ement of patients seems useful or even indispensable in view of potential mortality.Amaranthus tricolor has been reported to contain some antimicrobial compounds, such as alkaloids, polyphenols, and terpenoids. However, its effect on Staphylococcus aureus has been less well researched. Therefore, this study was designed to evaluate the antimicrobial activity and possible mechanism of action of the Amaranthus tricolor crude extract (ATCE) against S. aureus and potential application in cooked meat. The antimicrobial activity against S. aureus was assessed by disk diffusion, minimum inhibitory concentration (MIC) determinations, and growth curve. The changes of bacterial membrane potential, intracellular pH (pHin), content of bacterial protein and DNA, and cell morphology were measured to indicate its antimicrobial mechanism of action. The effects of different concentrations of ATCE on bacterial counts, pH, and color of lean cooked pork during 6 d storage were assessed. The results showed that the diameter of inhibition zone (DIZ) and ****of ATCE against S. aureus were 12.63 ± 0.34 to 12.94 ± 0.43 mm and 80 mg/mL, respectively. The mechanism of action of ATCE against S. aureus was associated with cell membrane depolarization, reduction of pHin, decrease of bacterial protein content, cleavage of cell DNA, and leakage of cytoplasm. Besides, ATCE resulted in a reduction of 1.02 log CFU/g from 3 log CFU/g in S. aureus-inoculated lean cooked pork. The pH values of lean cooked pork treated with ATCE did not show significant changes as the storage time increased, but there was a slight and significant decrease seen with the application of 1 and 2 ****of ATCE. After treating with ATCE, the color of lean cooked pork showed less lightness (L*), more redness (a∗), similar yellowness (b*), stronger chroma (C*), and weaker hue angle (h*) during 6 days of storage. Therefore, these findings indicate that ATCE has antimicrobial activities against S. aureus and possesses latent energy to become a natural preservative to maintain the quality of lean cooked pork.Single-cell analysis enables detailed molecular characterization of cells in relation to cell type, genotype, cell state, temporal variations, and microenvironment. These studies often include the analysis of individual genes and networks of genes. The total amount of RNA also varies between cells due to important factors, such as cell type, cell size, and cell cycle state. However, there is a lack of simple and sensitive methods to quantify the total amount of RNA, especially mRNA. Here, we developed a method to quantify total mRNA levels in single cells based on global reverse transcription followed by quantitative PCR. Standard curve analyses of diluted RNA and sorted cells showed a wide dynamic range, high reproducibility, and excellent sensitivity. Single-cell analysis of three sarcoma cell lines and human fibroblasts revealed cell type variations, a lognormal distribution of total mRNA levels, and up to an eight-fold difference in total mRNA levels among the cells. The approach can easily be combined with targeted or global gene expression profiling, providing new means to study cell heterogeneity at an individual gene level and at a global level. This method can be used to investigate the biological importance of variations in the total amount of mRNA in healthy as well as pathological conditions.BACKGROUND Gastric cancer is currently the third leading cause of cancer-related deaths worldwide, usually diagnosed at late stages. The development of new biomarkers to improve its prevention and patient management is critical for disease control. piRNAs are small regulatory RNAs important for gene silencing mechanisms, mainly associated with the silencing of transposable elements. piRNA pathways may also be involved in gene regulation and the deregulation of piRNAs may be an important factor in carcinogenic processes. Thus, several studies suggest piRNAs as potential cancer biomarkers. Translational studies suggest that piRNAs may regulate key genes and pathways associated with gastric cancer progression, though there is no functional annotation in piRNA databases. The impacts of genetic variants in piRNA genes and their influence in gastric cancer development remains elusive, highlighting the gap in piRNA regulatory mechanisms knowledge. Here, we discuss the current state of understanding of piRNA-mediated regulation and piRNA functions and suggest that genetic alterations in piRNA genes may affect their functionality, thus, it may be associated with gastric carcinogenesis. CONCLUSIONS In the era of precision medicine, investigations about genetic and epigenetic mechanisms are essential to further comprehend gastric carcinogenesis and the role of piRNAs as potential biomarkers for translational research.The crosstalk between actin and actin-related proteins (Arps), namely Arp2 and Arp3, plays a central role in facilitating actin polymerization in the cytoplasm and also in the nucleus. Nuclear F-actin is required for transcriptional regulation, double-strand break repair, and nuclear organization. The formation of nuclear F-actin is highly dynamic, suggesting the involvement of positive and negative regulators for nuclear actin polymerization. While actin assembly factors for nuclear F-actin have been recently described, information about inhibitory factors is still limited. https://www.selleckchem.com/GSK-3.html The actin-related protein Arp4 which is predominantly localized in the nucleus, has been previously identified as an integral subunit of multiple chromatin modulation complexes, where it forms a heterodimer with monomeric actin. Therefore, we tested whether Arp4 functions as a suppressor of nuclear F-actin formation. The knockdown of Arp4 (Arp4 KD) led to an increase in nuclear F-actin formation in NIH3T3 cells, and purified Arp4 potently inhibited F-actin formation in mouse nuclei transplanted into Xenopus laevis oocytes. Consistently, Arp4 KD facilitated F-actin-inducible gene expression (e.g., OCT4) and DNA damage repair. Our results suggest that Arp4 has a critical role in the formation and functions of nuclear F-actin.
ement of patients seems useful or even indispensable in view of potential mortality.Amaranthus tricolor has been reported to contain some antimicrobial compounds, such as alkaloids, polyphenols, and terpenoids. However, its effect on Staphylococcus aureus has been less well researched. Therefore, this study was designed to evaluate the antimicrobial activity and possible mechanism of action of the Amaranthus tricolor crude extract (ATCE) against S. aureus and potential application in cooked meat. The antimicrobial activity against S. aureus was assessed by disk diffusion, minimum inhibitory concentration (MIC) determinations, and growth curve. The changes of bacterial membrane potential, intracellular pH (pHin), content of bacterial protein and DNA, and cell morphology were measured to indicate its antimicrobial mechanism of action. The effects of different concentrations of ATCE on bacterial counts, pH, and color of lean cooked pork during 6 d storage were assessed. The results showed that the diameter of inhibition zone (DIZ) and MIC of ATCE against S. aureus were 12.63 ± 0.34 to 12.94 ± 0.43 mm and 80 mg/mL, respectively. The mechanism of action of ATCE against S. aureus was associated with cell membrane depolarization, reduction of pHin, decrease of bacterial protein content, cleavage of cell DNA, and leakage of cytoplasm. Besides, ATCE resulted in a reduction of 1.02 log CFU/g from 3 log CFU/g in S. aureus-inoculated lean cooked pork. The pH values of lean cooked pork treated with ATCE did not show significant changes as the storage time increased, but there was a slight and significant decrease seen with the application of 1 and 2 MIC of ATCE. After treating with ATCE, the color of lean cooked pork showed less lightness (L*), more redness (a∗), similar yellowness (b*), stronger chroma (C*), and weaker hue angle (h*) during 6 days of storage. Therefore, these findings indicate that ATCE has antimicrobial activities against S. aureus and possesses latent energy to become a natural preservative to maintain the quality of lean cooked pork.Single-cell analysis enables detailed molecular characterization of cells in relation to cell type, genotype, cell state, temporal variations, and microenvironment. These studies often include the analysis of individual genes and networks of genes. The total amount of RNA also varies between cells due to important factors, such as cell type, cell size, and cell cycle state. However, there is a lack of simple and sensitive methods to quantify the total amount of RNA, especially mRNA. Here, we developed a method to quantify total mRNA levels in single cells based on global reverse transcription followed by quantitative PCR. Standard curve analyses of diluted RNA and sorted cells showed a wide dynamic range, high reproducibility, and excellent sensitivity. Single-cell analysis of three sarcoma cell lines and human fibroblasts revealed cell type variations, a lognormal distribution of total mRNA levels, and up to an eight-fold difference in total mRNA levels among the cells. The approach can easily be combined with targeted or global gene expression profiling, providing new means to study cell heterogeneity at an individual gene level and at a global level. This method can be used to investigate the biological importance of variations in the total amount of mRNA in healthy as well as pathological conditions.BACKGROUND Gastric cancer is currently the third leading cause of cancer-related deaths worldwide, usually diagnosed at late stages. The development of new biomarkers to improve its prevention and patient management is critical for disease control. piRNAs are small regulatory RNAs important for gene silencing mechanisms, mainly associated with the silencing of transposable elements. piRNA pathways may also be involved in gene regulation and the deregulation of piRNAs may be an important factor in carcinogenic processes. Thus, several studies suggest piRNAs as potential cancer biomarkers. Translational studies suggest that piRNAs may regulate key genes and pathways associated with gastric cancer progression, though there is no functional annotation in piRNA databases. The impacts of genetic variants in piRNA genes and their influence in gastric cancer development remains elusive, highlighting the gap in piRNA regulatory mechanisms knowledge. Here, we discuss the current state of understanding of piRNA-mediated regulation and piRNA functions and suggest that genetic alterations in piRNA genes may affect their functionality, thus, it may be associated with gastric carcinogenesis. CONCLUSIONS In the era of precision medicine, investigations about genetic and epigenetic mechanisms are essential to further comprehend gastric carcinogenesis and the role of piRNAs as potential biomarkers for translational research.The crosstalk between actin and actin-related proteins (Arps), namely Arp2 and Arp3, plays a central role in facilitating actin polymerization in the cytoplasm and also in the nucleus. Nuclear F-actin is required for transcriptional regulation, double-strand break repair, and nuclear organization. The formation of nuclear F-actin is highly dynamic, suggesting the involvement of positive and negative regulators for nuclear actin polymerization. While actin assembly factors for nuclear F-actin have been recently described, information about inhibitory factors is still limited. https://www.selleckchem.com/GSK-3.html The actin-related protein Arp4 which is predominantly localized in the nucleus, has been previously identified as an integral subunit of multiple chromatin modulation complexes, where it forms a heterodimer with monomeric actin. Therefore, we tested whether Arp4 functions as a suppressor of nuclear F-actin formation. The knockdown of Arp4 (Arp4 KD) led to an increase in nuclear F-actin formation in NIH3T3 cells, and purified Arp4 potently inhibited F-actin formation in mouse nuclei transplanted into Xenopus laevis oocytes. Consistently, Arp4 KD facilitated F-actin-inducible gene expression (e.g., OCT4) and DNA damage repair. Our results suggest that Arp4 has a critical role in the formation and functions of nuclear F-actin.
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