Prevalence of Dry Eye Disease in Laser-Assisted In Situ Keratomileusis Candidates.

OBJECTIVE: To evaluate the prevalence of dry eye disease (DED) in laser-assisted in situ keratomileusis (LASIK) candidates. METHODS: A chart review of consecutive LASIK candidates who underwent full ocular surface work-up was performed, including ocular surface disease index (OSDI), noninvasive tests (noninvasive tear breakup time [ni-TBUT], tear meniscus height, lipid layer thickness, and meibography), and invasive tests (Schirmer test I, fluorescein TBUT, corneal staining, and meibomian gland [MG] expressibility). The prevalence of DED was calculated according to the Dry Eye Workshop II (DEWS II), and Japanese and Asia Dry Eye Society (JDES/ADES) criteria. RESULTS: In total, 135 patients (270 eyes) were evaluated. The mean age was 32.6±8.3 years, and 62.9% were women (n=85); 19 patients (15.4%) wore contact lenses, and 31 patients (23.8%) used artificial tears. The mean OSDI was 18.2±16.9, which was abnormal in 54.1% (n=62). Inferior lid MG dropout was the sign with the highest percentage of abnormal results (61.5%; n=83). There were no differences between men and women in any test except for ni-TBUT (6.3±0.3 and 7.2±0.2, respectively; P=0.002). Dry eye disease prevalence was 25.9% and 53.3%, according to JDES/ADES and DEWS II criteria, respectively. The only significant risk factor for DED was artificial tear use for both DEWS II (odds ratio [OR]=3.5, confidence interval [CI] [1.35-9.39]) and JDES/ADES (OR=2.58, CI [1.03-6.48]). CONCLUSIONS: This study found a high prevalence of DED and abnormalities in LASIK candidates and highlights the importance of ocular surface evaluation before photorefractive surgery.

Systematic exploration of a decade of publications on psychiatric genetics in Latin America.

Psychiatric disorders have a great impact in terms of mortality, morbidity, and disability across the lifespan. Considerable effort has been devoted to understanding their complex and heterogeneous genetic architecture, including diverse ancestry populations. Our aim was to review the psychiatric genetics research published with Latin American populations from 2010 to 2019, and classify it according to country of origin, type of analysis, source of funding, and other variables. We found that most publications came from Brazil, Mexico, and Colombia. Also, local funds are generally not large enough for genome-wide studies in Latin America, with the exception of Brazil and Mexico; larger studies are often done in collaboration with international partners, mostly funded by US agencies. In most of the larger studies, the participants are individuals of Latin American ancestry living in the United States, which limits the potential for exploring the complex gene-environment interaction. Family studies, traditionally strong in Latin America, represent about 30% of the total research publications. Scarce local resources for research in Latin America have probably been an important limitation for conducting bigger and more complex studies, contributing to the reduced representation of these populations in global psychiatric genetics studies. Increasing diversity must be a goal to improve generalizability and applicability in clinical settings.

Characterization of Adherent-Invasive Escherichia coli (AIEC) Outer Membrane Proteins Provides Potential Molecular Markers to Screen Putative AIEC Strains.

Adherent-invasive E. coli (AIEC) is a pathotype associated with the etiopathogenesis of Crohn's disease (CD), albeit with an as-yet unclear role. The main pathogenic mechanisms described for AIEC are adherence to epithelial cells, invasion of epithelial cells, and survival and replication within macrophages. A few virulence factors have been described as participating directly in these phenotypes, most of which have been evaluated only in AIEC reference strains. To date, no molecular markers have been identified that can differentiate AIEC from other E. coli pathotypes, so these strains are currently identified based on the phenotypic characterization of their pathogenic mechanisms. The identification of putative AIEC molecular markers could be beneficial not only from the diagnostic point of view but could also help in better understanding the determinants of AIEC pathogenicity. The objective of this study was to identify molecular markers that contribute to the screening of AIEC strains. For this, we characterized outer membrane protein (OMP) profiles in a group of AIEC strains and compared them with the commensal E. coli HS strain. Notably, we found a set of OMPs that were present in the AIEC strains but absent in the HS strain. Moreover, we developed a PCR assay and performed phylogenomic analyses to determine the frequency and distribution of the genes coding for these OMPs in a larger collection of AIEC and other E. coli strains. As result, it was found that three genes (chuA, eefC, and fitA) are widely distributed and significantly correlated with AIEC strains, whereas they are infrequent in commensal and diarrheagenic E. coli strains (DEC). Additional studies are needed to validate these markers in diverse strain collections from different geographical regions, as well as investigate their possible role in AIEC pathogenicity.

Design of Chitosan Nanocapsules with Compritol 888 ATO® for Imiquimod Transdermal Administration. Evaluation of Their Skin Absorption by Raman Microscopy.

PURPOSE: Design imiquimod-loaded chitosan nanocapsules for transdermal delivery and evaluate the depth of imiquimod transdermal absorption as well as the kinetics of this absorption using Raman Microscopy, an innovative strategy to evaluate transdermal absorption. This nanovehicle included Compritol 888ATO®, a novel excipient for formulating nanosystems whose administration through the skin has not been studied until now. METHODS: Nanocapsules were made by solvent displacement method and their physicochemical properties was measured by DLS and laser-Doppler. For transdermal experiments, newborn pig skin was used. The Raman spectra were obtained using a laser excitation source at 532 nm and a 20/50X oil immersion objective. RESULTS: The designed nanocapsules, presented nanometric size (180 nm), a polydispersity index <0.2 and a zeta potential +17. The controlled release effect of Compritol was observed, with the finding that half of the drug was released at 24 h in comparison with control (p < 0.05). It was verified through Raman microscopy that imiquimod transdermal penetration is dynamic, the nanocapsules take around 50 min to penetrate the stratum corneum and 24 h after transdermal administration, the drug was in the inner layers of the skin. CONCLUSIONS: This study demonstrated the utility of Raman Microscopy to evaluate the drugs transdermal penetration of in the different layers of the skin. Graphical Abstract New imiquimod nanocapsules: evaluation of their skin absorption by Raman Microscopy and effect of the compritol 888ATO® in the imiquimod release profile.

Underlying domains of anxiety trait in a Costa Rican sample: preliminary results.

BACKGROUND: Imprecision of the psychiatric phenotype might partially explain the failure of genetic research to identify genes that contribute to susceptibility of anxiety disorders. Previous research concluded two underlying constructs, worry and rumination, might explain anxiety sub-syndromic symptoms in Costa Rican patients with history of mania. The goal of the current study is to explore the presence of latent constructs for quantitative anxiety in a group of subjects with a wide diagnostic phenotype and non-affected individuals. METHODS: We conducted an exploratory factor analysis of anxiety trait in 709 subjects. Our sample was comprised by 419 subjects with psychiatric disorders and 290 non-affected individuals. We used principal factors extraction method with squared multiple correlations of the STAI (trait subscale). RESULTS: We found the following preliminary results: a three-factor solution with a good simple structure and statistical adequacy was obtained with a KMO of 0.92 (>0.6) and Bartlett's Test of Sphericity of 5644,44 (p<0.05). The STAI items were grouped into three factors: anxiety-absent, worry and rumination based on the characteristics of the symptoms. CONCLUSION: Two underlying constructs, worry and rumination may explain anxiety sub-syndromic symptoms in Costa Rican subjects. Our proposed underlying structure of subsyndromal anxiety in individuals should be considered as an important factor in defining better phenotypic characterizations on a broader diagnostic concept. Worry and rumination as a phenotypic characterization may assist in genotyping; however, its predictive value on actual illness outcome still requires more research. The Genome-Wide QTL analysis for anxiety trait in the same sample is ongoing.

Immunoproteomic analysis to identify Shiga toxin-producing Escherichia coli outer membrane proteins expressed during human infection.

Shiga-toxin producing Escherichia coli (STEC) is the etiologic agent of acute diarrhea, dysentery, and hemolytic-uremic syndrome (HUS). There is no approved vaccine for STEC infection in humans, and antibiotic use is contraindicated, as it promotes Shiga toxin production. In order to identify STEC-associated antigens and immunogenic proteins, outer membrane proteins (OMPs) were extracted from STEC O26:H11, O103, O113:H21, and O157:H7 strains, and commensal E. coli strain HS was used as a control. SDS-PAGE, two-dimensional-PAGE analysis, Western blot assays using sera from pediatric HUS patients and controls, and matrix-assisted laser desorption ionization-tandem time of flight analyses were used to identify 12 immunogenic OMPs, some of which were not reactive with control sera. Importantly, seven of these proteins have not been previously reported to be immunogenic in STEC strains. Among these seven proteins, OmpT and Cah displayed IgG and IgA reactivity with sera from HUS patients. Genes encoding these two proteins were present in a majority of STEC strains. Knowledge of the antigens produced during infection of the host and the immune response to those antigens will be important for future vaccine development.

Escherichia coli isolates from inflammatory bowel diseases patients survive in macrophages and activate NLRP3 inflammasome.

Crohn's disease (CD) is a multifactorial pathology associated with the presence of adherent-invasive Escherichia coli (AIEC) and NLRP3 polymorphic variants. The presence of intracellular E. coli in other intestinal pathologies (OIP) and the role of NLRP3-inflammasome in the immune response activated by these bacteria have not been investigated. In this study, we sought to characterize intracellular strains isolated from patients with CD, ulcerative colitis (UC) and OIP, and analyze NLRP3-inflammasome role in the immune response and bactericidal activity induced in macrophages exposed to invasive bacteria. For this, intracellular E. coli isolation from ileal biopsies, using gentamicin-protection assay, revealed a prevalence and CFU/biopsy of E. coli higher in biopsies from CD, UC and OIP patients than in controls. To characterize bacterial isolates, pulsed-field gel electrophoresis (PFGE) patterns, virulence genes, serogroup and phylogenetic group were analyzed. We found out that bacteria isolated from a given patient were closely related and shared virulence factors; however, strains from different patients were genetically heterogeneous. AIEC characteristics in isolated strains, such as invasive and replicative properties, were assessed in epithelial cells and macrophages, respectively. Some strains from CD and UC demonstrated AIEC properties, but not strains from OIP. Furthermore, the role of NLRP3 in pro-inflammatory cytokines production and bacterial elimination was determined in macrophages. E. coli strains induced IL-1ß through NLRP3-dependent mechanism; however, their elimination by macrophages was independent of NLRP3. Invasiveness of intracellular E. coli strains into the intestinal mucosa and IL-1ß production may contribute to CD and UC pathogenesis.

Cuidado bucal en mayores dependientes de un programa de cuidados domiciliarios / Oral care of dependent elderly in a domiciliary care program

Propósito Determinar las características de los cuidadores, su capacitación y rol en el cuidado de la higiene bucal del adulto mayor dependiente del Programa de cuidados domiciliarios de la comuna de Providencia. Método Se realizó un estudio descriptivo transversal, seleccionándose una muestra de 75 cuidadores mediante un muestreo aleatorizado simple del listado de pacientes adultos mayores pertenecientes al Programa de cuidados domiciliarios de la comuna de Providencia, Santiago, Chile. Se evaluó mediante un cuestionario las características del cuidador, entrenamiento en higiene bucal, frecuencia y dificultades a la hora de realizar la higiene bucal y quién realiza la higiene bucal según el nivel de dependencia de la persona mayor (índice de Katz). Resultados La mayoría de los cuidadores no ha recibido entrenamiento en el cuidado bucal (73,4%). El 57,3% ha realizado alguna vez la higiene bucal de la persona mayor y la mayoría ha tendido dificultades (82,7%), principalmente que el adulto mayor no quiere que otra persona lo realice. Todos los dependientes leves realizan por sí mismos el cuidado de su higiene bucal, pero la mayoría de los dependientes severos requiere de ayuda (7,9%), o que definitivamente el cuidador la realice (76,3%). Conclusiones Resulta fundamental mejorar la instrucción sobre higiene bucal en los ciudadores de adultos mayores dependientes domiciliarios. A mayor nivel de dependencia, el rol del cuidador en la higiene bucal cobra mayor importancia.

Purpose To determine the characteristics of the caregivers, their training and role in oral health care practices of the dependent elderly in a Domiciliary Care Program, Commune of Providencia. Method A cross-sectional study was conducted on a randomised sample of elderly individuals, who were part of the Domiciliary Care Program in the Commune of Providencia, Santiago, Chile. A questionnaire was used to determine the caregiver characteristics, their training in oral health care, frequency of oral hygiene, and difficulties in carrying it out, as well as who performs the hygiene according to the level of dependency of the elderly person (Katz Index). Results Most (73.4%) of the caregivers have not received any training for oral health care. More than half (57,3%) of the caregivers have carry out the oral hygiene of an elder and the majority (82.7%) had difficulties to complete it, mostly the elder doesn’t allow other people do the hygiene. All elders with mild dependency perform their own oral hygiene but the majority of elders with severe dependency need help (7.9%) or the caregiver must do it (76.3%). Conclusions It is fundamental to improve caregiver training on oral health care of domiciliary dependent elderly people. When dependency increases, the role of the caregiver in oral hygiene is most important.

Excision of an unstable pathogenicity island in Salmonella enterica serovar Enteritidis is induced during infection of phagocytic cells.

The availability of the complete genome sequence of several Salmonella enterica serovars has revealed the presence of unstable genetic elements in these bacteria, such as pathogenicity islands and prophages. This is the case of Salmonella enterica serovar Enteritidis (S. Enteritidis), a bacterium that causes gastroenteritis in humans and systemic infection in mice. The whole genome sequence analysis for S. Enteritidis unveiled the presence of several genetic regions that are absent in other Salmonella serovars. These regions have been denominated "regions of difference" (ROD). In this study we show that ROD21, one of such regions, behaves as an unstable pathogenicity island. We observed that ROD21 undergoes spontaneous excision by two independent recombination events, either under laboratory growth conditions or during infection of murine cells. Importantly, we also found that one type of excision occurred at higher rates when S. Enteritidis was residing inside murine phagocytic cells. These data suggest that ROD21 is an unstable pathogenicity island, whose frequency of excision depends on the environmental conditions found inside phagocytic cells.

Deletion of a prophage-like element causes attenuation of Salmonella enterica serovar Enteritidis and promotes protective immunity.

Salmonella enterica serovar Enteritidis (S. Enteritidis) is a wide host range serovar belonging to the S. enterica genus. Worldwide, it is one of the most frequent causes of food borne disease. Similar to S. Typhimurium, some virulence genes of S. Enteritidis are located in pathogenicity islands and prophages. In this study we have generated a mutant strain of S. Enteritidis lacking a prophage-like element, denominated varphiSE12. The resulting mutant strain was attenuated and promoted protective immunity in infected mice. Although S. Enteritidis strains lacking the complete prophage varphiSE12 remained capable of surviving inside phagocytic cells, they showed a significantly reduced capacity to colonize internal organs and failed to cause lethal disease in mice. Consistent with these data, infection with S. Enteritidis strains lacking prophage varphiSE12 promoted the production of anti-Salmonella IgG antibodies and led to protection against a challenge with virulent strains of S. Enteritidis. These results suggest that strains lacking this prophage can induce a protective immunity in mice and be considered as potential attenuated vaccines against S. Enteritidis.