A real-time reverse transcription-polymerase chain reaction (RT-PCR) assay was developed to rapidly detect the severe acute respiratory syndrome-associated coronavirus (SARS-CoV). The assay, based on ...multiple primer and probe sets located in different regions of the SARS-CoV genome, could discriminate SARS-CoV from other human and animal coronaviruses with a potential detection limit of <10 genomic copies per reaction. The real-time RT-PCR assay was more sensitive than a conventional RT-PCR assay or culture isolation and proved suitable to detect SARS-CoV in clinical specimens. Application of this assay will aid in diagnosing SARS-CoV infection.
Summary
The severity of infections caused by Salmonella enterica serovar Typhimurium varies depending on the host species. Numerous virulence genes have been identified in S. Typhimurium, largely ...from studies in mice, but their roles in infections of other species remain unclear. In the most comprehensive survey of its kind, through the use of signature‐tagged mutagenesis of S. Typhimurium we have identified mutants that were unable to colonize calf intestines, mutants unable to colonize chick intestines and mutants unable to colonize both species. The type three secretion systems encoded on Salmonella pathogenicity islands (SPIs) 1 and 2 were required for efficient colonization of cattle. However, disruption of these secretion systems only caused a minor defect in S. Typhimurium colonization of chicks. Transposon insertions in SPI‐4 compromised S. Typhimurium colonization of cattle, but not chicks. This is the first data confirming a role for SPI‐4 in pathogenesis. We have also been able to ascribe a role in colonization for cell surface polysaccharides, cell envelope proteins, and many ‘housekeeping’ genes and genes of unknown function. We conclude that S. Typhimurium uses different strategies to colonize calves and chicks. This has major implications for vaccine design.
Familial adenomatous polyposis (FAP) is a hereditary colorectal cancer syndrome, which results in the development of hundreds of adenomatous polyps carpeting the gastrointestinal tract. NSAIDs have ...reduced polyp burden in patients with FAP and synthetic rexinoids have demonstrated the ability to modulate cytokine-mediated inflammation and WNT signaling. This study examined the use of the combination of an NSAID (sulindac) and a rexinoid (bexarotene) as a durable approach for reducing FAP colonic polyposis to prevent colorectal cancer development. Whole transcriptomic analysis of colorectal polyps and matched normal mucosa in a cohort of patients with FAP to identify potential targets for prevention in FAP was performed. Drug-dose synergism of sulindac and bexarotene in cell lines and patient-derived organoids was assessed, and the drug combination was tested in two different mouse models. This work explored mRNA as a potential predictive serum biomarker for this combination in FAP. Overall, transcriptomic analysis revealed significant activation of inflammatory and cell proliferation pathways. A synergistic effect of sulindac (300 μmol/L) and bexarotene (40 μmol/L) was observed in FAP colonic organoids with primary targeting of polyp tissue compared with normal mucosa. This combination translated into a significant reduction in polyp development in
and
mice. Finally, the reported data suggest miRNA-21 could serve as a predictive serum biomarker for polyposis burden in patients with FAP. These findings support the clinical development of the combination of sulindac and bexarotene as a treatment modality for patients with FAP. PREVENTION RELEVANCE: This study identified a novel chemopreventive regimen combining sulindac and bexarotene to reduce polyposis in patients with FAP using
tools,
, and
models. This investigation provides the essential groundwork for moving this drug combination forward into a clinical trial.
A protective vaccine against hepatitis C virus (HCV) remains an unmet clinical need. HCV infects millions of people worldwide and is a leading cause of liver cirrhosis and hepatocellular cancer. ...Animal challenge experiments, immunogenetics studies, and assessment of host immunity during acute infection highlight the critical role that effective T cell immunity plays in viral control. In this first-in-man study, we have induced antiviral immunity with functional characteristics analogous to those associated with viral control in natural infection, and improved upon a vaccine based on adenoviral vectors alone. We assessed a heterologous prime-boost vaccination strategy based on a replicative defective simian adenoviral vector (ChAd3) and modified vaccinia Ankara (MVA) vector encoding the NS3, NS4, NS5A, and NS5B proteins of HCV genotype 1b. Analysis used single-cell mass cytometry and human leukocyte antigen class I peptide tetramer technology in healthy human volunteers. We show that HCV-specific T cells induced by ChAd3 are optimally boosted with MVA, and generate very high levels of both CD8(+) and CD4(+) HCV-specific T cells targeting multiple HCV antigens. Sustained memory and effector T cell populations are generated, and T cell memory evolved over time with improvement of quality (proliferation and polyfunctionality) after heterologous MVA boost. We have developed an HCV vaccine strategy, with durable, broad, sustained, and balanced T cell responses, characteristic of those associated with viral control, paving the way for the first efficacy studies of a prophylactic HCV vaccine.
The ability to perceive the world is not merely a passive process but depends on sensorimotor loops and interactions that guide and actively bias our sensory systems. Understanding which and how ...cognitive processes participate in this active sensing is still an open question. In this context, the auditory system presents itself as an attractive model for this purpose as it features an efferent control network that projects from the cortex to subcortical nuclei and even to the sensory epithelium itself. This efferent system can regulate the cochlear amplifier sensitivity through medial olivocochlear (MOC) neurons located in the brainstem. The ability to suppress irrelevant sounds during selective attention to visual stimuli is one of the functions that have been attributed to this system. MOC neurons are also directly activated by sounds through a brainstem reflex circuit, a response linked to the ability to suppress auditory stimuli during visual attention. Human studies have suggested that MOC neurons are also recruited by other cognitive functions, such as working memory and predictability. The aim of this research was to explore whether cognitive processes related to delayed responses in a visual discrimination task were associated with MOC function. In this behavioral condition, chinchillas held their responses for more than 2.5 s after visual stimulus offset, with and without auditory distractors, and the accuracy of these responses was correlated with the magnitude of the MOC reflex. We found that the animals' performance decreased in presence of auditory distractors and that the results observed in MOC reflex could predict this performance. The individual MOC strength correlated with behavioral performance during delayed responses with auditory distractors, but not without them. These results in chinchillas, suggest that MOC neurons are also recruited by other cognitive functions, such as working memory.
A trifunctional approach: 3‐hydroxy‐4‐pyridinones that contain phenol groups for antioxidant functionality are further elaborated with pendant glucosyl moieties for improved blood–brain barrier ...targeting (see structure; R=phenyl, 4‐hydroxyphenyl). Glycosidase removal of the carbohydrate substituents gives ligands that are ready to passivate excess metal ions, especially copper and zinc, in the brain. These molecules are potential prodrugs for treatment of neurodegenerative diseases, including Alzheimer's disease.
Materials capable of releasing reactive oxygen species (ROS) can display antibacterial and anticancer activity, and may also have anti-oxidant capacity if they suppress intracellular ROS (
e.g.
...nitric oxide, NO) resulting in anti-inflammatory activity. Herein we report silver phosphate (Ag
3
PO
4
)/polyindole (Pln) nanocomposites which display antibacterial, anticancer and anti-inflammatory activity, and have therefore potential for a variety of biomedical applications.
Materials capable of releasing reactive oxygen species (ROS) can display antibacterial and anticancer activity, and may also have antioxidant capacity if they suppress intracellular ROS (
e.g.
nitric oxide, NO) resulting in anti-inflammatory activity.
OBJECTIVES: To assess variation in the association between blood pressure (BP) and risk for dementia across a spectrum of older ages and to examine BP changes before dementia onset.
DESIGN: ...Prospective cohort study.
SETTING: A large health maintenance organization in Seattle, Washington.
PARTICIPANTS: A cohort of 2,356 members of a large health maintenance organization aged 65 and older who were initially without dementia.
MEASUREMENTS: Dementia diagnosis was assessed biennially, and systolic (SBP) and diastolic BP (DBP) were measured at baseline and at four follow‐up assessments. Cox proportional hazards models were used to estimate hazard ratios (HRs) for dementia and Alzheimer's disease (AD) associated with baseline BP in different age groups.
RESULTS: Within the youngest age group (65–74 at enrollment) a greater risk for dementia was found in participants with high SBP (≥160 mmHg) (hazard ratio (HR)=1.60, 95% confidence interval (CI)=1.01–2.55) or borderline‐high DBP (80–89 mmHg) (HR=1.59, 95% CI=1.07–2.35) than for those with normal BP (SBP <140 mmHg and DBP <80 mmHg). The dementia risk associated with SBP declined with increasing age (SBP‐by‐age interaction, P=.01). SBP declined similarly with aging in subjects who developed dementia and those who did not. Thus, in this sample, the association between SBP and dementia risk was not dependent on when BP was measured in relation to onset of dementia.
CONCLUSION: High SBP was associated with greater risk of dementia in the young elderly (<75) but not in older subjects. Adequate control of hypertension in early old age may reduce the risk for dementia.
Silica accumulation in the Yangtze Sea during the Ordovician–Silurian (O–S) transition appears to have coincided with global climatic fluctuations, widespread upwelling, and volcanism. There is a ...need to further evaluate their respective contributions to silica deposition and potential relationships among these factors. The current study selected siliceous deposits in the Wufeng and Longmaxi Formations from four sections spanning the inner to outer Yangtze Sea, South China, to gain a deeper understanding of the climatic and oceanographic evolution associated with silica enrichment. Al/(Al + Fe + Mn) values, the presence of radiolarians, and Si isotope values of samples recovered from the investigated shale successions offer compelling evidence that the silica is largely of biogenic origin with some terrigenous contributions. Further, various productivity and redox proxies suggest that biogenic silica (BSi) accumulated under conditions of enhanced marine productivity and anoxic bottom water conditions. Hg/TOC and Zr/Al2O3 profiles suggest intermittent volcanism during the BSi deposition in the Yangtze Sea. However, the lack of correlation between BSi and Hg/TOC values indicates that volcanic iron fertilization was not responsible for BSi accumulation. Instead, most BSi-rich samples are dominated by low MnEF × CoEF values (<0.5), consistent with BSi deposited in modern upwelling settings. Hydrographic reconstruction based on Mo–U covariation indicates a more open water setting in the outer Yangtze Sea, while the coeval inner Yangtze Sea was relatively restricted. Therefore, upwelling events appear to have been more vigorous in the outer Yangtze Sea. Published and new Chemical Index of Alteration (CIA), BSi, and MnEF × CoEF data for the Wufeng and Longmaxi Formations across the inner to outer Yangtze Sea demonstrate that temporal and spatial variations of BSi were controlled by climate-driven upwelling. In particular, cool-water upwelling contemporaneous with Hirnantian glaciation may have been responsible for the establishment of the cool-water fauna of the shallow-water Guanyinqiao Bed and enhanced silica deposition in deeper water. Moreover, a moderate negative relationship between compiled CIA and BSi contents suggests that enhanced upwelling driving BSi accumulation appears to have been favored during cooling events. Integrated analysis of BSi deposits of the Laurentia and Baltica continental margins further suggests that BSi accumulation on continental margins during the O–S transition was primarily influenced by global cooling. Therefore, we suggest that wind patterns or/and thermohaline circulation, influenced by climate fluctuations, induced widespread cold water upwelling events during the O–S transition. Moreover, elevated BSi production diluted accumulating OM resulting in the observed parabolic relationship of BSi and TOC.
•The O–S siliceous deposits derived from enhanced BSi productivity.•Upwelling was the first-order constraint on BSi distribution.•The upwelling events appeared to correlate with the global cooling.
, a tier 1 select agent and the etiological agent of melioidosis, transitions from soil and aquatic environments to infect a variety of vertebrate and invertebrate hosts. During the transition from ...an environmental saprophyte to a mammalian pathogen,
encounters and responds to rapidly changing environmental conditions. Environmental sensing systems that control cellular levels of cyclic di-GMP promote pathogen survival in diverse environments. Cyclic di-GMP controls biofilm production, virulence factors, and motility in many bacteria. This study is an evaluation of cyclic di-GMP-associated genes that are predicted to metabolize and interact with cyclic di-GMP as identified from the annotated genome of
1026b. Mutants containing transposon disruptions in each of these genes were characterized for biofilm formation and motility at two temperatures that reflect conditions that the bacteria encounter in the environment and during the infection of a mammalian host. Mutants with transposon insertions in a known phosphodiesterase (
) and a predicted hydrolase (Bp1026b_I2285) gene exhibited decreased motility regardless of temperature. In contrast, the phenotypes exhibited by mutants with transposon insertion mutations in a predicted diguanylate cyclase gene (Bp1026b_II2523) were strikingly influenced by temperature and were dependent on a conserved GG(D/E)EF motif. The transposon insertion mutant exhibited enhanced biofilm formation at 37°C but impaired biofilm formation at 30°C. These studies illustrate the importance of studying behaviors regulated by cyclic di-GMP under varied environmental conditions in order to better understand cyclic di-GMP signaling in bacterial pathogens.
This report evaluates predicted cyclic di-GMP binding and metabolic proteins from
1026b, a tier 1 select agent and the etiologic agent of melioidosis. Transposon insertion mutants with disruptions in each of the genes encoding these predicted proteins were characterized in order to identify key components of the
cyclic di-GMP-signaling network. A predicted hydrolase and a phosphodiesterase that modulate swimming motility were identified, in addition to a diguanylate cyclase that modulates biofilm formation and motility in response to temperature. These studies warrant further evaluation of the contribution of cyclic di-GMP to melioidosis in the context of pathogen acquisition from environmental reservoirs and subsequent colonization, dissemination, and persistence within the host.
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