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1.
Trends Immunol ; 44(10): 754-756, 2023 10.
Article in English | MEDLINE | ID: mdl-37690961

ABSTRACT

Research on human leukocyte antigen (HLA) molecules in coronavirus disease 2019 (COVID-19) raised high expectations but has yielded limited results. Augusto et al.'s recent study in Nature unveils a strong association of HLA-B*15:01 with asymptomatic COVID-19, representing an important contribution to genetics in COVID-19.


Subject(s)
COVID-19 , Humans , COVID-19/genetics , Alleles , Histocompatibility Antigens Class I/genetics , HLA Antigens/genetics , HLA-B Antigens/genetics
2.
Proc Natl Acad Sci U S A ; 119(15): e2109617119, 2022 04 12.
Article in English | MEDLINE | ID: mdl-35353605

ABSTRACT

α-Synuclein (α-syn) phosphorylation at serine 129 (pS129­α-syn) is substantially increased in Lewy body disease, such as Parkinson's disease (PD) and dementia with Lewy bodies (DLB). However, the pathogenic relevance of pS129­α-syn remains controversial, so we sought to identify when pS129 modification occurs during α-syn aggregation and its role in initiation, progression and cellular toxicity of disease. Using diverse aggregation assays, including real-time quaking-induced conversion (RT-QuIC) on brain homogenates from PD and DLB cases, we demonstrated that pS129­α-syn inhibits α-syn fibril formation and seeded aggregation. We also identified lower seeding propensity of pS129­α-syn in cultured cells and correspondingly attenuated cellular toxicity. To build upon these findings, we developed a monoclonal antibody (4B1) specifically recognizing nonphosphorylated S129­α-syn (WT­α-syn) and noted that S129 residue is more efficiently phosphorylated when the protein is aggregated. Using this antibody, we characterized the time-course of α-syn phosphorylation in organotypic mouse hippocampal cultures and mice injected with α-syn preformed fibrils, and we observed aggregation of nonphosphorylated α-syn followed by later pS129­α-syn. Furthermore, in postmortem brain tissue from PD and DLB patients, we observed an inverse relationship between relative abundance of nonphosphorylated α-syn and disease duration. These findings suggest that pS129­α-syn occurs subsequent to initial protein aggregation and apparently inhibits further aggregation. This could possibly imply a potential protective role for pS129­α-syn, which has major implications for understanding the pathobiology of Lewy body disease and the continued use of reduced pS129­α-syn as a measure of efficacy in clinical trials.


Subject(s)
Amyloid , Lewy Body Disease , Parkinson Disease , Protein Aggregation, Pathological , alpha-Synuclein , Amyloid/metabolism , Humans , Lewy Body Disease/genetics , Lewy Body Disease/metabolism , Parkinson Disease/genetics , Parkinson Disease/metabolism , Phosphorylation , Protein Aggregates , Protein Aggregation, Pathological/genetics , Protein Aggregation, Pathological/metabolism , Serine/metabolism , alpha-Synuclein/genetics , alpha-Synuclein/metabolism
3.
Crit Rev Biochem Mol Biol ; 57(5-6): 492-538, 2022.
Article in English | MEDLINE | ID: mdl-36877487

ABSTRACT

Understanding how Nature accomplishes the reduction of inert nitrogen gas to form metabolically tractable ammonia at ambient temperature and pressure has challenged scientists for more than a century. Such an understanding is a key aspect toward accomplishing the transfer of the genetic determinants of biological nitrogen fixation to crop plants as well as for the development of improved synthetic catalysts based on the biological mechanism. Over the past 30 years, the free-living nitrogen-fixing bacterium Azotobacter vinelandii emerged as a preferred model organism for mechanistic, structural, genetic, and physiological studies aimed at understanding biological nitrogen fixation. This review provides a contemporary overview of these studies and places them within the context of their historical development.


Subject(s)
Azotobacter vinelandii , Nitrogen Fixation , Azotobacter vinelandii/genetics , Azotobacter vinelandii/metabolism , Nitrogenase/chemistry , Nitrogenase/genetics , Nitrogenase/metabolism , Ammonia , Nitrogen
4.
J Bacteriol ; 206(4): e0045223, 2024 Apr 18.
Article in English | MEDLINE | ID: mdl-38551342

ABSTRACT

The wobble bases of tRNAs that decode split codons are often heavily modified. In bacteria, tRNAGlu, Gln, Asp contains a variety of xnm5s2U derivatives. The synthesis pathway for these modifications is complex and fully elucidated only in a handful of organisms, including the Gram-negative Escherichia coli K12 model. Despite the ubiquitous presence of mnm5s2U modification, genomic analysis shows the absence of mnmC orthologous genes, suggesting the occurrence of alternate biosynthetic schemes for the conversion of cmnm5s2U to mnm5s2U. Using a combination of comparative genomics and genetic studies, a member of the YtqA subgroup of the radical Sam superfamily was found to be involved in the synthesis of mnm5s2U in both Bacillus subtilis and Streptococcus mutans. This protein, renamed MnmL, is encoded in an operon with the recently discovered MnmM methylase involved in the methylation of the pathway intermediate nm5s2U into mnm5s2U in B. subtilis. Analysis of tRNA modifications of both S. mutans and Streptococcus pneumoniae shows that growth conditions and genetic backgrounds influence the ratios of pathway intermediates owing to regulatory loops that are not yet understood. The MnmLM pathway is widespread along the bacterial tree, with some phyla, such as Bacilli, relying exclusively on these two enzymes. Although mechanistic details of these newly discovered components are not fully resolved, the occurrence of fusion proteins, alternate arrangements of biosynthetic components, and loss of biosynthetic branches provide examples of biosynthetic diversity to retain a conserved tRNA modification in Nature.IMPORTANCEThe xnm5s2U modifications found in several tRNAs at the wobble base position are widespread in bacteria where they have an important role in decoding efficiency and accuracy. This work identifies a novel enzyme (MnmL) that is a member of a subgroup of the very versatile radical SAM superfamily and is involved in the synthesis of mnm5s2U in several Gram-positive bacteria, including human pathogens. This is another novel example of a non-orthologous displacement in the field of tRNA modification synthesis, showing how different solutions evolve to retain U34 tRNA modifications.


Subject(s)
Escherichia coli K12 , RNA, Transfer , Humans , RNA, Transfer/genetics , Escherichia coli K12/genetics , Bacteria/genetics , Methylation , Gram-Positive Bacteria/genetics
5.
J Oral Pathol Med ; 53(6): 341-357, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38782020

ABSTRACT

BACKGROUND: Head and neck cancer encompasses neoplasms affecting the oral cavity, pharynx, larynx, and thyroid. Identifying factors that modulate the carcinogenesis process can aid in identifying subgroups at higher risk of developing the disease, enabling implementation of prevention programs. Vitamin D receptor polymorphisms can affect the carcinogenesis of various tumors by altering vitamin D metabolism and cellular response. METHODS: To elucidate the role of vitamin D receptor polymorphisms in head and neck cancer, a systematic review was performed, searching the Embase, PubMed, Scopus, and Lilacs databases. A total of 19 articles met the inclusion criteria. The frequency of vitamin D receptors polymorphism alleles (FokI, ApaI, BsmI, TaqI, Cdx2, rs2107301, rs2238135) was recorded and pooled to calculate the odds ratio in a meta-analysis using the Review Manager software. RESULTS: Subgroup analysis demonstrated significant associations in the anatomical site of cancer (oral cancer in ApaI and BsmI, and unspecified subsites of head and neck cancer in TaqI), genotyping method (FokI and BsmI), and continent of the study (ApaI, FokI, and BsmI). CONCLUSION: Our findings were heterogeneous, as with other evidence available in the literature. Therefore, more clinical studies with larger sample sizes are needed to obtain more accurate results on the relationship between vitamin D receptor polymorphism and head and neck cancer.


Subject(s)
Genetic Predisposition to Disease , Head and Neck Neoplasms , Polymorphism, Genetic , Receptors, Calcitriol , Receptors, Calcitriol/genetics , Humans , Head and Neck Neoplasms/genetics , Risk Factors , Genotype
6.
BMC Musculoskelet Disord ; 25(1): 172, 2024 Feb 24.
Article in English | MEDLINE | ID: mdl-38402150

ABSTRACT

OBJECTIVE: To analyze the effects of using foam roller on pain intensity in individuals with chronic and acute musculoskeletal pain. METHODS: This systematic review was registered in the National Institute for Health Research's prospective online registry of systematic reviews (PROSPERO) under CRD42023456841. The databases Pubmed, Medline (via Ovid), Embase, BVS, and PEDro (Physiotherapy Evidence Database) were consulted to carry out this systematic review. Notably, the records of clinical trials characterized as eligible were manually searched. The search terms were: (foam rolling OR foam rolling vibration) AND (acute musculoskeletal pain) AND (chronic musculoskeletal pain). The search was performed until August 22, 2023. For the analysis of the methodological quality, the PEDro scale was used for each of the manuscripts included in the systematic review. Due to the heterogeneity in the studies included in this systematic review, performing a meta-analysis of the analyzed variables was impossible. RESULTS: Only six manuscripts were eligible for data analysis. The type of FR used was non-vibrational, being applied by a therapist in only one of the manuscripts. With an application time ranging from at least 45 s to 15 min, the non-vibrational FR was applied within a day up to six weeks. Using the PEDro scale, scores were assigned that varied between 4 and 8 points, with an average of 6 ± 1.29 points. Only two randomized clinical trials found a significant benefit in pain intensity of adding FR associated with a therapeutic exercise protocol in individuals with patellofemoral pain syndrome and chronic neck pain. CONCLUSION: The results of this systematic review do not elucidate or reinforce the clinical use of FR in pain intensity in individuals with chronic and acute musculoskeletal pain.


Subject(s)
Acute Pain , Chronic Pain , Musculoskeletal Pain , Humans , Musculoskeletal Pain/diagnosis , Musculoskeletal Pain/therapy , Pain Measurement , Prospective Studies , Randomized Controlled Trials as Topic , Systematic Reviews as Topic , Acute Pain/diagnosis , Acute Pain/therapy , Chronic Pain/diagnosis , Chronic Pain/therapy
7.
Biomed Chromatogr ; 38(2): e5781, 2024 Feb.
Article in English | MEDLINE | ID: mdl-37994231

ABSTRACT

Sulfamethazine (SMZ), trimethoprim (TMP) and doxycycline (DOXY) are drugs of choice used in the treatment of intestinal and respiratory infections that affect poultry and swine. The aim of this study was develop and validate a simple, sensitive and fast method for the simultaneous determination of SMZ, TMP and DOXY in veterinary formulations by high-performance liquid chromatography. The separation was performed on a Macherey-Nagel C8 analytical column (4 × 125 mm, 5 µm), with a flow rate of 0.5 ml min-1 and detection at 268, 270 and 350 nm, for SMZ, TMP and DOXY, respectively. All measurements were performed in acetonitrile-water (45:55 v/v; pH 3.0). The analytical curves were linear (r > 0.9997) in the concentration range of 5.0-35.0 µg ml-1 for SMZ, 1.0-7.0 µg ml-1 for TMP and 7.0-13.0 µg ml-1 for DOXY. The method proved to be precise, robust, accurate and selective. In accelerated stability, the sample was analyzed for 6 months, with no major variations observed in organoleptic analysis and pH. Therefore, the developed method was proved to be suitable for routine quality control analyses for the simultaneous determination of SMZ, TMP and DOXY in pharmaceutical formulations.


Subject(s)
Sulfamethazine , Trimethoprim , Animals , Swine , Trimethoprim/analysis , Chromatography, High Pressure Liquid/methods , Sulfamethazine/analysis , Doxycycline , Water
8.
Neurocrit Care ; 2024 Mar 05.
Article in English | MEDLINE | ID: mdl-38443709

ABSTRACT

BACKGROUND: Early posttraumatic brain injury (TBI) tranexamic acid (TXA) may reduce blood-brain barrier (BBB) permeability, but it is unclear if this effect is fixed regardless of dose. We hypothesized that post-TBI TXA demonstrates a dose-dependent reduction of in vivo penumbral leukocyte mobilization, BBB microvascular permeability, and enhancement of neuroclinical recovery. METHODS: CD1 male mice (n = 40) were randomly assigned to TBI by controlled cortical impact (injury [I]) or sham TBI (S), followed by intravenous bolus of either saline (placebo [P]) or TXA (15, 30, or 60 mg/kg). At 48 h, in vivo pial intravital microscopy visualized live penumbral BBB microvascular leukocytes and albumin leakage. Neuroclinical recovery was assessed by Garcia Neurological Test scores and animal weight changes at 24 h and 48 h after injury. RESULTS: I + TXA60 reduced live penumbral leukocyte rolling compared with I + P (p < 0.001) and both lower TXA doses (p = 0.017 vs. I + TXA15, p = 0.012 vs. I + TXA30). Leukocyte adhesion was infrequent and similar across groups. Only I + TXA60 significantly reduced BBB permeability compared with that in the I + P (p = 0.004) group. All TXA doses improved Garcia Test scores relative to I + P at both 24 h and 48 h (p < 0.001 vs. I + P for all at both time points). Mean 24-h body weight loss was greatest in the I + P (- 8.7 ± 1.3%) group and lowest in the I + TXA15 (- 4.4 ± 1.0%, p = 0.051 vs. I + P) group. CONCLUSIONS: Only higher TXA dosing definitively abrogates penumbral leukocyte mobilization, preserving BBB integrity post TBI. Some neuroclinical recovery is observed, even with lower TXA dosing. Better outcomes with higher dose TXA after TBI may occur secondary to blunting of leukocyte-mediated penumbral cerebrovascular inflammation.

9.
Int J Mol Sci ; 25(10)2024 May 12.
Article in English | MEDLINE | ID: mdl-38791312

ABSTRACT

Glioblastomas (GBM) are the most common primary malignant brain tumors, comprising 2% of all cancers in adults. Their location and cellular and molecular heterogeneity, along with their highly infiltrative nature, make their treatment challenging. Recently, our research group reported promising results from a prospective phase II clinical trial involving allogeneic vaccination with dendritic cells (DCs). To date, six out of the thirty-seven reported cases remain alive without tumor recurrence. In this study, we focused on the characterization of infiltrating immune cells observed at the time of surgical resection. An analytical model employing a neural network-based predictive algorithm was used to ascertain the potential prognostic implications of immunological variables on patients' overall survival. Counterintuitively, immune phenotyping of tumor-associated macrophages (TAMs) has revealed the extracellular marker PD-L1 to be a positive predictor of overall survival. In contrast, the elevated expression of CD86 within this cellular subset emerged as a negative prognostic indicator. Fundamentally, the neural network algorithm outlined here allows a prediction of the responsiveness of patients undergoing dendritic cell vaccination in terms of overall survival based on clinical parameters and the profile of infiltrated TAMs observed at the time of tumor excision.


Subject(s)
Brain Neoplasms , Dendritic Cells , Glioblastoma , Immunotherapy , Humans , Dendritic Cells/immunology , Glioblastoma/therapy , Glioblastoma/immunology , Glioblastoma/mortality , Glioblastoma/pathology , Immunotherapy/methods , Brain Neoplasms/immunology , Brain Neoplasms/therapy , Brain Neoplasms/mortality , Brain Neoplasms/pathology , Male , Female , Middle Aged , B7-H1 Antigen/metabolism , Prognosis , Adult , Tumor-Associated Macrophages/immunology , Tumor-Associated Macrophages/metabolism , Aged , Lymphocytes, Tumor-Infiltrating/immunology , Lymphocytes, Tumor-Infiltrating/metabolism
10.
Am J Orthod Dentofacial Orthop ; 165(3): 256-261, 2024 Mar.
Article in English | MEDLINE | ID: mdl-38149955

ABSTRACT

Molar incisor hypomineralization (MIH) is a systemic developmental qualitative defect of the enamel that affects 1-4 first permanent molars with or without incisor involvement. Enamel hypomineralization is identified visually as a demarcated opacity with a clear border with varying extensions and can be white, yellow, or brown. This opacity is characterized by a reduced quality of enamel that is of normal thickness but not fully mineralized. It is very common for the affected teeth to present with posteruptive breakdown, making them susceptible to caries, leading to their subsequent loss. Thus, MIH is a significant dental problem with clinical, economic, and psychosocial implications. The planned extraction of compromised teeth is a valid alternative to complex restorative treatment. Orthodontists' involvement is essential in developing effective treatment strategies for children affected by MIH, contributing to their oral health and well-being.


Subject(s)
Dental Enamel Hypoplasia , Molar Hypomineralization , Child , Humans , Dental Enamel Hypoplasia/therapy , Orthodontists , Prevalence , Dental Enamel
11.
Article in English | MEDLINE | ID: mdl-38935005

ABSTRACT

INTRODUCTION: This study evaluated the probability of developing malocclusions in mixed dentition. METHODS: A longitudinal study was conducted with 598 children (aged 5 years) in deciduous dentition. The children were followed for 3 years until mixed dentition (aged 8 years). Overjet, overbite, and transversal relations were evaluated. Bayesian models were used to analyze the data and estimate the parameters. RESULTS: The parameter θ was used for the distributions, indicating the probability of presenting a given condition with a credibility index (ICr) of 95%. After 3 years of follow-up, 121 children were reevaluated. The results showed that children have a high probability of malocclusion in mixed dentition. There was a higher probability of developing an increased overjet in the mixed dentition of 20.5% (ICr 95%, 13.6-28.4) to 48.3% (ICr 95%, 39.1-57.7) and a higher probability of having a normal overbite in the deciduous dentition and a lower probability in the mixed dentition (ICr 95%, 9.2-21.3). CONCLUSIONS: Considering the probabilistic model of Bayesian analysis, children with normal overjet in the deciduous dentition may show an increased overjet in the mixed dentition. Concerning overbite, children may present an anterior open bite during the transition between deciduous and mixed dentition, as well as self-correction of deep overbite in mixed dentition. Furthermore, they may present a posterior crossbite during the mixed dentition when there is a normal transverse relationship in the deciduous dentition.

12.
Environ Geochem Health ; 46(6): 202, 2024 May 02.
Article in English | MEDLINE | ID: mdl-38696051

ABSTRACT

Determining the origin and pathways of contaminants in the natural environment is key to informing any mitigation process. The mass magnetic susceptibility of soils allows a rapid method to measure the concentration of magnetic minerals, derived from anthropogenic activities such as mining or industrial processes, i.e., smelting metals (technogenic origin), or from the local bedrock (of geogenic origin). This is especially effective when combined with rapid geochemical analyses of soils. The use of multivariate analysis (MVA) elucidates complex multiple-component relationships between soil geochemistry and magnetic susceptibility. In the case of soil mining sites, X-ray fluorescence (XRF) spectroscopic data of soils contaminated by mine waste shows statistically significant relationships between magnetic susceptibility and some base metal species (e.g., Fe, Pb, Zn, etc.). Here, we show how qualitative and quantitative MVA methodologies can be used to assess soil contamination pathways using mass magnetic susceptibility and XRF spectra of soils near abandoned coal and W/Sn mines (NW Portugal). Principal component analysis (PCA) showed how the first two primary components (PC-1 + PC-2) explained 94% of the sample variability, grouped them according to their geochemistry and magnetic susceptibility in to geogenic and technogenic groups. Regression analyses showed a strong positive correlation (R2 > 0.95) between soil geochemistry and magnetic properties at the local scale. These parameters provided an insight into the multi-element variables that control magnetic susceptibility and indicated the possibility of efficient assessment of potentially contaminated sites through mass-specific soil magnetism.


Subject(s)
Environmental Monitoring , Soil Pollutants , Spectrometry, X-Ray Emission , Soil Pollutants/analysis , Spectrometry, X-Ray Emission/methods , Multivariate Analysis , Environmental Monitoring/methods , Mining , Portugal , Principal Component Analysis , Soil/chemistry , Tin/analysis , Magnetic Phenomena , Coal Mining , Coal
13.
J Contemp Dent Pract ; 25(4): 295-302, 2024 Apr 01.
Article in English | MEDLINE | ID: mdl-38956842

ABSTRACT

AIM: The purpose of this study was to evaluate the mandibular growth and/or projection following maxillary incisor proclination, overbite correction, and maxillary dentoalveolar expansion without the use of any class II mechanics, in growing class II division 2 patients treated with clear aligners. MATERIALS AND METHODS: Before and after treatment cone-beam computed tomographic (CBCT) generated lateral and posteroanterior cephalograms of thirty-two patients with skeletal class II division 2, 16 in the treatment group and 16 in the untreated group, were reviewed to evaluate treatment-related changes. Upper incisors were proclined and protruded, as well as upper arch expansion and overbite correction were performed as part of their regular treatment. Cephalometric analysis was performed to evaluate skeletal and dental changes. Unpaired statistical t-tests were performed to determine if significant skeletal class II correction was achieved in the treatment group. RESULTS: In the treatment group, after treatment, the upper incisors became more proclined and protruded, and the inter-molar width increased while the overbite was reduced compared to the control group. An increase in skeletal mandibular growth and forward projection was also observed, thus contributing to an improvement of the sagittal skeletal relationship as evidenced by ANB and Wits values compared to the control group. CONCLUSION: A combination of upper incisor proclination, correction of deep overbite, and maxillary dentoalveolar expansion using clear aligners appears to contribute to an improvement of the skeletal class II relationship in growing patients with class II division 2. CLINICAL SIGNIFICANCE: This study shows that unlocking the mandible by correcting a deep overbite, proclining upper incisors, and expanding the upper arch in growing class II division 2 patients can improve skeletal class II using clear aligners. How to cite this article: Mirzasoleiman P, El-Bialy T, Wiltshire WA, et al. Evaluation of Mandibular Projection in Class II Division 2 Subjects Following Orthodontic Treatment Using Clear Aligners. J Contemp Dent Pract 2024;25(4):295-302.


Subject(s)
Cephalometry , Cone-Beam Computed Tomography , Malocclusion, Angle Class II , Mandible , Humans , Malocclusion, Angle Class II/therapy , Malocclusion, Angle Class II/diagnostic imaging , Mandible/growth & development , Mandible/diagnostic imaging , Female , Male , Adolescent , Child , Incisor/diagnostic imaging , Overbite/therapy , Orthodontics, Corrective/methods , Orthodontics, Corrective/instrumentation
14.
Pain Pract ; 24(1): 211-230, 2024 Jan.
Article in English | MEDLINE | ID: mdl-37661339

ABSTRACT

PURPOSE: The aim of this review was to compare the heart rate variability (HRV) responses at rest of adults with chronic musculoskeletal pain against healthy controls. METHODS: The PubMed, Scopus, Web of Science (Science and Social Science Citation Index), and CINAHL databases were searched, with no date restrictions. Two independent reviewers selected observational studies that characterized the HRV responses at rest in adults with chronic musculoskeletal pain compared with those of healthy controls. Methodological quality was assessed using the Downs and Black checklist. RESULTS: This study followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement. HRV in adults with chronic musculoskeletal pain was evaluated systematically. Of the 4893 studies screened, 20 of poor-to-moderate quality met the inclusion criteria. Most studies used electrocardiography and at least one time and/or frequency domain index. Studies were found that investigated HRV in adults with temporomandibular disorders, neck pain, whiplash, low back pain, and fibromyalgia. The heterogeneity of the studies in relation to painful conditions, parameters or position for HRV analysis precluded a meta-analysis. In general, these studies seem to show increased sympathetic and decreased parasympathetic modulation in adults with musculoskeletal pain when compared to controls. CONCLUSIONS: Adults with musculoskeletal pain exhibited a decline in HRV compared to controls. However, definitive conclusions cannot be drawn since the evidence is heterogeneous and of moderate quality. Further high-quality research with standardized measurements is needed.


Subject(s)
Chronic Pain , Fibromyalgia , Musculoskeletal Pain , Adult , Humans , Heart Rate/physiology , Neck Pain
15.
Clin Infect Dis ; 77(Suppl 7): S588-S596, 2023 12 20.
Article in English | MEDLINE | ID: mdl-38118017

ABSTRACT

BACKGROUND: Strengthening external quality assessment (EQA) services across the One Health sector supports implementation of effective antimicrobial resistance (AMR) control strategies. Here we describe and compare 2 different approaches for conducting virtual laboratory follow-up assessments within an EQA program to evaluate quality management system (QMS) and procedures for pathogen identification and antimicrobial susceptibility testing (AST). METHODS: During the coronavirus disease 2019 (COVID-19) pandemic in 2021 and 2022, 2 laboratory assessment approaches were introduced: virtual-based and survey-based methodologies. The evaluation of 2 underperforming Animal Health laboratories through a virtual-based approach occurred between May and August 2021. This evaluation encompassed the utilization of 3 online meetings and document reviews, performed subsequent to the execution of EQA procedures. Within a distinct group of laboratories, the survey-based assessment was implemented from December 2021 to February 2022, also following EQA procedures. This phase encompassed the dissemination of an online survey to 31 participating laboratories, alongside a sole online consultation meeting involving 4 specific underperforming laboratories. RESULTS: The virtual-based assessment post-EQA aimed to identify gaps and areas for improvement in the laboratory's practices for pathogen identification and AST. This approach was, however, time-intensive, and, hence, only 2 laboratories were assessed. In addition, limited interactions in virtual platforms compromised the assessment quality. The survey-based post-EQA assessment enabled evaluation of 31 laboratories. Despite limitations for in-depth analysis of each procedure, gaps in QMS across multiple laboratories were identified and tailored laboratory-specific recommendations were provided. CONCLUSIONS: Reliable internet and plans for efficient time management, post-EQA virtual laboratory follow-up assessments are an effective alternative when conducting onsite evaluation is infeasible as observed during the COVID-19 pandemic, although the successful implementation of remediation plans will likely require in person assessments. We advocate application of hybrid approaches (both onsite and virtual) for targeted capacity building of AMR procedures with the ability to implement and oversee the process.


Subject(s)
Anti-Infective Agents , COVID-19 , One Health , Humans , Quality Control , Laboratories , Pandemics/prevention & control , Asia , Quality Assurance, Health Care , COVID-19 Testing
16.
J Am Chem Soc ; 145(6): 3443-3453, 2023 02 15.
Article in English | MEDLINE | ID: mdl-36689349

ABSTRACT

The generation of enantiodivergent biocatalysts for C-H oxyfunctionalizations is ever more important in modern synthetic chemistry. Here, we have applied the FuncLib algorithm based on phylogenetic and Rosetta calculations to design a diverse repertoire of active, stable, and enantiodivergent fungal peroxygenases. 24 designs, each carrying 4-5 mutations in the catalytic core, were expressed functionally in yeast and benchmarked against characteristic model compounds. Several designs were active and stable in a range of temperature and pH, displaying unprecedented enantiodivergence, changing regioselectivity from alkyl to aromatic hydroxylation, and increasing catalytic efficiencies up to 10-fold, with 15-fold improvements in total turnover numbers over the parental enzyme. We find that this dramatic functional divergence stems from beneficial epistasis among the mutations and an extensive reorganization of the heme channel. Our work demonstrates that FuncLib can rapidly design highly functional libraries enriched in enantioselective peroxygenases not seen in nature for a range of biotechnological applications.


Subject(s)
Mixed Function Oxygenases , Saccharomyces cerevisiae , Phylogeny , Mixed Function Oxygenases/chemistry , Catalysis , Catalytic Domain , Saccharomyces cerevisiae/metabolism
17.
Mol Med ; 29(1): 111, 2023 08 18.
Article in English | MEDLINE | ID: mdl-37596531

ABSTRACT

BACKGROUND: Soluble oligomeric forms of alpha-synuclein (aSyn-O) are believed to be one of the main toxic species in Parkinson's disease (PD) leading to degeneration. aSyn-O can induce Ca2+ influx, over activating downstream pathways leading to PD phenotype. Calcineurin (CN), a phosphatase regulated by Ca2+ levels, activates NFAT transcription factors that are involved in the regulation of neuronal plasticity, growth, and survival. METHODS: Here, using a combination of cell toxicity and gene regulation assays performed in the presence of classical inhibitors of the NFAT/CN pathway, we investigate NFAT's role in neuronal degeneration induced by aSyn-O. RESULTS: aSyn-O are toxic to neurons leading to cell death, loss of neuron ramification and reduction of synaptic proteins which are reversed by CN inhibition with ciclosporin-A or VIVIT, a NFAT specific inhibitor. aSyn-O induce NFAT nuclear translocation and transactivation. We found that aSyn-O modulates the gene involved in the maintenance of synapses, synapsin 1 (Syn 1). Syn1 mRNA and protein and synaptic puncta are drastically reduced in cells treated with aSyn-O which are reversed by NFAT inhibition. CONCLUSIONS: For the first time a direct role of NFAT in aSyn-O-induced toxicity and Syn1 gene regulation was demonstrated, enlarging our understanding of the pathways underpinnings synucleinopathies.


Subject(s)
Parkinson Disease , alpha-Synuclein , Humans , alpha-Synuclein/genetics , T-Lymphocytes , Homeostasis , Apoptosis , Calcineurin
18.
Ann Bot ; 131(4): 697-721, 2023 04 28.
Article in English | MEDLINE | ID: mdl-36821492

ABSTRACT

BACKGROUND AND AIMS: The most species-rich and ecologically diverse plant radiation on the Canary Islands is the Aeonium alliance (Crassulaceae). In island radiations like this, speciation can take place either within islands or following dispersal between islands. Aiming at quantifying intra- and inter-island speciation events in the evolution of Aeonium, and exploring their consequences, we hypothesized that (1) intra-island diversification resulted in stronger ecological divergence of sister lineages, and that (2) taxa on islands with a longer history of habitation by Aeonium show stronger ecological differentiation and produce fewer natural hybrids. METHODS: We studied the biogeographical and ecological setting of diversification processes in Aeonium with a fully sampled and dated phylogeny inferred using a ddRADseq approach. Ancestral areas and biogeographical events were reconstructed in BioGeoBEARS. Eleven morphological characters and three habitat characteristics were taken into account to quantify the morphological and ecological divergence between sister lineages. A co-occurrence matrix of all Aeonium taxa is presented to assess the spatial separation of taxa on each island. KEY RESULTS: We found intra- and inter-island diversification events in almost equal numbers. In lineages that diversified within single islands, morphological and ecological divergence was more pronounced than in lineages derived from inter-island diversification, but only the difference in morphological divergence was significant. Those islands with the longest history of habitation by Aeonium had the lowest percentages of co-occurring and hybridizing taxon pairs compared with islands where Aeonium arrived later. CONCLUSIONS: Our findings illustrate the importance of both inter- and intra-island speciation, the latter of which is potentially sympatric speciation. Speciation on the same island entailed significantly higher levels of morphological divergence compared with inter-island speciation, but ecological divergence was not significantly different. Longer periods of shared island habitation resulted in the evolution of a higher degree of spatial separation and stronger reproductive barriers.


Subject(s)
Crassulaceae , Ecosystem , Genetic Speciation , Phylogeny , Islands
19.
BMC Geriatr ; 23(1): 151, 2023 03 20.
Article in English | MEDLINE | ID: mdl-36941534

ABSTRACT

BACKGROUND: The COVID-19 pandemic primarily impacted long-term care facilities by restricting visiting and circulation of visitors, affecting the quality of life (QoL) of older adults living in these institutions. Volunteer activities, essential for older adults' daily life, were also interrupted and potentially negatively impacted the QoL of older adults, volunteers themselves, and also employees in these institutions. In this context, this study aims to evaluate the impact of the return of volunteer-led activities in a long-term care institution on the QoL of older adult residents, employees, and volunteers. METHODS: This study used a pre-test and post-test design within the same group. The first round of data collection was conducted before volunteer-led activities returned and the second round after 1 month of return. The instrument used to assess QoL was the EUROHIS-QoL-8 scale. This study was conducted within a nursing home in São Paulo, Brazil, created in 1937 by members of the Israeli community living in Brazil. Volunteer-led activities were part of residents' daily life before the COVID-19 pandemic, when these activities were interrupted for about 20 months. A total of 79 individuals participated in both rounds (pre and post), of which: 29 residents, 27 volunteers, and 23 employees of the long-term care institution. RESULTS: Using a Wilcoxon signed-rank test, the analyses indicated improvements after the 1 month return in different QoL aspects for the three groups. Volunteers improved their personal relationships (Z - 2.332, p < .05), residents their overall health (Z - 2.409, p < .05) and employees in their overall QoL perception (Z - 2.714, p < .05). Influencing factors for residents were the number of activities (3 or more), gender (male), and education (undergraduate/graduate). For employees, those who assumed additional activities due to the volunteer-led activities interruption had a significant impact on their overall QoL. CONCLUSIONS: Evidence from this study suggests that volunteers' return positively impacted different QoL aspects for volunteers, residents, and employees.


Subject(s)
COVID-19 , Long-Term Care , Humans , Male , Aged , Quality of Life , Cohort Studies , Pandemics , Brazil , COVID-19/epidemiology , Volunteers
20.
BMC Musculoskelet Disord ; 24(1): 574, 2023 Jul 14.
Article in English | MEDLINE | ID: mdl-37452316

ABSTRACT

BACKGROUND: Assessment instruments play an essential role in the management of knee osteoarthritis. This study aimed to verify the clinimetric properties and validate the short version of WOMAC's (SV-WOMAC) knee with two domains, pain (four items) and physical function (eight items) in individuals with knee osteoarthritis (KO). METHODS: Reliability and internal consistency Construct, criterion validity, Ceiling, and floor effects analyses were performed. In addition to the SV-WOMAC, the following instruments were used: the numerical rating scale (NRPS), International Knee Documentation Committee (IKDC), the Short Form Health Survey (SF-36), and WOMAC's original version. Spearman's correlation coefficient (rho) was used to determine the magnitude of the correlation between the AFAQ and the other instruments. Moreover, the test-retest reliability and internal consistency were assessed using the intraclass correlation coefficient (ICC) and Cronbach's alpha, respectively. In addition, standard error of measurement (SEM) and minimum detectable change (MDC) were calculated. RESULTS: One hundred and thirteen subjects with KO were included for validity analysis, and a subsample of 53 subjects was used for test-retest reliability. Adequate reliability and internal consistency were observed with ICC ≥ 0.76, SEM ≤ 1.85, MDC ≥ 5.1, and Cronbach's alpha ≥ 0.84. Regarding construct validity, correlations greater than 0.50 were observed with the IKDC, NRPS, and functional domains of the SF-36. The SV-WOMAC showed a correlation > 0.70 with the original version and did not show ceiling and floor effects. CONCLUSION: The SV-WOMAC knee has adequate measurement properties to analyze pain and physical function in Brazilian individuals with KO.


Subject(s)
Osteoarthritis, Knee , Humans , Osteoarthritis, Knee/diagnosis , Ontario , Reproducibility of Results , Universities , Pain , Severity of Illness Index , Surveys and Questionnaires
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