Antimicrobial Sensitivity Profile in Psittacine Birds at an Avian Teaching Hospital: A Retrospective Study, 2015-2022.

Veterinary hospitals house patient populations with diverse infectious statuses, microbiota, and histories of prior antibiotic therapy. Choanal swabs are commonly used for assessing the upper respiratory tract of birds for bacterial disease, with the samples submitted for cytologic testing and/or culture and antimicrobial sensitivity testing. The aim of this retrospective study was to identify and quantify bacteria isolated from choanal swabs collected from psittacine patients at a veterinary teaching hospital in Mexico City, Mexico. Data regarding bacterial isolates from choanal swabs were obtained from the medical records of companion psittacines suspected of upper respiratory bacterial disease that presented between November 2015 and December 2022. A total of 47.8% (175 of 366) of the bacterial isolates were from specimens obtained from red-lored Amazons (Amazona autumnalis). Gram-negative bacteria predominated, with 27 different genera identified. Klebsiella, Staphylococcus, and Escherichia were the most frequently isolated genera. A total of 90.4% (331 of 366) of the isolates were resistant to at least 1 antibiotic tested in the sensitivity panel, and a single Klebsiella isolate was resistant to 13 different antibiotics. Gentamicin had a high percentage of efficacy (79.5%; 182 of 229) against the bacterial isolates, whereas isolates tested against sulfonamide-trimethoprim (46.7%, 98 of 210), streptomycin (43.8%; 88 of 201), and clindamycin (12.9%; 15 of 116) had susceptibilities <50%. This is the first study to report common bacterial isolates and their antimicrobial susceptibility patterns from choanal swab samples collected from companion psittacines suspected of upper respiratory disease in Mexico. Clinicians can use the information presented in this study as a guide for therapeutic decision-making when managing upper respiratory bacterial infections in companion psittacine patients.

A potent candicidal peptide designed based on an encrypted peptide from a proteinase inhibitor.

Antimicrobial peptides (AMP) represent an alternative in the treatment of fungal infections associated with countless deaths. Here, we report a new AMP, named KWI-19, which was designed based on a peptide encrypted in the sequence of an Inga laurina Kunitz-type inhibitor (ILTI). KWI-19 inhibited the growth of Candida species and acted as a fungicidal agent from 2.5 to 20 µmol L-1, also showing synergistic activity with amphotericin B. Kinetic assays showed that KWI-19 killed Candida tropicalis cells within 60 min. We also report the membrane-associated mechanisms of action of KWI-19 and its interaction with ergosterol. KWI-19 was also characterized as a potent antibiofilm peptide, with activity against C. tropicalis. Finally, non-toxicity was reported against Galleria mellonella larvae, thus strengthening the interest in all the bioactivities mentioned above. This study extends our knowledge on how AMPs can be engineered from peptides encrypted in larger proteins and their potential as candicidal agents.

Synthetic angiotensin II peptide derivatives confer protection against cerebral and severe non-cerebral malaria in murine models.

Malaria can have severe long-term effects. Even after treatment with antimalarial drugs eliminates the parasite, survivors of cerebral malaria may suffer from irreversible brain damage, leading to cognitive deficits. Angiotensin II, a natural human peptide hormone that regulates blood pressure, has been shown to be active against Plasmodium spp., the etiologic agent of malaria. Here, we tested two Ang II derivatives that do not elicit vasoconstriction in mice: VIPF, a linear tetrapeptide, which constitutes part of the hydrophobic portion of Ang II; and Ang II-SS, a disulfide-bridged derivative. The antiplasmodial potential of both peptides was evaluated with two mouse models: an experimental cerebral malaria model and a mouse model of non-cerebral malaria. The latter consisted of BALB/c mice infected with Plasmodium berghei ANKA. The peptides had no effect on mean blood pressure and significantly reduced parasitemia in both mouse models. Both peptides reduced the SHIRPA score, an assay used to assess murine health and behavior. However, only the constrained derivative (Ang II-SS), which was also resistant to proteolytic degradation, significantly increased mouse survival. Here, we show that synthetic peptides derived from Ang II are capable of conferring protection against severe manifestations of malaria in mouse models while overcoming the vasoconstrictive side effects of the parent peptide.

Spatially resolved multiomics of human cardiac niches.

The function of a cell is defined by its intrinsic characteristics and its niche: the tissue microenvironment in which it dwells. Here we combine single-cell and spatial transcriptomics data to discover cellular niches within eight regions of the human heart. We map cells to microanatomical locations and integrate knowledge-based and unsupervised structural annotations. We also profile the cells of the human cardiac conduction system1. The results revealed their distinctive repertoire of ion channels, G-protein-coupled receptors (GPCRs) and regulatory networks, and implicated FOXP2 in the pacemaker phenotype. We show that the sinoatrial node is compartmentalized, with a core of pacemaker cells, fibroblasts and glial cells supporting glutamatergic signalling. Using a custom CellPhoneDB.org module, we identify trans-synaptic pacemaker cell interactions with glia. We introduce a druggable target prediction tool, drug2cell, which leverages single-cell profiles and drug-target interactions to provide mechanistic insights into the chronotropic effects of drugs, including GLP-1 analogues. In the epicardium, we show enrichment of both IgG+ and IgA+ plasma cells forming immune niches that may contribute to infection defence. Overall, we provide new clarity to cardiac electro-anatomy and immunology, and our suite of computational approaches can be applied to other tissues and organs.

Outbreak report of polymyxin-carbapenem-resistant Klebsiella pneumoniae causing untreatable infections evidenced by synergy tests and bacterial genomes.

Polymyxin-carbapenem-resistant Klebsiella pneumoniae (PCR-Kp) with pan (PDR)- or extensively drug-resistant phenotypes has been increasingly described worldwide. Here, we report a PCR-Kp outbreak causing untreatable infections descriptively correlated with bacterial genomes. Hospital-wide surveillance of PCR-Kp was initiated in December-2014, after the first detection of a K. pneumoniae phenotype initially classified as PDR, recovered from close spatiotemporal cases of a sentinel hospital in Rio de Janeiro. Whole-genome sequencing of clinical PCR-Kp was performed to investigate similarities and dissimilarities in phylogeny, resistance and virulence genes, plasmid structures and genetic polymorphisms. A target phenotypic profile was detected in 10% (12/117) of the tested K. pneumoniae complex bacteria recovered from patients (8.5%, 8/94) who had epidemiological links and were involved in intractable infections and death, with combined therapeutic drugs failing to meet synergy. Two resistant bacterial clades belong to the same transmission cluster (ST437) or might have different sources (ST11). The severity of infection was likely related to patients' comorbidities, lack of antimicrobial therapy and predicted bacterial genes related to high resistance, survival, and proliferation. This report contributes to the actual knowledge about the natural history of PCR-Kp infection, while reporting from a time when there were no licensed drugs in the world to treat some of these infections. More studies comparing clinical findings with bacterial genetic markers during clonal spread are needed.

Single-cell transcriptomics for the assessment of cardiac disease.

Cardiovascular disease is the leading cause of death globally. An advanced understanding of cardiovascular disease mechanisms is required to improve therapeutic strategies and patient risk stratification. State-of-the-art, large-scale, single-cell and single-nucleus transcriptomics facilitate the exploration of the cardiac cellular landscape at an unprecedented level, beyond its descriptive features, and can further our understanding of the mechanisms of disease and guide functional studies. In this Review, we provide an overview of the technical challenges in the experimental design of single-cell and single-nucleus transcriptomics studies, as well as a discussion of the type of inferences that can be made from the data derived from these studies. Furthermore, we describe novel findings derived from transcriptomics studies for each major cardiac cell type in both health and disease, and from development to adulthood. This Review also provides a guide to interpreting the exhaustive list of newly identified cardiac cell types and states, and highlights the consensus and discordances in annotation, indicating an urgent need for standardization. We describe advanced applications such as integration of single-cell data with spatial transcriptomics to map genes and cells on tissue and define cellular microenvironments that regulate homeostasis and disease progression. Finally, we discuss current and future translational and clinical implications of novel transcriptomics approaches, and provide an outlook of how these technologies will change the way we diagnose and treat heart disease.

Complexation of a Polypeptide-Polyelectrolytes Bioparticle as a Biomaterial of Antibacterial Activity.

The development of biomaterials to enable application of antimicrobial peptides represents a strategy of high and current interest. In this study, a bioparticle was produced by the complexation between an antimicrobial polypeptide and the biocompatible and biodegradable polysaccharides chitosan-N-arginine and alginate, giving rise to a colloidal polyelectrolytic complex of pH-responsive properties. The inclusion of the polypeptide in the bioparticle structure largely increases the binding sites of complexation during the bioparticles production, leading to its effective incorporation. After lyophilization, detailed evaluation of colloidal structure of redispersed bioparticles evidenced nano or microparticles with size, polydispersity and zeta potential dependent on pH and ionic strength, and the dependence was not withdrawn with the polypeptide inclusion. Significant increase of pore edge tension in giant vesicles evidenced effective interaction of the polypeptide-bioparticle with lipid model membrane. Antibacterial activity against Aeromonas dhakensis was effective at 0.1% and equal for the isolated polypeptide and the same complexed in bioparticle, which opens perspectives to the composite material as an applicable antibacterial system.

Uncovering Pseudogenes and Intergenic Protein-coding Sequences in TriTryps' Genomes.

Trypanosomatids belong to a remarkable group of unicellular, parasitic organisms of the order Kinetoplastida, an early diverging branch of the phylogenetic tree of eukaryotes, exhibiting intriguing biological characteristics affecting gene expression (intronless polycistronic transcription, trans-splicing, and RNA editing), metabolism, surface molecules, and organelles (compartmentalization of glycolysis, variation of the surface molecules, and unique mitochondrial DNA), cell biology and life cycle (phagocytic vacuoles evasion and intricate patterns of cell morphogenesis). With numerous genomic-scale data of several trypanosomatids becoming available since 2005 (genomes, transcriptomes, and proteomes), the scientific community can further investigate the mechanisms underlying these unusual features and address other unexplored phenomena possibly revealing biological aspects of the early evolution of eukaryotes. One fundamental aspect comprises the processes and mechanisms involved in the acquisition and loss of genes throughout the evolutionary history of these primitive microorganisms. Here, we present a comprehensive in silico analysis of pseudogenes in three major representatives of this group: Leishmania major, Trypanosoma brucei, and Trypanosoma cruzi. Pseudogenes, DNA segments originating from altered genes that lost their original function, are genomic relics that can offer an essential record of the evolutionary history of functional genes, as well as clues about the dynamics and evolution of hosting genomes. Scanning these genomes with functional proteins as proxies to reveal intergenic regions with protein-coding features, relying on a customized threshold to distinguish statistically and biologically significant sequence similarities, and reassembling remnant sequences from their debris, we found thousands of pseudogenes and hundreds of open reading frames, with particular characteristics in each trypanosomatid: mutation profile, number, content, density, codon bias, average size, single- or multi-copy gene origin, number and type of mutations, putative primitive function, and transcriptional activity. These features suggest a common process of pseudogene formation, different patterns of pseudogene evolution and extant biological functions, and/or distinct genome organization undertaken by those parasites during evolution, as well as different evolutionary and/or selective pressures acting on distinct lineages.

Multiple contiguous spinous process fractures, a case report and literature review.

Contiguous spinous process fractures are an extremely rare event with only a few cases described in the literature. They are usually stable lesions treated with analgesic medication, immobilization, physical activity restriction for 4 to 6 weeks and close surveillance. These fractures appear to have a significant risk of nonunion, despite the good reported results. We report a case of a 61 years-old male patient who suffered eight contiguous spinous process fractures following a tractor accident.

The synthetic antimicrobial peptide IKR18 displays anti-infectious properties in Galleria mellonella in vivo model.

Antimicrobial peptides (AMPs) are promising tools for developing new antibiotics. We described the design of IKR18, an AMP designed with the aid of computational tools. IKR18 showed antimicrobial activity against Gram-negative and Gram-positive bacteria, including methicillin-resistant Staphylococcus aureus (MRSA). CD studies revealed that IKR18 assumes an alpha-helical structure in the membrane-mimetic environment. The action mechanism IKR18 involves damage to the bacteria membrane, as demonstrated by Sytox green uptake. Furthermore, IKR18 displayed synergic and additive effects in combination with antibiotics ciprofloxacin and vancomycin. The peptide showed anti-biofilm activity in concentration and efficiency compared with commercial antibiotics, involving the direct death of bacteria, as confirmed by scanning electron microscopy. The anti-infective activity of IKR18 was demonstrated in the Galleria mellonella model infected with S. aureus, MRSA, and Acinetobacter baumannii. The novel bioinspired peptide, IKR18, proved to be effective in the control of bacterial infection, opening opportunities for the development of further assays, including preclinical models.

Bio-Strings: A Relational Database Data-Type for Dealing with Large Biosequences.

DNA sequencers output a large set of very long biological data strings that we should persist in databases rather than basic text file systems. Many different data models and database management systems (DBMS) may deal with both storage and efficiency issues regarding genomic datasets. Specifically, there is a need for handling strings with variable sizes while keeping their biological meaning. Relational database management systems (RDBMS) provide several data types that could be further explored for the genomics context. Besides, they enforce integrity, consistency, and enable good abstractions for more conventional data. We propose the relational text data type to represent and manipulate biological sequences and their derivatives. We present a logical schema for representing the core biological information, which may be inferred from a given biological conceptual data schema and the corresponding function manipulations. We implement and evaluate these stored functions into an actual RDBMS for both efficacy and efficiency. We show that it is possible to enforce basic and complex requirements for the genomic domain. We claim that the well-established relational text data type in RDBMS may appropriately handle the representation and persistency of biological sequences. We base our approach on the idea of domain-specific abstract data types that can store data with semantically defined functions while hiding those details from non-technical end-users.

Pathogenic variants damage cell composition and single cell transcription in cardiomyopathies.

Pathogenic variants in genes that cause dilated cardiomyopathy (DCM) and arrhythmogenic cardiomyopathy (ACM) convey high risks for the development of heart failure through unknown mechanisms. Using single-nucleus RNA sequencing, we characterized the transcriptome of 880,000 nuclei from 18 control and 61 failing, nonischemic human hearts with pathogenic variants in DCM and ACM genes or idiopathic disease. We performed genotype-stratified analyses of the ventricular cell lineages and transcriptional states. The resultant DCM and ACM ventricular cell atlas demonstrated distinct right and left ventricular responses, highlighting genotype-associated pathways, intercellular interactions, and differential gene expression at single-cell resolution. Together, these data illuminate both shared and distinct cellular and molecular architectures of human heart failure and suggest candidate therapeutic targets.

Toxicity and Residual Activity of Insecticides against Diadegma insulare, a Parasitoid of the Diamondback Moth.

Plutella xylostella is the main pest of cruciferous crops worldwide. To reduce P. xylostella populations, better integration of natural control and chemical control (dominant tactic used) is needed. This work analyzed the compatibility of nine insecticides with the parasitoid Diadegma insulare, outlining them as complementary tools in an integrated pest management strategy. The acute toxicity of spinosad, imidacloprid, indoxacarb, flonicamid, naled, pyridalyl, emamectin benzoate, and spinetoram against the parasitoid was assessed. Residual activity (persistence) was also evaluated over time; the mortality of the parasitoid in contact with leaf tissue of plants treated with insecticides was analyzed. According to the International Organization of Biological Control, all nine insecticides were toxic to D. insulare; the lowest mortality was recorded with spirotetramat (64%) and pyridalyl (48%), while the rest of the insecticides caused 100% mortality at 72 h after application. In terms of persistence, by days 14, 16, 16, 17, 17, 21, and 22 after application, flonicamid, naled, spirotetramat, spinosad, piridalyl, imidacloprid, and indoxacarb caused mortality of less than 25%, respectively, so they were considered harmless (Category 1). Nonetheless, some insecticide toxicity and residual activity must be regarded within integrated pest management programs for conserving the role of D. insulare field populations.

Dentinal microcracks induced by endodontic procedures: A scientometric and bibliometric analysis.

Background: Due to the large number of publications relating the occurrence of dental microcracks to endodontic procedures, this bibliometric study evaluated the scientific pattern and trends in literature and provided an overview of scientific production in this context. Aim: To analyze, quantify, and characterize the scientific production and trends of published articles evaluating dentinal microcracks formation after endodontic treatment procedures between 2010 and 2020 using bibliometric indicators. Materials and Methods: Published articles were found by the search in the Medline (PubMed) and Scopus database using the combination of the following keywords: Dentinal crack OR Dentinal Microcrack OR Crack formation OR Dentin Defect AND Endodontic treatment OR Root canal preparation OR Canal Preparation OR Root canal treatment OR endodontic procedures. The search was also conducted in the Journal of Endodontics and International Endodontic Journal. After inclusion and exclusion criteria application, data from all studies included were collected. Results: Among the 556 results after the search, 45 studies were included and analyzed in this bibliometry. No trend was observed in terms of the increasing number of articles over time. Most of them used an in vitro design, compared the effect of different endodontic techniques/systems for root canal instrumentation on dentinal microcracks formation, and were conducted in Turkey, Brazil, and India. Journal of Endodontics and International Endodontic Journal were the main journals with a higher number of articles published. Interestingly, studies conducted with some funding did not lead to higher citation numbers. Moreover, a relevant proportion of studies did not consider the inclusion of control groups, baseline evaluation, or statistical analysis. Micro-CT was the main technique used to evaluate microcrack presence. Conclusion: Microcrack formation after the use of different endodontic techniques/systems has been constantly evaluated in the literature. There is a pattern of methodologies used, which may explain the concentration of these studies in specific journals and countries.

Clinical and Functional Results of Reverse Total Shoulder Arthroplasty and Postoperative Rehabilitation Protocol.

Purpose This retrospective study aims to analyze the clinical and functional results obtained over a seven-year period of performing reverse total shoulder arthroplasty (RTSA) and the subsequent postoperative rehabilitation protocol. Methods We analyzed data from 80 patients who were evaluated at a preoperative, as well as monthly postoperative outpatient consultation, until the discharge from the rehabilitation program, using Constant Score (CS). Results A comparison of preoperative and postoperative (after rehabilitation protocol) results revealed an improved functional score of absolute CS (20.8 increase), normal relative CS (29.1 increase), and individual relative CS (31.7 increase) with statistical significance (p<0.05). From the analysis of CS subscores, there was a positive evolution of the pain subscore, as well as flexion, abduction, and external rotation combined with abduction range of motion (ROM). Contrarily, there was a negative evolution of the combined internal rotation, extension, and adduction ROM, as well as deltoid muscle strength. No statistically significant correlations were found between age and postoperative CS, as well as between the time interval from surgery to the beginning of outpatient rehabilitation and CS evolution. Conclusion Our study demonstrates that RTSA is an effective therapeutic option that, if combined with a well-structured rehabilitation program, can improve pain, mobility, and upper limb functionality.

Efficacy of Hounsfield Units Measured by Lumbar Computer Tomography on Bone Density Assessment: A Systematic Review.

STUDY DESIGN: Systematic review. OBJECTIVE: This work aimed to compare the Hounsfield units (HU) value obtained from computed tomography and the t score of dual-energy x-ray absorptiometry (DXA) in the prediction of the lumbar spine bone mineral density (BMD). SUMMARY OF BACKGROUND DATA: Several reports have found a correlation between HU and BMD values based on DXA. Using HUs to infer bone quality has a thorough clinical relevance as it could triage patients at risk for osteoporotic and fragility fractures or modify surgical indications. METHODS: A systematic review in Cochrane Library, Medline, Scopus and Web of Science was performed, using the following query: "hounsfield units" AND ("osteoporosis" OR "spine" OR "bone mineral density" OR "dual x-ray absorptiometry"). We included 18 cohort studies that compared HU value obtained from computed tomography and t score of DXA for predicting regional BMD. RESULTS: A total of 18 studies were included, enrolling 5307 patients. The HU measurement was most frequently made at L1 (N = 3; 18.8%). The mean HU values differentiated based on BMD measured through DXA were reported in seven studies, with values from 54.7 to 130 for osteoporotic, 78.8 to 146 for osteopenic, and from 120.8 to 230 in normal patients. Eight studies identified thresholds for diagnosing osteoporosis through receiver-operating characteristic (ROC) curves, with values ranging from 0.66 to 0.96. Medium HU values reported as diagnostic of osteoporosis ranged between 110 and 150, after exclusion of the two papers presenting outlier values. We infer an HU interval value of 90.9 to 138.7 (95% CI, P  < 0.001) for the diagnosis osteoporosis. CONCLUSIONS: Present data evidence favorable results regarding the possibility of establishing a threshold value for osteoporosis diagnosis from CT measurements of HU. Prospective large-scale studies are needed to more robustly infer the possibility of quantifying BMD based on CT as a screening test and infer a prognostic value of the CT-based evaluation.Level of Evidence: 2.

Surgical risk factors associated with the development of adjacent segment pathology in the lumbar spine.

Adjacent segment pathology (ASP) is a major cause of disability, and the recognition of the surgical risk factors associated with the development of this condition is essential for its prevention.Different surgical approaches, from decompression without fusion to non-instrumented and instrumented fusion, have distinct contributions to the development of ASP.Although motion-preservation procedures could reduce the prevalence of ASP, these are also associated with a higher percentage of complications.Several risk factors associated with previous surgery, namely the chosen surgical approach and anatomical dissection, the choice of interbody fusion, the increment and length of the fusion, and the restoration of sagittal alignment, may influence the development of ASP. Cite this article: EFORT Open Rev 2021;6:966-972. DOI: 10.1302/2058-5241.6.210050.

Patient-Related Risk Factors for the Development of Lumbar Spine Adjacent Segment Pathology.

OBJECTIVES: Individual risk factors for the development of adjacent segment pathology (ASP) need to be investigated and identified to address possible modifiable factors in advance and improve outcomes and reduce medical costs. This study aimed to review the literature regarding patient-related risk factors and sagittal alignment parameters associated with ASP development. METHODS: The authors performed an extensive review of the literature addressing the objectives mentioned earlier. RESULTS: Certain patient factors such as age, gender, obesity, preexisting degeneration, osteoporosis, postmenopausal state, rheumatoid arthritis, and facet tropism may contribute to adjacent segment degeneration. Genetic influences, such as polymorphisms of the vitamin D receptor and collagen IX genes, can also be a potential cause for disc degeneration with consequent deterioration of the motion segment.The influence of sagittal imbalances, particularly after lumbar fusion, is a significant parameter to be taken into account as an independent risk factor for ASP development. CONCLUSIONS: Patient-specific risk factors, such as age, gender, obesity, preexisting degeneration, and genetic features increase the likelihood of developing ASP. On the other hand, sagittal alignment plays a significant role in the development of this condition.

Traumatic Isolated Thumb Carpometacarpal Joint Dislocation - Report of Two Clinical Cases.

Isolated thumb carpometacarpal joint dislocation is a rare lesion that accounts for less than 1% of all hand lesions. The authors present two cases of traumatic isolated thumb carpometacarpal joint dislocation. One of them was treated with closed reduction and cast immobilization, and the other was treated with closed reduction, Kirschner-wires pinning, and cast immobilization. The first patient had a good functional outcome and showed no signs of thumb carpometacarpal instability. The patient treated with Kirschner wires presented signs of clinical instability and radiological subluxation. Isolated thumb carpometacarpal dislocation is a rare lesion that can cause joint instability, which interferes with the normal function of the hand and can lead to articular degenerative changes. The best management of this lesion is still controversial, since there is lack of evidence in the literature showing superiority of one treatment over the other.

Comparison of Different Methods for Spongin-like Collagen Extraction from Marine Sponges (Chondrilla caribensis and Aplysina fulva): Physicochemical Properties and In Vitro Biological Analysis.

This study aimed to compare different protocols (Protocol 1: P1; Protocol 2: P2; Protocol 3: P3; Protocol 4: P4) for the extraction of spongin-like collagen (SC) from marine sponges. The SEM micrographs demonstrated a fibrillar structure for the extracts from Chondrilla caribensis and the nodular/particulate aggregates for Aplysina fulva. FTIR showed for all samples peaks similar to collagen for both species. For C. caribensis, the extracts obtained using P2, P3, and P4 protocols presented higher values of extraction yield, TPQ, and GAGs. P2 and P4 showed higher values of SC concentration and for antioxidant analysis. For A. fulva, P2, P3, and P4 provided a higher extraction yield besides an increase in the antioxidant assay. For both species, no difference was observed for Col quantification and TPQ analysis; also, higher values of GAGs were found using P2 and P4. Fibroblast proliferation observed for C. caribensis was lower for P1 on day 1 and for P2 and P3 on day 3 (for 50%) compared to the control group. There was a significant reduction in fibroblast cell proliferation for all A. fulva extracts evaluated. It can be concluded that protocols P2 and P4 were more efficient for extracting SC from C. caribensis.