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Recent research has revealed that Plasmodium falciparum (P. falciparum) possesses an Artemisinin-based Combination Therapy (ACT) resistance mechanism. To address this issue, new antimalarial chemicals are required. In vitro, in vivo, and in silico tests have been conducted on metabolite extracts of Streptomyces hygroscopicus subsp. hygroscopicus (S. hygroscopicus) and showed antimalarial effects through various mechanisms. The aim of this research was to determine the effectiveness and cytotoxicity of active fractions 38K, 38T, and 56 of S. hygroscopicus secondary metabolites as antimalarial. Fractionation procedure was performed using flash column chromatography BUCHI Reveleris® PREP Purification System. Effectiveness test was conducted on Plasmodium berghei (P. berghei) culture by parasite inhibition counts. The cytotoxicity effect of S. hygroscopicus was observed utilizing the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide Assay (MTT Assay) on Michigan Cancer Foundation-7 (MCF-7) breast cancer cell culture. Inhibition Concentration 50% (IC50) and Cytotoxicity Concentration 50% (CC50) was obtained from probit analysis. S. hygroscopicus fractions 38K, 38T, and 56 showed antimalarial activity against P. berghei, with fraction 38T having the strongest activity as antimalarial from selectivity index. Fraction 38T had the IC50 with the value of 11.66 µg/mL which included in the promising activity classification of IC50. Almost all asexual stages of parasite morphology were damaged by the fractions. MCF-7, a cell line used to study antimalarial cytotoxicity, was unaffected by any of the three fractions due to its CC50 exceeding 50 μg/mL. S. hygroscopicus fractions 38K, 38T, and 56 of the metabolite extracts contain non-toxic compounds that can damage the morphology of intra-erythrocytic stage and inhibit the growth of P. berghei in vitro. [ABSTRACT FROM AUTHOR]

Keloid is connective skin tissue overgrowth resulting from abnormal wound healing activated by transforming growth factor-β (TGF-β) through particularly TGF-β/Smad signaling pathway which translocate Smad2/3 into nucleus to induce type I collagen synthesis and normally regulated by inhibitory Smad7. The expression of Smad2/3 are increased while Smad7 is decreased in keloid fibroblast. The keloid fibroblast phosphorylated p38 (p-p38) increased and inhibited Smad7 expression. Those biomolecular findings cause keloid recurrence. Therefore, studies on keloid recurrence prevention must be developed. Quercetin is a flavonoid found in many fruits and vegetables. Previous studies showed that quercetin inhibited p-p38 which provided opportunity increasing Smad7, decreasing nuclear Smad2/3, and preventing keloid recurrence. This study aimed to determine the quercetin effect on p-p38, Smad7, Smad2/3, and collagen type I levels in human keloid fibroblasts. Four replicates three-passages primary human keloid fibroblast culture incubated with 5, 10, 20 µg/mL quercetin were conducted with culture media as control for 48 hours. The expressions of p-p38, Smad7, and Smad2/3 were measured using western blotting. The level of collagen type I was measured using enzyme-linked immunosorbent assay (ELISA). Mean comparisons between multiple groups were analyzed using one-way analysis of variance (ANOVA) or Kruskal-Wallis test. Mean comparison between two groups were analyzed using independent T-test or Mann-Whitney test. The results showed that 10 µg/mL quercetin group showed lower p-p38 expression than that of control group (P < 0.05). The 10 and 20 µg/mL quercetin groups showed lower Smad7 expression than that of control (P < 0.05). The 20 µg/mL quercetin group showed lower nuclear Smad2/3 expression than that of control (P < 0.05). All quercetin groups demonstrated lower nuclear Smad2/3 compared to cytoplasmic Smad2/3 expression (P < 0.05), while control group did not show nuclear and cytoplasmic Smad2/3 expression significant difference. This study confirmed that quercetin could regulate keloid fibroblast TGF-β/Smad pathway by inhibiting p-p38 and Smad2/3 nuclear translocation and could be used, as biomolecular basis, for keloid prevention in wound healing process. [ABSTRACT FROM AUTHOR]

Lipopolysaccharide (LPS) and lipooligosaccharide (LOS) are inflammatory response inducers triggering downstream inflammatory signals by binding to the TLR4-MD2 complex. LPS/LOS exposure through TLR4 receptor on dendritic cells (DC) causes an increase in interferon (IFN)-β expression via an increase in interferon regulatory factor (IRF)3 expression and a decrease in nuclear factor (NF)-κβ, which result in HIV replication inhibition. This study aimed to compare the binding affinity between Escherichia coli LPS and Neisseria gonorrhoeae LOS toward TLR4-MD2 in silico and the production of NF-κβ, IRF3, and IFN-β from naive-HIV monocyte-derived dendric cells (MDDCs) after LPS and LOS exposure in vitro, to find a deeper understanding of host immune response, disease pathogenesis, and trends of drug development in the future. Molecular docking and dynamics were analyzed using AutoDock Vina and YASARA, respectively. The naive-HIV DC culture was exposed to LPS at 50, 100, and 200 ng/mL, or LOS at concentrations of 2.5, 5, and 10% for 24-h. Levels of NF-κβ, IRF3, and IFN- β were measured by Enzyme-linked Immunosorbent Assay (ELISA). The results showed that LOS demonstrated higher binding affinity to TLR4-MD2 complex than LPS, although the docking between LOS and TLR4-MD2 complex involved fewer amino acids. Inversely, LPS exposure significantly increased IRF3 and IFN-β production (P < 0.01). Production of IFN-β was significantly increased with higher doses of LOS (P < 0.01). Different doses of LPS induced significant differences in NF-κβ and IFN-β levels (P < 0.01 and P < 0.05, respectively). Despite LOS showing higher binding affinity to TLR4- MD2 complex, LPS exposure induced higher production of IRF3 and IFN-β from MDDCs. These findings provided information for deeper apprehension of immune responses and disease pathogenesis during HIV co-infections. [ABSTRACT FROM AUTHOR]

The high mortality and morbidity rates of malaria remains to be a global problem, particularly in tropical regions. Cerebral malaria, the leading cause of death, possesses a complex pathogenesis involving host immune and parasite interactions. Lately, according to the gut-brain axis concept, the gut microbiota was identified to protect the host against systemic pathogen infection through the modulation of immune signaling pathways. In this study, we aimed to explore the effect of probiotic supplementation toward neurological outcomes of cerebral malaria in vivo. Twenty-eight male C57BL/6 mice were randomly divided into 5 groups after 2 weeks acclimatization period. All groups except the negative control were given oral probiotic supplementation with either Lactobacillus casei at 109 colony forming unit/100 µL (CFU/100 µL), Bifidobacterium longum at 109 CFU/100 µL, or the combination of each at 5x108 CFU/100 µL for five days before to six days after Plasmodium berghei infection. Neurological manifestations were recorded by using the SmithKline, Harwell, Imperial College, Royal Hospital, Phenotype Assessment (SHIRPA) score. The data was analyzed by using One-way ANOVA and post-hoc LSD. The results showed that there were significant differences of motor behavior (P < 0.01), muscle tone (P < 0.01), reflex and sensory (P < 0.05), and neuropsychiatric state (P < 0.05) between the control group and that receiving combination of L. casei and B. longum. The results suggested that probiotic supplementation showed promising effects in improving the neurological manifestations during cerebral malaria. Further studies are required to explicate the pathways involved in the immunological interplay. In the future, it is propitious that the mortalities by cerebral malaria could be mitigated through the optimal use of probiotics. [ABSTRACT FROM AUTHOR]

Malaria is a disease caused by Plasmodium infection. Of the 5 species that infect humans, Plasmodium vivax and Plasmodium falciparum are the two most common species found in Indonesia. Early and accurate diagnosis is crucial for effective treatment and disease management. Conventional diagnostic methods, microscopy, and rapid diagnostic tests (RDTs) have limitations in sensitivity and specificity. PCR-based assays have undergone significant enhancement such as the development of touchdown PCR and primer designing. However, the use of PCR-based assays in Plasmodium vivax infections still has several limitations including the formation of dimers and cross reactivity. Two isolated Plasmodium vivax DNA samples taken from malaria confirmed patients and stored in the Biomedical Laboratory, Faculty of Medicine, Universitas Brawijaya were tested with both standard Plasmodium vivax (rViv) primers and new (Pv-Rv) primer for Plasmodium vivax 18SSU RNA gene detection. Each pairs of primers underwent both regular nest 2-PCR and touchdown nest 2-PCR. The PCR products were checked by 3% agarose gel electrophoresis, and then quantified by using GelDoc Imaging System and ImageJ software. The paired t-test was used to analyze the results statistically. The results showed that, using rViv primers, there was no statistically significant difference in the ratio of band intensity to background intensity between regular nest 2-PCR and touchdown nest 2-PCR (P > 0.05). However, when the background intensity was analyzed, a statistically significant difference was discovered (P < 0.05). By using Pv-Rv primers, a significant difference in the background intensity as well as in the ratio of band intensity to background intensity between regular nest 2-PCR and touchdown nest 2-PCR was observed (P < 0.05). The results indicated that application of touchdown nest 2-PCR had an effect in reducing background noise by using either rViv primers (standard primer) or Pv-Rv primers (new primer). Further research and modification of Pv-Rv primers were required for Plasmodium vivax identification. [ABSTRACT FROM AUTHOR]

Background: Toxoplasma gondii is one of the zoonotic protozoa parasites. It can prevalently infect humans and warmblooded animals, causing human health problems and substantial economic losses to the livestock industry worldwide. Chicken is one of the potential sources of toxoplasmosis, but there is no report of the prevalence of toxoplasmosis and their genotypes in free-range chickens in Libya. Aim: This study aims to conduct a survey of molecular prevalence and identify the T. gondii genotype in free-range chickens and its association with the risk factors of age, gender, and region in Northeastern Libya. Methods: This study was conducted by examining a total of 315 free-range chicken organs (brain and heart) derived from three administrative districts in Northeastern Libya. The molecular prevalence was determined by PCR technique using B1 gene amplification. and the T. gondii genotype was determined by nested PCR-RFLP of GRA6 gene amplicon with restriction enzymes (MseI). Results: The overall molecular prevalence of T. gondii in free-range chicken in all three districts was 9.5% (30/315), and the highest (15.4%) was in the Al-Marj district (p = 0.01; x² = 9.238). The highest prevalence of T. gondii by age was in chickens aged more than 2 years (p = 0.001; x² = 15.530). The difference in T. gondii prevalence in male and female chickens was not significant (p = 0.372; x² = 0.798). The predominant genotype I (93.3%) had identified at position 544 and 194 bp at the GRA6 marker, and only two positives were from genotype II (6.7%) at 700 and 100 bp fragments. Conclusion: The molecular prevalence of toxoplasmosis in free-range chicken in three districts in Northeastern Libya was 9.5%, and the highest rate was shown in the Al Marj district. Chicken by age more than 2 years had more risk to transmit toxoplasmosis in human. There was no different infection risk by consuming male or female free-range chicken. It is the first report to determine the predominant genotype, which was genotype I. [ABSTRACT FROM AUTHOR]

This study aimed to obtain a microbial active compound as a novel antimalarial drug from Indonesian isolates. Target-based assays were used to screen for antimalarial activity against the parasite mitochondrial, Plasmodium falciparum malate:quinone oxidoreductase (PfMQO) enzyme. In total, 1600 crude extracts, composed from 800 fungi and 800 actinomycetes extracts, were screened against PfMQO, yielding six active extracts as primary hits. After several stages of stability tests, one extract produced by Aspergillus sp. BioMCC f.T.8501 demonstrated stable PfMQO inhibitory activity. Several purification stages, including OCC, TLC, and HPLC, were performed to obtain bioactive compounds from this active extract. All purification steps were followed by an assay against PfMQO. We identified the active compound as nornidulin based on its LC-MS and UV spectrum data. Nornidulin inhibited PfMQO activity at IC50 of 51 µM and P. falciparum 3D7 proliferation in vitro at IC50 of 44.6 µM, however, it had no effect on the growth of several mammalian cells. In conclusion, we isolated nornidulin from Indonesian Aspergillus sp. BioMCC f.T.8501 as a novel inhibitor of PfMQO, which showed inhibitory activity against the proliferation of P. falciparum 3D7 in vitro. [ABSTRACT FROM AUTHOR]

The high frequency of patient visits to the ER will disrupt the service process at the Emergency Department (IGD). One solution is the application of a triage system, which is the process of sorting patients according to their level of gravity. The accuracy of triage decisions has a significant impact on patient outcomes so that in this case nurses in the ER must have knowledge, understanding, and skills about triage. Analyze personal factors that influence emergency nurse decision-making. This study aimed to identify personal factors influencing triage decision-making among emergency nurses. Respondents were 30 male and female nurses who worked and were active up to now in the IGD Hospital of the University of Muhammadiyah Malang. Data on work experience, education, and training were taken from data on the characteristics of respondents. The knowledge questionnaire came from the Emergency nurses association/ENA while the skills questionnaire used the Triage Skill Questionnaire/TSQ. The Spearman correlation test shows that there is a significant relationship between skills and education with triage decision making (p=0.000; r=0.626 and p=0.039; r=0.378). Meanwhile, the variables of knowledge, work experience, and training do not correlate with triage decision making (p> 0.05) except that GELS training shows a significant relationship between GELS training and triage decision making (p=0.016; r=0.437). The results of linear regression analysis, skills, and education are factors that influence decision-making in conducting triage in the ER (45,4%). Skills and education are the factors that most influence triage decision-making. [ABSTRACT FROM AUTHOR]

Background: In Drug-Resistant Tuberculosis (DR-TB), high IL17 level was corelated with severe tissue damage while low level was associated with mortality. High levels of IL-17 and IL - 23 were found in patients with a high bacterial load. TB treatment is carried out by combination of anti-tuberculosis drugs for a period of 9 to 24 months. This study aimed to analyze the differences in IL-17 and IL-23 levels before and after treatment compared to healthy controls, and their relationship with conversion AFB smear. Methods: A prospective cohort study was conducted with total 38 subjects diagnosed as DR-TB using a molecular rapid test and 34 normal subjects as case and control groups. Examination of sputum AFB, IL-17 and IL-23 levels were carried out at the start of treatment and 30 days after treatment. Treatment was given in short term and individul drugs resistant regiments. Results: The average levels of IL-17 and IL 23 before treatment compared to those after treatment showed no significant difference (p=0.153; p= 0.328). When compare to healthy controls group, the mean levels of IL-17 and IL 23 before treatment was not significantly different (p= 0.883; p= 0.928), whereas after 1 month treatment were significantly different (p= 0.047; p=0.042). An increase of IL-17 levels followed by an increase in IL-23 levels (p=0,000 r=.0.872). TB treatment affected the conversion rate of 97.4% with linear association. Conclusion: The levels of IL-17 and IL-23 after treatment were increasing and significantly different with healthy group. These related with the high AFB conversion rate. [ABSTRACT FROM AUTHOR]

Background: Malaria screening for blood derived from any donors prior to transfusions is a standard procedure that should be performed; but, in fact, it is not routinely conducted. In case of the blood is infected with Plasmodium spp., the survival of parasites may be depending on, or even influencing, the profile of red blood cells (RBCs). Methods: This observational longitudinal study was conducted upon 55 bags of donor blood that randomly selected. Malaria infections were detected using Rapid Diagnostic Test/RDT with thin and thick blood smear confirmation. The changes of Plasmodium spp. viability and RBCs profiles, as well as other hematological parameters, were observed from the results of routine hematological examinations which were performed on days 1,7,14 and 21 of storage. Results: Among 55 blood samples, there were 17 and 38 bags, respectively, positive and negative for malaria, then used for analysis as the case and control groups. There were significant decreasing values (p< 0.05) of all routine blood examination parameters of donor blood, started from days 1, 7, 14, 21, and 28. There were no differences in decreasing profiles between those infected and non-infected donor blood (p> 0.05). On days 21 and 28 none of the positive samples still contained parasites. Conclusion: Erythrocytes profiles of donor blood significantly decreased with the duration of storage, but were not influenced by the presence of Plasmodium spp. [ABSTRACT FROM AUTHOR]

Introduction: Previous research has proven that ethyl acetate extract of Streptomyces hygroscopicus can inhibit Plasmodium growth in vivo and in vitro through inhibition of the Proteasome ubiquitin system. In this study, we analyzed the activity of several fractions of S. hygroscopicus toward the growth of P. falciparum culture with specific targets on the mitochondrial enzyme of P. falciparum; L-malate: quinone oxidoreductase (PfMQO) and dihydroorotate dehydrogenase (PfDHODH). Methodology: the fractionation process uses the Flash Column Chromography (FCC) BUCHI Reveleris PREP, analyzed using thin layer chromatography (TLC). The antimalarial activity of the fractions was tested on the culture of P. falciparum 3D7 using enzyme lactate dehydrogenase (PfLDH) assay and their activity toward PfMQO and PfDHODH target enzymes. The fractions analyzed using HPLC and compared to the standard dihydroeponemycin. The toxicity of the fractions, the WST-8 assay was conducted. Results: of the 30 fractions produced, 6 fractions were able to suppress the P. falciparum and two of them were able to inhibit more than 50% of P. falciparum growth. Both of these fractions were proven to inhibit the activity of the PfMQO enzyme but had no inhibition of the PfDHODH enzyme. Both fractions reduced human cell viability by less than 50% in toxicity test. HPLC analysis indicated that besides dehydroeponemycin, there were other compounds presences in the extract. Conclusions: fractions of S. hygroscopicus metabolite contains non-toxic compounds in addition to dihydroeponemycin that could suppress the growth of P. falciparum in vitro. This potential effect might be related to the inhibition of PfMQO enzyme. [ABSTRACT FROM AUTHOR]

This study aimed to reveal the role of ICAM-1 and HIF-1α in brain endothelial cell of cerebral malaria mice model after exposure to 2.4 absolute atmospheres (ATA) hyperbaric oxygen (HBO). Thirty-nine C57BL/6 mice were divided into three groups: control negative (normal mice without any exposure), control positive (Plasmodium berghei ANKA [PbA] infection without HBO exposure), and treatment (PbA infection and exposed to HBO for 10 sessions after the parasite grew). Parasitemia and clinical symptoms were observed every day. Brain tissues were isolated on day 13 post-infection for histopathological and immunohistochemical examination (observed at 400x magnification in 10 visual fields). HBO decreased HIF-1α and ICAM-1 expression in endothelial cells. There was a moderate correlation between HIF-1α and ICAM-1 expression. Ten HBO sessions prevented cerebral malaria, as denoted by the decreased expression of ICAM-1 and HIF-1α in brain vascular endothelial cells from the experimental mice. [ABSTRACT FROM AUTHOR]