Microansamycins J and K from <i>Micromonospora</i> sp. HK160111mas13OE

Microansamycins were novel pentaketide ansamycins isolated from Micromonospora sp. HK160111mas13OE with AHBA-C2-C2-C3-C3 skeleton and diverse post-PKS modifications. In this paper, two new congeners, namely microansamycins J (1) and K (2), were identified based on their NMR, HRESIMS data and compared with those of microansamycins F and E. Neither showed antibacterial activity against Staphy­lococcus aureus ATCC25923 and Escherichia coli at 40 µg/mL.</p

Construction, Model-Based Analysis, and Characterization of a Promoter Library for Fine-Tuned Gene Expression in <i>Bacillus subtilis</i>

Promoters are among the most-important and most-basic tools for the control of metabolic pathways. However, previous research mainly focused on the screening and characterization of some native promoters in <i>Bacillus subtilis</i>. To develop a broadly applicable promoter system for this important platform organism, we created a <i>de novo</i> synthetic promoter library (SPL) based on consensus sequences by analyzing the microarray transcriptome data of <i>B. subtilis</i> 168. A total of 214 potential promoters spanning a gradient of strengths was isolated and characterized by a green fluorescence assay. Among these, a detailed intensity analysis was conducted on nine promoters with different strengths by reverse-transcription polymerase chain reaction (RT-PCR) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Furthermore, reconstructed promoters and promoter cassettes (tandem promoter cluster) were designed via statistical model-based analysis and tandem dual promoters, which showed strength that was increased 1.2- and 2.77-fold compared to that of promoter P43, respectively. Finally, the SPL was employed in the production of inosine and acetoin by repressing and over-expressing the relevant metabolic pathways, yielding a 700% and 44% increase relative to the respective control strains. This is the first report of a <i>de novo</i> synthetic promoter library for <i>B. subtilis</i>, which is independent of any native promoter. The strategy of improving and fine-tuning promoter strengths will contribute to future metabolic engineering and synthetic biology projects in <i>B. subtilis</i>

Schematic diagram of persistent chronic inflammation-related HPV infection response contributes to oropharyngeal carcinogenesis.

<p>Schematic diagram of persistent chronic inflammation-related HPV infection response contributes to oropharyngeal carcinogenesis.</p

HPV infection correlates with the progression of oropharyngeal carcinogenesis.

<p>(A) Percentage of HPV 16 positive infection in patient tissues with normal oral mucosa, dysplasia, cancer in situ and cancer was shown. (B) Representative flow cytometry image of CD11b+ LIN- HLA-DR- CD33+ MDSCs in tissues of OPSCC patients with HPV-negative and HPV-positive. We first examined the percentage of LIN- HLA-DR- cells, and then screened the percentage of CD11b+ CD33+ cells in LIN- HLA-DR- cells. This image showed that the percentage of CD11b+ LIN- HLA-DR- CD33+ MDSCs in HPV-negative and HPV-positive OPSCC patients was 9.39% and 13.81%, respectively. (C) Representative immunohistochemical image (C3) of MPO in cancer tissues of OPSCC patients. C1 and C2 were H& E staining and C2 was amplification of C1.</p

Chronic inflammation correlates with the progression of oropharyngeal carcinogenesis.

<p>(A) The morphology of neutrophils (Black arrow) and lymphocytes (White arrow) in cell smear. (B) Percentage of different degrees of chronic inflammation in cell smear of indicated types of tissues were shown. The results showed that there was significantly difference between the percentage of moderate-severe inflammation in the cell smear of dysplasia and normal oral mucous (<i>P</i><0.001), and the inflammation degrees showed no different in the cell smear of dysplasia, cancer in situ and cancer (<i>P</i>>0.05). Error bars represent the mean±SD of triplicate experiments. (C) Representative image of normal oral mucosa, dysplasia, cancer in situ and cancer with mild inflammation. (D) Percentage of different degrees of chronic inflammation in indicated types of tissues are shown. The results showed that there was significantly difference between the percentage of moderate-severe inflammation in the tissues of dysplasia and normal oral mucous (<i>P</i><0.001), and the inflammation degrees showed no different in dysplasia, cancer in situ and cancer specimens (<i>P</i>>0.05). Error bars represent the mean±SD of triplicate experiments. (E) Transcription levels of IFN-γ, IL-10, IL-12, and IL-2 in normal oral mucosa, dysplasia, cancer in situ and cancer tissue, relative to GAPDH, determined by quantitative RT-PCR. Error bars represent the mean±SD of triplicate experiments.</p