Pengembangan Prototipe Rapid Test (ICT-S typhl) Berbasis Protein Adhesi Membran Luar Dan Fimbria Salmonella typhl Dari Penderita Demam Tifoid

Berdasarkan hasil penelitian ini dapat disimpulkan bahwa: 1. Seluruh area koding gen ompC dan fimH Salmonella typhi dapat diisolasi dengan teknik PCR. 2. Seluruh area koding gen ompC dan fimH Salmonella typhi yang terdapat pada plasmid rekombinan OmpC dan FimH dari penderits demam tifod dapat diakses di genbank dengan accession number AL627274, AE014613.1, Berdasarkan hasil penelitian ini dapat disimpulkan bahwa: 1. Seluruh area koding gen ompC dan fimH Salmonella typhi dapat diisolasi dengan teknik PCR. 2. Seluruh area koding gen ompC dan fimH Salmonella typhi yang terdapat pada plasmid rekombinan OmpC dan FimH dari penderits demam tifod dapat diakses di genbank dengan accession number AL627274, AE014613.1, M31424.1,FJ009676.1untuk nukleotida gen ompC dan AE014613.1 dan AL627267.1 untuk gen fimH. 3. Homologi sekuen gen ompC dari penderita demam tifoid adalah 99% dengan accession number AL627274, AE014613.1, M31424.1,FJ009676.1 . Sekuen seluruh area koding gen fimH dari penderits demam tifoid mempunyai homologi 94% dengan sekuen accession number AE014613.1 dan AL627267.1. 4. Protein rekombinan OmpC dan FimH yang dikode oleh plasmid rekombinan pOmpC dan pFimH dari penderita demam tifoid dapat diekspresikan dalam E.coli DH10{3 dengan induksi IPTG sehingga dapat menghasilkan protein rekombinan OmpC dan FimH yang dapat digunakan sebagai biomaterial untuk pengembangan diagnosis demam tifoid.31424.1,FJ009676.1untuk nukleotida gen ompC dan AE014613.1 dan AL627267.1 untuk gen fimH. 3. Homologi sekuen gen ompC dari penderita demam tifoid adalah 99% dengan accession number AL627274, AE014613.1, M31424.1,FJ009676.1 . Sekuen seluruh area koding gen fimH dari penderits demam tifoid mempunyai homologi 94% dengan sekuen accession number AE014613.1 dan AL627267.1. 4. Protein rekombinan OmpC dan FimH yang dikode oleh plasmid rekombinan pOmpC dan pFimH dari penderita demam tifoid dapat diekspresikan dalam E.coli DH10{3 dengan induksi IPTG sehingga dapat menghasilkan protein rekombinan OmpC dan FimH yang dapat digunakan sebagai biomaterial untuk pengembangan diagnosis demam tifoid

The Clinical Pattern and Prevalence of Streptococcus mutans and Streptococcus sobrinus among Adult and Children Patients with Dental Caries

Objective: To explore the clinical pattern, host factors, and presentation of Streptococcus mutans related to caries incidence among children and adults visiting Universitas Airlangga dental clinic. Material and Methods: This was an observational study with a cross-sectional approach with 50 patients in each group of carious children (6-12 years) and adults (18-35 years). Dental decay samples were taken by sterile excavator, put in a BHI’s transport medium, and directly incubated overnight at 37 ºC. The next day, they were sub-cultured microbiologically in Tryptone Yeast Cystine Sucrose Bacitracin (TYCSB) selective medium. Bacterial species and serogroups were examined by PCR. All patient’s data were collected from medical records and direct observation. Results: Caries were mostly media type in both children and adults. Oral hygiene (OHIS) in children was higher than in adults but not significantly different according to their DMFT. The highest scores for decay, missed and filled teeth were 16, 8 and 7, with an average of 6.82, 1.22 and 0.63, considered quite high. Conclusion: The prevalence of S. mutans was higher in children’s caries than in adults, but among the adult patients the co-incidence of S. mutans and S. sobrinus was associated with higher DMFT. The mutans serotypes e, f, and d were more prevalent among children than adults

Menaquinone-4 enhances testosterone production in rats and testis-derived tumor cells

<p>Abstract</p> <p>Background</p> <p>Vitamin K is essential for the posttranslational modification of various Gla proteins. Although it is widespread in several organs, including the testis, the function of vitamin K in these organs is not well characterized. In this study, we investigated the function of vitamin K in the testis and analyzed its role in steroidogenesis.</p> <p>Methods</p> <p>Eight-week-old male Wistar rats were fed a diet supplemented with menaquinone-4 (MK-4, 75 mg/kg diet), one of the predominant K₂vitamins present in the testis, for 5 weeks. <it>In vivo </it>testosterone levels of the rats' plasma and testes were measured by enzyme-linked immunosorbent assay, and <it>in vitro </it>testosterone levels of testis-derived tumor cells (I-10 cells) maintained in Ham's F-10 medium with 10% fetal bovine serum were measured following treatment with MK-4 (0 to 100 μM) at several time points. Testosterone and cellular protein levels were analyzed with respect to their effects on steroidogenesis.</p> <p>Results</p> <p>Testosterone levels in the plasma and testes of MK-4-fed rats were significantly increased compared to those of control rats, with no obvious differences in plasma luteinizing hormone levels. Secreted testosterone levels from I-10 cells were elevated by MK-4, but not by vitamin K₁, in a dose-dependent manner independent of cAMP treatment. Western blot analysis revealed that expression of CYP11A, the rate-limiting enzyme in steroidogenesis, and phosphorylation levels of protein kinase A (PKA) and the cAMP response element-binding protein were all stimulated by the presence of MK-4. Enhancement of testosterone production was inhibited by H89, a specific inhibitor of PKA, but not by warfarin, an inhibitor of γ-glutamylcarboxylation.</p> <p>Conclusions</p> <p>MK-4 stimulates testosterone production in rats and testis-derived tumor cells via activation of PKA. MK-4 may be involved in steroidogenesis in the testis, and its supplementation could reverse the downregulation of testosterone production in elders.</p

Antitumor effect of oral cancer vaccine with Bifidobacterium delivering WT1 protein to gut immune system is superior to WT1 peptide vaccine

Despite the revolutionary progress of immune checkpoint inhibitors (CPIs) for cancer immunotherapy, CPIs are effective only in a subset of patients. Combining CPIs and cancer vaccines to achieve better clinical outcomes is a reasonable approach since CPI enhances cancer vaccine-induced tumor-associated antigen (TAA) specific CTL. Among the various TAAs so far identified, WT1 protein is one of the most promising TAAs as a cancer vaccine target. Until now clinical trials of WT1 vaccine have demonstrated only modest clinical efficacy. These WT1 vaccines were based on peptides or dendritic cells (DCs), and there was no oral cancer vaccine. Recently, we developed a WT1 oral cancer vaccine using a recombinant Bifidobacterium displaying WT1 protein, which can efficiently deliver WT1 protein to the gut immune system, and we demonstrated that this oral cancer vaccine had a significant anti-tumor effect in a C1498-WT1 murine leukemia syngeneic tumor model. The WT1 protein displayed in this vaccine consists of about 70% of the WT1 amino acid sequence including multiple known CD4 and CD8 T-cell epitopes of WT1. In this commentary, we introduce our recent data indicating the superior anti-tumor effect of a WT1 oral cancer vaccine delivering WT1 protein to the gut immune system compared to a peptide vaccine

Dynamic Changes in Reactive Oxygen Species in the Shoot Apex Contribute to Stem Cell Death in Arabidopsis thaliana

In monocarpic plants, stem cells are fated to die. However, the potential mechanism of stem cell death has remained elusive. Here, we reveal that the levels of two forms of reactive oxygen species (ROS), superoxide anion free radical (O2·-) and hydrogen peroxide (H2O2), show dynamic changes in the shoot apex during the plant life cycle of Arabidopsis thaliana. We found that the level of O2·- decreased and disappeared at four weeks after bolting (WAB), while H2O2 appeared at 3WAB and showed a burst at 5 WAB. The timing of dynamic changes in O2·- and H2O2 was delayed for approximately three weeks in clv3-2, which has a longer lifespan. Moreover, exogenous application of H2O2 inhibited the expression of the stem cell determinant WUSCHEL (WUS) and promoted the expression of the developmentally programmed cell death (dPCD) marker gene ORESARA 1 (ORE1). These results indicate that H2O2 triggers an important signal inducing dPCD in stem cells. Given that O2·- plays roles in maintaining WUS expression and stem cell activity, we speculate that the dynamic shift from O2·- to H2O2 in the shoot apex results in stem cell death. Our findings provide novel insights for understanding ROS-mediated regulation during plant stem cell death

The Cell Differentiation of Idioblast Myrosin Cells: Similarities With Vascular and Guard Cells

Idioblasts are defined by abnormal shapes, sizes, and contents that are different from neighboring cells. Myrosin cells are Brassicales-specific idioblasts and accumulate a large amount of thioglucoside glucohydrolases (TGGs, also known as myrosinases) in their vacuoles. Myrosinases convert their substrates, glucosinolates, into toxic compounds when herbivories and pests attack plants. In this review, we highlight the similarities and differences between myrosin cells and vascular cells/guard cells (GCs) because myrosin cells are distributed along vascular cells, especially the phloem parenchyma, and myrosin cells share the master transcription factor FAMA with GCs for their cell differentiation. In addition, we analyzed the overlap of cell type-specific genes between myrosin cells and GCs by using published single-cell transcriptomics (scRNA-seq) data, suggesting significant similarities in the gene expression patterns of these two specialized cells

Histone Demethylases JMJ30 and JMJ32 Modulate the Speed of Vernalization Through the Activation of FLOWERING LOCUS C in Arabidopsis thaliana

Vernalization is the promotion of flowering after prolonged exposure to cold. In Arabidopsis thaliana, vernalization induces epigenetic silencing of the floral repressor gene FLOWERING LOCUS C (FLC). Among the repressive epigenetic marks, the trimethylation of lysine 27 on histone H3 proteins (H3K27me3) is a critical contributor to the epigenetic silencing of FLC. The deposition of H3K27me3 is mediated by Polycomb Repressive Complex 2 (PRC2). Conversely, the elimination of H3K27me3 is mediated by histone demethylases, Jumonji-C domain-containing protein JMJ30 and its homolog JMJ32. However, the role of JMJ30 and JMJ32 in vernalization is largely unknown. In this study, we found that cold treatment dramatically reduced the expression levels of JMJ30 and did not reduce those of JMJ32. Next, by using the genetic approach, we found that the flowering of jmj30 jmj32 was accelerated under moderate vernalized conditions. Under moderate vernalized conditions, the silencing of FLC occurred more quickly in jmj30 jmj32 than in the wild type. These results suggested that the histone demethylases JMJ30 and JMJ32 brake vernalization through the activation of FLC. Our study suggested that PRC2 and Jumonji histone demethylases act in an opposing manner to regulate flowering time via epigenetic modifications

Identification of a Devernalization Inducer by Chemical Screening Approaches in Arabidopsis thaliana

10.3389/fpls.2021.634068Frontiers in Plant Science1263406