A multi-scale map of cell structure fusing protein images and interactions.

The cell is a multi-scale structure with modular organization across at least four orders of magnitude1. Two central approaches for mapping this structure-protein fluorescent imaging and protein biophysical association-each generate extensive datasets, but of distinct qualities and resolutions that are typically treated separately2,3. Here we integrate immunofluorescence images in the Human Protein Atlas4 with affinity purifications in BioPlex5 to create a unified hierarchical map of human cell architecture. Integration is achieved by configuring each approach as a general measure of protein distance, then calibrating the two measures using machine learning. The map, known as the multi-scale integrated cell (MuSIC 1.0), resolves 69 subcellular systems, of which approximately half are to our knowledge undocumented. Accordingly, we perform 134 additional affinity purifications and validate subunit associations for the majority of systems. The map reveals a pre-ribosomal RNA processing assembly and accessory factors, which we show govern rRNA maturation, and functional roles for SRRM1 and FAM120C in chromatin and RPS3A in splicing. By integration across scales, MuSIC increases the resolution of imaging while giving protein interactions a spatial dimension, paving the way to incorporate diverse types of data in proteome-wide cell maps.

High-Resolution Plasma Metabolomics Identifies Alterations in Fatty Acid, Energy, and Micronutrient Metabolism in Adults Across the Leprosy Spectrum.

BACKGROUND: High-resolution metabolomics (HRM) is an innovative tool to study challenging infectious diseases like leprosy, where the pathogen cannot be grown with standard methods. Here, we use HRM to better understand associations between disease manifestations, nutrition, and host metabolism. METHODS: From 2018 to 2019, adults with leprosy and controls were recruited in Minas Gerais, Brazil. Plasma metabolites were detected using an established HRM workflow and characterized by accurate mass, mass to charge ratio m/z and retention time. The mummichog informatics package compared metabolic pathways between cases and controls and between multibacillary (MB) and paucibacillary (PB) leprosy. Additionally, select individual metabolites were quantified and compared. RESULTS: Thirty-nine cases (62% MB and 38% PB) and 25 controls were enrolled. We found differences (P < .05) in several metabolic pathways, including fatty acid metabolism, carnitine shuttle, retinol, vitamin D3, and C-21 steroid metabolism, between cases and controls with lower retinol and associated metabolites in cases. Between MB and PB, leukotrienes, prostaglandins, tryptophan, and cortisol were all found to be lower in MB (P < .05). DISCUSSION: Metabolites associated with several nutrient-related metabolic pathways appeared differentially regulated in leprosy, especially MB versus PB. This pilot study demonstrates the metabolic interdependency of these pathways, which may play a role in the pathophysiology of disease.

The use of Lentilactobacillus buchneri PJB1 and Lactiplantibacillus plantarum MTD1 on the ensiling of whole-plant corn silage, snaplage, and high-moisture corn.

Experiments were conducted over a 3-yr period to evaluate the effects of bacterial inoculants on the fermentation profile and aerobic stability of whole-plant corn silage (WPC), snaplage (SNA), and high-moisture corn (HMC). Whole-plant corn was inoculated with Lentilactobacillus buchneri PJB1 in combination with Lactiplantibacillus plantarum MTD1 or with Lpb. plantarum alone (experiments 1 and 2). Snaplage (experiment 3) and HMC (experiments 4 and 5) were inoculated with Len. buchneri in combination with Lpb. plantarum or with Len. buchneri alone. After inoculation, the feedstuffs were ensiled in 7.57-L silos and stored at 21 ± 2°C for 30 or 90 d. In experiment 5, silage was subjected to air stress for 2 h every 2 wk through 42 d and then for 2 h/wk until 90 d and had samples analyzed for their bacterial community composition by metagenomics. Overall, in all experiments, silages inoculated with Len. buchneri alone or in combination with Lpb. plantarum had more acetic acid and 1,2-propanediol and fewer yeasts than uninoculated silages. After 30 d of ensiling, inoculation with Len. buchneri alone or in combination with Lpb. plantarum did not affect the aerobic stability of SNA, but it slightly increased the stability of WPC and markedly improved the stability of HMC. After 90 d of ensiling, inoculation with Len. buchneri alone or in combination with Lpb. plantarum markedly improved the aerobic stability of WPC, SNA, and HMC. In experiment 5, inoculation increased the relative abundance (RA) of Lactobacillaceae and reduced the RA of Enterobacteriaceae and Leuconostocaceae in HMC at 30 and 90 d and the RA of Clostridiaceae in non-air-stressed HMC at 90 d. Air-stressed HMC inoculated with Len. buchneri had less lactic acid, more acetic acid and 1,2-propanediol, and markedly greater aerobic stability than uninoculated air-stressed HMC at 90 d. In conclusion, inoculation with Len. buchneri PJB1 alone or in combination with Lpb. plantarum MTD1 increased the production of acetic acid and 1,2-propanediol, inhibited yeasts development, and improved the aerobic stability of WPC, SNA, and HMC. In HMC, inoculation markedly improved aerobic stability as soon as after 30 d of ensiling, and after 90 d, inoculation improved stability even under air stress conditions.

Looking through Providers' Eyes: Pain in the Neonatal Intensive Care Unit.

OBJECTIVE: Evaluate the pain of critically ill newborns is a challenge because of the devices for cardiorespiratory support. This study aim to verify the adults' gaze when assessing the critically ill neonates' pain at bedside. STUDY DESIGN: Cross-sectional study in which pediatricians, nursing technicians, and parents evaluated critically ill neonates' pain at bedside, for 20 seconds with eye-tracking glasses. At the end, they answered whether the neonate was in pain or not. Visual tracking outcomes: number and time of visual fixations in four areas of interest (AOI) (face, trunk, and upper [UL] and lower [LL] limbs) were compared between groups and according to pain perception (present/absent). RESULTS: A total of 62 adults (21 pediatricians, 23 nursing technicians, 18 parents) evaluated 27 neonates (gestational age: 31.8 ± 4.4 weeks; birth weight: 1,645 ± 1,234 g). More adults fixed their gaze on the face (96.8%) and trunk (96.8%), followed by UL (74.2%) and LL (66.1%). Parents performed a greater number of fixations on the trunk than nursing technicians (11.0 vs. 5.5 vs. 6.0; p = 0.023). Controlled for visual tracking variables, each second of eye fixation in AOI (1.21; 95% confidence interval [CI]: 1.03-1.42; p = 0.018) and UL (1.07; 95% CI: 1.03-1.10; p < 0.001) increased the chance of perceiving the presence of pain. CONCLUSION: Adults, when assessing at bedside critically ill newborns' pain, fixed their eyes mainly on the face and trunk. The time spent looking at the UL was associated with the perception of pain presence. KEY POINTS: · Pain assessment in critically ill newborns is a challenge.. · To assess critically ill neonates' pain, adults mainly look at the face and trunk.. · Looking at the upper limbs also helps in assessing critically ill neonates' pain..

Changes to the microbiome of alfalfa during the growing season and after ensiling with Lentilactobacillus buchneri and Lentilactobacillus hilgardii inoculant.

AIMS: This study evaluated changes in epiphytic microbial population of alfalfa (Medicago sativa) during the growing season. First cut forage was harvested to study the effects of an inoculant combining two obligate heterofermentative lactic acid bacteria strains on the bacterial and fungal communities and the fermentation of alfalfa silage. METHODS AND RESULTS: The epiphytic microbiome of alfalfa was evaluated 10-times during the growing season. Alfalfa wilted to 395.0 g/kg was treated with water (Control) or with a combination of L. buchneri NCIMB 40788 and L. hilgardii CNCM-I-4785 (LBLH). Mini-silos were opened after 1, 4, 8, 16, 32, and 64 days of ensiling. The relative abundance (RA) of the epiphytic bacterial and fungal families varied during the growing season. After 1 day, Weissella was the most abundant genus and present at similar RA in the two treatments (average 80.4%). Compared with Control, LBLH had a higher RA of Lactobacillus at day 1, 16, 32, and 64, and a lower RA of Weissella from day 8 to 64. Control contained more bacteria belonging to the Enterobacteriales than LBLH up to day 16. Inoculated silage had more acetate than Control at day 32 and 64. The fungal population were similar between treatments. The enhanced development and dominance of Lactobacillus in inoculated silage led to greater accumulation of acetate and propionate, which reduced the numbers of culturable yeasts but did not markedly affect the fungal community structure. CONCLUSIONS: The bacterial community composition of alfalfa stands in the filed changed over time and was affected by cutting. For the ensiling trial, inoculation modified the composition of the bacterial community of alfalfa, increasing the RA of Lactobacillus while reducing the RA of Weissella and of Enterobacteriaceae. SIGNIFICANCE AND IMPACT OF STUDY: Inoculation increased the RA of Lactobacillus, hampering the dominance of Weissella in the early stages of ensiling, improving antifungal compounds production and reducing the numbers of culturable yeasts.

Effect of dry matter content on the microbial community and on the effectiveness of a microbial inoculant to improve the aerobic stability of corn silage.

Silage inoculants are commonly used as a tool to improve the fermentation and aerobic stability of corn silage fed to dairy cows. However, their effectiveness can be inconsistent. Our objective was to determine the effect of the dry matter (DM) content of freshly chopped whole-plant corn on its microbial community as affected by an inoculant containing Lentilactobacillus hilgardii, Lentilactobacillus buchneri, and Pediococcus pentosaceus on improving the aerobic stability of silage. Whole-plant corn was harvested at low (31.80%, LDM), medium (33.32%, MDM), or high (39.44%, HDM) DM content and treated with nothing (CTR) or an inoculant (INO) containing L. hilgardii CNCM I-4785 at 150,000 cfu/g fresh forage, L. buchneri NCIMB 40788 at 150,000 cfu/g fresh forage, P. pentosaceus NCIMB12455 at 100,000 cfu/g of fresh forage, ß-glucanase (5,750 IU/g), and xylanase (30,000 IU/g) and ensiled for 20 and 60 d. Data were analyzed as a completely randomized design in a 3-by-2 factorial arrangement of treatments. Fresh LDM forage had a higher concentration of reducing sugars, a less rich, diverse, and even bacterial community, and greater relative abundance of Saccharomycetales than MDM and HDM forages. Silages at 20 and 60 d, inoculated LDM had a more modest proliferation of culturable lactic acid bacteria than inoculated MDM. At 20 d, regardless of treatment, LDM had greater concentrations of lactic and acetic acids. Also at 20 d, LDM had lower numbers of culturable yeasts but greater relative abundance of Enterobacteriaceae than MDM and HDM. For silage at 20 d, HDM silage was more aerobically stable than LDM and MDM and inoculation improved aerobic stability 1.8-fold compared with CTR. For silage at 60 d, there was an interaction between DM content and inoculation. The improvements in stability by inoculation, compared with CTR, were greater in MDM (261 vs. 41 h) and HDM (320 vs. 66 h) silages than in LDM (85 vs. 46 h). The lower DM content and possible slower pH decline in LDM might have facilitated the development of undesirable bacteria and coupled with its greater concentration of reducing sugars and lactic and acetic acids, which are substrates for aerobic microorganisms, might explain the more modest improvements in aerobic stability from inoculation in LDM compared with MDM and HDM. Our findings suggest that the DM content of whole-plant corn affected its epiphytic microbial community and the effectiveness of the inoculant, which improved aerobic stability at all DM but to a greater extent in HDM and MDM than in LDM, especially after 60 d of ensiling.

Could an Internet-Based Foot-Ankle Therapeutic Exercise Program Modify Clinical Outcomes and Gait Biomechanics in People with Diabetic Neuropathy? A Clinical Proof-of-Concept Study.

Previous studies have shown the efficacy of foot-ankle exercises in people with diabetic peripheral neuropathy (DPN), but the quality of evidence is still low. This proof-of-concept study pursues preliminary evidence for potential clinical and gait biomechanical benefits from an internet-based foot-ankle therapeutic exercise program for people with DPN. We randomized 30 individuals with DPN (IWGDF risk category 1 or 2) into either the control group (CG) receiving the usual care or the intervention group (IG) receiving the usual care plus an internet-based foot-ankle exercise program, fully guided by the Sistema de Orientação ao Pé Diabético (SOPeD; translation: Diabetic Foot Guidance System) three times per week for 12 weeks. We assessed face-to-face clinical and biomechanical outcomes at baseline, 12 weeks, and 24 weeks (follow up). Participants had good adherence to the proposed intervention and it led to only mild adverse events. The IG showed improvements in the ankle and first metatarsophalangeal joint motion after 12 and 24 weeks, changed forefoot load absorption during foot rollover during gait after 24 weeks, reduced foot pain after 12 weeks, and improved foot function after 24 weeks. A 12-week internet-based foot-ankle exercise program using the SOPeD software (version 1.0) has the potential to reduce foot pain, improve foot function, and modify some important foot-ankle kinematic outcomes in people with DPN.

The effects of Lactobacillus hilgardii 4785 and Lactobacillus buchneri 40788 on the microbiome, fermentation, and aerobic stability of corn silage ensiled for various times.

We evaluated the ability of an inoculant containing a combination of Lactobacillus hilgardii and Lactobacillus buchneri to modify the microbiome and improve the aerobic stability of whole-plant corn silage after various lengths of ensiling. Chopped whole-plant corn at about 33% dry matter (DM) was uninoculated (CTR) or inoculated with L. hilgardii CNCM I-4785 and L. buchneri NCIMB 40788 at 200,000 cfu/g of fresh forage weight each (combined application rate of 400,000 cfu of lactic acid bacteria/g of fresh forage weight; LHLB), L. buchneri NCIMB 40788 at 400,000 cfu/g of fresh forage weight and Pediococcus pentosaceus NCIMB 12455 at 100,000 cfu/g of fresh forage weight, used as a positive control (LB500), L. hilgardii CNCM I-4785 at the application rate used in the LHLB formulation of 200,000 cfu/g of fresh forage weight (LH), or L. buchneri NCIMB 40788 at the application rate used in the LHLB formulation of 200,000 cfu/g of fresh forage weight (LB). Silos were opened after 34 and 99 d of ensiling and analyzed for nutrient composition, fermentation profile, microbiome, and aerobic stability. After 34 d of ensiling, the inoculated silages had greater numbers of culturable lactic acid bacteria, a bacterial community less rich and diverse, greater relative abundance of Lactobacillus, lower relative abundance of Klebsiella, and a greater concentration of propionic acid than uninoculated silages. Inoculation decreased the ratio of lactic acid to acetic acid, except for LB alone. Treatment LHLB resulted in silage with a greater concentration of 1,2-propanediol than LB500 and was the only treatment to have a lower relative abundance of Saccharomycetes compared with uninoculated silage. Treatments LHLB and LB500 improved the aerobic stability compared with CTR, but the individual LH and LB treatments applied at a low dose did not. Whereas LB500 was stable 34 h longer than CTR, LHLB was stable 91 h longer. After 99 d of ensiling, all inoculated silages had markedly greater aerobic stability than uninoculated silage and were stable for more than 360 h. The inoculant containing a combination of L. hilgardii and L. buchneri markedly improved the aerobic stability of corn silage after a relatively short period of ensiling, and such improvements were greater than the ones obtained from inoculation with the combination of L. buchneri and P. pentosaceus. Inoculating with the combination of L. hilgardii and L. buchneri may be helpful to producers that must feed silage shortly after ensiling.

Feasibility and Preliminary Efficacy of a Foot-Ankle Exercise Program Aiming to Improve Foot-Ankle Functionality and Gait Biomechanics in People with Diabetic Neuropathy: A Randomized Controlled Trial.

Foot-ankle strengthening and mobility exercises are part of international guideline recommendations for people at risk of diabetic foot disease. We examined the feasibility and preliminary efficacy of a 12-week foot-ankle exercise program on clinical, functional and biomechanical outcomes in people with diabetic neuropathy (DPN). We randomly allocated 30 people with DPN to usual care (control) or usual care plus a supervised exercise program (intervention). For feasibility, we assessed recruitment rate and participant adherence and satisfaction. For program efficacy, we assessed baseline to 12-week changes in daily physical activity level, gait speed, tactile sensitivity, ankle range of motion, DPN symptoms, quality of life, foot health and functionality, foot strength and plantar pressure during gait, using paired t-tests (p < 0.05). In 52 weeks, we recruited 45 eligible participants (0.90/week). Program adherence was 80% and participants' satisfaction had a mean (SD) of 4.57 (0.70) out of 5. The intervention group significantly improved on toes strength, contact time during gait and DPN symptoms, and peak forefoot pressures increased over time; controls showed significantly increased heel peak pressures and force. The exercise program was feasible, based on a moderate recruitment rate and an adherent and satisfied population, and the intervention showed several positive preliminary effects over time compared to usual care.

The Association Between Rearfoot Motion While Barefoot and Shod in Different Types of Running Shoes in Recreational Runners.

The rearfoot angle (RFA) is a biomechanical variable widely used to determine the rearfoot motion (RM). Shoe manufacturers began to develop running shoes with RM control that would supposedly alter foot-ground interaction mechanics and neutralize excessive pronation or supination; moreover, some studies have not shown differences in rearfoot motion in shod condition compared to barefoot. This study intended to answer three questions: Do the shoes runners wear correspond to their respective barefoot RM? Does the eversion angle change during shod running, regardless the shoes worn? Can footwear designed for a specific RM (supination, pronation, neutral) correct or neutralize the eversion angle of runners? One hundred and eleven runners (38.6 ± 9.7years; 74.9 ± 12.0kg; 1.74 ± 0.08 m), who ran an average of 32 ± 17km/week, were included in this cross-sectional study. They had their RFA measured by a motion capture system when running barefoot and wearing their habitual running shoes (shod condition). Chi-squared test was used to assess associations between barefoot and shod condition and RFA was compared between conditions using Wilcoxon tests (p = 0.05). There was no association between the type of running shoe and barefoot RM (p > 0.05). There was an association between RFA when barefoot and when shod (p < 0.05). Among all participants classified as neutral, 61% continued to exhibit a normal/neutral RFA when wearing their habitual shoes. Among the overpronators, 100% showed a change in the RM to either normal or supinator. Among the participants classified as supinators, 62% exhibited normal pronation when shod even without using the appropriate footwear, claimed by the manufacturer. Only 44.1% of the sample chose the correct running shoe for their barefoot RM. The majority of runners did not choose their shoes designed for their natural type of RM. The rearfoot eversion angle changed an average 4 degrees when running shod and the RM barefoot altered quite a lot when using a running shoe. The running shoes did not correct the pronation detected barefoot, as claimed by the manufacturers.

Implementation of Telehealth Resources in Primary Care in Brazil and Its Association with Quality of Care.

Background:Although several reviews on the relation between telemedicine and health care outcomes have pointed out some evidence, they have also underscored the need for further investigation.Introduction:Brazil has a national telehealth program implemented in 2007, involving teleconsulting and distance education actions in primary care.Objective:This study aims to describe the implementation of telehealth in primary care in Brazil and to identify if there is an association between telehealth and quality of care.Methods:A cross-sectional study analyzing data from interviews with 29,778 primary care team professionals in Brazil in 2014, which represents 92.5% of existing health care teams, on aspects of information technology, telehealth, and care was carried out. A multiple binary regression analysis was performed to study the associations between the extent of health care actions and using telehealth.Results:Of the 24,055 primary care units in Brazil where teams worked, only 50.1% (n: 12,055) had internet access, and 32.71% of teams claimed to be involved in telehealth actions in 2014. Between 2012 and 2014 there was an expansion in the use of telehealth resources in all regions of Brazil. The highest magnitude was observed for the association between employment of telehealth and improvement in child care (odds ratio [OR] = 2.09), followed by diabetes mellitus care (OR = 1.91), hypertension (OR = 1.89), and finally, women's health (OR = 1.86).

An evaluation of the effectiveness of a chemical additive based on sodium benzoate, potassium sorbate, and sodium nitrite on the fermentation and aerobic stability of corn silage.

We evaluated the effectiveness of an additive comprising sodium benzoate, potassium sorbate, and sodium nitrite (SSL) as active ingredients for its ability to improve the aerobic stability of corn silages made in North America. In experiment 1, treatment with SSL (1.5 and 2.0 L/t) on whole-plant corn (WPC) was compared with treatment with an additive containing buffered propionic acid and citric acid (BPA; 2 L/t) on corn harvested at 32 and 38% dry matter and ensiled for 120 d. Silage treated with BPA was higher in ammonia-N and propionic acid relative to other treatments. Treatments with all of the additives had numerically, but not statistically, fewer yeasts compared with untreated silage. Both application rates of SSL resulted in lower concentrations of ethanol compared with untreated and BPA silages. Treatment with BPA improved the aerobic stability of silages compared with untreated silage, but the effect from SSL was markedly greater. In experiment 2, WPC was untreated or treated with 2 or 3 L of SSL/t or a microbial inoculant containing Enterococcus faecium M74, Lactobacillus plantarum CH6072, and Lactobacillus buchneri LN1819 (final total lactic acid bacteria application rate of 150,000 cfu/g of fresh forage). Silages were air stressed for 24 h at 28 and 42 d of storage and ensiled for 49 d before opening. Inoculation had no effect on acid end products, ethanol, number of yeasts, or aerobic stability compared with other treatments. Treatment with SSL decreased the amount of ethanol, had no effect on number of yeasts, and improved aerobic stability in a dose-dependent manner compared with other treatments. In experiment 3, WPC was untreated or treated with 2 L of SSL/t and ensiled for 5, 15, and 30 d. Treatment with SSL resulted in silage with fewer yeasts and lower concentrations of ethanol after all times of ensiling compared with untreated silage. In addition, SSL improved aerobic stability after each period of ensiling, but the effect was more at 15 and 30 d compared with 5 d of storage. Treating WPC with SSL can improve the aerobic stability of corn silage made in North America, and the effect can be observed as soon as 5 d after ensiling.

Insights into the Effect of the PDMS-Layer on the Kinetics and Thermodynamics of Analyte Sorption onto the Matrix-Compatible Solid Phase Microextraction Coating.

The currently presented research investigated the performance of matrix compatible PDMS-overcoated fibers (PDMS-DVB/PDMS) as compared to unmodified PDMS/DVB coatings using aqueous samples and employing a wide range of analyte polarities, molecular weights, and functionalities. In the first part of the work, a kinetic approach was taken to investigate the effect of the PDMS outer layer on the uptake rate of analytes during the mass transfer process. In short, the results can be simplified into two models: (1) the rate-limiting step is the diffusion through the coating and (2) the rate-limiting step is the diffusion through the aqueous diffusional boundary layer. For polar compounds, according to the theoretical discussion, the rate-limiting step is the diffusion through the coating; therefore, the outer PDMS layer influences the uptake rate into the matrix compatible coatings. On the other hand, for nonpolar compounds, the rate-limiting step of the uptake process is diffusion through the aqueous diffusional boundary layer; as such, the overcoated PDMS does not affect uptake rate into the matrix-compatible coatings as compared to DVB/PDMS fibers. From a thermodynamic point of view, the calculated fiber constants further corroborate the hypothesis that the additional PDMS layer does not impair the extraction phase capacity.

Forensic entomotoxicology revisited-towards professional standardisation of study designs.

Forensic entomotoxicology is the use of insects as evidence of whether a toxicant is present in an environment such as a corpse, river or landscape. The earliest overtly forensic study was published in 1977, and since then, at least 63 papers have been published, most of them focused on the detection of toxicants in insects or on effects of toxicants on diverse insect indicator taxa. A comprehensive review of the published literature revealed various inconsistencies between studies that could be addressed by introducing standard protocols for such studies. These protocols could include selecting widespread and common model organisms (such as Lucilia sericata, Calliphora vicina, Chrysomya megacephala and Dermestes maculatus) and model toxicants (e.g. morphine and amitriptyline) to build up comparative databases; developing a standard matrix for use as a feeding substrate; setting guidelines for statistically adequate sample sizes; and deploying more sophisticated analytical methods from the general field of toxicology. Future studies should then be aimed at refining standardised protocols to improve experimental results, and make these results more comparable between studies.

Capturing Plant Metabolome with Direct-Immersion in Vivo Solid Phase Microextraction of Plant Tissues.

For the first time, an in vivo sampling mode of direct immersion-solid phase microextraction (DI-SPME) was employed to capture the metabolome of living plant specimens, using apple (Malus × domestica Borkh.) as a model system. Metabolites were extracted from apple tissues and introduced by thermal desorption into a comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry instrument. The feasibility of this sampling approach, based on exploitation of microextraction principles, including negligible depletion of free analyte concentrations, solventless sampling and sample preparation, and on-site compatibility, was determined in global metabolite analysis. Rather than adopting an approach of traditional sample preparation, requiring metabolism quenching and laborious sample preparation, the objective of the study was to capture the metabolome in vivo, evaluate the feasibility of the approach to provide unbiased extraction coverage, and compare analytical precision when different SPME sampling modes are employed. The potential of in vivo DI-SPME in quantitative plant metabolomics was assessed by evaluating changes in metabolic fingerprints in response to fruit maturation. The in vivo SPME sampling approach has been demonstrated as capable of sampling living systems with high reproducibility, considering that nearly 50% of hundreds of evaluated compounds included in the determination of analytical performance met the 15% RSD FDA criterion. Esters were extracted with high repeatability (% RSD for hexyl butanoate and butyl butanoate of 16.5 and 5.9, respectively, from 9 determinations in 3 apples) and found to be upregulated in response to apple fruit maturation.

Headspace versus direct immersion solid phase microextraction in complex matrixes: investigation of analyte behavior in multicomponent mixtures.

This work aims to investigate the behavior of analytes in complex mixtures and matrixes with the use of solid-phase microextraction (SPME). Various factors that influence analyte uptake such as coating chemistry, extraction mode, the physicochemical properties of analytes, and matrix complexity were considered. At first, an aqueous system containing analytes bearing different hydrophobicities, molecular weights, and chemical functionalities was investigated by using commercially available liquid and solid porous coatings. The differences in the mass transfer mechanisms resulted in a more pronounced occurrence of coating saturation in headspace mode. Contrariwise, direct immersion extraction minimizes the occurrence of artifacts related to coating saturation and provides enhanced extraction of polar compounds. In addition, matrix-compatible PDMS-modified solid coatings, characterized by a new morphology that avoids coating fouling, were compared to their nonmodified analogues. The obtained results indicate that PDMS-modified coatings reduce artifacts associated with coating saturation, even in headspace mode. This factor, coupled to their matrix compatibility, make the use of direct SPME very practical as a quantification approach and the best choice for metabolomics studies where wide coverage is intended. To further understand the influence on analyte uptake on a system where additional interactions occur due to matrix components, ex vivo and in vivo sampling conditions were simulated using a starch matrix model, with the aim of mimicking plant-derived materials. Our results corroborate the fact that matrix handling can affect analyte/matrix equilibria, with consequent release of high concentrations of previously bound hydrophobic compounds, potentially leading to coating saturation. Direct immersion SPME limited the occurrence of the artifacts, which confirms the suitability of SPME for in vivo applications. These findings shed light into the implementation of in vivo SPME strategies in quantitative metabolomics studies of complex plant-based systems.

Use of a Modelling Tool to Optimise the Organisation of the Demand for Primary Oral Health Care in the Brazilian Unified Health System.

PURPOSE: To describe the use of work process modelling to optimise the organisation of the demand for oral health treatment in primary care units in Brazil. MATERIALS AND METHODS: The oral health care routine was at first described as the "AS IS" model, which was evaluated by the oral team professionals, rearranged, and further described as the "TO BE" model described using a business process management modelling tool. The significant increase in the demand of patients due to restrictions offered by the dental service in addition to non-urgent treatments being avoided by patients during COVID-19 pandemic was also considered. RESULTS: Structuring the work processes in a visual way using modelling tools was useful to picture the entire treatment process and adjust when needed. The use of the managerial tool was useful to understand and reorganise the workflow of organising the demand and ultimately improve the efficiency of the resources. The use of such managerial tools helped oral health professionals to efficiently rearrange their tasks and set priorities to meet their needs. CONCLUSIONS: With the use of management tools, each unit can readjust its structures and ways of working, aiming to improve the quality of public health care services provided to patients.

The effects of the use of customized silicone digital orthoses on pre-ulcerative lesions and plantar pressure during walking in people with diabetic neuropathy: A study protocol for a randomized controlled trial.

Background: People with diabetes and diabetic peripheral neuropathy (DPN) often develop calluses due to toe misalignment and increased plantar pressure. Untreated, these issues can progress into ulcers, making early intervention crucial. This trial protocol aims to evaluate the efficacy and safety of customized silicone digital orthoses in preventing ulcers, pre-ulcerative lesions, and peak pressure during gait in people with DPN. Methods: In this superiority randomized controlled parallel trial with single-blind assessment, 60 participants will be allocated to the control group (CG) or the intervention group (IG). The CG will receive specialized nurse-administered foot care, including callus removal, nail care guidance, and self-care education. The IG will receive the same care plus a customized silicone orthosis for toe realignment for 6 months. Assessments will occur at baseline and 3 and 6 months for the primary outcomes (pre-ulcerative lesions and ulcer incidence) and secondary outcomes (pressure distribution, foot function and health, quality of life, safety, and comfort). Two-way ANOVAs (p < .05) will assess group, time, and group by time effects following an intention-to-treat approach. Conclusion: Although recommended for foot ulcer prevention, custom silicone orthosis adoption remains limited due to the low certainty of evidence. This trial seeks to provide more consistent evidence for the use of toe orthoses in preventing callus and ulcer formation for individuals with DPN. Trial registry: ClinicalTrials.gov (NCT05683106) "Effects of Customized Silicone Digital Orthoses in People with Diabetic Neuropathy" (registered on December 20, 2022).

Molecular Characterization and Genome Mechanical Features of Two Newly Isolated Polyvalent Bacteriophages Infecting Pseudomonas syringae pv. garcae.

Coffee plants have been targeted by a devastating bacterial disease, a condition known as bacterial blight, caused by the phytopathogen Pseudomonas syringae pv. garcae (Psg). Conventional treatments of coffee plantations affected by the disease involve frequent spraying with copper- and kasugamycin-derived compounds, but they are both highly toxic to the environment and stimulate the appearance of bacterial resistance. Herein, we report the molecular characterization and mechanical features of the genome of two newly isolated (putative polyvalent) lytic phages for Psg. The isolated phages belong to class Caudoviricetes and present a myovirus-like morphotype belonging to the genuses Tequatrovirus (PsgM02F) and Phapecoctavirus (PsgM04F) of the subfamilies Straboviridae (PsgM02F) and Stephanstirmvirinae (PsgM04F), according to recent bacterial viruses' taxonomy, based on their complete genome sequences. The 165,282 bp (PsgM02F) and 151,205 bp (PsgM04F) genomes do not feature any lysogenic-related (integrase) genes and, hence, can safely be assumed to follow a lytic lifestyle. While phage PsgM02F produced a morphogenesis yield of 124 virions per host cell, phage PsgM04F produced only 12 virions per host cell, indicating that they replicate well in Psg with a 50 min latency period. Genome mechanical analyses established a relationship between genome bendability and virion morphogenesis yield within infected host cells.

Long COVID Among Kidney Transplant Recipients Appears to Be Attenuated During the Omicron Predominance.

BACKGROUND: Omicron variant has been associated with milder cases of COVID-19 among kidney transplant recipients. However, little is known about postacute sequelae, referred to as Long COVID. METHODS: Prospective, single-center cohort study investigating prevalence and risk factors for Long COVID among kidney transplant recipients during the omicron predominance in Brazil. The analysis included adult patients with confirmed SARS-CoV-2 infection between January 5, 2022, and July 18, 2022, were alive, had a functioning kidney transplant 3 mo after symptom onset, and answered a telephonic survey about physical complains of Long COVID. RESULTS: From the 1529 eligible, 602 (39%) patients responded the survey. Sixteen percent reported a previous SARS-CoV-2 infection, and 85% had been fully vaccinated. The prevalence of Long COVID was 52%, with the most common complaints being weakness (46%), myalgia (41%), dizziness (33%), and headache (31%). Among employed patients, 94% were able to resume their normal work activities. In multivariable analysis, female gender (hazard ratio [HR], 2.14; 95% confidence interval [CI], 1.51-3.02; P < 0.0001), previous SARS-CoV-2 infection (HR, 3.55; 95% CI, 1.91-6.60; P < 0.0001), fatigue (HR, 2.32; 95% CI, 1.18-4.55; P = 0.014), myalgia (HR, 1.48; 95% CI, 1.03-2.15; P = 0.036) during the acute phase, and hospitalization because of COVID-19 (HR, 1.71; 95% CI, 1.06-2.76; P = 0.028) were independently associated with Long COVID. CONCLUSIONS: In the "omicron era," Long COVID among kidney transplant recipients exhibited milder characteristics and had a less significant impact on their ability to resume normal life activities. The risk factors for persistent symptoms were similar to those observed in the general population except for the vaccination status, underscoring the importance of closer monitoring in special subgroups.