Use of Schirmer strips and conjunctival swabs for virus detection on the ocular surface of adults: a scoping review / Fitas de Schirmer e swab conjunctival para detecção de vírus na superfície ocular de adultos: scoping review

ABSTRACT Schirmer strips and conjunctival swabs are used in ophthalmology for the collection of tears and fluids. One of the biggest challenges during the COVID-19 pandemic has been accurate diagnosis and, in some cases, ocular manifestations are among the first symptoms. In this context, this study aimed to collect evidence to support the use of Schirmer strips and conjunctival swabs as a method of sample collection for viral analysis. A literature search was conducted following the Scoping Review protocol defined by The Joanna Briggs Institute. Studies were analyzed regarding virus research, collection methods, and sample analysis. The findings support that viruses can be detected on the ocular surface through analysis of Schirmer strips and conjunctival swabs. However, additional studies with larger samples and time data are necessary to confirm these conclusions.

RESUMO A fita de Schirmer e o swab conjunctival são utilizados na oftalmologia como métodos de coleta para lágrimas e fluidos. Durante a pandemia da COVID-19, um dos desafios foi o diagnóstico correto e se sabe que, em alguns casos, as manifestações oculares podem ser um dos primeiros sintomas. Nesse contexto, este estudo tem como objetivo levantar evidência que destaque o uso de fitas de Schirmer e de swabs conjuntivais como método de coleta para análise viral. Conduziu-se uma revisão de literatura seguindo o protocolo para Scoping Review definido pelo Joanna Briggs Institute. Os pesquisadores analisaram os estudos em busca do vírus pesquisado, os métodos de coleta e os métodos de análise. Vírus podem ser detectados na superfície ocular através da análise de fitas de Schirmer e de swabs conjuntivais, entretanto novos estudos com populações maiores e com definições claras de tempo são necessários para conclusões mais assertivas no tema.

Advances in clinical application of mass spectrometry for protein analysis / 中华检验医学杂志

In recent years, mass spectrometry-based protein analysis has emerged as a powerful tool in clinical laboratories, offering more reliable assessments than conventional laboratory tools and providing benefits for disease diagnosis and treatment. The use of targeted protein quantitation has gained widespread acceptance, particularly in the detection of insulin-like growth factor I, thyroglobulin, and monoclonal antibody therapeutics. Meanwhile, significant advancements have been made in non-targeted protein analysis, such as the diagnosis of amyloidosis, monoclonal gammopathies, and membranous nephropathy. High-resolution mass spectrometry has enabled the identification and differentiation of pathogenic proteins in these diseases, as well as the discovery of new proteins involved in disease onset and progression. As a result, clinical diagnosis and disease differentiation have improved, as well as our understanding of these diseases.

Advances in Protein Characterization by High-Resolution Mass Spectrometry

High-resolution mass spectrometry has become an indispensable tool for a variety of biological research areas, includingprotein chemistry. The coupling of electrospray ionization to the MS has revolutionized the approaches for the identification ofnew proteins. Some examples of these applications include the identification of proteins involved in the virulence of pathogenicbacterial strains. MS played an important role in advancing protein folding studies, identification of new biomarkers for thedetection of diseases in early stages. A recent development in MS technique called fast photochemical oxidation of proteinssignificantly advanced the protein structural analysis.

Comparison of the Effectiveness between Sampling Methods for Protein Analysis of Tear Fluids

PURPOSE: Although various sampling methods of tears from conjunctival sac have been reported, no previous study compared their effectiveness or efficiency based on protein extraction. By comparing the compliance, volume and protein concentration of each tear sampling method, we searched for the most efficient tear collection method. METHODS: Resting tear samples of 14 eyes of normal subjects were collected using Schirmer paper, capillary tube, cellolose acetate rod and 3 different ophthalmic sponges made of different materials and density (Merocel(R), KeraCel(R) and Weck-Cel(R)). After centrifugation of the collected tear samples, the tear volume and protein concentration were measured for each method. Additionally, the compliance of each tear sampling method was analyzed by numerically representing the amount of discomfort experienced during resting tear collection. RESULTS: The average volume retrieved by each tear sampling method was 9.0 +/- 1.1 microL with no significant differences between groups. The average concentration of protein retrieved by each tear sampling method was 5.3 +/- 1.2 microg/microL. Merocel(R) retrieved 7.6 +/- 0.61 microg/microL, which was significantly higher than other sampling methods (p < 0.05). The compliance of Merocel(R) and the capillary tube were the highest, while KeraCel(R) showed the lowest compliance. CONCLUSIONS: Merocel(R) retrieved the highest amount of protein and showed high compliance and may be the most effective and easily applicable tear sampling method in clinical settings.

Establishment and evaluation of method to collect rat gingival crevicular fluid by using absorbent paper points / 吉林大学学报(医学版)

Objective To establish and evaluate the method to collect the rat gingival crevicular fluid (GCF)by using absorbent paper points, and to lay foundation for analysis on GCF.Methods 20 healthy male rats were selected and randomly divided into GCF group and saliva group.The GCF of the right upper molar gingival trough of the rats in GCF group and the saliva of the rats in saliva group were collected by using 1 5# absorbent paper points.The SDS-PAGE analysis and abundance detection were applied to analyze the protein bands of the samples in two groups.Results The SDS-PAGE analysis identified the proteins at 77 000,66 000,55 000,51 000,and 28 000,especially 66 000 in GCF group.While saliva group had lower brightness protein bands at 66 000,60 000, and 48 000.The data of protein abundance of 66 000 between two groups had statistically significant difference (P<0.05).Conclusion The number and types of the protein bands are different between GCF Group and saliva group,so using 15# absorbent paper points can collect the rat GCF successfully.

Study on the relationship between the plasma levels of ET-1, TAT, and hs-CRP and slow coronary flow syndrome / 中国医师杂志

Objective To investigate the relationship between the plasma levels of ET-1,TAT,and hs-CRP and slow coronary flow syndrome (SCFS),and explore effects of coronary endothelial function,coagulation function,and inflammatory reaction on blood flow of coronary artery.Methods A total of 400 cases with normal blood flow of coronary artery by coronary angiogram was randomly selected.The coronary flow patterns were determined by corrected thrombolysis in myocardial infarction frame count method (cT-FC).Among them,45 cases whose average cTFC more than 27 were assigned as SCFS group,the other 45 cases no SCFS.Plasma levels of ET-1,TAT and hs-CRPwere examined with enzyme-linked immunosorbent assay (ELISA),and were compared between two groups.Moreover,multivariate analysis evaluating predictors of SCFS was performed with regression test.Results No statistical difference was found between two groups concerning the gender,history of hypertension,diabetes mellitus,and cigarette alcohol percentage..The plasma level of HDL in SCFS group was lower than that of no SCFS [(1.22 ± 0.42) mmol/L vs (1.44±0.34) mmol/L,t =-2.731,P ＜0.01],but the plasma level of glucose in the former was higher than that of the latter [(5.68 ±0.62) mmol/L vs (5.10 ±0.84) mmol/L,t =3.727,P ＜0.01].However,Plasma levels of ET-1,TAT and hs-CRP in SCFS were higher than that of no SCFS [(94.3 ± 16.78) ng/Lvs (83.5±12.53) ng/L,t =3.051,P ＜0.01;(12.96±3.24)μg/Lvs (8.76 ±2.64)μg/L,t =5.945,P ＜ 0.01 ; (2.48 ± 0.35) μg/L vs (1.38 ± 0.46) μg/L,t =11.259,P ＜ 0.01].Furthermore,Logistic regression analysis showed that ET-1,TAT and hs-CRP were risk factors for SCFS (OR ＞ 1.22).Conclusions Due to coronary endothelial dysfunction,endothelial inflammatory reaction,and activated coagulation function,slow coronary flow of coronary artery occurs.

Tear Proteomics in Young Malays with Dry Eyes

Changes in tear protein concentrations may reflect ocular surface health. This study analyzes changes in tear protein concentrations of young Malays with dry eye (DE) and determines its association with the clinical findings. Methods: Subjects were screened using McMonnies questionnaire (MDEQ) and flourescein tear break up time (TBUT). Total tear protein concentration (TTPC) was determined using Bradford's technique and specific tear protein (sIgA, lysozyme, lactoferrin and human serum albumin (HSA)) concentrations were determined using SDS-PAGE. Parametric and nonparametric tests were used to compare means between groups. Spearman correlation was used to determine the association between variables measured. Results: A total of 42 subjects (21 DE and 21 NDE) were included. Mean MDEQ score for DE was 16.00±1.48 and NDE was 8.47±3.47. Mean TBUT for DE was 3.47±0.47s and NDE was 4.98±0.43s. Mean TTPC for DE and NDE was 9.84±2.40mg/ml and 8.96±1.84mg/ml respectively. Mean sIgA, lysozyme, lactoferrin and HSA for DE was 0.54±0.10mg/ml, 1.68±0.17mg/ml, 1.47±0.25mg/ml, 0.06±0.03mg/ml and for NDE was 0.57±0.09mg/ ml, 2.04±0.19mg/ml, 1.75±0.23mg/ml, 0.06±0.03mg/ml accordingly. Significant differences were noted in MDEQ score (p=0.01), TBUT (p=0.01), lactoferrin (p=0.01) and lysozyme (p=0.01) but not in TTPC (p=0.19), HSA (p=0.74) and sIgA (p=0.24) between groups. Significant correlations were noted between TBUT with lactoferrin (r=0.02, p=0.02) and lysozyme (r=0.63, p=0.01) and between MDEQ score with lactoferrin (r=-0.34, p=0.02) and lysozyme (r=-0.64, p=0.01). Conclusions: There are changes in specific tear protein in dry eye patients, which correlate well with clinical results. Tear protein analysis may play an important role in the diagnosis of the dry eye.

Characterization and lytic activity of Pseudomonas fluorescens phages from sewage

Pseudomonas fluorescens phages from sewage were tested against P. fluorescens isolates of soil and sewage. The phages were characterized as to host range, morphology, structural proteins and genome fingerprint. Of the seven phages isolated, one was found to be abundant in sewage (5.9×10(7) pfu/mL), having broad host range, and distinct protein and DNA profile when compared to the other six phages. DNA restriction and protein profiles of the phages and their morphology indicate the diversity in the sewage environment. None of the isolates from the rhizosphere regions of various cultivated soils were susceptible to phages isolated from sewage.

Métodos e estratégias em proteômica e suas aplicações na área vegetal / Methods and strategies in proteomics and their applications in plants

A implementação da espectrometria de massa (MS) para as análises de peptídeos (MS) e de aminoácidos (MS em tandem ou MS/MS) tornou possível a identificação de centenas de proteínas em experimentos únicos. Uma grande variedade de estratégias está disponível atualmente para o fracionamento e a purificação de amostras, a identificação de proteínas, a quantificação, a análise de modificações pós-traducionais (MPT's) e os estudos de interação. Dessa forma, a proteômica abre novas perspectivas na biologia de plantas com ênfase nos estudos de variabilidade genética, estresses fisiológicos e desenvolvimento de plantas.

The implementation of mass spectrometry (MS) for peptides (MS) and amino acids (tandem MS or MS/MS) analysis allowed the identification of hundreds of proteins in single experiments. A number of different strategies are current available for sample fractioning and purification, proteins identification, quantification, post-translational modifications (PTM) and interaction analyses. In this way, the proteomics open up new perspectives in plant biology with emphasis on studies of genetic variability, physiological stresses and plant development.

SDS-PAGE analysis of differentially expressed proteins from Anopheles stephensi hemolymph produced during the Plasmodium yoelii oocysts melanization / 第三军医大学学报

Objective To explore primarily the differentially expressed proteins in the hemolymph from adult female Anopheles stephensi ( An stephensi ) infected with Plasmodium yoelii ( P yoelii ) after being fed with sucrose solution containing nitroquine or not at different time points Methods Hemolymph of 2 groups of adult female An stephensi was collected with the expulsion method from the first day to the fifth day after the feeding Hemolymph samples were examined with SDS PAGE The protein gels were visualized by either Coomassie brilliant blue or silver staining, scanned and automatically analyzed by the BioRad1000 gel image analysis system for differential proteins bands Results On the second day of feeding with nitroquine, a few oocysts were partially melanized Furthermore, during the period from the fifth day to the ninth day, the number of mosquitoes with malanized oocysts and the number of melanized oocysts gradually increased The number of hemolymph protein binds in the treatment group was markedly more than that in the control Many different bands, mainly located at the molecular weight of (20～40)?10 3 and (60～80)?10 3, were visualized in the 2 groups The number of protein bands stained by the silver staining was more than that by the Coomassie brilliant blue staining Conclusion There are differentially expressed proteins in the hemolymph in An stephensi infected with P yoelii after being fed with sucrose solution containing nitroquine These differential proteins may be the melanization engaging proteins

Effect of human vitreous fluid on the morphology and expression of cytoskeleton of cultured retinal pigment epithelial cells / 中华眼底病杂志

Objective To describe cultured human retinal pigment epithelial (RPE) cells transdifferentiation and investigate the effects of human vitreous fluid on the morphologic and cytoskeleton changes of RPE cells in vitro. Methods Cytoskeleton characteristics in the 2 nd, 5 th, 8 th passage of RPE cells in normal culture, which included cytokeratin 18 (CK18) and ?-smooth muscle actin (?-SMA) were analyzed by Western blot. RPE cells were cultured in human vitreous-conditioned medium (VCM) at the concentration of 1∶4 for 6 days, morphologic changes were examined by light and electron microscopy, and cytoskeleton characteristics were analyzed by imunocytochemistry and Western blot. Results During culture in vitro, RPE cells lost epithelial characteristics and aquired fibroblast-like phenotype. The expression of CK18 was the highest at the 5 th passage, and it decreased in the following passage, but ?-SMA increased gradually. The morphologic transdifferentiation from epithelial to fibroblast-like cells of RPE was accelerated by VCM. Ultrastructural changes such as decreased microvilli and gradually increased rough endoplasmic reticulum and Golgi complex were found during the cultivation. CK18 produced by RPE cells decreased in VMC (P

Search for Primary Focus of Staphylococcus aureus Peritonitis in Continuous Ambulatory Peritoneal Dialysis (CAPD) Patients / 대한내과학회지

OBJECTIVES: Peritonitis is the most important complication of CAPD, often leading to failure of the technique and recourse to hemodialysis. Staphylococci is the most common organism in bacterial peritonitis associated with CAPD. The importance of the skin as a source of peritonitis causing isolate is suggested. We investigated the importance of anterior nares, hands and catheter exit-site skin as a source of peritonitis in CAPD patients by comparing the plasmid analysis with the bacterial protein analysis. METHODS: Thirty patients were suffered by peritonitis which was caused by S. aureus were studied. At presentation with an episode of S. aureus peritonitis, peritoneal dialysates, anterior nares, hands and catheter exit-site skin cultures were obtained. Antibiotics-sensitivity tests was performed and antibiogram of S. aureus which was cultured from peritoneal dialysates was compared with that from the skin. The similar antibiogram was identified in sixteen patients. The isolates were typed by rapid plasmid screen analysis and by means of visual comparison of autoradiographs of 35S-methionine staphylococcal protein analysis. RESULTS: The same plasmid analysis pattern of S. aureus isolated from the skin as that from the peritoneal dialysate was observed in 7patients and bacterial protein analysis pattern in 3patients. In seven patients who had the same plasmid analysis patients, three patients had the same plasmid analysis pattern of S. aureus from peritoneal dialysate as that from anterior nares and four patients had the same plasmid analysis pattern as that from the isolates of the exit-site skin. CONCLUSION: This study confirms the epidemiological link between carriage of S. aureus and peritonitis in CAPD patients and clinical usefulness of plasmid analysis for the delineation of focus of infection.