Prawiro, Sumarno Reto
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The Effect of Low Power Ultrasonic Wave Exposure to Suppress Methicillin-Resistant Staphylococcus aureus (MRSA) In Vitro Mansyur, Mas; Yudaningtyas, Erni; Prawiro, Sumarno Reto; Widjajanto, Edi
Journal of Tropical Life Science Vol 8, No 2 (2018)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.08.02.07

Abstract

The incidence of methicillin-resistant Staphylococcus aureus (MRSA) infection keeps increasing in every part of the world. Currently, the infection prevalence of MRSA has reached 70% in Asia. In Indonesia in 2006 the prevalence was 23.5%; the infection prevalence of MRSA in RS Atmajaya Jakarta reached 47%, in RSUP Dr. Moh. Husin Palembang reached 46%, and RSUD Abdul Moeloek Lampung in 2013 reached 38.4%. MRSA is multiresistant to antibiotics and is hard to kill compared to most other negative gram bacteria. The purpose of this research is to find the lethal power and exposure of ultrasonic waves to kill MRSA, monitoring its ef-fects via changes in shape, size, structure and Gram staining as indicators. The observations were done mac-roscopically by culturing the MRSA in a petri dish filled with Chromagar MRSA medium, while the morpho-logical observations of MRSA were done by SEM, changes in the structure using TEM, and changes in the color of MRSA cells using Gram staining. Ultrasonic wave exposure, at a lethal power = 8.432 watt, killed a significant percentage of MRSA over the control (p = 0.000). The death indicators of the MRSA due to expo-sure to ultrasonic waves of various power were: changes in shape of MRSA affected by ultrasonic power (p = 0.005), changes in size is not affected by ultrasonic power (p= 0.470), the stain of MRSA cell staining from purple to pink affected by ultrasonic power (p = 0.000), all compared with the control. MRSA died due to ne-crosis, with physical evidence of the MRSA death such as mechanical stress marked by swollen MRSA cell, shift cell wall, crack and tears, cavitation marked by pieces of MRSA cell in the field of view due to explosions inside the cell, change to an irregular cell shape, and changes in color from black to transparent.
Cross Reaction among Antibody Pili sub unit Hemagglutinin Proteins and Outer Membrane sub unit Hemagglutinin Proteins of Shigella flexneri Fitrianingsih, Avin Ainur; Rachma, Lailia Nur; Milliana, Alvi; Hernowati, Tinny Endang; Aulanni'am, Aulanni'am; Santoso, Sanarto; Prawiro, Sumarno Reto
Journal of Tropical Life Science Vol 7, No 1 (2017)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.07.01.01

Abstract

Shigella  flexneri is the most common causal agent of shigellosis. Its pili are composed of pili protein subunits. Adhesion molecules can be found on the pili and outer membrane proteins (Omp). A hemagglutination reaction can be used for screening of adhesion molecules. Objectives: The purpose of this study was to determine the molecular weight of the pili protein subunits and outer membrane proteins of S. flexneri that act as hemagglutinin proteins, and to prove whether there is a cross-reaction between antibodies against hemagglutinin pili protein subunits and outer membrane proteins of S. flexneri. Methods: Pili protein subunits were isolated using pili bacteria cutters, and the outer membrane proteins were solubilized and obtained using sodium dodecyl sulfate 0.05% as detergent for Omp isolation.  The hemagglutination reaction used mice erythrocytes. The cross reactions between subunit pili proteins were conducted by Western blot and Dot blot. Results:. Antibodies against hemagglutinin sub unit pili protein 18 kDa responded to pili protein subunits 18 kDa; 23 kDa; 34 kDa; and 53 kDa; and Omp 23 kDa and 27 kDa. Omp and subunit pili proteins S. flexneri consists of several identical epitopes  that were responsible for the similarity of the response profile in the cross-reactions of antibodies. 
Group A β-hemolytic Streptococcus Detection Using Anti-Outer Membrane Protein (OMP) Immunoglobulin G (IgG) Alluza, Hamid Hunaif Dhofi; Mayasari, Ema Dianita; Prawiro, Sumarno Reto; Winarsih, Sri
Journal of Tropical Life Science Vol 7, No 1 (2017)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.07.01.02

Abstract

Streptococcal pharyngitis sequel such as Rheumatic Fever (RF) or Rheumatic Heart Disease (RHD) is an autoimmune response mediated by T cells and IgG. Since it is an autoimmune process, the result of bacterial culture as the gold standard of diagnosis often shows negative results. IgG against the 33 kDa OMP is considered as an important mediator in the process of these autoimmune diseases, so its presence in blood serum can be used as a diagnostic tool. The purpose of this study is to prove that 33 kDa OMP is one of the immunogenic parts of the Streptococcus Group A β-hemolytic, so it is expected that IgG anti-33 kDa OMP can recognize and respond the bacteria and to support the probability of the Streptococcus Group A β-hemolytic infection. This study was a laboratory experimental study with a control group design. Animal used was RattusNovergicus immunized with whole cell bacteria or 33 kDa OMP mixed with Complete Freund’s Adjuvant or Incomplete Freund’s Adjuvant. Polyclonal IgG was obtained by drawing blood serum from the animals after immunization with Streptococcus Group A β-hemolytic for 4 weeks (A; n = 5) and 8 weeks (B; n = 5) or immunization with OMP 33 kDa for 4 weeks (C; n = 5) and 8 weeks (D; n = 5) and also negative control group (E; n = 5). Immunological tests were done using Dot Blot assay, ELISA, and immunocytochemical examination. The data obtained was then evaluated with statistical tests Kruskal-Wallis, Mann-Whitney and Repeated ANOVA (p < 0.05). The result showed that there was a difference in humoral immune response (IgG) between the groups albeit the difference was not significant (p > 0.05). Dot Blot and immunocytochemical tests indicated that IgG anti-33 kDa OMP were able to recognize and respond the Streptococcus Group A β-hemolytic antigen. This study concluded that 33 kDa OMP was the immunogenic part of the bacteria and that IgG anti-33 kDa OMP could recognize and respond the Streptococcus Group A β-hemolytic bacteria.Â