Brucella melitensis VjbR and C12-HSL regulons: contributions of the N-dodecanoyl homoserine lactone signaling molecule and LuxR homologue VjbR to gene expression

<p>Abstract</p> <p>Background</p> <p>Quorum sensing is a communication system that regulates gene expression in response to population density and often regulates virulence determinants. Deletion of the <it>luxR </it>homologue <it>vjbR </it>highly attenuates intracellular survival of <it>Brucella melitensis </it>and has been interpreted to be an indication of a role for QS in <it>Brucella </it>infection. Confirmation for such a role was suggested, but not confirmed, by the demonstrated <it>in vitro </it>synthesis of an auto-inducer (AI) by <it>Brucella </it>cultures. In an effort to further delineate the role of VjbR to virulence and survival, gene expression under the control of VjbR and AI was characterized using microarray analysis.</p> <p>Results</p> <p>Analyses of wildtype <it>B. melitensis </it>and isogenic Δ<it>vjbR </it>transciptomes, grown in the presence and absence of exogenous <it>N</it>-dodecanoyl homoserine lactone (C₁₂-HSL), revealed a temporal pattern of gene regulation with variances detected at exponential and stationary growth phases. Comparison of VjbR and C₁₂-HSL transcriptomes indicated the shared regulation of 127 genes with all but 3 genes inversely regulated, suggesting that C₁₂-HSL functions via VjbR in this case to reverse gene expression at these loci. Additional analysis using a Δ<it>vjbR </it>mutant revealed that AHL also altered gene expression in the absence of VjbR, up-regulating expression of 48 genes and a <it>luxR </it>homologue <it>blxR </it>93-fold at stationary growth phase. Gene expression alterations include previously un-described adhesins, proteases, antibiotic and toxin resistance genes, stress survival aids, transporters, membrane biogenesis genes, amino acid metabolism and transport, transcriptional regulators, energy production genes, and the previously reported <it>fliF </it>and <it>virB </it>operons.</p> <p>Conclusions</p> <p>VjbR and C₁₂-HSL regulate expression of a large and diverse number of genes. Many genes identified as virulence factors in other bacterial pathogens were found to be differently expressed, suggesting an important contribution to intracellular survival of <it>Brucella</it>. From these data, we conclude that VjbR and C₁₂-HSL contribute to virulence and survival by regulating expression of virulence mechanisms and thus controlling the ability of the bacteria to survive within the host cell. A likely scenario occurs via QS, however, operation of such a mechanism remains to be demonstrated.</p

Correlative Gene Expression to Protective Seroconversion in Rift Valley Fever Vaccinates

Rift Valley fever Virus (RVFV), a negative-stranded RNA virus, is the etiological agent of the vector-borne zoonotic disease, Rift Valley fever (RVF). In both humans and livestock, protective immunity can be achieved through vaccination. Earlier and more recent vaccine trials in cattle and sheep demonstrated a strong neutralizing antibody and total IgG response induced by the RVF vaccine, authentic recombinant MP-12 (arMP-12). From previous work, protective immunity in sheep and cattle vaccinates normally occurs from 7 to 21 days after inoculation with arMP-12. While the serology and protective response induced by arMP-12 has been studied, little attention has been paid to the underlying molecular and genetic events occurring prior to the serologic immune response. To address this, we isolated RNA from whole blood of vaccinated calves over a time course of 21 days before and after vaccination with arMP-12. The time course RNAs were sequenced by RNASeq and bioinformatically analyzed. Our results revealed time-dependent activation or repression of numerous gene ontologies and pathways related to the vaccine induced immune response and its regulation. Additional bioinformatic analyses identified a correlative relationship between specific host immune response genes and protective immunity prior to the detection of protective serum neutralizing antibody responses. These results contribute an important proof of concept for identifying molecular and genetic components underlying the immune response to RVF vaccination and protection prior to serologic detection.The open access fee for this work was funded through the Texas A&M University Open Access to Knowledge (OAK) Fund

Global Rsh-dependent transcription profile of Brucella suis during stringent response unravels adaptation to nutrient starvation and cross-talk with other stress responses

International audienceBackground: In the intracellular pathogen Brucella spp., the activation of the stringent response, a global regulatorynetwork providing rapid adaptation to growth-affecting stress conditions such as nutrient deficiency, is essential forreplication in the host. A single, bi-functional enzyme Rsh catalyzes synthesis and hydrolysis of the alarmone (p)ppGpp, responsible for differential gene expression under stringent conditions.Results: cDNA microarray analysis allowed characterization of the transcriptional profiles of the B. suis 1330 wildtypeand Δrsh mutant in a minimal medium, partially mimicking the nutrient-poor intramacrophagic environment.A total of 379 genes (11.6% of the genome) were differentially expressed in a rsh-dependent manner, of which 198were up-, and 181 were down-regulated. The pleiotropic character of the response was confirmed, as the genesencoded an important number of transcriptional regulators, cell envelope proteins, stress factors, transport systems,and energy metabolism proteins. Virulence genes such as narG and sodC, respectively encoding respiratory nitratereductase and superoxide dismutase, were under the positive control of (p)ppGpp, as well as expression of thecbb3-type cytochrome c oxidase, essential for chronic murine infection. Methionine was the only amino acid whosebiosynthesis was absolutely dependent on stringent response in B. suis.Conclusions: The study illustrated the complexity of the processes involved in adaptation to nutrient starvation,and contributed to a better understanding of the correlation between stringent response and Brucella virulence.Most interestingly, it clearly indicated (p)ppGpp-dependent cross-talk between at least three stress responsesplaying a central role in Brucella adaptation to the host: nutrient, oxidative, and low-oxygen stress

Use of simplified claustrophobia questionnaire in predicting adherence to positive airway pressure (PAP) therapy in obstructive sleep apnea (OSA) patients

TITLE: Use of simplified claustrophobia questionnaire in predicting adherence to continuous positive airway pressure (CPAP) in obstructive sleep apnea (OSA) patients. Introduction: Claustrophobia could affect adherence to CPAP in sleep apnea patients. High score in a claustrophobia questionnaire containing 12 restriction and 14 suffocation items was associated to poor CPAP adherence in previous research. The restriction and suffocation items were equivalent on predicting CPAP adherence which allowed to limit the survey to only the suffocation questionnaire. The goal of this study is to find the predictability of CPAP adherence for each question of the suffocation questionnaire. Methods: We performed a prospective chart review of 114 patients with newly diagnosed OSA using home sleep apnea testing (HSAT). On initial consultation, patients were provided with a suffocation claustrophobia questionnaire. Patients’ demographics, home sleep study data and objective adherence data were collected within the first 3 months of usage.Results: Items 2 (OR =1.67, p-value = 0.049), 3 (OR=1.60, p-value = 0.021), and 13 (OR= 1.52, p-value = 0.056) are most promising in association with non-adherence. Results indicate that higher scores on item 2 is associated with higher odds of non-adherence to CPAP. Specifically, each point increase on item 2 was associated with a 67% increase in odds of non-adherence. However, these items alone do not show a large effect in providing accurate classification of adherence. Of these items, item 3 had the lower rate of missing data, suggesting that it may be the most patient-friendly item. Conclusions: Based on these results, the 3 questions with highest predictability for CPAP adherence will be studied in the clinical arena to address feasibility and predictability.https://scholarlycommons.henryford.com/merf2019clinres/1013/thumbnail.jp

Exploring the radio-loudness of SDSS quasars with spectral stacking

© 2024 The Author(s). Published by Oxford University Press on behalf of Royal Astronomical Society. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY), https://creativecommons.org/licenses/by/4.0/We use new 144 MHz observations over 5634 deg 2 from the LOFAR (Low Frequency Array) Two-metre Sky Survey (LoTSS) to compile the largest sample of uniformly selected, spectroscopically confirmed quasars from the 14th data release of the Sloan Digital Sky Survey (SDSS-DR14). Using the classical definition of radio loudness, R = log (L 1.4GHz/L i), we identify 3697 radio-loud (RL) and 111 132 radio-quiet (RQ) sources at 0.6 < z < 3.4. To study their properties, we develop a new rest-frame spectral stacking algorithm, designed with forthcoming massively multiplexed spectroscopic surveys in mind, and use it to create high signal-to-noise composite spectra of each class, matched in redshift and absolute i-band magnitude. We show that RL quasars have redder continuum and enhanced [O II] emission than their RQ counterparts. These results persist when additionally matching in black hole mass, suggesting that this parameter is not the defining factor in making a quasi-stellar object (QSO) RL. We find that these features are not gradually varying as a function of radio loudness, but are maintained even when probing deeper into the RQ population, indicating that a clear-cut division in radio loudness is not apparent. Upon examining the star formation rates (SFRs) inferred from the [O II] emission line, with the contribution from active galactic nucleus removed using the [Ne V] line, we find that RL quasars have a significant excess of star formation relative to RQ quasars out to z = 1.9 at least. Given our findings, we suggest that RL sources either preferably reside in gas-rich systems with rapidly spinning black holes, or represent an earlier obscured phase of QSO evolution.Peer reviewe

Molecular rationale for the use of PI3K/AKT/mTOR pathway inhibitors in combination with crizotinib in ALK-mutated neuroblastoma

Mutations in the ALK tyrosine kinase receptor gene represent important therapeutic targets in neuroblastoma, yet their clinical translation has been challenging. The ALKF1174L mutation is sensitive to the ALK inhibitor crizotinib only at high doses and mediates acquired resistance to crizotinib in ALK-translocated cancers. We have shown that the combination of crizotinib and an inhibitor of downstream signaling induces a favorable response in transgenic mice bearing ALKF1174L/MYCN-positive neuroblastoma. Here, we investigated the molecular basis of this effect and assessed whether a similar strategy would be effective in ALK-mutated tumors lacking MYCN overexpression. We show that in ALK-mutated, MYCN-amplified neuroblastoma cells, crizotinib alone does not affect mTORC1 activity as indicated by persistent RPS6 phosphorylation. Combined treatment with crizotinib and an ATP-competitive mTOR inhibitor abrogated RPS6 phosphorylation, leading to reduced tumor growth and prolonged survival in ALKF1174L/MYCN-positive models compared to single agent treatment. By contrast, this combination, while inducing mTORC1 downregulation, caused reciprocal upregulation of PI3K activity in ALK-mutated cells expressing wild-type MYCN. Here, an inhibitor with potency against both mTOR and PI3K was more effective in promoting cytotoxicity when combined with crizotinib. Our findings should enable a more precise selection of molecularly targeted agents for patients with ALK-mutated tumors

Mars Sample Handling and Requirements Panel (MSHARP)

In anticipation of the return of samples from Mars toward the end of the first decade of the next century, NASA's Office of Space Sciences chartered a panel to examine how Mars samples should be handled. The panel was to make recommendations in three areas: (1) sample collection and transport back to Earth; (2) certification of the samples as nonhazardous; and (3) sample receiving, curation, and distribution. This report summarizes the findings of that panel. The samples should be treated as hazardous until proven otherwise. They are to be sealed within a canister on Mars, and the canister is not to be opened until within a Biosafety Hazard Level 4 (BSL-4) containment facility here on Earth. This facility must also meet or exceed the cleanliness requirements of the Johnson Space Center (JSC) facility for curation of extraterrestrial materials. A containment facility meeting both these requirements does not yet exist. Hazard assessment and life detection experiments are to be done at the containment facility, while geochemical characterization is being performed on a sterilized subset of the samples released to the science community. When and if the samples are proven harmless, they are to be transferred to a curation facility, such as that at JSC

Radio emission and mass loss rate limits of four young solar-type stars

Aims. Observations of free-free continuum radio emission of four young main-sequence solar-type stars (EK Dra, p1UMa, ?1Ori, and ?1Cet) are studied to detect stellar winds or at least to place upper limits on their thermal radio emission, which is dominated by the ionized wind. The stars in our sample are members of The Sun in Time programme and cover ages of ~0.1-0.65 Gyr on the main-sequence. They are similar in magnetic activity to the Sun and thus are excellent proxies for representing the young Sun. Upper limits on mass loss rates for this sample of stars are calculated using their observational radio emission. Our aim is to re-examine the faint young Sun paradox by assuming that the young Sun was more massive in its past, and hence to find a possible solution for this famous problem. Methods. The observations of our sample are performed with the Karl G. Jansky Very Large Array (VLA) with excellent sensitivity, using the C-band receiver from 4-8 GHz and the Ku-band from 12-18 GHz. Atacama Large Millimeter/Submillitmeter Array (ALMA) observations are performed at 100 GHz. The Common Astronomy Software Application (CASA) package is used for the data preparation, reduction, calibration, and imaging. For the estimation of the mass loss limits, spherically symmetric winds and stationary, anisotropic, ionized winds are assumed. We compare our results to 1) mass loss rate estimates of theoretical rotational evolution models; and 2) to results of the indirect technique of determining mass loss rates: Lyman-a absorption. Results. We are able to derive the most stringent direct upper limits on mass loss so far from radio observations. Two objects, EK Dra and ?1Ori, are detected at 6 and 14 GHz down to an excellent noise level. These stars are very active and additional radio emission identified as non-thermal emission was detected, but limits for the mass loss rates of these objects are still derived. The emission of ?1Ori does not come from the main target itself, but from its M-dwarf companion. The stars p1UMa and ?1Cet were not detected in either C-band or in Ku-band. For these objects we give upper limits to their radio free-free emission and calculate upper limits to their mass loss rates. Finally, we reproduce the evolution of the Sun and derive an estimate for the solar mass of the Sun at a younger age

Systems Biology Analysis of Brucella Infected Peyers Patch Reveals Rapid Invasion with Modest Transient Perturbations of the Host Transcriptome

Brucella melitensis causes the most severe and acute symptoms of all Brucella species in human beings and infects hosts primarily through the oral route. The epithelium covering domed villi of jejunal-ileal Peyer’s patches is an important site of entry for several pathogens, including Brucella. Here, we use the calf ligated ileal loop model to study temporal in vivo Brucella-infected host molecular and morphological responses. Our results document Brucella bacteremia occurring within 30 min after intraluminal inoculation of the ileum without histopathologic traces of lesions. Based on a system biology Dynamic Bayesian Network modeling approach (DBN) of microarray data, a very early transient perturbation of the host enteric transcriptome was associated with the initial host response to Brucella contact that is rapidly averted allowing invasion and dissemination. A detailed analysis revealed active expression of Syndecan 2, Integrin alpha L and Integrin beta 2 genes, which may favor initial Brucella adhesion. Also, two intestinal barrier-related pathways (Tight Junction and Trefoil Factors Initiated Mucosal Healing) were significantly repressed in the early stage of infection, suggesting subversion of mucosal epithelial barrier function to facilitate Brucella transepithelial migration. Simultaneously, the strong activation of the innate immune response pathways would suggest that the host mounts an appropriate protective immune response; however, the expression of the two key genes that encode innate immunity anti-Brucella cytokines such as TNF-a and IL12p40 were not significantly changed throughout the study. Furthermore, the defective expression of Toll-Like Receptor Signaling pathways may partially explain the lack of proinflammatory cytokine production and consequently the absence of morphologically detectable inflammation at the site of infection. Cumulatively, our results indicate that the in vivo pathogenesis of the early infectious process of Brucella is primarily accomplished by compromising the mucosal immune barrier and subverting critical immune response mechanisms.The open access fee for this work was funded through the Texas A&M University Open Access to Knowledge (OAK) Fund