Genotypic characterization and antimicrobial susceptibility of human Campylobacter jejuni isolates in Southern Spain.

Campylobacter jejuni is the main cause of bacterial gastroenteritis and a public health problem worldwide. Little information is available on the genotypic characteristics of human C. jejuni in Spain. This study is based on an analysis of the resistome, virulome, and phylogenetic relationship, antibiogram prediction, and antimicrobial susceptibility of 114 human isolates of C. jejuni from a tertiary hospital in southern Spain from October 2020 to June 2023. The isolates were sequenced using Illumina technology, and a bioinformatic analysis was subsequently performed. The susceptibility of C. jejuni isolates to ciprofloxacin, tetracycline, and erythromycin was also tested. The resistance rates for each antibiotic were 90.3% for ciprofloxacin, 66.7% for tetracycline, and 0.88% for erythromycin. The fluoroquinolone resistance rate obtained is well above the European average (69.1%). CC-21 (n = 23), ST-572 (n = 13), and ST-6532 (n = 13) were the most prevalent clonal complexes (CCs) and sequence types (STs). In the virulome, the cadF, ciaB, and cdtABC genes were detected in all the isolates. A prevalence of 20.1% was obtained for the genes wlaN and cstIII, which are related to the pathogenesis of Guillain-Barré syndrome (GBS). The prevalence of the main antimicrobial resistance markers detected were CmeABC (92.1%), RE-cmeABC (7.9%), the T86I substitution in gyrA (88.9%), blaOXA-61 (72.6%), tet(O) (65.8%), and ant (6)-Ia (17.1%). High antibiogram prediction rates (>97%) were obtained, except for in the case of the erythromycin-resistant phenotype. This study contributes significantly to the knowledge of C. jejuni genomics for the prevention, treatment, and control of infections caused by this pathogen.IMPORTANCEDespite being the pathogen with the greatest number of gastroenteritis cases worldwide, Campylobacter jejuni remains a poorly studied microorganism. A sustained increase in fluoroquinolone resistance in human isolates is a problem when treating Campylobacter infections. The development of whole genome sequencing (WGS) techniques has allowed us to better understand the genotypic characteristics of this pathogen and relate them to antibiotic resistance phenotypes. These techniques complement the data obtained from the phenotypic analysis of C. jejuni isolates. The zoonotic transmission of C. jejuni through the consumption of contaminated poultry supports approaching the study of this pathogen through "One Health" approach. In addition, due to the limited information on the genomic characteristics of C. jejuni in Spain, this study provides important data and allows us to compare the results with those obtained in other countries.

The integrated genomic surveillance system of Andalusia (SIEGA) provides a One Health regional resource connected with the clinic.

The One Health approach, recognizing the interconnectedness of human, animal, and environmental health, has gained significance amid emerging zoonotic diseases and antibiotic resistance concerns. This paper aims to demonstrate the utility of a collaborative tool, the SIEGA, for monitoring infectious diseases across domains, fostering a comprehensive understanding of disease dynamics and risk factors, highlighting the pivotal role of One Health surveillance systems. Raw whole-genome sequencing is processed through different species-specific open software that additionally reports the presence of genes associated to anti-microbial resistances and virulence. The SIEGA application is a Laboratory Information Management System, that allows customizing reports, detect transmission chains, and promptly alert on alarming genetic similarities. The SIEGA initiative has successfully accumulated a comprehensive collection of more than 1900 bacterial genomes, including Salmonella enterica, Listeria monocytogenes, Campylobacter jejuni, Escherichia coli, Yersinia enterocolitica and Legionella pneumophila, showcasing its potential in monitoring pathogen transmission, resistance patterns, and virulence factors. SIEGA enables customizable reports and prompt detection of transmission chains, highlighting its contribution to enhancing vigilance and response capabilities. Here we show the potential of genomics in One Health surveillance when supported by an appropriate bioinformatic tool. By facilitating precise disease control strategies and antimicrobial resistance management, SIEGA enhances global health security and reduces the burden of infectious diseases. The integration of health data from humans, animals, and the environment, coupled with advanced genomics, underscores the importance of a holistic One Health approach in mitigating health threats.

Use of Knotless Barbed Sutures in Laparoscopic Inguinal Hernioplasty in Horses: 40 Cases.

Inguinal hernias (IHs) and ruptures are a relatively common condition in horses, occurring in foals (congenital) and adult (acquired) animals. A retrospective observational analysis was conducted on 40 cases that underwent laparoscopic surgery to close the VRs using barbed sutures alone or combined with other techniques. Signalment, clinical presentation, surgery, and follow-up data were obtained. In total, fifty-nine VRs were closed using barbed sutures (alone or in combination with other methods), with six cases performed prophylactically and forty-four due to acquired IH. Of the forty-four cases with IH, four were non-strangulated hernias, while thirty presented with strangulated small intestines (twenty-eight acquired and two congenital). The results obtained in this study suggest that laparoscopic hernioplasty with barbed sutures is an effective and safe surgical procedure that could be recommended as a standard practice for managing inguinal hernias in horses, particularly when sparing testicles or preserving reproductive capabilities is a priority.

Molecular Detection of Cryptosporidium spp. and Microsporidia in Human and Animal Stool Samples.

Cryptosporidium spp. and Microsporidia are opportunistic microorganisms with remarkable zoonotic transmission potential due to their capacity to infect humans and animals. The aim of this study was to evaluate the prevalence of these microorganisms in stool samples of animal and human origin. In total, 369 stool samples (205 from human patients with diarrhea and 164 of animal origin) were included in the study. Cryptosporidium spp. and Microsporidia presence were determined by using multiplex nested PCR. Positive results were analyzed by using Sanger sequencing of the amplicon, utilizing BLASTN and ClustalX software to confirm identification. Cryptosporidium spp. were found in 0.97% and 4.26% of human and animal samples, respectively. Enterocytozoon bieneusi was detected in human and animal stools in 6.82% and 3.05% of the samples, respectively. No associations were found when analyzing the presence of Cryptosporidium spp. and E. bieneusi and the demographic and clinical variables of patients and animals. This study demonstrates the presence of these microorganisms in human and animal samples from different species, and the most interesting findings are the detection of Cryptosporidium spp. in pets (e.g., rodents) that are not usually included in this type of study, and the identification of E. bieneusi in patients with diarrhea without underlying disease.

Conversational agents for depression screening: A systematic review.

OBJECTIVE: This work explores the advances in conversational agents aimed at the detection of mental health disorders, and specifically the screening of depression. The focus is put on those based on voice interaction, but other approaches are also tackled, such as text-based interaction or embodied avatars. METHODS: PRISMA was selected as the systematic methodology for the analysis of existing literature, which was retrieved from Scopus, PubMed, IEEE Xplore, APA PsycINFO, Cochrane, and Web of Science. Relevant research addresses the detection of depression using conversational agents, and the selection criteria utilized include their effectiveness, usability, personalization, and psychometric properties. RESULTS: Of the 993 references initially retrieved, 36 were finally included in our work. The analysis of these studies allowed us to identify 30 conversational agents that claim to detect depression, specifically or in combination with other disorders such as anxiety or stress disorders. As a general approach, screening was implemented in the conversational agents taking as a reference standardized or psychometrically validated clinical tests, which were also utilized as a golden standard for their validation. The implementation of questionnaires such as Patient Health Questionnaire or the Beck Depression Inventory, which are used in 65% of the articles analyzed, stand out. CONCLUSIONS: The usefulness of intelligent conversational agents allows screening to be administered to different types of profiles, such as patients (33% of relevant proposals) and caregivers (11%), although in many cases a target profile is not clearly of (66% of solutions analyzed). This study found 30 standalone conversational agents, but some proposals were explored that combine several approaches for a more enriching data acquisition. The interaction implemented in most relevant conversational agents is text-based, although the evolution is clearly towards voice integration, which in turns enhances their psychometric characteristics, as voice interaction is perceived as more natural and less invasive.

Development and Validation of a Highly Sensitive Multiplex Immunoassay for SARS-CoV-2 Humoral Response Monitorization: A Study of the Antibody Response in COVID-19 Patients with Different Clinical Profiles during the First and Second Waves in Cadiz, Spain.

There is still a long way ahead regarding the COVID-19 pandemic, since emerging waves remain a daunting challenge to the healthcare system. For this reason, the development of new preventive tools and therapeutic strategies to deal with the disease have been necessary, among which serological assays have played a key role in the control of COVID-19 outbreaks and vaccine development. Here, we have developed and evaluated an immunoassay capable of simultaneously detecting multiple IgG antibodies against different SARS-CoV-2 antigens through the use of Bio-PlexTM technology. Additionally, we have analyzed the antibody response in COVID-19 patients with different clinical profiles in Cadiz, Spain. The multiplex immunoassay presented is a high-throughput and robust immune response monitoring tool capable of concurrently detecting anti-S1, anti-NC and anti-RBD IgG antibodies in serum with a very high sensitivity (94.34-97.96%) and specificity (91.84-100%). Therefore, the immunoassay proposed herein may be a useful monitoring tool for individual humoral immunity against SARS-CoV-2, as well as for epidemiological surveillance. In addition, we show the values of antibodies against multiple SARS-CoV-2 antigens and their correlation with the different clinical profiles of unvaccinated COVID-19 patients in Cadiz, Spain, during the first and second waves of the pandemic.

Molecular characterization of consecutive isolates of OXA-48-producing Klebsiella pneumoniae: changes in the virulome using next-generation sequencing (NGS).

Little is known about the clonality of consecutive OXA-48 producing-Klebsiella pneumoniae isolates from the same patient and the possibility of changes in their virulomes over time. We studied the molecular characteristics of twenty OXA-48-producing K. pneumoniae consecutive isolates from six patients using whole-genome sequencing. The genomes were screened for antimicrobial resistance and virulence factor genes and for replicon groups. MLST and SNPs analysis was performed. MLST analysis found 3 STs: ST11 (n = 13; 65.0%); ST4975 (n = 5, 25.0%); ST307 (n = 2; 10.0%). AcrAb efflux pump, siderophore enterobactin and rcsAB capsule synthesis regulator were detected in all sequenced isolates. The regulator of mucoid phenotype A (rmpA) and rmpA2 were not detected. Isolates also carried type 3 fimbriae (n = 19; 95.0%), yersiniabactin (n = 15; 75.0%) and type 1 fimbriae (7; 35.0%). Type 3 fimbriae and yersiniabactin were lost and recovered in consecutive isolates of two patients, probably acquired by horizontal gene transfer. Our findings reveal that recurrent infections are due to the same isolate, with an average of 2.69 SNPs per month, with different virulence profiles, and that the acquisition of virulence factor genes over time is possible.

Correction: Iridium-(κ2-NSi) catalyzed dehydrogenation of formic acid: effect of auxiliary ligands on the catalytic performance.

Correction for 'Iridium-(κ2-NSi) catalyzed dehydrogenation of formic acid: effect of auxiliary ligands on the catalytic performance' by Alejandra Gomez-España et al., Dalton Trans., 2023, 52, 6722-6729, https://doi.org/10.1039/d3dt00744h.

Synthesis and catalytic activity of well-defined Co(I) complexes based on NHC-phosphane pincer ligands.

A new methodology for the preparation of Co(I)-NHC (NHC = N-heterocyclic carbene) complexes, namely, [Co(PCNHCP)(CO)2][Co(CO)4] (1) and [Co(PCNHCP)(CO)2]BF4 (2), has been developed (PCNHCP = 1,3-bis(2-(diphenylphosphanyl)ethyl)-imidazol-2-ylidene). Both complexes can be straightforwardly prepared by direct reaction of their parent imidazolium salts with the Co(0) complex Co2(CO)8. Complex 1 efficiently catalyses the reductive amination of furfural and levulinic acid employing silanes as reducing agents under mild conditions. Furfural has been converted into a variety of secondary and tertiary amines employing dimethyl carbonate as the solvent, while levulinic acid has been converted into pyrrolidines under solventless conditions. Dehydrocoupling of the silane to give polysilanes has been observed to occur as a side reaction of the hydrosilylation process.

Assessment of validity and comparison of two Spanish versions of the Geriatric Depression Scale.

Introduction: The Geriatric Depression Scale is an instrument used to identify depression in people of an older age. The original English version of this scale has been translated into Spanish (GDS-VE); two shorter versions of 5- (GDS-5) and 15-items (GDS-15) have been developed. Aim of the study: To assess the validity and compare the 5- and 15-item Spanish versions of the GDS among the Spanish population. Materials and methods: 573 Galicia residents aged >50 years participated in this study. The following instruments were applied: the 19-item Control, Autonomy, Self-Realization and Pleasure scale, the Subjective Memory Complaints Questionnaire, the Mini-Mental State Examination test, the GDS-5, and the GDS-15. Results: We found differences in total score between GDS-5 and GDS-15 regarding the variable sex. Internal reliability for GDS-5 and GDS-15 was 0.495 and 0.715, respectively. Sensitivity and specificity for GDS-5 - with a cut-off value of 1 - was 0.517 and 0.650, respectively; for GDS-15 - with a cut-off value of 3 points - sensitivity was 0.755 and specificity 0.668. GDS-5 has a ROC curve of 0.617 and GDS-15 of 0.764. Conclusion: GDS-15, and to a greater extent GDS-5, should be revised or even reformulated to improve their diagnostic usefulness by choosing higher discriminative ability items or even include new items with greater sensitivity that consider currently prevailing psychosocial factors.

Iridium-(κ2-NSi) catalyzed dehydrogenation of formic acid: effect of auxiliary ligands on the catalytic performance.

The iridium(III) complexes [Ir(H)(Cl)(κ2-NSitBu2)(κ2-bipyMe2)] (2) and [Ir(H)(OTf)(κ2-NSitBu2)(κ2-bipyMe2)] (3) (NSitBu2 = {4-methylpyridine-2-yloxy}ditertbutylsilyl) have been synthesized and characterized including X-ray studies of 3. A comparative study of the catalytic activity of complexes 2, 3, [Ir(H)(OTf)(κ2-NSitBu2)(coe)] (4), and [Ir(H)(OTf)(κ2-NSitBu2)(PCy3)] (5) (0.1 mol%) as catalysts precursors for the solventless formic acid dehydrogenation (FADH) in the presence of Et3N (40 mol%) at 353 K has been performed. The highest activity (TOF5 min ≈ 3260 h-1) has been obtained with 3 at 373 K. However, at that temperature the FTIR spectra show traces of CO together with the desired products (H2 and CO2). Thus, the best performance was achieved at 353 K (TOF5 min ≈ 1210 h-1 and no observable CO). Kinetic studies at variable temperature show that the activation energy of the 3-catalyzed FADH process is 16.76 kcal mol-1. Kinetic isotopic effect (5 min) values of 1.6, 4.5, and 4.2 were obtained for the 3-catalyzed dehydrogenation of HCOOD, DCOOH, and DCOOD, respectively, at 353 K. The strong KIE found for DCOOH and DCOOD evidenced that the hydride transfer from the C-H bond of formic acid to the metal is the rate-determining step of the process.

A systematic review evaluating loneliness assessment instruments in older adults.

Introduction and objectives: The experiences and changes that come along with old age may lead to a feeling of loneliness, usually followed by negative physical and mental manifestations. In this systematic review, we evaluated the existing tools to assess loneliness in older adults. Methods: We performed a literature search in the Web of Science, Medline, and PsycINFO, following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. After, we examined the psychometric properties of the instruments with a focus on reliability, validity, and main conclusions. Results: We included 27 articles published between 1996 and 2021. Conclusion: To date, there are few instruments to assess loneliness in older adults. In general, they present adequate psychometric properties, although it is true that some scales show somewhat low levels of reliability and validity.

Correction: Iridium-(κ2-NSi) catalyzed dehydrogenation of formic acid: effect of auxiliary ligands on the catalytic performance.

Correction for 'Iridium-(κ2-NSi) catalyzed dehydrogenation of formic acid: effect of auxiliary ligands on the catalytic performance' by Alejandra Gomez-España et al., Dalton Trans., 2023, https://doi.org/10.1039/d3dt00744h.

Cardiovascular-related proteomic changes in ECFCs exposed to the serum of COVID-19 patients.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection significantly affects the cardiovascular system, causing vascular damage and thromboembolic events in critical patients. Endothelial dysfunction represents one of the first steps in response to COVID-19 that might lead to cardiovascular complications and long-term sequelae. However, despite the enormous efforts in the last two years, the molecular mechanisms involved in such processes remain poorly understood. Herein, we analyzed the protein changes taking place in endothelial colony forming cells (ECFCs) after the incubation with the serum from individuals infected with COVID-19, whether asymptomatic or critical patients, by application of a label free-quantitative proteomics approach. Specifically, ECFCs from healthy individuals were incubated ex-vivo with the serum of either COVID-19 negative donors (PCR-/IgG-, n:8), COVID-19 asymptomatic donors at different infective stages (PCR+/ IgG-, n:8and PCR-/IgG+, n:8), or hospitalized critical COVID-19 patients (n:8), followed by proteomics analysis. In total, 590 proteins were differentially expressed in ECFCs in response to all infected serums. Predictive analysis highlighted several proteins like CAPN5, SURF4, LAMP2 or MT-ND1, as highly discriminating features between the groups compared. Protein changes correlated with viral infection, RNA metabolism or autophagy, among others. Remarkably, the angiogenic potential of ECFCs in response to the infected serums was impaired, and many of the protein alterations in response to the serum of critical patients were associated with cardiovascular-related pathologies.

Expression profile of microRNAs related with viral infectivity, inflammatory response, and immune activation in people living with HIV.

Objective: To evaluate the serum expression of microRNAs (miRNAs) with ability to modulate the human immunodeficiency (HIV) replication or inflammatory status in people living with HIV (PLWH). Methods: Forty healthy controls and two groups of PLWH were evaluated: (a) Group 1 (n = 30), patients with detectable viral load at inclusion, analyzed before receiving antiretroviral therapy (ART) and 12 months after initiating it; (b) Group 2 (n = 55), PLWH with prolonged undetectable viral load. Intestinal barrier disruption (I-FABP) and bacterial translocation (16S rDNA) markers, inflammatory markers such as interleukin (IL)-6 and sCD163, immune activation and expression of specific miRNAs were evaluated. Results: Serum concentrations of I-FABP, 16S rDNA, IL-6, sCD163 and activated T lymphocytes were increased in PLWH. Serum miR-34a was overexpressed at inclusion and remained elevated after ART. The expression of the remaining miRNAs that modulate HIV infectivity (miR-7, mir-29a, miR-150, and miR-223) was similar in PLWH and controls. Related to miRNAs implicated in inflammation (miR-21, miR-155, and miR-210), significant overexpression were observed in miR-21 and miR-210 levels in untreated PLWH, but levels were restored in those patients treated for a long period. Conclusion: A sustained overexpression of miR-34a was detected even after prolonged HIV controlled replication. miR-21 and miR-210 can be considered new markers of inflammation with high sensitivity to its modifications.

Perspectives of Therapeutic Drug Monitoring of Biological Agents in Non-Infectious Uveitis Treatment: A Review.

Biological drugs, especially those targeting anti-tumour necrosis factor α (TNFα) molecule, have revolutionized the treatment of patients with non-infectious uveitis (NIU), a sight-threatening condition characterized by ocular inflammation that can lead to severe vision threatening and blindness. Adalimumab (ADA) and infliximab (IFX), the most widely used anti-TNFα drugs, have led to greater clinical benefits, but a significant fraction of patients with NIU do not respond to these drugs. The therapeutic outcome is closely related to systemic drug levels, which are influenced by several factors such as immunogenicity, concomitant treatment with immunomodulators, and genetic factors. Therapeutic drug monitoring (TDM) of drug and anti-drug antibody (ADAbs) levels is emerging as a resource to optimise biologic therapy by personalising treatment to bring and maintain drug concentration within the therapeutic range, especially in those patients where a clinical response is less than expected. Furthermore, some studies have described different genetic polymorphisms that may act as predictors of response to treatment with anti-TNFα agents in immune-mediated diseases and could be useful in personalising biologic treatment selection. This review is a compilation of the published evidence in NIU and in other immune-mediated diseases that support the usefulness of TDM and pharmacogenetics as a tool to guide clinicians' treatment decisions leading to better clinical outcomes. In addition, findings from preclinical and clinical studies, assessing the safety and efficacy of intravitreal administration of anti-TNFα agents in NIU are discussed.

Simultaneous and divergent evolution of resistance to cephalosporin/ß-lactamase inhibitor combinations and imipenem/relebactam following ceftazidime/avibactam treatment of MDR Pseudomonas aeruginosa infections.

OBJECTIVES: To describe and characterize the emergence of resistance to ceftolozane/tazobactam, ceftazidime/avibactam and imipenem/relebactam in a patient receiving ceftazidime/avibactam treatment for an MDR Pseudomonas aeruginosa CNS infection. METHODS: One baseline (PA1) and two post-exposure (PA2 and PA3) isolates obtained before and during treatment of a nosocomial P. aeruginosa meningoventriculitis were evaluated. MICs were determined by broth microdilution. Mutational changes were investigated through WGS. The impact on ß-lactam resistance of mutations in blaPDC and mexR was determined through cloning experiments and complementation assays. RESULTS: Isolate PA1 showed baseline resistance mutations in DacB (I354A) and OprD (N142fs) conferring resistance to conventional antipseudomonals but susceptibility to ceftazidime/avibactam, ceftolozane/tazobactam and imipenem/relebactam. Post-exposure isolates showed two divergent ceftazidime/avibactam-resistant phenotypes associated with distinctive mutations affecting the intrinsic P PDC ß-lactamase (S254Ins) (PA2: ceftolozane/tazobactam and ceftazidime/avibactam-resistant) or MexAB-OprM negative regulator MexR in combination with modification of PBP3 (PA3: ceftazidime/avibactam and imipenem/relebactam-relebactam-resistant). Cloning experiments demonstrated the role of PDC modification in resistance to ceftolozane/tazobactam and ceftazidime/avibactam. Complementation with a functional copy of the mexR gene in isolate PA3 restored imipenem/relebactam susceptibility. CONCLUSIONS: We demonstrated how P. aeruginosa may simultaneously develop resistance and compromise the activity of new ß-lactam/ß-lactamase inhibitor combinations when exposed to ceftazidime/avibactam through selection of mutations leading to PDC modification and up-regulation of MexAB-OprM-mediated efflux.

Cobalt-catalysed nucleophilic fluorination in organic carbonates.

The novel P-N ligand 1-((diphenylphosphaneyl)methyl)-1H-benzo-1,2,3-triazole (1), based on a benzotriazole scaffold, has been prepared. The reaction of 1 with [CoCp*(CH3CN)3][BF4]2 and [CoCp*(I)2]2 (Cp* = pentamethylcyclopentadienyl) affords the chelate complexes [CoCp*(CH3CN)(P-N)][BF4]2 (2) and [CoCp*(I)(P-N)]I (3), respectively. Complexes 2 and 3 were studied as catalysts in the fluorination of aromatic and aliphatic acyl chlorides in CH2Cl2, with 3 showing notably higher activities than 2. Subsequently, organic carbonates (dimethyl carbonate and propylene carbonate) were also employed as solvents, which led to shorter reaction times and to the broadening of the substrate scope to a variety of aliphatic halides. Comparative studies between 3 and the analogous complex [CoCp*(I)2(PMePh2)], which features a monodentate phosphane ligand, showed that higher yields were obtained in the case of the former. DFT calculations and experimental studies were performed in order to shed light on the reaction mechanism, which entails the formation of a cobalt fluoride species that reacts via nucleophilic attack with the substrate to afford the corresponding fluorinated compounds.