Homeostasis of the gut microbiota critically influences host health and aging. Developing genetically engineered probiotics holds great promise as a new therapeutic paradigm to promote healthy aging. ...Here, through screening 3,983 Escherichia coli mutants, we discovered that 29 bacterial genes, when deleted, increase longevity in the host Caenorhabditis elegans. A dozen of these bacterial mutants also protect the host from age-related progression of tumor growth and amyloid-beta accumulation. Mechanistically, we discovered that five bacterial mutants promote longevity through increased secretion of the polysaccharide colanic acid (CA), which regulates mitochondrial dynamics and unfolded protein response (UPRmt) in the host. Purified CA polymers are sufficient to promote longevity via ATFS-1, the host UPRmt-responsive transcription factor. Furthermore, the mitochondrial changes and longevity effects induced by CA are conserved across different species. Together, our results identified molecular targets for developing pro-longevity microbes and a bacterial metabolite acting on host mitochondria to promote longevity.
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•Systematic analysis of longevity-promoting microbial genetic variations•Colanic acid as a pro-longevity natural compound effective in different species•Bacterial metabolites regulate host mitochondrial dynamics and UPRmit
The genetic composition of gut microbes controls the production of metabolites that impact host longevity.
Sensitive and specific visualization of small biomolecules in living systems is highly challenging. We report stimulated Raman-scattering imaging of alkyne tags as a general strategy for studying a ...broad spectrum of small biomolecules in live cells and animals. We demonstrate this technique by tracking alkyne-bearing drugs in mouse tissues and visualizing de novo synthesis of DNA, RNA, proteins, phospholipids and triglycerides through metabolic incorporation of alkyne-tagged small precursors.
Microorganisms and their hosts share the same environment, and microbial metabolic molecules (metabolites) exert crucial effects on host physiology. Environmental factors not only shape the ...composition of the host's resident microorganisms, but also modulate their metabolism. However, the exact molecular relationship among the environment, microbial metabolites and host metabolism remains largely unknown. Here, we discovered that environmental methionine tunes bacterial methyl metabolism to regulate host mitochondrial dynamics and lipid metabolism in Caenorhabditis elegans through an endocrine crosstalk involving NR5A nuclear receptor and Hedgehog signalling. We discovered that methionine deficiency in bacterial medium decreases the production of bacterial metabolites that are essential for phosphatidylcholine synthesis in C. elegans. Reductions of diundecanoyl and dilauroyl phosphatidylcholines attenuate NHR-25, a NR5A nuclear receptor, and release its transcriptional suppression of GRL-21, a Hedgehog-like protein. The induction of GRL-21 consequently inhibits the PTR-24 Patched receptor cell non-autonomously, resulting in mitochondrial fragmentation and lipid accumulation. Together, our work reveals an environment-microorganism-host metabolic axis regulating host mitochondrial dynamics and lipid metabolism, and discovers NR5A-Hedgehog intercellular signalling that controls these metabolic responses with critical consequences for host health and survival.
Purpose To examine the relationship between pre-corneal and pre-contact lens tear film stability (TFS), and to determine whether pre-corneal TFS is a reliable predictor of subsequent pre-lens TFS ...after a contact lens is placed on the eye. Methods 667 records met inclusion criteria and were extracted from a soft contact lens multi-study database. Multivariable linear mixed effects models were fit to examine the association between pre-corneal and pre-lens TFS, adjusting for potential confounders and accounting for repeated measures. Receiver Operating Characteristic (ROC) analysis was employed to assess the predictive performance of pre-corneal TFS for subsequent pre-lens TFS. TFS was quantified for this analysis as the non-invasive tear breakup time (NITBUT). Results Pre-corneal NITBUT was significantly related to the pre-lens NITBUT at both 10 min (p<0.001) and 2-6 hrs (p<0.001) post-lens insertion. However, the sensitivities of pre-corneal NITBUT for predicting symptom-associated thresholds of pre-lens NITBUT ranged from 50-65%, and specificities ranged from 57-72%, suggesting poor-to-moderate diagnostic performance. Conclusions Despite the association of pre-corneal and pre-lens TFS, the inherent lability and sensitivity to environmental exposures of the tear film introduce significant variability into NITBUT measurements. Using pre-corneal NITBUT to identify likely successful contact lens candidates prior to fitting is thus not sufficiently accurate to be relied upon in the clinical setting.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Glutathione plays many important roles in biological processes; however, the dynamic changes of glutathione concentrations in living cells remain largely unknown. Here, we report a reversible ...reaction-based fluorescent probe-designated as RealThiol (RT)-that can quantitatively monitor the real-time glutathione dynamics in living cells. Using RT, we observe enhanced antioxidant capability of activated neurons and dynamic glutathione changes during ferroptosis. RT is thus a versatile tool that can be used for both confocal microscopy and flow cytometry based high-throughput quantification of glutathione levels in single cells. We envision that this new glutathione probe will enable opportunities to study glutathione dynamics and transportation and expand our understanding of the physiological and pathological roles of glutathione in living cells.
Current efforts in the proteolysis targeting chimera (PROTAC) field mostly focus on choosing an appropriate E3 ligase for the target protein, improving the binding affinities towards the target ...protein and the E3 ligase, and optimizing the PROTAC linker. However, due to the large molecular weights of PROTACs, their cellular uptake remains an issue. Through comparing how different warhead chemistry, reversible noncovalent (RNC), reversible covalent (RC), and irreversible covalent (IRC) binders, affects the degradation of Bruton's Tyrosine Kinase (BTK), we serendipitously discover that cyano-acrylamide-based reversible covalent chemistry can significantly enhance the intracellular accumulation and target engagement of PROTACs and develop RC-1 as a reversible covalent BTK PROTAC with a high target occupancy as its corresponding kinase inhibitor and effectiveness as a dual functional inhibitor and degrader, a different mechanism-of-action for PROTACs. Importantly, this reversible covalent strategy is generalizable to improve other PROTACs, opening a path to enhance PROTAC efficacy.
PURPOSE:Women have a higher dry eye disease prevalence compared with men, although only relatively minor differences in the ocular surface have been observed. Interestingly, a sex difference in pain ...sensitivity is known, and recent research suggests that pain sensitivity is associated with dry eye symptoms. This study attempts to discern whether the association between pain sensitivity and dry eye symptoms varies between women and men.
METHODS:In this prospective cross-sectional study, subjects were seen for one visit where they were asked to fill out a set of questionnaires consisting of the Pain Sensitivity Questionnaire, Ocular Surface Disease Index (OSDI), and other dry eye questionnaires. This was followed by an ocular surface assessment on both eyes.
RESULTS:Two hundred eighty-seven subjects (194 women, 93 men) completed the study. Intersex differences in the ocular surface were noted. Even after accounting for these differences, an interaction effect between sex and Pain Sensitivity Questionnaire-minor score on dry eye symptoms was observed, with only women noting increased symptoms on the OSDI (P < 0.005) and other dry eye questionnaires (P values ranging from 0.01 to <0.005) with greater pain sensitivity. After controlling for other variables, women with the highest pain sensitivity had a 17-point higher OSDI score and greater symptoms, as reported by all the other dry questionnaires compared with their male counterparts.
CONCLUSIONS:The role of pain sensitivity on dry eye symptoms appears to vary between women and men. This difference provides insight into why women have a significantly higher dry eye disease prevalence than men.
Assess potential risk factors for severe Meibomian gland atrophy (SMGA) in a young adult population.
Cross-sectional study using medical history and ocular surface examination to evaluate ...relationships with study outcomes: SMGA, tear lipid layer (TLL) thickness, non-invasive (NITBUT) and fluorescein (FTBUT) tear breakup times, and symptoms using the Standard Patient Evaluation of Eye Dryness (SPEED) questionnaire.
One hundred one participants (101; 202 eyes; Age: mean±SD = 22.3±4.0 years) completed the study. Hormonal birth control (HBC) use was the only significant risk factor for SMGA (p = 0.028). Female HBC users had 4.8 times greater odds of having SMGA compared to female HBC non-users (p = 0.028), but the odds of having SMGA was similar between female HBC non-users and males (p = 0.885). Multivariable analysis suggested that the relationship between SMGA and TLL thickness was dependent on HBC use. Compared to female HBC non-users without SMGA, TLL thickness for HBC users was estimated to be 10 nm thinner if SMGA was absent (p = 0.007) and 21 nm thinner if SMGA was present (p<0.001). SMGA status had no significant impact on TLL thickness among female HBC non-users (p = 0.552). The effect of TLL thickness on FTBUT was small but significant (p = 0.026). TLL thickness was not significantly associated with NITBUT (p = 0.349). Neither FTBUT nor NITBUT was significantly associated with the SPEED score.
HBC use may be associated with SMGA, supporting the hypothesis that SMGA could lead to thinner TLL. However, less evidence was present to support that thin TLL could lead to clinically detectable tear film instability and subsequently to increased ocular dryness symptoms. Further investigation with a larger sample size is warranted to confirm these findings.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
This review summarizes the current state of knowledge regarding interfacial properties of very complex biological colloids, specifically, human meibum and tear lipids, and their interactions with ...proteins similar to the proteins found in aqueous part of human tears. Tear lipids spread as thin films over the surface of tear-film aqueous and play crucial roles in tear-film stability and overall ocular-surface health. The vast majority of papers published to date report interfacial properties of meibum-lipid monolayers spread on various aqueous sub-phases, often containing model proteins, in Langmuir trough. However, it is well established that natural human ocular tear lipids exist as multilayered films with a thickness between 30 and 100nm, that is very much disparate from 1 to 2nm thick meibum monolayers. We employed sessile-bubble tensiometry to study the dynamic interfacial and rheological properties of reconstituted multilayered human tear-lipid films. Small amounts (0.5–1μg) of human tear lipids were deposited on an air-bubble surface to produce tear-lipid films in thickness range 30–100nm corresponding to ocular lipid films. Thus, we were able to overcome major Langmuir-trough method limitations because ocular tear lipids can be safely harvested only in minute, sub-milligram quantities, insufficient for Langmuir through studies. Sessile-bubble method is demonstrated to be a versatile tool for assessing conventional synthetic surfactants adsorption/desorption dynamics at an air-aqueous solution interface. (Svitova T., Weatherbee M., Radke C.J. Dynamics of surfactant sorption at the air/water interface: continuous-flow tensiometry. J. Colloid Interf. Sci. 2003;261:1170–179). The augmented flow-sessile-bubble setup, with step-strain relaxation module for dynamic interfacial rheological properties and high-precision syringe pump to generate larger and slow interfacial area expansions-contractions, was developed and employed in our studies. We established that this method is uniquely suitable for examination of multilayered lipid-film interfacial properties. Recently it was compellingly proven that chemical composition of human tear lipids extracted from whole tears is substantially different from that of meibum lipids. To be exact, healthy human tear lipids contain 8–16% of polar lipids, similar to lung lipids, and they are mostly double-tailed phospholipids, with C16 and longer alkyl chains. Rationally, one would assume that the results obtained for meibum lipids, devoid of surface-active components such as phospholipids, and, above all, in a form of monolayers, are not pertinent or useful for elucidating behavior and stability of an averaged 60-nm thick ocular tear-lipid films in vivo. The advantage of sessile-bubble technique, specifically, using a small amount of lipids required to attain multilayered films, unlocks the prospect of evaluating and comparing the interfacial properties of human tear lipids collected from a single individual, typically 100–150μg. This is in sharp contrast with several milligrams of lipids that would be required to build equally thick films for Langmuir-trough experiments. The results of our studies provided in-depth understanding of the mechanisms responsible for properties and stability of human tear-lipid films in vivo. Here we summarize recent publications and our latest findings regarding human tear-lipid interfacial properties, their chemical composition, and their interaction with model proteins mimicking the proteins found in human tear-aqueous phase.
Graphic abstract
Dynamic interfacial properties of human tear lipids and other tear constituents: Sessile bubble tensiometry studies Tatyana F. Svitova, Meng C. Lin Display omitted
•Human tear films are extremely complex biochemical systems remaining not well studied and understood.•Tear films physico-chemical and dynamic interfacial properties affect human eye health and vision clarity.•Tear film stability is largely dependent on lipid films interfacial dynamics.•Dynamic interfacial properties of tear-lipid films depend on film thickness, subjects’ race and presence of proteins.•Knowledge of tear-lipids interfacial dynamics provides new understanding of tear-film stability and dry-eye syndrome.
To utilize the Pain Sensitivity Questionnaire (PSQ) to assess the influence of pain sensitivity on perceptions of ocular discomfort and dryness.
Subjects completed a battery of questionnaires, ...including history of ocular and general health, contact lens wear history, the Ocular Surface Disease Index (OSDI) questionnaire, visual analog scale (VAS) 100-point rating scales to assess severity and frequency of average and end of day (EOD) discomfort and dryness, and the PSQ to assess pain sensitivity level. Masked subjects were then instructed to wear one inverted and one normally oriented soft contact lens contralaterally for 30 minutes to induce an inter-eye difference in comfort and dryness sensations. Subjects rated comfort and dryness in each eye on VAS every 5 minutes during contact lens wear. A slit lamp examination was performed to evaluate ocular surface health and to assess contact lens fit.
One hundred and fifty-three subjects (111 females, 42 males) completed the study. In separate models, a higher PSQ score was significantly associated with higher OSDI score (p = 0.002), lower average and EOD comfort (p = 0.005 and 0.001, respectively), and greater EOD dryness (p = 0.04). The minimum (0.14) and maximum (7.14) PSQ scores observed in our subject cohort (i.e., from the subjects who were the least and most sensitive to pain, respectively) corresponded to an estimated difference of 11 points on the OSDI, 20 points on the VAS scale for average comfort, 31 points for EOD comfort and 17 points for EOD dryness. In a mixed effects model, a higher PSQ score was significantly associated with a greater inter-eye difference in comfort (p = 0.013) and dryness (p = 0.010) during CL wear.
Pain sensitivity influences perceptions of ocular discomfort and dryness, and should be taken into account when evaluating subjective assessments of these symptoms.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
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