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“…It was vortexed and re-centrifuged for 30 s at 12,000 rpm, and the supernatant was obtained. The supernatant from this process produced nucleotide extraction results and was stored at −80 °C before performing PCR analysis [ 22 ].…”
Section: Methodsmentioning
confidence: 99%
“…This protocol is optimized for the CFX Connect System (USA) real-time PCR machine instrument. The protocol was adjusted with the instrument by changing the dye dilution according to the manual instructions and following the manufacturer's recommended instrument for the RT-PCR cycle program [ 22 , 23 ].…”
“…It was vortexed and re-centrifuged for 30 s at 12,000 rpm, and the supernatant was obtained. The supernatant from this process produced nucleotide extraction results and was stored at −80 °C before performing PCR analysis [ 22 ].…”
Section: Methodsmentioning
confidence: 99%
“…This protocol is optimized for the CFX Connect System (USA) real-time PCR machine instrument. The protocol was adjusted with the instrument by changing the dye dilution according to the manual instructions and following the manufacturer's recommended instrument for the RT-PCR cycle program [ 22 , 23 ].…”
“…The identification of genetic factors that affect the natural risk regarding infectious disease can give new insight about the defence mechanisms towards infectious disease. The resistance or the vulnerability to intracellular pathogenic infection like Salmonella and Mycobacterium are controlled by the Natural Resistance associated Macrophage Protein (NRAMP 1) located in chromosome 1, inside macrophage [ 17 , 18 ].…”
Background
Miana (
Coleus Scutellariodes
[L] Benth) inhibits growth of bacterial pathogen inside macrophage.
Objective
The aim of this study is to determine the protein level of Natural Resistance Associated Macrophage Protein 1 (NRAMP1), after administration of Miana extracts in BALB/c mice induced
Klebsiella pneumoniae
.
Methods
This is an experimental study using animal model with post test-only controlled group design. Twenty healthy adult male BALB/c mice were randomly divided into four groups, negative control group (distilled water), Levofloxacin 100 mg/kg, injection intraperitoneal, first treatment group (Miana leaves extract/MLE 510 mg/kg) and second treatment group (Miana + levofloxacin). MLE were administered via gastric gavage for ten consecutive days. The blood was drawn from each mice on the first day, on the eight day of experiment (2 h after treatment), and at 10 days. The blood sample was examined by ELISA to determine the NRAMP1 protein level. Analysis of the number of lung tissue bacteria used Plate count agar to see the growth of
Klebsiella pneumonia
.
Results
NRAMP1 protein level in BALB/c mice after administration of Miana extract was increased significantly in after 10 days treatment (p < 0.0001). The highest increasing in protein levels was found in treatment group (Miana + levofloxacin) with an increase before treatment 3036,07 to 10010,30 pg/ml after treatment p < 0,0001.
Conclusion
NRAMP1 protein level in BALB/c mice were highest increasing in protein levels after administration of Miana extract and Levofloxacin compared Miana or Levofloxacin only and clinical impact proved a comparable effect on suppressing
Klebsiella pneumoniae
growth.
The institutional protocol number of this study is 1010/UN4.6.4.5.31/PP36/2019.
“…In addition, the NRAMP-1 gene functions as a metal ion transporter that regulates cellular levels which may limit intracellular pathogen replication by changing the phagolisosomal environment [15,16,17]. Recently have been reported the association of NRAMP-1 in several infectious disease such as typhoid fever, tuberculosis, leprosy, buruli ulcer [18,19,20].…”
Section: Introductionmentioning
confidence: 99%
“…There is an urgent need for adequate and efficient detection methods for the establishment of acute recurrence state of typhoid fever, perhaps based on host susceptibility of NRAMP-1 gene [18]. Due to the important role of NRAMP1 in immune response against S. typhi infection and the high incidence of recurrent acute in typhoid fever, research related to host susceptibility is needed.…”
Background and Aim: The molecular pathogenesis of typhoid fever is complex and the host susceptibility mechanisms such as Natural Resistance Associated Macrophages Protein-1 (NRAMP-1) are poorly understood. This study explores the expression of NRAMP-1 in the serum of Acute Recurrence State (ARS) of typhoid fever compared with typhoid fever patients and healthy persons.
Methods: Thirty of ARS of typhoid fever and 30 typhoid fever patients were collect from several Primary Health Care and Hospitals in the endemic area. Diagnosis of typhoid fever based on clinical symptoms and confirmed by blood culture. Healthy persons were collect from the Blood Transfusion Unit, Makassar, Indonesia. The expression of NRAMP-1 was determined using Enzyme-linked immunosorbent assay (ELISA).
Results: The mean of NRAMP-1 expression in 30 ARS of typhoid fever and 30 typhoid fever were found 10.941,56 pg/mL and 11.027,65 pg/mL, respectively. However, the mean of NRAMP-1 expression in healthy persons was found 21.103,91 pg/mL.
Conclusion: No different the expression of NRAMP-1 in ARS of typhoid fever compared with typhoid fever patients. However, expression of NRAMP-1 in both ARS of typhoid fever and typhoid fever showed significantly lower than healthy persons. Future study is needed to explore the other molecular factors involved to become ARS.
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