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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]

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]

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]