scholarly journals Rsp Activates Expression of the Cnt System in Staphylococcus aureus

2020 ◽  
Author(s):  
Laura Vinué ◽  
David C. Hooper
Keyword(s):  
Gene Expression ◽  
Parental Strain ◽  
Double Mutant ◽  
Statistical Significance ◽  
Essential Metals ◽  
Band Shift ◽  
Fold Reduction ◽  
Specific Band ◽  
The Relationship ◽  
Candidate Regulator

Abstract Background: The Cnt system is crucial for the optimal import of essential metals in metal-limiting conditions and contributes to virulence in S. aureus. In a screen for regulators of efflux pumps in a phage-based ultra-high-density transposon library, we identified Rsp as a candidate regulator of the cntE gene. Results: A two-fold decrease in expression of all genes of the cnt operon was observed by RT-qPCR in the rsp mutant compared to the parental strain, indicating that Rsp acts as an activator of the cnt operon. To determine whether the Rsp activation depends on iron, we compared mutant and parent cnt expression under varying metal conditions. A 2-fold reduction in cnt gene expression was detected in the rsp mutant in TSB, and a slightly smaller decrease (1.9, 1.7, and 1.5-fold changes for cntK, cmtA, and cntE respectively) was observed after addition of dipyridyl. The greatest decrease was seen with addition of FeSO4 (4.1, 5.3 and 6.3-fold changes for cntK, cmtA and cntE respectively). These findings suggest that Rsp activates the cnt operon in low and high iron conditions. To study the relationship between Rsp and the cnt repressors Fur and Zur, we created single and double mutants. Both fur and zur single mutants had significant increases in cnt gene expression compared to the parental strain, as did the fur rsp double mutant. The zur rsp double mutant also had a significant increase in cntK expression and a trend in increases in cntA and cntE expression just below statistical significance. Thus, the ability of Fur and Zur to repress cnt gene expression are not eliminated by the presence of Rsp. However, there were significantly smaller increases in cnt gene expression in the double mutants compared to single mutants, suggesting that Rsp activation can still occur in the absence of these repressors. To determine if Rsp directly modulates expression of cnt genes, incubation of purified Rsp caused a DNA-specific band shift for the cntK and cntA promoters. Conclusions: Rsp activation may act to maintain basal cellular levels of staphylopine to scavenge free metals when needed, in addition to metal dependent regulation by Fur and Zur.

scholarly journals Rsp activates expression of the Cnt system in Staphylococcus aureus

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Laura Vinué ◽  
David C. Hooper
Keyword(s):  
Gene Expression ◽  
Parental Strain ◽  
Double Mutant ◽  
Statistical Significance ◽  
Essential Metals ◽  
Band Shift ◽  
Fold Reduction ◽  
Specific Band ◽  
The Relationship ◽  
Candidate Regulator

Abstract Background The Cnt system is crucial for the optimal import of essential metals in metal-limiting conditions and contributes to virulence in S. aureus. In a screen for regulators of efflux pumps in a phage-based ultra-high-density transposon library, we identified Rsp as a candidate regulator of the cntE gene. Results A two-fold decrease in expression of all genes of the cnt operon was observed by RT-qPCR in the rsp mutant compared to the parental strain, indicating that Rsp acts as an activator of the cnt operon. To determine whether the Rsp activation depends on iron, we compared mutant and parent cnt expression under varying metal conditions. A 2-fold reduction in cnt gene expression was detected in the rsp mutant in TSB, and a slightly smaller decrease (1.9, 1.7, and 1.5-fold changes for cntK, cmtA, and cntE respectively) was observed after addition of dipyridyl. The greatest decrease was seen with addition of FeSO4 (4.1, 5.3 and 6.3-fold changes for cntK, cmtA and cntE respectively). These findings suggest that Rsp activates the cnt operon in low and high iron conditions. To study the relationship between Rsp and the cnt repressors Fur and Zur, we created single and double mutants. Both fur and zur single mutants had significant increases in cnt gene expression compared to the parental strain, as did the fur rsp double mutant. The zur rsp double mutant also had a significant increase in cntK expression and a trend in increases in cntA and cntE expression just below statistical significance. Thus, the ability of Fur and Zur to repress cnt gene expression are not eliminated by the presence of Rsp. However, there were significantly smaller increases in cnt gene expression in the double mutants compared to single mutants, suggesting that Rsp activation can still occur in the absence of these repressors. To determine if Rsp directly modulates expression of cnt genes, incubation of purified Rsp caused a DNA-specific band shift for the cntK and cntA promoters. Conclusions Rsp activation may act to maintain basal cellular levels of staphylopine to scavenge free metals when needed, in addition to metal dependent regulation by Fur and Zur.

scholarly journals Rsp Activates Expression of the Cnt System in Staphylococcus aureus

2020 ◽  
Author(s):  
Laura Vinué ◽  
David C. Hooper
Keyword(s):  
Gene Expression ◽  
Parental Strain ◽  
Double Mutant ◽  
Statistical Significance ◽  
Essential Metals ◽  
Band Shift ◽  
Fold Reduction ◽  
Specific Band ◽  
The Relationship ◽  
Candidate Regulator

Abstract Background: The Cnt system is crucial for the optimal import of essential metals in metal-limiting conditions and contributes to virulence in S. aureus. In a screen for regulators of efflux pumps in a phage-based ultra-high-density transposon library, we identified Rsp as a candidate regulator of the cntE gene. Results: A two-fold decrease in expression of all genes of the cnt operon was observed by RT-qPCR in the rsp mutant compared to the parental strain, indicating that Rsp acts as an activator of the cnt operon. To determine whether the Rsp activation depends on iron, we compared mutant and parent cnt expression under varying metal conditions. A 2-fold reduction in cnt gene expression was detected in the rsp mutant in TSB, and a slightly smaller decrease (1.9, 1.7, and 1.5-fold changes for cntK, cmtA, and cntE respectively) was observed after addition of dipyridyl. The greatest decrease was seen with addition of FeSO4 (4.1, 5.3 and 6.3-fold changes for cntK, cmtA and cntE respectively). These findings suggest that Rsp activates the cnt operon in low and high iron conditions. To study the relationship between Rsp and the cnt repressors Fur and Zur, we created single and double mutants. Both fur and zur single mutants had significant increases in cnt gene expression compared to the parental strain, as did the fur rsp double mutant. The zur rsp double mutant also had a significant increase in cntK expression and a trend in increases in cntA and cntE expression just below statistical significance. Thus, the ability of Fur and Zur to repress cnt gene expression are not eliminated by the presence of Rsp. However, there were significantly smaller increases in cnt gene expression in the double mutants compared to single mutants, suggesting that Rsp activation can still occur in the absence of these repressors. To determine if Rsp directly modulates expression of cnt genes, incubation of purified Rsp caused a DNA-specific band shift for the cntK and cntA promoters. Conclusions: Rsp activation may act to maintain basal cellular levels of staphylopine to scavenge free metals when needed, in addition to metal dependent regulation by Fur and Zur.

scholarly journals Rsp Activates Expression of the Cnt System in Staphylococcus aureus

2020 ◽  
Author(s):  
Laura Vinué ◽  
David C. Hooper
Keyword(s):  
Gene Expression ◽  
Parental Strain ◽  
Double Mutant ◽  
Statistical Significance ◽  
Essential Metals ◽  
Band Shift ◽  
Fold Reduction ◽  
Specific Band ◽  
The Relationship ◽  
Candidate Regulator

Abstract Background: The Cnt system is crucial for the optimal import of essential metals in metal-limiting conditions and contributes to virulence in S. aureus. In a screen for regulators of efflux pumps in a phage-based ultra-high-density transposon library, we identified Rsp as a candidate regulator of the cntE gene. Results: A two-fold decrease in expression of all genes of the cnt operon was observed by RT-qPCR in the rsp mutant compared to the parental strain, indicating that Rsp acts as an activator of the cnt operon. To determine whether the Rsp activation depends on iron, we compared mutant and parent cnt expression under varying metal conditions. A 2-fold reduction in cnt gene expression was detected in the rsp mutant in TSB, and a slightly smaller decrease (1.9, 1.7, and 1.5-fold changes for cntK, cmtA, and cntE respectively) was observed after addition of dipyridyl. The greatest decrease was seen with addition of FeSO4 (4.1, 5.3 and 6.3-fold changes for cntK, cmtA and cntE respectively). These findings suggest that Rsp activates the cnt operon in low and high iron conditions. To study the relationship between Rsp and the cnt repressors Fur and Zur, we created single and double mutants. Both fur and zur single mutants had significant increases in cnt gene expression compared to the parental strain, as did the fur rsp double mutant. The zur rsp double mutant also had a significant increase in cntK expression and a trend in increases in cntA and cntE expression just below statistical significance. Thus, the ability of Fur and Zur to repress cnt gene expression are not eliminated by the presence of Rsp. However, there were significantly smaller increases in cnt gene expression in the double mutants compared to single mutants, suggesting that Rsp activation can still occur in the absence of these repressors. To determine if Rsp directly modulates expression of cnt genes, incubation of purified Rsp caused a DNA-specific band shift for the cntK and cntA promoters. Conclusions: Rsp activation may act to maintain basal cellular levels of staphylopine to scavenge free metals when needed, in addition to metal dependent regulation by Fur and Zur.

scholarly journals Rsp Activates Expression of the Cnt System in Staphylococcus aureus

2020 ◽  
Author(s):  
Laura Vinué ◽  
David C. Hooper
Keyword(s):  
Gene Expression ◽  
Parental Strain ◽  
Double Mutant ◽  
Statistical Significance ◽  
Band Shift ◽  
Fold Reduction ◽  
Specific Band ◽  
The Relationship ◽  
Twofold Decrease ◽  
Candidate Regulator

Abstract Background: The Cnt system is crucial for the optimal import of essential metals in metal-limiting conditions and contributes to virulence in S. aureus. In a screen for regulators of efflux pumps in a phage-based ultra-high-density transposon library, we identified Rsp as a candidate regulator of the cntE gene. Results: A twofold decrease in expression of all genes of the cnt operon was observed by RT-qPCR in the rsp mutant compared to the parental strain, indicating that Rsp acts as an activator of the cnt operon. To determine whether the Rsp activation depends on iron, we compared mutant and parent cnt expression under varying metal conditions. A 2-fold reduction in cnt gene expression was detected in the rsp mutant in TSB, and a slightly smaller decrease (1.9, 1.7, and 1.5-fold changes for cntK, cmtA, and cntE respectively) was observed after addition of dipyridyl. The greatest decrease was seen with addition of FeSO4 (4.1, 5.3 and 6.3-fold changes for cntK, cmtA and cntE respectively). These findings suggest that Rsp activates the cnt operon in low and high iron conditions. To study the relationship between Rsp and the cnt repressors Fur and Zur, we created single and double mutants. Both fur and zur single mutants had significant increases in cnt gene expression compared to the parental strain, as did the fur rsp double mutant. The zur rsp double mutant also had a significant increase in cntK expression and a trend in increases in cntA and cntE expression just below statistical significance. Thus, the ability of Fur and Zur to repress cnt gene expression are not eliminated by the presence of Rsp. However, there were significantly smaller increases in cnt gene expression in the double mutants compared to single mutants, suggesting that Rsp activation can still occur in the absence of these repressors. To determine if Rsp directly modulates expression of cnt genes, incubation of purified Rsp caused a DNA-specific band shift for the cntK and cntA promoters. Conclusions: Rsp activation may act to maintain basal cellular levels of staphylopine to scavenge free metals when needed, in addition to metal dependent regulation by Fur and Zur.

scholarly journals Rsp Activates Expression of the Cnt System in Staphylococcus aureus

2020 ◽  
Author(s):  
Laura Vinué ◽  
David C. Hooper
Keyword(s):  
Gene Expression ◽  
Staphylococcus Aureus ◽  
Parental Strain ◽  
Essential Metals ◽  
Band Shift ◽  
High Iron ◽  
Fold Reduction ◽  
Specific Band ◽  
The Relationship ◽  
Candidate Regulator

Abstract Background The Cnt system is crucial for the optimal import of essential metals in metal-limiting conditions and contributes to virulence in S. aureus. In a screen for regulators of efflux pumps and other exporters in a phage-based ultra-high-density transposon library, we identified Rsp as a candidate regulator of the cntE exporter gene. Results A two-fold decrease in expression of all genes of the cnt operon was observed by RT-qPCR in the rsp mutant compared to the parental strain, indicating that Rsp acts as an activator of the cnt operon. To determine whether the Rsp activation depends on iron, we compared mutant and parent cnt expression under varying metal conditions. A 2-fold reduction in cnt gene expression was detected in the rsp mutant in TSB, and a slightly smaller decrease (1.9, 1.7, and 1.5-fold changes for cntK, cmtA, and cntE respectively) was observed after addition of dipyridyl. The greatest decrease was seen with addition of FeSO4 (4.1, 5.3 and 6.3-fold changes for cntK, cmtA and cntE respectively). These findings suggest that Rsp activates the cnt operon in both low and high iron conditions. To study the relationship between Rsp and the cnt repressors Fur and Zur, we created single and double mutants. A decrease in the expression of cntK (1.5-fold) and cntA (1.6-fold) was seen for Δfur Δrsp compared to Δfur strain. Similarly decreased expression of cntK (1.3-fold), cntA (1.6-fold) and cntE (1.6-fold) genes was seen comparing Δzur Δrsp and Δzur mutants but statistically not significant, indicating that Rsp is dependent on Fur and Zur. To determine if Rsp directly modulates expression of cnt genes, incubation of purified Rsp caused a DNA-specific band shift for both the cntK and cntA promoters. Conclusions Rsp activation may act to maintain basal cellular levels of staphylopine to scavenge free metals when needed, in addition to metal dependent regulation by Fur and Zur.

Study of the infl uence of information and communication tools on the psychophysiological state of schoolchildren on the basis of subjective evaluation

2020 ◽  
pp. 47-55
Author(s):  
Aleksandra Rakhmanova ◽  
Georgiy Loginov ◽  
Vladimir Dolich ◽  
Nataliya Komleva ◽  
Galina Rakhmanova
Keyword(s):  
High School ◽  
Emotional State ◽  
Statistical Significance ◽  
Subjective Evaluation ◽  
Educational Process ◽  
The Body ◽  
Leningrad Region ◽  
Communication Tools ◽  
Information And Communication ◽  
The Relationship

The relevance of the article is determined by the existence of contradictions between the need to introduce innovative technologies into the educational process at school, as an integral attribute of modern education, and the negative influence of factors on the physical and psycho-emotional state of health of students related to the use of information and communication tools (computers, phones, headphones). The goal of the study was to assess the relationship between the timing of the use of information and communication tools and the frequency of functional and psycho-emotional complaints in groups of middle and high school schoolchildren. 400 schoolchildren of the Saratov Region, the Moscow Region, Leningrad Region and the Republic of Dagestan were surveyed, who made up two groups of research: middle-school schoolchildren (grades 5–6) and high-school schoolchildren (grades 10–11 The survey was carried out by means of the standardized formalized cards which included the questions considering usage time of computers and mobile phones, complaints to a headache, hands pain, other pain and/or feeling of discomfort from visual organ and the organs of hearing, as well as a psycho-emotional state. Statistical analysis of the data was performed using the STATISTICA application software program by StatSoft Inc (USA). To compare the frequencies of a binary feature, a fourfold table of absolute frequencies was constructed and the level of statistical significance for the exact Fisher’s two-tailed test criterion was determined. The study was conducted according to the requirements of bioethics, after signing informed consent statement by teenagers and their parents. The study examined the relationship between the timing of the use of information and communication tools and the frequency of complaints in groups of schoolchildren. The results of the study should be taken into account when developing and implementing preventive measures to prevent negative effects of computers and mobile devices on the body of students.

The Relationship Between The Perception of Elderly’s Health Status and Health Behaviors in Nakhon Nayok Province, Thailand

2019 ◽  
Vol 12 (1) ◽  
pp. 420-423
Author(s):  
Prapada Watcharanat ◽  
Prasong Tanpichai ◽  
Ravee Sajjasophon
Keyword(s):  
Regression Analysis ◽  
Health Status ◽  
Health Behaviors ◽  
Multiple Regression Analysis ◽  
Multiple Regression ◽  
Statistical Significance ◽  
The Elderly ◽  
Cross Sectional ◽  
Congenital Diseases ◽  
The Relationship

Purpose: This research aims to study the relationship between perception of elderly’s health and health behaviors in Nakhon Nayok province, Thailand Methods: This research was a cross-sectional study. The questionnaire was used to collect the data. This research was conducted in Nakhon Nayok province. The sample size was 270 which applied Taro Yamane's formula at a significant level 0.05. The descriptive statistics was implemented to describe the variables by presenting the frequency, percentage, mean and standard deviation. Furthermore, multiple regression analysis was applied to analyze the relationship between perception of elderly’s health and health behaviors. The statistical significance was considered to reject Hypothesis-null at < 0.05. Results: From a total of 270 people, more than 58.22% of the elderly perceived that they had moderate health conditions. Most elderly had congenital diseases (62.2%). The multiple regression analysis results showed that health status perception and health status perception when compared to their cohort related significantly to health behavior. Conclusion: The government should support the elderly on participation, trust, engagement, and cultural concern of the people in the community, which can contribute to promoting the physical, mental and social condition of the elderly.

scholarly journals Endolysosomal Cation Channels and MITF in Melanocytes and Melanoma

Biomolecules ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 1021
Author(s):  
Carla Abrahamian ◽  
Christian Grimm
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Transcription Factor ◽  
Convincing Evidence ◽  
Signaling Cascades ◽  
Cation Channels ◽  
Pore Channel ◽  
Different Types ◽  
Canonical Wnt ◽  
The Relationship

Microphthalmia-associated transcription factor (MITF) is the principal transcription factor regulating pivotal processes in melanoma cell development, growth, survival, proliferation, differentiation and invasion. In recent years, convincing evidence has been provided attesting key roles of endolysosomal cation channels, specifically TPCs and TRPMLs, in cancer, including breast cancer, glioblastoma, bladder cancer, hepatocellular carcinoma and melanoma. In this review, we provide a gene expression profile of these channels in different types of cancers and decipher their roles, in particular the roles of two-pore channel 2 (TPC2) and TRPML1 in melanocytes and melanoma. We specifically discuss the signaling cascades regulating MITF and the relationship between endolysosomal cation channels, MAPK, canonical Wnt/GSK3 pathways and MITF.

scholarly journals Correlation between L-Lactate Concentrations in Beef Cattle, Obtained Using a Hand-Held Lactate Analyzer and a Lactate Assay Colorimetric Kit

Animals ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 926
Author(s):  
Daniela M. Meléndez ◽  
Sonia Marti ◽  
Luigi Faucitano ◽  
Derek B. Haley ◽  
Timothy D. Schwinghamer ◽  
...  
Keyword(s):  
Statistical Significance ◽  
Road Transportation ◽  
Long Distance ◽  
Suitable Alternative ◽  
Blood Samples ◽  
Time Points ◽  
Pooled Data ◽  
Red Angus ◽  
Relationship Of ◽  
The Relationship

Lactate is a product of anaerobic glycolysis, used in animal research as an indicator of muscle fatigue. Therefore, it has been used as an indicator of cattle response to long distance transportation. The aim of this study was to assess the relationship of L-lactate concentrations measured using a Lactate Scout+ analyzer and a traditional lactate assay colorimetric kit. Blood samples were collected by venipuncture from 96 steers (Black or Red Angus × Hereford/Simmental and Black or Red Angus × Charolais; 247 ± 38.2 kg BW) prior to loading (LO1) and after 36 h of transport, and prior to reloading and after an additional 4 h of road transportation, and on d 1, 2, 3, 5, 14, and 28 after transport. The Lactate Scout+ analyzer strip was dipped in blood at the time of sampling, while blood samples were collected into sodium fluoride tubes for use in the colorimetric analysis. Pearson correlations were calculated to assess the strength of the relationship between the experimental methods for the quantification of L-lactate concentrations. The magnitude and direction of the correlation, and the level of statistical significance varied over the observed time points, ranging from r = −0.03 (p = 0.75; LO1) to r = 0.75 (p < 0.0001; d 3). The correlation for the pooled data was weak but statistically significant (r = 0.33, p < 0.0001). Based on the low magnitude of the correlation due to variability across sampling time points in this study, the Lactate Scout+ analyzer is not a suitable alternative to a lab-based assay (considered the gold standard) for measuring L-lactate in transported cattle.

scholarly journals PSVII-3 Correlations between L-lactate concentrations obtained using a handheld lactate analyser and a lactate assay colorimetric kit in beef cattle transported by road

2020 ◽  
Vol 98 (Supplement_4) ◽  
pp. 296-297
Author(s):  
Daniela M Meléndez ◽  
Sonia Marti ◽  
Luigi Faucitano ◽  
Derek B Haley ◽  
Timothy D Schwinghamer ◽  
...  
Keyword(s):  
Statistical Significance ◽  
Cost Effective ◽  
Road Transportation ◽  
Long Distance ◽  
Suitable Alternative ◽  
Blood Samples ◽  
Handheld Device ◽  
Time Points ◽  
Relationship Of ◽  
The Relationship

Abstract Blood metabolites are used to assess a variety of animal conditions for veterinary diagnosis and research. Concentration of metabolites in blood can be measured using a commercially-available lab-based assay or in real-time using a handheld device developed to be more time- and cost-effective than the lab-based method. Lactate is a product of anaerobic glycolysis, used in animal research as an indicator of muscle fatigue. Therefore, it has been used as an indicator of cattle response to long distance transportation. The aim of this study was to assess the relationship of L-lactate concentrations measured using a Lactate Scout+ analyzer (Lactate Scout, EFK Diagnostics, Barleben, Germany) and a lactate assay colorimetric kit (Lactate Assay Kit, Cell Biolabs Inc., San Diego, CA). Blood samples were collected by venipuncture from 96 steers (245 ± 35.7 kg BW) prior to (L1) and after 36 h, and prior to and after an additional 4 h of road transportation, and on d 1, 2, 3, 5, 14, and 28 after transport. The Lactate Scout+ analyzer strip was dipped in blood at the time of sampling, while blood samples were collected into sodium fluoride tubes for use in colorimetric analysis. Pearson correlations were calculated to determine the relationship between the experimental methods for the quantification of L-lactate concentrations. The strengths and levels of statistical significance of the correlation varied over the observed time points, r = -0.03, P = 0.75 (L1) to r = 0.75, P = &lt; 0.0001 (d 3). The correlation for the pooled data was weak but statistically significant (r = 0.33, P &lt; 0.001). Based on the experimental results, the Lactate Scout+ analyzer is not a suitable alternative to a lab-based assay for measuring L-lactate in transported cattle, due to variability across sampling time points and weak correlation with the traditional enzymatic method.

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