Collagen organization and structure in FBLN5-/- mice using label-free microscopy: implications for pelvic organ prolapse.

Pelvic organ prolapse (POP) is a gynecological disorder described by the descent of superior pelvic organs into or out of the vagina as a consequence of disrupted muscles and tissue. A thorough understanding of the etiology of POP is limited by the availability of clinically relevant samples, restricting longitudinal POP studies on soft-tissue biomechanics and structure to POP-induced models such as fibulin-5 knockout (FBLN5-/- ) mice. Despite being a principal constituent in the extracellular matrix, little is known about structural perturbations to collagen networks in the FBLN5-/- mouse cervix. We identify significantly different collagen network populations in normal and prolapsed cervical cross-sections using two label-free, nonlinear microscopy techniques. Collagen in the prolapsed mouse cervix tends to be more isotropic, and displays reduced alignment persistence via 2-D Fourier transform analysis of images acquired using second harmonic generation microscopy. Furthermore, coherent Raman hyperspectral imaging revealed elevated disorder in the secondary structure of collagen in prolapsed tissues. Our results underscore the need for in situ multimodal monitoring of collagen organization to improve POP predictive capabilities.

Daily disposable soft lens sagittal depth and other lens parameter changes measured at room temperature (20 °C) and on-eye temperature (34 °C).

PURPOSE: To investigate the differences in contact lens sagittal depth (CL-SAG), base curve (BC), and overall lens diameter (OAD) of daily disposable (DD) SCLs at room temperature (20 °C) and on-eye (34 °C) temperature. METHODS: Twelve different spherical daily SCLs were analyzed (labeled power -3.00 D) at 20 °C and 34 °C. The posterior CL-SAG, BC, and OAD were measured directly using a spectral domain OCT-based instrument (Optimec is830). RESULTS: The CL-SAG range for the daily disposable lenses measured at 20 °C was 3468-4016 µm. At 34 °C, the DD CL-SAG of all lenses individually decreased compared to 20 °C (t = 7.78, p < 0.001), ranging from 3385 to 3913 µm. On average at the higher temperature, the measured overall lens diameter was smaller (t = 8.18, p < 0.001) and the BC was marginally steeper (t = 2.11, p = 0.058). CONCLUSIONS: Soft lens parameters change when equilibrated from room temperature (20 °C) to on-eye temperature (34 °C). All lenses shrink when going to on-eye temperature (both lens diameter and CL-SAG decreases), compared to lenses measured at room temperature by ISO standards. Higher water content materials tend to have greater changes to the lens parameters compared to lower water content material lenses.

Hepatic mitochondrial reductive stress in the pathogenesis and treatment of steatotic liver disease.

Steatotic liver diseases (SLDs) affect one-third of the population, but the pathogenesis underlying these diseases is not well understood, limiting the available treatments. A common factor in SLDs is increased hepatic mitochondrial reductive stress, which occurs as a result of excessive lipid and alcohol metabolism. Recent research has also shown that genetic risk factors contribute to this stress. This review aims to explore how these risk factors increase hepatic mitochondrial reductive stress and how it disrupts hepatic metabolism, leading to SLDs. Additionally, the review will discuss the latest clinical studies on pharmaceutical treatments for SLDs, specifically peroxisome proliferator-activated receptor gamma (PPAR-γ) agonists, thyroid hormone receptor (THR) agonists, acetyl-CoA carboxylase (ACC) inhibitors, and mitochondrial uncouplers. These treatments have a common effect of decreasing hepatic mitochondrial reductive stress, which has been largely overlooked.

Collagen organization and structure in FLBN5-/- mice using label-free microscopy: implications for pelvic organ prolapse.

Pelvic organ prolapse (POP) is a gynecological disorder described by the descent of superior pelvic organs into or out of the vagina as a consequence of disrupted muscles and tissue. A thorough understanding of the etiology of POP is limited by the availability of clinically relevant samples, restricting longitudinal POP studies on soft-tissue biomechanics and structure to POP-induced models such as fibulin-5 knockout (FBLN5-/-) mice. Despite being a principal constituent in the extracellular matrix, little is known about structural perturbations to collagen networks in the FBLN5-/- mouse cervix. We identify significantly different collagen network populations in normal and prolapsed cervical cross-sections using two label-free, nonlinear microscopy techniques. Collagen in the prolapsed mouse cervix tends to be more isotropic, and displays reduced alignment persistence via 2-D Fourier Transform analysis of images acquired using second harmonic generation microscopy. Furthermore, coherent Raman hyperspectral imaging revealed elevated disorder in the secondary structure of collagen in prolapsed tissues. Our results underscore the need for in situ multimodal monitoring of collagen organization to improve POP predictive capabilities.

Concerning the presumptive identification of Candida kefyr on Uriselect™4 agar.

OBJECTIVE: Non-albicans Candida species, such as Candida kefyr, are emerging pathogens. Chromogenic media are highly useful for the diagnosis of urinary tract infections (UTIs). The aim was to describe the behavior of this specie on a non-specific chromogenic medium. METHODS: A retrospective study of cases of candiduria detected in the Microbiology laboratory of the Virgen de las Nieves Hospital in Granada (Spain) between 2016 and 2021 (N=2,130). Urine samples were quantitatively seeded on non-selective UriSelect™4 chromogenic agar. RESULTS: Between 2016 and 2021, C. kefyr was the seventh most frequent Candida species responsible for candiduria in our setting (n=15). The macroscopic appearance of C. kefyr colonies, punctiform and bluish, allowed the direct identification of these microorganisms. CONCLUSIONS: This study provides the first description of the specific behavior of C. kefyr on UriSelect™4 agar, which differentiates it from other Candida species based on its enzymatic characteristics.

Effects of dietary forage-to-concentrate ratio and forage type on milk phospholipids and fatty acid composition of polar lipids.

The effects of grass silage and red clover silage on milk fatty acid (FA) composition are extensively studied, but little is known of their effects on minor lipid constituents of milk fat globule membrane. We investigated the effects of forage:concentrate (FC) ratio in grass silage-based diets and forage type (grass silage vs. red clover silage) on selected molecular species of milk phospholipids (PL) and the FA composition of PL. Ten multiparous Nordic Red cows were offered following dietary treatments: grass silage-based diets containing 70:30 (HG) or 30:70 (LG) FC ratio or a red clover silage-based diet (RC) comprising 50:50 FC ratio on a dry matter basis. The most abundant molecular species within the phosphatidylcholines was 16:0-18:1 phosphatidylcholine that was increased by 18% in HG compared with LG milk. Dietary treatments did not affect the relative proportion of 18:1-18:1+18:0-18:2 phosphatidylethanolamine that was the most prevalent species (ca. 44%-45%) in that class. We identified the d18:1-22:0 sphingomyelin as the most abundant sphingomyelin species that tended to increase in HG milk compared with LG. The FC ratio did not affect the relative proportions of saturated FA nor monounsaturated FA in PL, but the proportion of cis-9 18:1 was elevated in HG versus LG milk, whereas the proportion of 18:2n-6 was 50% higher in LG versus HG milk. The RC diet increased monounsaturated FA and 18:3n-3 levels in PL compared with grass silage-based diets and decreased the relative proportion of saturated FA. However, the RC diet did not affect the relative proportion of polyunsaturated FA in PL, although red clover silage typically increases the proportion of polyunsaturated FA in milk fat. This study provides valuable knowledge of the minor lipid components in milk on species level in relation to common feeding strategies in high-forage systems.

Prevention of incisional hernia with retrorectus synthetic mesh versus biological mesh following loop ileostomy closure (Preloop trial).

BACKGROUND: The rate of incisional hernia after closure of a temporary loop ileostomy is significant. Synthetic meshes are still commonly avoided in contaminated wounds. The Preloop trial was a multicentre RCT designed to evaluate the benefits of synthetic mesh in incisional hernia prevention, and its safety for use in a contaminated surgical site compared with biological mesh. METHODS: Study patients who underwent closure of a loop ileostomy after anterior resection for rectal cancer were assigned to receive either retrorectus synthetic or biological mesh to prevent incisional hernia. The primary outcomes were surgical-site infections within 30 days, and clinical or radiological incisional hernia incidence at 10 months. Secondary outcomes were reoperation rate, operating time, duration of hospital stay, other complications within 30 days of surgery, 5-year quality of life measured by RAND-36, and incisional hernia incidence within 5 years of follow-up. RESULTS: Between November 2018 and September 2021, 102 patients were randomised, of whom 97 received the intended allocation. At 10-month follow-up, 90 patients had undergone clinical evaluation and 88 radiological evaluation. One patient in each group (2 per cent) had a clinical diagnosis of incisional hernia (P = 0.950) and one further patient in each group had a CT-confirmed incisional hernia (P = 0.949). The number of other complications, reoperation rate, operating time, and duration of hospital stay did not differ between the study groups. CONCLUSION: Synthetic mesh appeared comparable to biological mesh in efficacy and safety for incisional hernia prevention at the time of loop ileostomy closure. REGISTRATION NUMBER: NCT03445936 (http://www.clinicaltrials.gov).

ESYT1 tethers the ER to mitochondria and is required for mitochondrial lipid and calcium homeostasis.

Mitochondria interact with the ER at structurally and functionally specialized membrane contact sites known as mitochondria-ER contact sites (MERCs). Combining proximity labelling (BioID), co-immunoprecipitation, confocal microscopy and subcellular fractionation, we found that the ER resident SMP-domain protein ESYT1 was enriched at MERCs, where it forms a complex with the outer mitochondrial membrane protein SYNJ2BP. BioID analyses using ER-targeted, outer mitochondrial membrane-targeted, and MERC-targeted baits, confirmed the presence of this complex at MERCs and the specificity of the interaction. Deletion of ESYT1 or SYNJ2BP reduced the number and length of MERCs. Loss of the ESYT1-SYNJ2BP complex impaired ER to mitochondria calcium flux and provoked a significant alteration of the mitochondrial lipidome, most prominently a reduction of cardiolipins and phosphatidylethanolamines. Both phenotypes were rescued by reexpression of WT ESYT1 and an artificial mitochondria-ER tether. Together, these results reveal a novel function for ESYT1 in mitochondrial and cellular homeostasis through its role in the regulation of MERCs.

Importancia del papel de Haemophilus no ducreyi en el tracto genital femenino y su relación con la clínica / Importance of the role of Haemophilus no ducreyi isolates in female genital tract and its clinical relationship

Antecedentes: Cada vez son más frecuentes los informes microbiológicos con agentes emergentes en episodios clínicos del aparato genital de sujetos con sospecha de infección, como son las especies de Haemophilus no ducreyi (HND). El objetivo de este trabajo es analizar la importancia clínica del aislamiento de estas especies en el tracto genital del sexo femenino. Pacientes y métodos: Se realizó un estudio observacional descriptivo y retrospectivo en un hospital universitario del sudeste español, donde se evalúan los aislamientos de HND en muestras de exudados genitales femeninos procedentes de atención sanitaria especializada entre 2016 y 2019. Se analizaron variables clínicas, epidemiológicas y microbiológicas de los episodios infecciosos de mujeres adultas y niñas. Resultados: Se encontraron 45 (25 mujeres y 20 niñas) aislamientos de HND, correspondiendo al 1% del total, siendo la especie más frecuente Haemophilus influenzae (64,4%). En mujeres predominaron la leucorrea y el dolor abdominal, y en el 72% hubo aislamiento polimicrobiano. En niñas se aisló frecuentemente de forma aislada, con presencia de eritema vulvovaginal, flujo patológico y prurito local. Destacó la alta tasa de resistencia de Haemophilus parainfluenzae a azitromicina (72,7%) y cotrimoxazol (18,2%) en mujeres adultas, y la resistencia a azitromicina en niñas (25%). Conclusiones: H. influenzae y H.parainfluenzae deben tenerse en cuenta como posible agente etiológico en casos de vaginitis y cervicitis en mujeres adultas, así como en sospecha de enfermedad pélvica inflamatoria. En niñas, H.influenzae representa uno de los agentes microbiológicos de las infecciones vulvovaginales. La tasa de resistencia a azitromicina de H.parainfluenzae y a cotrimoxazol de ambas especies se debe tener presente.(AU)

Background: The isolation of new pathogens in clinical samples from the genital tract of subjects with suspected infection, such as Haemophilus no ducreyi (HND) species, is becoming more frequent. The objective of this work is to analyze the pathogenic role and the clinical importance of the isolation of these species in female genital tract. Patients and methods: We carried out an observational, descriptive, and retrospective study from a Hospital in Granada (Spain). HND isolates in female genital samples between 2016 and 2019 from specialized care were studied. Clinical, epidemiological, and microbiological variables of clinical episodes of adult women and girls were analyzed. Results: Forty-five (25 women and 20 girls) isolates of HND were found, corresponding to 1%; the most frequent specie was Haemophilus influenzae (64.4%). In women, leukorrhea and abdominal pain was frequent and in 72% there was a polymicrobial isolate. In girls, it was frequently in isolation, with the presence of vulvovaginal erythema, pathological discharge, and local itching. We highlight the high rate of resistance of Haemophilus parainfluenzae to azithromycin (72.7%) and cotrimoxazole (18.2%) in adult women, in contrast to resistance to azithromycin in girls (25%). Conclusions: H. influenzae and H. parainfluenzae should be considered as a possible etiological agent in cases of vaginitis and cervicitis in adult women, as well as in suspected pelvic inflammatory disease. In girls, H.influenzae represents one of the microbiological agents within the etiologies of vulvovaginal infections. We highlight the rate of resistance to azithromycin in H.parainfluenzae and to cotrimoxazole in both species.(AU)

Decrease in suicide rates in Brazil during the COVID-19 pandemic.

Natural disasters such as public health epidemics may potentially affect suicide rates. The global COVID-19 pandemic poses an unprecedented challenge for healthcare systems and general populations worldwide. In this retrospective ecological study, we aimed to examine any changes in the suicide rates during the COVID-19 pandemic and to assess the relationship between COVID-19 death rates and deaths by suicide in Brazil. Data on suicide and COVID-19 case numbers were extracted from the Ministry of Health agencies and grouped weekly. We performed a time series analysis of suicide rates, a comparison of mean suicide rates between the pre-COVID-19 period and the COVID-19 period, and conducted a Poisson regression to examine the relationship between deaths due to COVID-19 and suicide rates. Our results showed decreased suicide rates during the COVID-19 pandemic. We also found that deaths owing to COVID-19 impact those owing to suicide after 10 weeks in the upward direction; however, we did not observe for enough time to see a change in the suicide rate curve. These findings are fundamental to understand suicidal behaviors in epidemic situations. However, the field needs more studies evaluating the impact of significant public health events on suicidality, incorporating extended follow-up periods.

[Tuberculous otitis media: A case presentation and review of european literature]. / Otitis media tuberculosa: Presentación de un caso y revisión de la literatura europea.

Tuberculous otitis media (TOM) is a rare affectation in our environment that represents a challenge in its diagnosis due to the non-specific symptoms that it usually presents. This paper presents our experience in the diagnosis of a case of TOM in a 66-year-old woman with hearing loss and chronic otorrhea of more than 6 months of evolution that did not respond to conventional treatments. In addition, a review of the cases published in the last 20 years (2000-2022) in countries of the European Union (EU) is carried out. The most common symptoms were otorrhea (n=43; 100%), hearing loss (n=37; 86.05%), eardrum perforation (n=19; 44.18%), facial paralysis (n=12, 27,91%) and ear pain (n=13; 30,23%). The most used sample for diagnosis was the biopsy obtained by mastoidectomy (n=34; 79.06%). All patients were given antituberculous therapy for a mean duration of 8.11 months (range, 6-12 months). The most frequent aftereffect was hearing loss (n=28; 65.12%). TOM should be included in the differential diagnosis of chronic suppurative otitis, since early diagnosis and treatment reduce the probability of suffering irreversible sequelae.

[Antibiotic resistance changes in episodes of symptomatic bacteriospermia: development in a health area of southeast Spain]. / Cambios en la resistencia antibiótica en episodios de bacteriospermia sintomática: Evolución en un área de salud del sudeste español.

OBJECTIVE: Chronic bacterial prostatitis (CBP) is an entity of difficult clinical diagnosis and treatment, being the microbiological study of semen the main diagnostic test. This study aimed to determine the etiology and antibiotic resistance in patients with symptomatic bacteriospermia (SBP) in our environment. METHODS: A cross-sectional and retrospective descriptive study has been carried out from a Regional Hospital of the Spanish Southeast. The participants were patients assisted in the consultations of the Hospital with clinic compatible with CBP, between 2016 and 2021. The interventions were collection and analysis of the results derived from the microbiological study of the semen sample. The main determinations were the etiology and rate of antibiotic resistance of BPS episodes are analyzed. RESULTS: The main isolated microorganism is Enterococcus faecalis (34.89%), followed by Ureaplasma spp. (13.74%) and Escherichia coli (10.98%). The rate of antibiotic resistance of E. faecalis to quinolones (11%) is lower than previous studies, while for E. coli it has been higher (35%). The low rate of resistance shown by E. faecalis and E. coli to fosfomycin and nitrofurantoin stands out. CONCLUSIONS: In the SBP, gram-positive and atypical bacteria are established as the main causative agents of this entity. This forces us to rethink the therapeutic strategy used, which will avoid the increase in antibiotic resistance, recurrences, and chronicity of this pathology.

Synthetic mesh versus biological mesh to prevent incisional hernia after loop-ileostomy closure: a randomized feasibility trial.

BACKGROUND: Incisional hernia is a frequent complication after loop-ileostomy closure, rationalizing hernia prevention. Biological meshes have been widely used in contaminated surgical sites instead of synthetic meshes in fear of mesh related complications. However, previous studies on meshes does not support this practice. The aim of Preloop trial was to study the safety and efficacy of synthetic mesh compared to a biological mesh in incisional hernia prevention after loop-ileostomy closure. METHODS: The Preloop randomized, feasibility trial was conducted from April 2018 until November 2021 in four hospitals in Finland. The trial enrolled 102 patients with temporary loop-ileostomy after anterior resection for rectal cancer. The study patients were randomized 1:1 to receive either a light-weight synthetic polypropylene mesh (Parietene Macro™, Medtronic) (SM) or a biological mesh (Permacol™, Medtronic) (BM) to the retrorectus space at ileostomy closure. The primary end points were rate of surgical site infections (SSI) at 30-day follow-up and incisional hernia rate during 10 months' follow-up period. RESULTS: Of 102 patients randomized, 97 received the intended allocation. At 30-day follow-up, 94 (97%) patients were evaluated. In the SM group, 1/46 (2%) had SSI. Uneventful recovery was reported in 38/46 (86%) in SM group. In the BM group, 2/48 (4%) had SSI (p > 0.90) and in 43/48 (90%) uneventful recovery was reported. The mesh was removed from one patient in both groups (p > 0.90). CONCLUSIONS: Both a synthetic mesh and biological mesh were safe in terms of SSI after loop-ileostomy closure. Hernia prevention efficacy will be published after the study patients have completed the 10 months' follow-up.

[Shigellosis attended in the emergency department of a regional hospital in southeastern Spain: from their presence to multiresistance]. / Shigelosis atendidas en el servicio de urgencias de un hospital regional del sudeste de España: Desde su presencia a la multirresistencia.

OBJECTIVE: In the bacterial etiology of severe acute infectious diarrhea, except that caused by Clostridioides difficile, most of them have an invasive character and antibiotic treatment will be necessary in specific situations. Shigella is a classic pathogen, in which it is crucial to know the sensitivity to different classic and alternative antimicrobials. The objective of this work was to analyze the presence of shigellosis and the rate of antibiotic resistance. METHODS: A descriptive-retrospective study of the reports of shigellosis of stool cultures issued between January 2016 and April 2022 was conducted. RESULTS: A total of 34 episodes (16 -47.1%- by Shigella sonnei) were observed, as of 2018. There were only 2 pediatric cases. The overall resistance rate to azithromycin, trimethoprim-sulfamethoxazole and ciprofloxacin was 52.9%, 64.7% and 44.1%, respectively. 26.5% were resistant to the 3 groups of antibiotics. There was a higher rate of resistance for S. sonnei. The emergence of resistance to cephalosporins in recent years stands out. Episodes of multidrug-resistant shigellosis were detected between 2020 (1 by S. flexneri) and 2022 (4 by S. sonnei). CONCLUSIONS: The episodes of shigellosis are emerging in our environment with a higher rate of multi-resistance. In this context, current empirical treatments for acute enteroinvasive enteritis are at risk of failure, if necessary.

Identification of Proximity Interactors of Mammalian Nucleoid Proteins by BioID.

Mitochondrial nucleoids are compact nucleoprotein complexes, in which mtDNA is located, replicated, and transcribed. Several proteomic approaches have been previously employed to identify nucleoid proteins; however, a consensus list of nucleoid-associated proteins has not been generated. Here we describe a proximity-biotinylation assay, BioID, which allows identification of proximity interactors of mitochondrial nucleoid proteins. It uses a promiscuous biotin ligase fused to a protein of interest which covalently attaches biotin to lysine residues of its proximal neighbors. Biotinylated proteins can be further enriched by a biotin-affinity purification and identified by mass-spectrometry. BioID can identify transient and weak interactions and can be used to identify changes in the interactions upon different cellular treatments, for different protein isoforms or for pathogenic variants.

Loss of mitochondrial Chchd10 or Chchd2 in zebrafish leads to an ALS-like phenotype and Complex I deficiency independent of the mitochondrial integrated stress response.

Mutations in CHCHD10 and CHCHD2, encoding two paralogous mitochondrial proteins, have been identified in cases of amyotrophic lateral sclerosis, frontotemporal lobar degeneration, and Parkinson's disease. Their role in disease is unclear, though both have been linked to mitochondrial respiration and mitochondrial stress responses. Here, we investigated the biological roles of these proteins during vertebrate development using knockout (KO) models in zebrafish. We demonstrate that loss of either or both proteins leads to motor impairment, reduced survival and compromised neuromuscular junction integrity in larval zebrafish. Compensation by Chchd10 was observed in the chchd2-/- model, but not by Chchd2 in the chchd10-/- model. The assembly of mitochondrial respiratory chain Complex I was impaired in chchd10-/- and chchd2-/- zebrafish larvae, but unexpectedly not in a double chchd10-/- and chchd2-/- model, suggesting that reduced mitochondrial Complex I cannot be solely responsible for the observed phenotypes, which are generally more severe in the double KO. We observed transcriptional activation markers of the mitochondrial integrated stress response (mt-ISR) in the double chchd10-/- and chchd2-/- KO model, suggesting that this pathway is involved in the restoration of Complex I assembly in our double KO model. The data presented here demonstrates that the Complex I assembly defect in our single KO models arises independently of the mt-ISR. Furthermore, this study provides evidence that both proteins are required for normal vertebrate development.

[Microbial etiology of bacterial chronic prostatitis: systematic review]. / Etiología microbiana de la prostatitis crónica bacteriana: revisión sistemática.

OBJECTIVE: The importance of Gram-positive microorganisms and atypical bacteria in chronic bacterial prostatitis (CBP) has recently been described. For this reason, this study analyzes the etiology of CBP, as well as the evolution of antibiotic resistance through a systematic review. METHODS: A systematic review of studies obtained through the MEDLINE (PubMed) database, related to the etiology and antibiotic resistance profile of CBP, published up July 1, 2021. RESULTS: The most frequent isolated microorganisms that we have found in publications are Enterococcus faecalis (46.90%), Staphylococcus spp. (22.30%), Escherichia coli (15.09%) and atypical bacteria (6.04%). CONCLUSIONS: CBP is undergoing and unprecedented change of paradigm. Gram-positive bacteria and atypical bacteria are the main pathogens involved in the aetiology of this entity. This forces us to rethink the therapeutic strategy used, since it is necessary to use antibiotics that assume the etiological change and the profile of antibiotic resistance described.