Conformational change as a mechanism for toxin activation in bacterial toxin-antitoxin systems.

Toxin/antitoxin (TA) systems are present in nearly every prokaryotic genome and play the important physiological roles of phage inhibition by reducing metabolism (this includes persistence for the extreme case of complete cessation of metabolism), genetic element stabilization, and biofilm formation. TA systems have also been incorporated into other cell systems, such as CRISPR-Cas and phage quorum sensing. For the simplest and best-studied case, proteinaceous toxins and antitoxins (i.e., type II), toxin activity is masked by direct binding of the antitoxin. A long-standing, unresolved question in the TA field is how toxins are activated when bound to antitoxins at nanomolar affinity. The current paradigm envisions preferential degradation of the antitoxin by a protease, but this is highly unlikely in that a protease cannot discriminate between bound toxin and bound antitoxin because both are highly structured. Strikingly, recent results from several studies show one likely mechanism for toxin activation is conformational changes in the TA complex that result in the release or activation of the toxin as a result of a protein trigger, such as that from phages, and as a result of thermally-driven refolding dynamics.

Monitoring System for Operating Variables in Incubators in the Neonatology Service of a Highly Complex Hospital through the Internet of Things (IoT).

BACKGROUND: Around 15 million premature babies are born annually, requiring specialized care. Incubators are vital for maintaining their body temperature, which is crucial for their well-being. Ensuring optimal conditions in incubators, including constant temperature, oxygen control, and comfort, is essential for improving the care and survival rates of these infants. METHODS: To address this, an IoT-based monitoring system was developed in a hospital setting. The system comprised hardware components such as sensors and a microcontroller, along with software components including a database and a web application. The microcontroller collected data from the sensors, which was then transmitted to a broker via WiFi using the MQTT protocol. The broker validated and stored the data in the database, while the web application provided real-time access, alerts, and event recording. RESULTS: Two certified devices were created, employing high quality components. The system was successfully implemented and tested in both the biomedical engineering laboratory and the neonatology service of the hospital. The results of the pilot test supported the concept of IoT-based technology, demonstrating satisfactory responses in temperature, humidity, and sound variables within the incubators. CONCLUSIONS: The monitoring system facilitated efficient record traceability, allowing access to data over various timeframes. It also captured event records (alerts) related to variable problems, providing information on duration, date, hour, and minutes. Overall, the system offered valuable insights and enhanced monitoring capabilities for neonatal care.

Engineering anaerobic digestion via optimizing microbial community: effects of bactericidal agents, quorum sensing inhibitors, and inorganic materials.

Anaerobic digestion of sewage sludge (SS) is one of the effective ways to reduce the waste generated from human life activities. To date, there are many reports to improve or repress methane production during the anaerobic digestion of SS. In the anaerobic digestion process, many microorganisms work positively or negatively, and as a result of their microbe-to-microbe interaction and regulation, methane production increases or decreases. In other words, understanding the complex control mechanism among the microorganisms and identifying the strains that are key to increase or decrease methane production are important for promoting the advanced production of bioenergy and beneficial compounds. In this mini-review, the literature on methane production in anaerobic digestion has been summarized based on the results of antibiotic addition, quorum sensing control, and inorganic substance addition. By optimizing the activity of microbial groups in SS, methane or acetate can be highly produced. KEY POINTS: • Bactericidal agents such as an antibiotic alter microbial community for enhanced CH4 production. • Bacterial interaction via quorum sensing is one of the key points for biofilm and methane production. • Anaerobic digestion can be altered in the presence of several inorganic materials.

Characterization of gallium resistance induced in a Pseudomonas aeruginosa cystic fibrosis isolate.

The repurposing of gallium nitrate as an antibacterial, a drug used previously for the treatment of hypercalcemia, is a plausible alternative to combat infections by Pseudomonas aeruginosa, since it has antipseudomonal properties in vitro and in vivo in animal models and in human lung infections. Furthermore, gallium nitrate tolerance in clinical isolates is very rare. Nevertheless, studies on the reference strains PA14 and PAO1 show that resistance against gallium nitrate is achieved by decreasing gallium intracellular levels by increasing the production of pyocyanin. In this work, we induced resistance in a cystic fibrosis P. aeruginosa isolate and explored its resistance mechanisms. This isolated strain, INP-58M, was not a pyocyanin producer, and its pyoverdine levels remained unchanged upon gallium addition. However, it showed higher activities of NADPH-producing enzymes and the antioxidant enzyme SOD when gallium was added, which suggests a better antioxidant response. Remarkably, gallium intracellular levels in the resistant isolate were higher than those of the parental strain at 20 h but lower after 24 h of culture, suggesting that this strain is capable of gallium efflux.

Wnt-induced activation of glucose metabolism mediates the in vivo neuroprotective roles of Wnt signaling in Alzheimer disease.

Dysregulated Wnt signaling is linked to major neurodegenerative diseases, including Alzheimer disease (AD). In mouse models of AD, activation of the canonical Wnt signaling pathway improves learning/memory, but the mechanism for this remains unclear. The decline in brain function in AD patients correlates with reduced glucose utilization by neurons. Here, we test whether improvements in glucose metabolism mediate the neuroprotective effects of Wnt in AD mouse model. APPswe/PS1dE9 transgenic mice were used to model AD, Andrographolide or Lithium was used to activate Wnt signaling, and cytochalasin B was used to block glucose uptake. Cognitive function was assessed by novel object recognition and memory flexibility tests. Glucose uptake and the glycolytic rate were determined using radiotracer glucose. The activities of key enzymes of glycolysis such as hexokinase and phosphofructokinase, Adenosine triphosphate (ATP)/Adenosine diphosphate (ADP) levels and the pentose phosphate pathway and activity of glucose-6 phosphate dehydrogenase were measured. Wnt activators significantly improved brain glucose utilization and cognitive performance in transgenic mice. Wnt signaling enhanced glucose metabolism by increasing the expression and/or activity of hexokinase, phosphofructokinase and AMP-activated protein kinase. Inhibiting glucose uptake partially abolished the beneficial effects of Wnt signaling on learning/memory. Wnt activation also enhanced glucose metabolism in cortical and hippocampal neurons, as well as brain slices derived from APPswe/PS1E9 transgenic mice. Combined, these data provide evidence that the neuroprotective effects of Wnt signaling in AD mouse models result, at least in part, from Wnt-mediated improvements in neuronal glucose metabolism.

Ascorbic acid-cyclodextrin complex alters the expression of genes associated with lipid metabolism in bovine in vitro produced embryos.

Ascorbic acid (AC) used as antioxidant in embryo culture is very sensitive and degrades unavoidably in aqueous solution. Methyl-ß-cyclodextrin (CD) improved the stability of AC in solution to elevated temperature, light, humidity and oxidation. The aim of this study was to evaluate the effect of the complex AC-CD during in vitro maturation (IVM) or in vitro culture (IVC) on oocyte developmental competence and subsequent embryo development and quality. AC-CD (100 µM) was added to IVM media, and maturation level and embryo development were examined. Matured oocytes, their cumulus cells and produced blastocysts were snap-frozen for gene expression analysis by RT-qPCR. Besides, in vitro-produced zygotes were cultured with 100 µM of AC-CD and blastocysts were as well snap-frozen for gene expression analysis. A group without AC-CD (control- ) and other with CD (control+ ) were included. No differences were found on maturation, cleavage or blastocyst rates. However, in matured oocytes, AC-CD downregulated BAX, GPX1 and BMP15. In cumulus cells, AC-CD downregulated BAX/BCL2 and GSTA4 while upregulated BCL2 and CYP51A1. The expression of SL2A1, FADS1, PNPLA and MTORC1 was downregulated in blastocysts derived from oocytes matured with AC-CD, while in blastocysts derived from zygote cultured with AC-CD, CYP51A1 and IGF2R were downregulated and PNPLA2 was upregulated. In conclusion, AC-CD in both IVM and IVC media may reduce accumulated fat by increasing lipolysis and suppressing lipogenesis in blastocysts derived from both oocytes and zygotes cultured with AC-CD, suggesting that CD improves the quality of embryos and bioavailability of AC during IVM and IVC.

In Vitro Effectiveness of a Terpenic Denture Cleanser on Old Biofilm Surfaces.

PURPOSE: To assess the effects of terpenic denture cleanser on denture biofilm removal using scanning electron microscopy (SEM). MATERIALS AND METHODS: The internal surface biofilm of four maxillary dentures was elucidated with Caristop-revelador Dual Tone, and 40 blue-stained specimens (0.6 cm × 0.4 cm × 2 mm) were obtained. These specimens were randomly assigned to one of the following four groups of 10 specimens each: control, Eci Clean, Fitty Dent, and terpenic denture cleanser. The period of immersion in each solution was 12 hours. Biofilm removal was evaluated using SEM, and morphologically varying areas of the SEM images were quantified with Imaris software. The data were analyzed using Kolmogorov-Smirnov, t-tests, ANOVA, and Tamhane's tests (p = 0.05). RESULTS: Data revealed that terpenic denture cleanser removed significantly more biofilm than any other treatment examined in this study. The t-tests revealed significant differences in the clean area that resulted from the use of the terpenic cleanser compared with the clean area that resulted from the use of Eci Clean (p = 0.013). Fitty Dent was the least effective and left dirty acrylic resin. The average areas with few removed layers were 59.3%, 43.3%, and 9.5% in Fitty Dent, Eci Clean, and terpenic cleanser groups, respectively. Tamhane's tests indicated that the Eci Clean and Fitty Dent groups were significantly different from the 0.5% terpenic cleanser group (p = 0.008). CONCLUSION: The terpenic denture cleanser was effective in removing denture biofilm.

Emerging Synaptic Molecules as Candidates in the Etiology of Neurological Disorders.

Synapses are complex structures that allow communication between neurons in the central nervous system. Studies conducted in vertebrate and invertebrate models have contributed to the knowledge of the function of synaptic proteins. The functional synapse requires numerous protein complexes with specialized functions that are regulated in space and time to allow synaptic plasticity. However, their interplay during neuronal development, learning, and memory is poorly understood. Accumulating evidence links synapse proteins to neurodevelopmental, neuropsychiatric, and neurodegenerative diseases. In this review, we describe the way in which several proteins that participate in cell adhesion, scaffolding, exocytosis, and neurotransmitter reception from presynaptic and postsynaptic compartments, mainly from excitatory synapses, have been associated with several synaptopathies, and we relate their functions to the disease phenotype.

Morphophysiological and cuticular chemical alterations caused by Xenos entomophagus endoparasites in the social wasp Polistes ferreri (Hymenoptera, Vespidae).

Social wasps can face many challenges during their colony cycle, including the presence of parasites. The order Strepsiptera is among the main parasites of the wasp genus Polistes. The aim of this study was to evaluate the effect of an endoparasite species on the host Polistes ferreri, with the hypothesis that females of this social wasp would undergo morphophysiological alterations as well as changes in their cuticular chemical profile caused by the obligate endoparasite. On average, parasitism was found in 10% of the colonies studied. All the parasitized females showed filamentous ovarioles without developing oocytes, which indicates a physiological castration. Moreover, the endoparasites present in the gaster of females caused its volume to increase, and the presence of endoparasites changed the cuticular chemical profiles of females, confirming our hypothesis. It is likely that this parasitism effect could hamper the maintenance of wasp colonies.

Beneficial knockouts in Escherichia coli for producing hydrogen from glycerol.

Glycerol is an inexpensive and abundant source for biofuel production on a large scale. Escherichia coli is a robust bacterium for producing hydrogen; however, its hydrogen productivity from glycerol is low. In this study, we conducted random transposon mutagenesis to identify uncharacterized genes whose inactivation is beneficial for hydrogen production from glycerol. Through screening, four mutant strains were found that are able to have from 1.3- to 1.6-fold higher hydrogen productivity (µmol H2/mg protein) than that of their parent strain (p < 0.05). These mutations were identified as aroM, gatZ, ycgR, and yfgI. The hydrogen yield (mol H2/mol glycerol consumed) of the aroM, gatZ, ycgR, and yfgI strains was 1.7-, 1.4-, 2.4-, and 2.1-fold higher than that of their parent strain, respectively. Moreover, a single disruption in these genes resulted in a faster cell growth and glycerol consumption under anaerobic conditions. In E. coli, AroM is predicted to be involved in the shikimate pathway, GatZ is tagatose-1,6-bisphosphate aldolase 2 which converts dihydroxyacetone phosphate to 1,6-biphosphate, and YcgR acts as a molecular brake limiting the swimming speed and ATP consumption. So far, the function of YfgI in general and in hydrogen production in particular remains unknown.

A new type V toxin-antitoxin system where mRNA for toxin GhoT is cleaved by antitoxin GhoS.

Among bacterial toxin-antitoxin systems, to date no antitoxin has been identified that functions by cleaving toxin mRNA. Here we show that YjdO (renamed GhoT) is a membrane lytic peptide that causes ghost cell formation (lysed cells with damaged membranes) and increases persistence (persister cells are tolerant to antibiotics without undergoing genetic change). GhoT is part of a new toxin-antitoxin system with YjdK (renamed GhoS) because in vitro RNA degradation studies, quantitative real-time reverse-transcription PCR and whole-transcriptome studies revealed that GhoS masks GhoT toxicity by cleaving specifically yjdO (ghoT) mRNA. Alanine substitutions showed that Arg28 is important for GhoS activity, and RNA sequencing indicated that the GhoS cleavage site is rich in U and A. The NMR structure of GhoS indicates it is related to the CRISPR-associated-2 RNase, and GhoS is a monomer. Hence, GhoT-GhoS is to our knowledge the first type V toxin-antitoxin system where a protein antitoxin inhibits the toxin by cleaving specifically its mRNA.

Capturing methane with recombinant soluble methane monooxygenase and recombinant methyl-coenzyme M reductase.

Methane capture via oxidation is considered one of the 'Holy Grails' of catalysis (Tucci and Rosenzweig, 2024). Methane is also a primary greenhouse gas that has to be reduced by 1.2 billion metric tonnes in 10 years to decrease global warming by only 0.23°C (He and Lidstrom, 2024); hence, new technologies are needed to reduce atmospheric methane levels. In Nature, methane is captured aerobically by methanotrophs and anaerobically by anaerobic methanotrophic archaea; however, the anaerobic process dominates. Here, we describe the history and potential of using the two remarkable enzymes that have been cloned with activity for capturing methane: aerobic capture via soluble methane monooxygenase and anaerobic capture via methyl-coenzyme M reductase. We suggest these two enzymes may play a prominent, sustainable role in addressing our current global warming crisis.

Implications of lytic phage infections inducing persistence.

Phage therapy holds much promise as an alternative to antibiotics for fighting infection. However, this approach is no panacea as recent results show that a small fraction of cells survives lytic phage infection due to both dormancy (i.e. formation of persister cells) and resistance (genetic change). In this brief review, we summarize evidence suggesting phages induce the persister state. Therefore, it is predicted that phage cocktails should be combined with antipersister compounds to eradicate bacterial infections.

Image-guided programming deep brain stimulation improves clinical outcomes in patients with Parkinson's disease.

Deep brain stimulation (DBS) is an effective treatment for patients with Parkinson's disease (PD). However, some patients may not respond optimally to clinical programming adjustments. Advances in DBS technology have led to more complex and time-consuming programming. Image-guided programming (IGP) could optimize and improve programming leading to better clinical outcomes in patients for whom DBS programming is not ideal due to sub-optimal response. We conducted a prospective single-center study including 31 PD patients with subthalamic nucleus (STN) DBS and suboptimal responses refractory to clinical programming. Programming settings were adjusted according to the volumetric reconstruction of the stimulation field using commercial postoperative imaging software. Clinical outcomes were assessed at baseline and at 3-month follow-up after IGP, using motor and quality of life (QoL) scales. Additionally, between these two assessment points, follow-up visits for fine-tuning amplitude intensity and medication were conducted at weeks 2, 4, 6, and 9. After IGP, twenty-six patients (83.9%) experienced motor and QoL improvements, with 25.8% feeling much better and 38.7% feeling moderately better according to the patient global impression scale. Five patients (16.1%) had no clinical or QoL changes after IGP. The MDS-UPDRS III motor scale showed a 21.9% improvement and the DBS-IS global score improved by 41.5%. IGP optimizes STN-DBS therapy for PD patients who are experiencing suboptimal clinical outcomes. These findings support using IGP as a standard tool in clinical practice, which could save programming time and improve patients' QoL.

DRD4 gene polymorphism and impulse control disorder induced by dopamine agonists in Parkinson's disease.

Impulse control disorders and their consequences display variability among individuals, indicating potential involvement of environmental and genetic factors. In this retrospective study, we analyzed a cohort of Parkinson's disease patients treated with dopamine agonists and investigated the influence of the dopamine D4 receptor gene polymorphism, DRD4 7R+, which is linked to psychiatric disorders, impulsive traits, and addictive behaviors. We found that DRD4 7R+ is a significant genetic risk factor associated with the severity of ICD.

Formation of Golgi-derived active zone precursor vesicles.

Vesicular trafficking of presynaptic and postsynaptic components is emerging as a general cellular mechanism for the delivery of scaffold proteins, ion channels, and receptors to nascent and mature synapses. However, the molecular mechanisms leading to the selection of cargos and their differential transport to subneuronal compartments are not well understood, in part because of the mixing of cargos at the plasma membrane and/or within endosomal compartments. In the present study, we have explored the cellular mechanisms of active zone precursor vesicle assembly at the Golgi in dissociated hippocampal neurons of Rattus norvegicus. Our studies show that Piccolo, Bassoon, and ELKS2/CAST exit the trans-Golgi network on a common vesicle that requires Piccolo and Bassoon for its proper assembly. In contrast, Munc13 and synaptic vesicle proteins use distinct sets of Golgi-derived transport vesicles, while RIM1α associates with vesicular membranes in a post-Golgi compartment. Furthermore, Piccolo and Bassoon are necessary for ELKS2/CAST to leave the Golgi in association with vesicles, and a core domain of Bassoon is sufficient to facilitate formation of these vesicles. While these findings support emerging principles regarding active zone differentiation, the cellular and molecular analyses reported here also indicate that the Piccolo-Bassoon transport vesicles leaving the Golgi may undergo further changes in protein composition before arriving at synaptic sites.

[Internet as an information source for health in primary care patients and its influence on the physician-patient relationship]. / Internet como fuente de información sobre salud en pacientes de atención primaria y su influencia en la relación médico-paciente.

OBJECTIVE: To describe the use of the Internet by primary care patients to seek health related information, understand how they are influenced by this information, and evaluate its impact on the doctor-patient relationship. DESIGN: Cross sectional study, through self-administered survey. SETTING: One urban health center in Madrid. PARTICIPANTS: A total of 323 questionnaires were collected from patients between 14 and 75 years old who attended a physician's office for any reason, excluding illiterate patients and those with neurological or psychiatric problems preventing them from completing the survey. MEASUREMENTS: Internet usage, ability of the internet to clarify doubts regarding health issues, patient lifestyle changes, socio-demographic variables, and physician's receptivity to the use of internet by patients. RESULTS: 61% (CI95%: 56%-67%) of patients used the Internet as a source of health information: Internet queries were able to address health doubts in 92.4% of users, 53.5% reported that the Internet changed their thinking about their health in at least one instance, 30% made behavioral changes (of which 60.1% discussed these changes with their physician), 44.3% had more questions at the physician's office, and 80.8% believe that the doctor would be willing to talk about the information found on the internet. CONCLUSIONS: Using the Internet to find information about health is very common, with positive influence on physician-patient relationship. This may be useful for achieving behavioral changes in patients and can be used as a tool in medical practice.

Cells immersed in collagen matrices show a decrease in plasma membrane fluidity as the matrix stiffness increases.

Cells are constantly adapting to maintain their identity in response to the surrounding media's temporal and spatial heterogeneity. The plasma membrane, which participates in the transduction of external signals, plays a crucial role in this adaptation. Studies suggest that nano and micrometer areas with different fluidities at the plasma membrane change their distribution in response to external mechanical signals. However, investigations linking fluidity domains with mechanical stimuli, specifically matrix stiffness, are still in progress. This report tests the hypothesis that the stiffness of the extracellular matrix can modify the equilibrium of areas with different order in the plasma membrane, resulting in changes in overall membrane fluidity distribution. We studied the effect of matrix stiffness on the distribution of membrane lipid domains in NIH-3 T3 cells immersed in matrices of varying concentrations of collagen type I, for 24 or 72 h. The stiffness and viscoelastic properties of the collagen matrices were characterized by rheometry, fiber sizes were measured by Scanning Electron Microscopy (SEM) and the volume occupied by the fibers by second harmonic generation imaging (SHG). Membrane fluidity was measured using the fluorescent dye LAURDAN and spectral phasor analysis. The results demonstrate that an increase in collagen stiffness alters the distribution of membrane fluidity, leading to an increasing amount of the LAURDAN fraction with a high degree of packing. These findings suggest that changes in the equilibrium of fluidity domains could represent a versatile and refined component of the signal transduction mechanism for cells to respond to the highly heterogeneous matrix structural composition. Overall, this study sheds light on the importance of the plasma membrane's role in adapting to the extracellular matrix's mechanical cues.

Validation of the PEG Scale in Spanish (PEG-S) Among Adults Receiving Care for Pain in US Primary Care.

This study sought to evaluate the psychometric properties of a Spanish version of the PEG scale (PEG-S, whose items assess Pain intensity and pain interference with Enjoyment of life and General activity) in a sample of Spanish-speaking adults receiving care for pain at primary care clinics in the Northwestern United States. We evaluated the PEG-S's 1) internal consistency, 2) convergent validity, and 3) discriminant validity. All participants (n = 200, mean age = 52 years [SD = 15], 76% women, mean PEG-S score = 5.7 [SD = 2.5]) identified as having Hispanic or Latino ethnicity, and detailed ethnic origin was predominantly Mexican or Chicano (70%). The PEG-S's internal consistency (Cronbach's alpha, .82) was good. Correlations between the PEG-S scale scores and established measures of pain intensity and interference ranged from .68 to .79, supporting the measure's convergent validity. The correlation between the PEG-S scale score and the Patient Health Questionnaire-9 (r = .53) was weaker than those between the PEG-S scale and measures of pain intensity and interference, supporting the measure's discriminant validity. The findings support reliability and validity of the PEG-S for assessing a composite score of pain intensity and interference among Spanish-speaking adults. PERSPECTIVE: We present evidence supporting the reliability and validity of the PEG scale in Spanish (PEG-S) in a sample of adults receiving pain care at primary care clinics in the Northwestern United States. This 3-item composite measure of pain intensity and interference can help clinicians and researchers assess pain among Spanish-speaking adults.

Division of labor in colonies of the eusocial wasp, Mischocyttarus consimilis.

The division of labor between castes and the division of labor in workers according to age (temporal polyethism) in social wasps are crucial for maintaining social organization. This study evaluated the division of labor between castes, and the temporal polyethism in workers of Mischocyttarus consimilis Zikán (Hymenoptera: Vespidae). To describe the behavioral repertory of this species, observations were made of 21 colonies, with 100 hours of observations. In order to observe temporal polyethism, each newly emerged wasp was marked with colored dots on the upper area of the thorax. This allowed the observation of behavioral acts performed by each worker from the time of emergence to its death. Through hybrid multidimensional scaling, a clear division between queens and workers could be identified, in which the behaviors of physical dominance and food solicitation characterized the queen caste; while behaviors such as adult-adult trophallaxis, destruction of cells, alarm, foraging for prey, foraging for nectar, and unsuccessful foraging characterized the worker caste. Hybrid multidimensional scaling characterized two groups, with intra-nest activities preferentially accomplished by younger workers, while extra-nest activities such as foraging were executed more frequently by older workers.