi Vol. 29 No. 1 (2021): April
Publisher : Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20473/mog.V29I12021.14-17

Abstract

Objectives: To identify the success rates of pregnancy on the third and fifth day embryo transfer at Graha Amerta Hospital, Surabaya, Indonesia.Materials and Methods: This study used comparative cross sectional design. Data were taken from medical record of IVF participants who met the inclusion and exclusion criteria at Graha Amerta Hospital for the period of January 2016 - December 2016.Results: Successful pregnancy rates were found in this research. The embryo transfer on the third day and the fifth day were 35% and 49.3% respectively. In other words, the rates of pregnancy success were not affected by embryo transfer on the third day and the fifth day in the medical record sample as it had p value of 0.090.Conclusion: Embryo transfer on the third and fifth days had the same rates of pregnancy success in IVF participants at Graha Amerta Hospital, Surabaya, Indonesia. 
Pengaruh Rendaman Air Terhadap Kualitas DNA pada Sperma dengan STR-CODIS D13S317 dan D21S1 Abdul Hadi Furqoni; Ahmad Yudianto; Puspa Wardhani
Jurnal Biosains Pascasarjana Vol. 19 No. 1 (2017): JURNAL BIOSAINS PASCASARJANA
Publisher : Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (984.572 KB) | DOI: 10.20473/jbp.v19i1.2017.41-54

Abstract

AbstrakKriminalitas yang terjadi akibat kejahatan seksual banyak terjadi di mana-mana. Kejahatan ini bisa menimpa beberapa orang dan dari berbagai umur.  Dalam kasus ini pasti akan di temukan barang bukti di tempat kejadian perkara. Salah satu bukti akibat kejahatanseksual adalah bercak sperma. Bercak sperma dapat di temukan seperti pada pakaian yang digunakan korban atau pelaku. Dengan keterbatasan waktu yang dimiliki oleh polisi tentunyaperlu  cara  untuk  pengidentifikasi  siapa  pelaku  dari  kejahatan  tersebut.  Sampel  bercak sperma akan di isolasi DNA.   Isolasi DNA adalah memisahkan DNA yang ada pada sel sperma dengan komponen yang lainnya. Setelah DNA di dapatkan akan di ketahui kadar dankemurniannya. Hasil tersebut bisa di pengaruhi oleh media bercak sperma berada. Faktor- faktor tersebut salah satunya media air dan kain katun. Sifat kain katun adalah mempunyaikemampuan menyerap yang tinggi walaupun dalam keadaan basah sekalipun.  Setelah isolasi DNA, tahapan selanjutnya adalah amplifikasi. Proses amplifikasi dalam penelitian ini adalah menggunakaan STR dengan lokus D13S317 dan D21S11.Kata kunci : Kriminal, Sperma, DNA, STR-CODIS
INHIBITORY ACTIVITY OF COBALT(II)–MORIN COMPLEX AGAINST THE REPLICATION OF DENGUE VIRUS TYPE 2 Teguh Hari Sucipto; Siti Churrotin; Harsasi Setyawati Setyawati; Kris Cahyo Mulyatno; Ilham Harlan Amarullah; Shuhai Ueda; Tomohiro Kotaki; Sri Sumarsih; Puspa Wardhani; Sri Subekti Bendryman; Aryati Aryati; Soegeng Soegijanto; Masanori Kameoka
Indonesian Journal of Tropical and Infectious Disease Vol. 6 No. 6 (2017)
Publisher : Institute of Topical Disease Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (402.971 KB) | DOI: 10.20473/ijtid.v6i6.6126

Abstract

Dengue virus (DENV) is a significant pathogen emerging worldwide as a cause of infectious disease. Antidengue treatments are urgently required to control the emergence of dengue. DENV is a mosquito-borne disease responsible for acute systemic diseases and serious health conditions. DENVs were distributed in the tropical and sub-tropical areas and transmitted to humans by Aedes agypty and Aedes albopictus. Dengue vaccine or antiviral has not yet been clinically approved for humans, even though there have been great efforts toward this end. Antiviral activity against DENV is an important alternative for the characterization and development of drugs. Metal–organic compounds were reported to exhibit fungicidal, bactericidal, and antiviral activities its inhibitory activity was not significant, at high concentration it was more toxic to replicating cells than to stationary cell monolayers of Vero cells. The aim of this study is to investigate the antiviral effects of Cobalt(II)–Morin complex. This compound was further investigated for its inhibitory effect on the replication of DENV-2 in Vero cells. The replication of DENV was measured by enzyme-linked immunosorbent assay and the value of selectivity index (SI). SI was determined as the ratio of the 50% cytotoxic concentration (CC50) to the 50% inhibitory concentration (IC50). The IC50 value of the Cobalt(II)–Morin complex for DENV-2 was 3.08 µg/ml, and the CC50 value of the complex for Vero cells was 3.36 µg/ml; thus, the SI value was 1.09. The results of this study demonstrate the antidengue serotype 2 inhibitory activity of Cobalt(II)–Morin complex and its high toxicity in Vero cells. Further studies are not required before Co(II)–Morin can be applied in the treatment of DENV-2 infections.
ANTI-DENGUE TYPE 2 VIRUS ACTIVITIES OF ZINC (II) COMPLEX COMPOUNDS WITH 2-(2,4 -DIHYDROXYPHENYL)-3,5,7-TRIHYDROXYCROMEN-4-ONE LIGANDS IN VERO CELLS Teguh Hari Sucipto; Harsasi Setyawati; Siti Churrotin; Ilham Harlan Amarullah; Sri Sumarsih; Puspa Wardhani; Aryati Aryati; Soegeng Soegijanto
Indonesian Journal of Tropical and Infectious Disease Vol. 7 No. 5 (2019)
Publisher : Institute of Topical Disease Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (777.006 KB) | DOI: 10.20473/ijtid.v7i5.10851

Abstract

Dengue virus (DENV) is a disease that is transmitted through Aedes aegypti and Aedes albopictus mosquitoes, and is spread in tropical and sub-tropical regions. Now, dengue or antiviral vaccines for humans do not yet exist, but there are great efforts to achieve this goal. Complex compounds are reported to fungicidal, bactericidal and antiviral activity. Antiviral activity against DENV is an important alternative to the characterization and development of drugs candidate. The purpose of this study was to study zinc(II) compounds with 2-(2,4-dihydroxyphenyl)-3,5,7-trihydroxycromen-4-one ligand on DENV-2 replication in Vero cells. Vero cell lines (African green monkey kidney) was used in this study, maintained and propagated in Minimum Essential Eagle Medium containing 10% fetal bovine serum at 37°C in 5% CO2. The activity of dengue virus was carried out by enzyme-immunosorbent assay (ELISA) method and CellTiter96® Non-Radioactive Proliferation. The value of activity inhibition (IC50) of complex compounds with variations of mol metal: ligand 1:2, 1:3, and 1:4 against dengue virus type 2 (DENV2) was 2.44 μg/ml, 2.75 μg/ml, respectively and 2.00 μg/ml, also the toxicity value (CC50) of complex compounds with variation mol metal: ligand 1:4 for Vero cells is 3.59 μg/ml. The results of this study were indicate that these properties have been shown to inhibit anti-dengue type 2 virus (DENV-2), but are also toxic in Vero cells. Including previous study about complex compound interaction with dengue virus type 2 activity, Zn(II) more reactive compound then Cu(II), and Co(II). The comparison with Cu(II) complex compound, it has been revealed that Co(II) and Zn(II) is more toxic, was found to be nontoxic to human erythrocyte cells even at a concentration of 500 μg/ml.
Antimalarial Activity of Ethanol Extract of Kelakai Leaves (Stenochlaena palustris) to Parasitemia and Splenomegaly in BALB/c Mice Infected with Plasmodium berghei ANKA Laily Nur Azizah; Puspa Wardhani; Heny Arwati
JUXTA: Jurnal Ilmiah Mahasiswa Kedokteran Universitas Airlangga Vol. 13 No. 1 (2022): Jurnal Ilmiah Mahasiswa Kedokteran Universitas Airlangga
Publisher : Faculty of Medicine Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20473/juxta.V13I12022.1-5

Abstract

Introduction: Malaria is one of global health problems. Splenomegaly is one of malaria symptoms. Antimalarial drug resistance had been reported. Alternative treatment is by using traditional medicinal plants such as kelakai (Stenochlaena palustris). Kelakai contains alkaloid and flavonoid which had been reported to have antimalarial activity. The aim of this study was to discover antimalarial activity of ethanol extract of kelakai leaves to parasitemia and splenomegaly of Plasmodium berghei ANKA in infected BALB/c mice.Methods: This study was based on a modified Peter test using BALB/c mice infected with P. berghei ANKA treated with ethanol extract of kelakai leaves, with chloroquine diphosphate as a positive control. The negative control was P. berghei ANKA infected mice without any additional treatment. Administration of ethanol extract of kelakai leaves was performed for 4 days with a serial doses of 100, 10, and 1 mg/kg body weight. The positive control was given chloroquine diphosphate 20 mg/kg body weight. Parasitemia was observed daily prior to the calculation of the percentage of parasite growth and parasite growth inhibition. At the end of the test, the mice were sacrificed and spleens were isolated to measure their sizes. Probit analysis was performed to obtain ED50 to find the effect of extract in parasite killing by 50%. Spearman test was performed to analyze the correlation of doses of extract and splenomegaly.Results: Parasitemia growth inhibition was directly proportional to the dose. Higher parasitemia inhibition was obtained at higher doses and vice versa. Result of probit analysis showed an ED50 was 77.05 mg/kg body weight. Statistical analysis resulted in insignificant correlation between doses and splenomegaly p = 1.0 (significancy < 0.05).Conclusion: Ethanol extract of kelakai leaves possessed good antimalarial activity and there was no correlation between extract doses and splenomegaly in Plasmodium berghei ANKA-infected mice.
Genetic Diversity of Plasmodium falciparum Glutamate Rich Protein in Patients Attending the Merauke Hospital in Papua Province, Indonesia Thomas Tandi Manu; Puspa Wardhani; Heny Arwati; Aryati Aryati
INDONESIAN JOURNAL OF CLINICAL PATHOLOGY AND MEDICAL LABORATORY Vol 27, No 2 (2021)
Publisher : Indonesian Association of Clinical Pathologist and Medical laboratory

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24293/ijcpml.v27i2.1662

Abstract

Malaria remains an important health problem in Indonesia with the highest transmission in Papua Province, an easternpart of this country. The genetic diversity of malaria parasites is the main problem in understanding several aspects ofmalaria infections and the dynamics of their transmission, which also play a role in the development of a vaccine.Plasmodium falciparum is the deadliest of the human malaria parasites. Plasmodium falciparum glutamate-rich protein(Pfglurp) is one of the many erythrocytic stages antigens currently under development for a vaccine. The Pfglurp gene hasbeen extensively used as a marker to investigate the genetic diversity, Multiplicity of Infection (MOI), the level of malariatransmission, immunity against malaria, as well as a discriminatory instrument to distinguish new from recrudescentinfections of the field parasite population. Thus, this genotyping study aimed to find out the genetic population ofP.falciparum at the Merauke District, Province of Papua, Indonesia. DNA samples were isolated from Dried Blood Spots(DBS) obtained from P.falciparum infected patients in the Regional Public Hospital of Merauke, Province of Papua, Indonesiaduring May 2019-July 2019. The isolated DNAs were then amplified for nested Polymerase Chain Reaction (PCR) prior toPfglurp genotyping. The glurp gene was identified in all 51 DBS samples of P.falciparum-infected patients, and 18 variants ofallele were found. Among them, 45.10% were found to bear multigenotype infections. The size of the dominant allele(12.5%) was 701-750 bp. The MOI was 1.58. The genetic population of P.falciparum in Merauke Hospital has contained ahigher percentage of multigenotypes compared with monogenotypes indicating the high transmission of malaria in thestudied area.
Description of Fecal Culture Results in Diarrhea Patients Due To Antibiotic Use Suci Tresna; I.G.A.A Putri Sri Rejeki; Puspa Wardhani
INDONESIAN JOURNAL OF CLINICAL PATHOLOGY AND MEDICAL LABORATORY Vol 26, No 2 (2020)
Publisher : Indonesian Association of Clinical Pathologist and Medical laboratory

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24293/ijcpml.v26i2.1448

Abstract

Diarrhea infection is common in developing countries and causes death of around 3 million people every year. Diarrheais also the second leading cause of death in infants. Riskesdas in 2013 showed 30,775 cases of diarrhea. Causes were such asbacterial infections Salmonella, Shigella, Vibrio, Entamoeba, and Yersinia. Other influences can occur due to viral andfungal infections. Diarrhea is a nosocomial infection that is common in hospitalized patients due to the long-term use ofantibiotics caused by Clostridium difficile. This study was a follow-up study of diarrhea patients who received antibiotictherapy for more than two days with the results of C.difficile negative toxin, then continued with fecal culture examination.This study aimed to look at the description causes of diarrhea other than C.difficile in patients who received long-termantibiotic therapy. This research is an observational study. Samples were taken from 30 diarrhea patients with 2 x 24 hours ofantibiotic use who were hospitalized in the ICU, Dr. Soetomo Hospital Surabaya from August 2017 to May 2018. Sampleswith negative C.difficiletoxin results were then followed by fecal culture examination using conventional methods. Theresults of culture examination from 30 samples showed three samples with positive culture results extended-spectrumβ lactamase producing E.coli, two samples positive culture just E.coli, and 25 other samples showed negative culture results.The results of the fecal culture examination showed a description of causes of diarrhea in patients who received antibiotictherapy was pathogenic E.coli (ESBL). The possibility of other causes that cannot be detected from the culture such as viraland fungal infections, still requires further research.