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.

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.

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.

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.

Acalculous cholecystitis associated with levodopa-carbidopa intestinal infusion therapy: A case report.

Continuous intra jejunal infusion of levodopa-carbidopa intestinal gel (LCIG) is one of the primary therapies for improving advanced Parkinson's disease symptoms. Placement of the jejunal catheter through the abdominal wall for drug administration requires a percutaneous interventional procedure called percutaneous endoscopic gastrostomy (PEG). PEG is considered a safe and straightforward procedure, and it is performed very commonly in clinical practice. In the context of LCIG treatment, severe adverse events have been identified, such as intestinal bleeding and acute abdomen [1], but acute acalculous cholecystitis (AAC) has never been reported.

Optimization of lignocellulolytic bacterial inoculum and substrate mix for lignocellulose degradation and product quality on co-composting of green waste with food waste.

The present study evaluates the effect of the mixing ratio of substrates and inoculation with lignocellulolytic bacteria on green waste (GW) and food waste (FW) co-composting. A Box-Behnken design was used to simultaneously optimize the lignocellulose degradation (%LD) and end-product quality. The best operational conditions were 4.85*105 CFU g-1 of Bacillus sp. F3X3 and 1.44*106 CFU g-1 of Paenibacillus sp. F1A5 with a substrate mixture containing 50% GW, 32.5% unprocessed FW, 2.5% processed FW, 13% sawdust, and 2% phosphate rock; with a C/N ratio of 27. Under these conditions, the %LD was 33% and the end-product has pH 8.3, TOC 22,4%, TN 1,7%, and a germination index of 103%. Therefore, the product complies with quality standards for organic fertilizers. The results of this study allow the identification of appropriate strategies to optimize GW composting, increasing the degradation of lignocellulose and improving the end-product quality.

Impact of 5-fluorouracil on anaerobic digestion using sewage sludge.

The role of bacterial interaction is vital to control bacterial functions; however, it has not been fully understood in microbial consortia (including anaerobic digestion). In this study, fluorouracil (FU), which is an anticancer agent and a quorum sensing (QS) inhibitor to some of the Gram-negative bacteria was found to inhibit methane production from sewage sludge under anaerobic conditions, as shown in a result where methane production in the presence of FU was eight times lower than the control (sewage sludge without FU). Whereas FU did not influence the hydrolysis process, in the acidogenesis/acetogenesis process, butyrate, and acetate accumulated in samples with FU. Also, in the methanogenesis process, FU remarkably inhibited methane production by acetoclastic methanogens rather than by the hydrogenotrophic ones. This result agreed with the result that growth and methane production of Methanosarcina acetivorans C2A was inhibited in the presence of FU. However, the inhibitory effect of FU was high in the condition that both bacteria and archaea were active. It indicates that FU influences methanogens and bacteria in the process of methane fermentation. The analyses of microbial communities (bacteria and archaea) showed that the abundance ratio of the Firmicutes phyla is high, and hydrogenotrophic methanogens become dominant in the presence of FU. Conversely, the abundance of Spirochaetes significantly decreased under FU. The inhibition of methane production by FU was due to the growth inhibition of methanogenic archaea and the changes in the composition of the bacterial population.

Selective Surface and Intraluminal Localization of Wnt Ligands on Small Extracellular Vesicles Released by HT-22 Hippocampal Neurons.

The Wnt signaling pathway induces various responses underlying the development and maturation of the nervous system. Wnt ligands are highly hydrophobic proteins that limit their diffusion through an aqueous extracellular medium to a target cell. Nevertheless, their attachment to small extracellular vesicles-like exosomes is one of the described mechanisms that allow their transport under this condition. Some Wnt ligands in these vehicles are expected to be dependent on post-translational modifications such as acylation. The mechanisms determining Wnt loading in exosomes and delivery to the target cells are largely unknown. Here, we took advantage of a cell model that secret a highly enriched population of small extracellular vesicles (sEVs), hippocampal HT-22 neurons. First, to establish the cell model, we characterized the morphological and biochemical properties of an enriched fraction of sEVs obtained from hippocampal HT-22 neurons that express NCAM-L1, a specific exosomal neuronal marker. Transmission electron microscopy showed a highly enriched fraction of exosome-like vesicles. Next, the exosomal presence of Wnt3a, Wnt5a, and Wnt7a was confirmed by western blot analysis and electron microscopy combined with immunogold. Also, we studied whether palmitoylation is a necessary post-translational modification for the transport Wnt in these vesicles. We found that proteinase-K treatment of exosomes selectively decreased their Wnt5a and Wnt7a content, suggesting that their expression is delimited to the exterior membrane surface. In contrast, Wnt3a remained attached, suggesting that it is localized within the exosome lumen. On the other hand, Wnt-C59, a specific inhibitor of porcupine O-acyltransferase (PORCN), decreased the association of Wnt with exosomes, suggesting that Wnt ligand acylation is necessary for them to be secreted by exosomes. These findings may help to understand the action of the Wnt ligands in the target cell, which could be defined during the packaging of the ligands in the secretory cell sEVs.

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.

Implementation of strategies to optimize the co-composting of green waste and food waste in developing countries. A case study: Colombia.

Green waste (GW) management is a key issue due to its high production rate and its variety of physical properties and chemical composition. Composting is a promising alternative for GW treatment and valorization. However, the presence of recalcitrant components such as lignin and cellulose increase the processing time. Strategies such as addition of co-substrates and operative modifications have improved the processing time and compost quality. Therefore, in this study, three strategies have been implemented (i) addition of unprocessed food (UF) and processed foods (PF) as co-substrates for GW to improve the nutrients composition of the substrates at the beginning of the process, (ii) addition of phosphate rock (PR) to improve product quality, and (iii) the use of two-stage composting (TSC) to accelerate the degradation. For this purpose, three treatments with the same mixture (48% GW + 21% UF + 18% PF + 13% sawdust (SW)) were conducted: (i) TA (TSC + 15% PR), (ii) TB (traditional composting +15% PR), and (iii) TC (traditional composting). TSC did not show significant differences compared with TC regarding the process and compost quality, while the addition of PR increased the phosphorus content of the product. However, TC produced the compost with the highest quality according to the Colombian legislation for soil amendment.

Canonical Wnt Signaling Modulates the Expression of Pre- and Postsynaptic Components in Different Temporal Patterns.

Wnt ligands play critical roles in neuronal development, synapse formation, synaptic activity, and plasticity. Synaptic plasticity requires molecular remodeling of synapses, implying the expression of key synaptic components. Some studies have linked Wnt signaling activity to changes in synaptic protein levels. However, the presynaptic and postsynaptic gene expression profiles of hippocampal neurons exposed to Wnt proteins have not been studied. Hence, we treated rat cultured hippocampal neurons with recombinant Wnt3a, lithium, and the Wnt inhibitor Dkk-1 for different treatment durations and measured the mRNA and protein levels of pre- and postsynaptic components. The ligand Wnt3a promoted the differential temporal expression of genes encoding presynaptic and postsynaptic proteins. Gene expression of the presynaptic proteins Rim1, piccolo (Pclo), Erc2, Ctbp1 and Rimbp2 increased in a specific temporal pattern. Simultaneously, the mRNA and protein levels of postsynaptic components showed a different temporal expression pattern, e.g., the mRNAs for postsynaptic scaffolding components such as postsynaptic density protein-95 (PSD-95/Dlg4), Homer1 and Shank1 were temporally regulated by both Wnt3a and lithium. On the other hand, the mRNA levels of the gene encoding the protein calcium/calmodulin-dependent protein kinase IV (Camk4), canonically upregulated by Wnt, were increased. Our results suggest that Wnt signaling orchestrates expressional changes in genes encoding presynaptic and postsynaptic components, probably as part of a synaptic plasticity mechanism in neurons.

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.

Serine-Arginine Protein Kinase SRPK2 Modulates the Assembly of the Active Zone Scaffolding Protein CAST1/ERC2.

Neurons release neurotransmitters at a specialized region of the presynaptic membrane, the active zone (AZ), where a complex meshwork of proteins organizes the release apparatus. The formation of this proteinaceous cytomatrix at the AZ (CAZ) depends on precise homo- and hetero-oligomerizations of distinct CAZ proteins. The CAZ protein CAST1/ERC2 contains four coiled-coil (CC) domains that interact with other CAZ proteins, but also promote self-assembly, which is an essential step for its integration during AZ formation. The self-assembly and synaptic recruitment of the Drosophila protein Bruchpilot (BRP), a partial homolog of CAST1/ERC2, is modulated by the serine-arginine protein kinase (SRPK79D). Here, we demonstrate that overexpression of the vertebrate SRPK2 regulates the self-assembly of CAST1/ERC2 in HEK293T, SH-SY5Y and HT-22 cells and the CC1 and CC4 domains are involved in this process. Moreover, the isoform SRPK2 forms a complex with CAST1/ERC2 when co-expressed in HEK293T and SH-SY5Y cells. More importantly, SRPK2 is present in brain synaptic fractions and synapses, suggesting that this protein kinase might control the level of self-aggregation of CAST1/ERC2 in synapses, and thereby modulate presynaptic assembly.

Presymptomatic Treatment With Andrographolide Improves Brain Metabolic Markers and Cognitive Behavior in a Model of Early-Onset Alzheimer's Disease.

Alzheimer's disease (AD) is the most common type of dementia. The onset and progression of this pathology are correlated with several changes in the brain, including the formation of extracellular aggregates of amyloid-beta (Aß) peptide and the intracellular accumulation of hyperphosphorylated tau protein. In addition, dysregulated neuronal plasticity, synapse loss, and a reduction in cellular energy metabolism have also been described. Canonical Wnt signaling has also been shown to be downregulated in AD. Remarkably, we showed previously that the in vivo inhibition of Wnt signaling accelerates the appearance of AD markers in transgenic (Tg) and wild-type (WT) mice. Additionally, we found that Wnt signaling stimulates energy metabolism, which is critical for the ability of Wnt to promote the recovery of cognitive function in AD. Therefore, we hypothesized that activation of canonical Wnt signaling in a presymptomatic transgenic animal model of AD would improve some symptoms. To explore the latter, we used a transgenic mouse model (J20 Tg) with mild AD phenotype expression (high levels of amyloid aggregates) and studied the effect of andrographolide (ANDRO), an activator of canonical Wnt signaling. We found that presymptomatic administration of ANDRO in J20 Tg mice prevented the reduction in cellular energy metabolism markers. Moreover, treated animals showed improvement in cognitive performance. At the synaptic level, J20 Tg animals showed severe deficiencies in presynaptic function as determined by electrophysiological parameters, all of which were completely restored to normal by ANDRO administration. Finally, an analysis of hippocampal synaptosomes by electron microscopy revealed that the length of synapses was restored with ANDRO treatment. Altogether, these data support the idea that the activation of canonical Wnt signaling during presymptomatic stages could represent an interesting pharmacological strategy to delay the onset of AD.

Modulating Wnt signaling at the root: Porcupine and Wnt acylation.

Communication between cells occurs through secreted molecules, among which Wnt ligands play a critical role in balancing cell proliferation, differentiation and cellular homeostasis. The action of Wnt signaling can be modulated at several levels, including posttranslational modification of the Wnt ligands, whose acylation is critical for biological activity. At least three enzymes are necessary for Wnt acylation/deacylation: stearoyl CoA desaturase (SCD), porcupine (PORCN) and Notum. At the endoplasmic reticulum (ER), SCD provides the monounsaturated fatty acid to PORCN, which adds it to the Wnt ligand; at the extracellular matrix, the fatty acid is removed by Notum. Obviously, the interplay between these enzymes will define Wnt signaling ligand secretion and activity. Excessive activation of Wnt signaling has been observed in a variety of solid tumors, which has led the pharmaceutical industry to develop specific inhibitors for this pathway that mainly target PORCN, some of which are in early clinical trials. In the central nervous system (CNS), Wnt signaling activation has been shown to have a neuroprotective effect, and conversely, its inhibition induces neurodegeneration, which implies that the inhibition of PORCN in cancer therapies should be used with caution, and the cognitive performance of the patient should be monitored during treatment. This review collects information about the PORCN enzyme in relation to its role in the Wnt pathway through the acylation of Wnt ligands, its inhibition by drugs in the treatment of some cancers, and its putative modulation in the treatment of neurodegenerative diseases.

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.

The Exocyst Component Exo70 Modulates Dendrite Arbor Formation, Synapse Density, and Spine Maturation in Primary Hippocampal Neurons.

Neurons are highly polarized cells displaying an elaborate architectural morphology. The design of their dendritic arborization and the distribution of their synapses contribute importantly to information processing in the brain. The growth and complexity of dendritic arbors are driven by the formation of synapses along their lengths. Synaptogenesis is augmented by the secretion of factors, like BDNF, Reelin, BMPs, or Wnts. Exo70 is a component of the exocyst complex, a protein complex that guides membrane addition and polarized exocytosis. While it has been linked to cytokinesis and the establishment of cell polarity, its role in synaptogenesis is poorly understood. In this report, we show that Exo70 plays a role in the arborization of dendrites and the development of synaptic connections between cultured hippocampal neurons. Specifically, while the overexpression of Exo70 increases dendritic arborization, synapse number, and spine density, the inhibition of Exo70 expression reduces secondary and tertiary dendrite formation and lowers synapse density. Moreover, increasing Exo70 expression augmented synaptic vesicle recycling as evaluated by FM4-64 dye uptake and the inverse was observed with downregulation of endogenous Exo70. Monitoring the formation of dendritic spines by super-resolution microscopy, we also observed that mRFP-Exo70 accumulates at the tip of EGFP-ß-actin-positive filopodia. Together, these results suggest that Exo70 is essentially involved in the formation of synapses and neuronal dendritic morphology.