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Arterial thrombosis contributes to some of the most frequent causes of mortality globally, such as myocardial infarction and stroke. Platelets are essential mediators of physiological haemostasis and pathological thrombosis. Platelet activation is controlled by a multitude of signalling pathways. Upon activation, platelets shed platelet-derived extracellular vesicles (pEVs). In this Special Issue: Extracellular Vesicles, Moon et al. investigate the impact of various platelet agonists (thrombin, ADP, collagen) on the proteome of pEVs. The study demonstrates that pEVs exhibit an agonist-dependent altered proteome compared to their parent cells, with significant variations in proteins related to coagulation, complement, and platelet activation. The study observes the rapid generation of pEVs following agonist stimulation with specific proteome alterations that underscore an active packaging process. This commentary highlights the implications of their findings and discusses the role of pEV cargo in cardiovascular disease with potential novel therapeutic and diagnostic opportunities.
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Plaquetas , Vesículas Extracelulares , Ativação Plaquetária , Proteoma , Humanos , Vesículas Extracelulares/metabolismo , Vesículas Extracelulares/efeitos dos fármacos , Plaquetas/metabolismo , Plaquetas/efeitos dos fármacos , Proteoma/metabolismo , Ativação Plaquetária/efeitos dos fármacos , Proteômica/métodos , Trombina/metabolismo , Trombina/farmacologiaRESUMO
Reflective writing develops meta-cognition among students. Therefore, it is of interest to compare effectiveness of post lecture reflective writing to didactic lecture between individual and group reflective writing. Hence, we included 124 first-year students from AIIMS Bhopal, India and divided them in two groups of 62 students. Both groups took a pre-test using a reflection questionnaire. Students were taught reflective writing. Both groups attended physiology lectures on two different topics. First lecture on body fluids where Group A wrote reflections individually and Group B did so in sub-groups (B1 to B6). After another lecture on Pathophysiology of oedema, Group A wrote reflections in groups and Group B wrote individually (A1 to A6). Both groups took a test in the form of MCQ about reflective writing on lectures. After intervention both groups took a post-test using a reflection questionnaire. Mean and standard deviation of Pre-test is 3.86 ± 0.86 and Post-test is 7.58 ± 1.01, respectively. The Mean and standard deviation of reflection who wrote individually is 38.05 ± 4.41 and in group is 27.45 ± 3.93, respectively with p-value < 0.05. Evaluation of students who wrote reflection in groups after second lecture the mean and standard deviation of reflection who wrote individually is 38.22 ± 4.64 and in group is 27.03 ± 2.87 respectively with p-value < 0.05. The performance of students who wrote reflection in groups is not satisfactory as compared to students who wrote their reflection individually.
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A rapid, straightforward, and sensitive approach to quantifying enantiomeric barbiturates in serum was developed by integrating ultrasound-assisted dispersive liquid-liquid microextraction (UA-DLLME) with large-volume sample stacking (LVSS) in capillary electrophoresis (CE). UA-DLLME was employed for sample preparation, and on-column preconcentration by using LVSS with polarity switching was implemented to enhance sensitivity. We thoroughly investigated and optimized various parameters influencing extraction and stacking to achieve optimal detection performance with the highest enrichment efficiencies. Under optimal extraction conditions (injection of a mixed solution containing 40 µL of CHCl3 and 200 µL of tetrahydrofuran into 1 mL of a sample solution at pH 10.0), LVSS was performed using 600 mM Tris-boric acid (pH 9.5) containing 35 mM hydroxypropyl-ß-cyclodextrin and sodium taurodeoxycholate hydrate. A voltage of 20 kV was applied and a preinjection water plug was loaded at a height of 25 cm for 10 s. Subsequently, the sample solution was injected at a height of 25 cm for 480 s, after which a voltage of -20 kV was applied and the sample stacking was initiated. The stacking process was completed when 95 % of the separation current was attained. Under optimized conditions, the contraction folds of the four barbiturate analytes (R, S-Secobarbital, R, S-pentobarbital) were improved by approximately 6400-fold, achieving detection limits of 0.1 ng/mL. The limits of quantification for all analyte enantiomers were 0.5-50 ng/mL, demonstrating good linearity (r > 0.997). Migration times exhibited a relative standard deviation of less than 1.7 %, whereas peak areas for the four analytes exhibited a deviation of 8.7 %. Finally, the established method was effectively applied to the analysis of human serum samples.
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Barbitúricos , Eletroforese Capilar , Limite de Detecção , Microextração em Fase Líquida , Eletroforese Capilar/métodos , Microextração em Fase Líquida/métodos , Estereoisomerismo , Humanos , Barbitúricos/sangue , Barbitúricos/química , Reprodutibilidade dos TestesRESUMO
In cases of sexual assault, the interpretation of biological traces on clothing, and particularly undergarments, may be complex. This is especially so when the complainant and defendant interact socially, for instance as (ex-)partners or by co-habitation. Here we present the results from a study where latent male DNA on female worn undergarments is recovered in four groups with different levels of male-female social interaction. The results conform to prior expectation, in that less interaction tend to result in less male DNA on undergarments. We explore the use of these experimental data for evaluative reporting given activity level propositions in a mock case scenario. We show how the selection of different populations to represent the social interaction between complainant and defendant may affect the strength of the evidence. We further show how datasets of limited size can be used for robust activity level evaluative reporting.
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Vestuário , Impressões Digitais de DNA , DNA , Humanos , Feminino , Masculino , DNA/análise , Delitos Sexuais , Conjuntos de Dados como Assunto , Interação Social , Funções VerossimilhançaRESUMO
In addition to transmitting and carrying genetic information, RNA plays an important abiotic role in the world of nanomaterials. RNA is a natural polyanionic biomacromolecule, and its ability to promote osteogenesis by binding with other inorganic materials as an osteogenic induction agent was discovered only recently. However, whether it can promote osseointegration on implants has not been reported. Here, we investigated the effect of the RNA-containing coating materials on peri-implant osseointegration. Total RNA extracted from rat muscle tissue was used as an osteogenic induction agent, and hyaluronic acid (HA) was used to maintain its negative charge. In simulated body fluids (SBF), in vitro studies demonstrated that the resulting material encouraged calcium salt deposition. Cytological experiments showed that the RNA-containing coating induced greater cell adhesion and osteogenic differentiation in comparison to the control. The results of animal experiments showed that the RNA-containing coating had osteoinductive and bone conduction activities, which are beneficial for bone formation and osseointegration. Therefore, the RNA-containing coatings are useful for the surface modification of titanium implants to promote osseointegration.
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Fluid biopsy technology, characterized by its minimally invasive nature, speed, and continuity, has become a rapidly advancing and widely applied real-time diagnostic technique. Among various biomarkers, proteins represent the most abundant class of disease indicators. The sensitive and accurate detection of protein markers in bodily fluids is significantly influenced by the control exerted by recognition ligands. Aptamers, which are structurally dynamic functional oligonucleotides, exhibit high affinity, specific recognition of targets, and notable characteristics of high editability and modularity. These features make aptamer universal "recognition-capture" components, contribute to a significant leap in their applications within the biosensor domain. In this context, we provide a comprehensive review of the extensive application of aptamer-based biosensors in fluid biopsy. We systematically compile the characteristics and construction strategies of aptamer-based biosensors tailored for fluid biopsy, including aptamer sequences, affinity (KD), fluid background, sensing technologies, sensor construction strategies, incubation time, detection performance, and influencing factors. Furthermore, a comparative analysis of their advantages and disadvantages was conducted. In conclusion, we delineate and deliberate on prospective research trajectories and challenges that lie ahead in the realm of aptamer-based biosensors for fluid biopsy.
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Aptâmeros de Nucleotídeos , Biomarcadores , Técnicas Biossensoriais , Humanos , Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/métodos , Biomarcadores/análise , Biópsia Líquida/métodos , Proteínas/análise , Proteínas/química , Líquidos Corporais/químicaRESUMO
Background: Sterile body locations are usually associated with clinical urgency and life-threatening illnesses, and they are typically contaminated with diverse bacterial etiologies. If the bacteria acquire resistance to antimicrobial drugs, the public health crisis will only worsen. In developing countries, drug-resistant bacteria are common because of poor surveillance, diagnostic capacity, and control measures. Early diagnosis, and assessing the drug resistance and factors associated with infection are important to combat the drug resistance and treatment. This study aimed to assess the bacterial etiologies, antimicrobial susceptibility pattern, and possible associated factors among patients suspected of sterile body sites. Methods: A hospital-based cross-sectional study was conducted from June 2022 to August 2022 at Debre Markos Comprehensive Specialized Hospital in Amhara regional state, Ethiopia. One hundred seven study participants were selected using consecutive convenient sampling techniques. A structured questionnaire was used to collect socio-demographic and clinical data. Gram stain was done for a preliminary report and inoculated into blood agar, MacConkey agar, and chocolate agar and incubated aerobically and micro aerobically at 37°C for 24 h. Antimicrobial susceptibility testing was done by the modified Kirby Bauer's disk diffusion method. Data were analyzed using bivariate and multivariate logistic regression was used. A p-value less than 0.05 is considered as statistically significant. Results: The overall magnitude of sterile body site infection among study participants was 7.5% (14/187). The majority of the isolates were Gram-negative bacteria with the predominant species Enterobacter cloacae accounting for 28.57% (4/14). Among isolates 78.57%(11/14) of them were multidrug-resistant isolates. Being inpatient, co-morbidity, and alcohol consumption were significantly associated with sterile body site infection. Conclusion: In our study, Gram-negative bacteria were the predominant bacteria that infects sterile body fluid. The prevalence of multi-drug resistance bacteria isolates was significantly high. Therefore, before prescribing an empirical treatment, a medical professional should identify the bacterial etiology of sterile body fluids and the susceptibility of microbes to the drug.
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Aim: Urine as a biofluid has been rarely used as a diagnostic fluid in oral diseases. The article aims to systematically review the utility of human urinary carcinogen metabolites as an approach for obtaining important information about tobacco and cancer. Materials and Methods: The following article reviews the use of urine and its metabolites as biomarkers in various lesions of the oral cavity including oral squamous cell carcinoma and as a screening method in evaluating tobacco and its components. A bibliographic comprehensive search was carried out in the main databases: PUBMED, SciELO, Google Scholar, VHL, and LILACS for articles that were published from 1985 to 2020. The inclusion criteria were "urinary metabolites," "oral cancer/HNSCC," "body fluids," "tobacco," and "metabolomics." A total of 55 articles were collected which included laboratory studies, systematic reviews, and literature of urinary metabolites in tobacco users. Results: Most of the studies carried out show accurate results with high sensitivity of urinary metabolite biomarkers in individuals with tobacco-based habits and lesions caused by them. Conclusion: The review indicates that urinary metabolite analysis demonstrates its applicability for the diagnosis and prognosis of disease. Urine is a remarkable and useful biofluid for routine testing and provides an excellent resource for the discovery of novel biomarkers, with an advantage over tissue biopsy samples due to the ease and less invasive nature of collection.
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Noninvasive and effortless diagnosis of Alzheimer's disease (AD) remains challenging. Here we report the multiplexed profiling of extracellular vesicle (EV) surface proteins at the single EV level in five types of easily accessible body fluids using a proximity barcoding assay (PBA). A total of 183 surface proteins were detected on the EVs from body fluids collected from APP/PS1 transgenic mice and patients with AD. The AD-associated differentially expressed EV proteins could discriminate between the control and AD/AD model samples with high accuracy. Based on machine learning predictive models, urinary EV proteins exhibited the highest diagnostic potential compared to those on other biofluid EVs, both in mice and humans. Single EV analysis further revealed AD-associated EV subpopulations in the tested body fluids, and a urinary EV subpopulation with the signature proteins PLAU, ITGAX and ANXA1 could diagnose patients with AD in blinded datasets with 88% accuracy. Our results suggest that EVs and their subpopulations from noninvasive body fluids, particularly urine, are potential diagnostic biomarkers for AD.
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Doença de Alzheimer , Líquidos Corporais , Vesículas Extracelulares , Humanos , Camundongos , Animais , Doença de Alzheimer/diagnóstico , Doença de Alzheimer/metabolismo , Vesículas Extracelulares/metabolismo , Biomarcadores/metabolismo , Camundongos Transgênicos , Proteínas de Membrana/metabolismo , Líquidos Corporais/metabolismoRESUMO
Background: Exposure to splash of body fluids is one of the common ways of transmitting blood-borne infections from patients to healthcare practitioners. Globally, there is a paucity of evidence on exposure to splash of body fluids among hospital housekeepers. This study, therefore, investigated splash of body fluid and its predisposing factors among healthcare support staff in the Greater Accra region, Ghana. Methods: An analytic cross-sectional survey was conducted among support staff in 10 major hospitals between 30 January and 31 May 2023. A multi-stage sampling procedure was the overarching technique employed, and study participants were recruited through simple random and probability proportional-to-size sampling techniques. The data analyses were conducted using STATA 15 software. The preliminary association between exposure to splash of body fluids and predisposing factors was established through Chi-square, Fisher's exact, and Mann-Whitney U tests. Log-binomial regression analyses were employed to validate the factors related to splash of body fluids at a significance level of p-value < 0.05. Results: The investigation was conducted among 149 healthcare support staff. The exposure to splash of body fluids over the past 1 year was 53.7% (95% CI: 45.3%-61.9%). The types of body fluids that were mostly encountered through these splash exposures were amniotic fluids (36.3%) and urine (23.8%). Several factors were found to be significantly associated with splash of body fluids, namely: employed as a healthcare assistant [APR = 1.61 (1.16, 2.22)], holding a supervisory position [APR = 0.24 (0.11, 0.51)], having a system in place for reporting body fluid splashes [APR = 0.61 (0.44, 0.85)], male healthcare support staff [APR = 0.62 (0.41, 0.93)], and adherence to standard precautions most of the time [APR = 1.66 (1.11, 2.48)]. Conclusion: Healthcare support staff were highly exposed to splash of body fluids. Gender, supervisory role, category of worker, reporting systems, and adherence to standard precautions were associated with exposure to splash of body fluids. Facility managers are advised to enhance the efficiency of reporting systems.
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Background: The surface properties of dental and orthopedic implants are directly related to their osseointegration rate. Coating and/or modifying the implant surface might reduce the time of healing. In this study, we aimed to examine the effects of a hybrid surface consisting of a brushite surface coating and cross-linked water-soluble eggshell membrane protein on the osseointegration of titanium (Ti) screws under in vivo conditions. Methods: Twenty Ti alloy screws were implanted monocortically in anteromedial regions of New Zealand rabbit tibiae. Ten screws were untreated and used as controls. The remaining 10 screws were coated with calcium phosphate and following cross-linked with ostrich eggshell membrane protein. All rabbits were sacrificed six weeks after the surgery. Peri-screw tissues were evaluated by micro-computed tomography (µ-CT), histological and histomorphometrical methods. Results: The µ-CT assessments indicated that the experimental group had significantly higher mean bone surface area (BSA) and trabeculae number (TbN) than those of the control group (p Ë 0.05). Bone surface area (BV), trabecular separation (TbSp), trabecular thickness (TbTh), and bone mineral density (BMD) scores of the control and experimental groups were quite similar (p > 0.05). The vascularization score of the experimental group was significantly higher than the control group (4.29 vs. 0.92%). No sign of the graft-versus-host reaction was observed. Conclusion: Our findings reveal that coating Ti alloy implants with calcium phosphate cross-linked with ostrich eggshell membrane protein increases the osseointegration of Ti alloy screws by increasing the bone surface area, number of trabeculae and vascularization in the implant site.
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Osseointegração , Titânio , Coelhos , Animais , Titânio/farmacologia , Água , Ligas/farmacologia , Microtomografia por Raio-X , Casca de Ovo , Materiais Revestidos Biocompatíveis/farmacologia , Fosfatos de Cálcio/farmacologia , Proteínas de Membrana , Propriedades de SuperfícieRESUMO
Blood is critical for health, supporting key functions like immunity and oxygen transport. While studies have found links between common blood clinical indicators and COVID-19, they cannot provide causal inference due to residual confounding and reverse causality. To identify indicators affecting COVID-19, we analyzed clinical data (n = 2,293, aged 18-65 years) from Guangzhou Medical University's first affiliated hospital (2022-present), identifying 34 significant indicators differentiating COVID-19 patients from healthy controls. Utilizing bidirectional Mendelian randomization analyses, integrating data from over 2.46 million participants from various large-scale studies, we established causal links for six blood indicators with COVID-19 risk, five of which is consistent with our observational findings. Specifically, elevated Troponin I and Platelet Distribution Width levels are linked with increased COVID-19 susceptibility, whereas higher Hematocrit, Hemoglobin, and Neutrophil counts confer a protective effect. Reverse MR analysis confirmed four blood biomarkers influenced by COVID-19, aligning with our observational data for three of them. Notably, COVID-19 exhibited a positive causal relationship with Troponin I (Tnl) and Serum Amyloid Protein A, while a negative association was observed with Plateletcrit. These findings may help identify high-risk individuals and provide further direction on the management of COVID-19.
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COVID-19 , Análise da Randomização Mendeliana , Humanos , Troponina I , Estudo de Associação Genômica AmplaRESUMO
Ostarine, also known as MK-2866 or enobosarm, is a selective androgen receptor modulator (SARM). It has anabolic properties and as such is widely used in doping, accounting in 2021 for 25 % of the adverse analytical findings (AAF) among the class S1.2 "Other anabolic agents" of products banned by the World Anti-Doping Agency, to which it belongs. But in some cases, it can be responsible for an AAF following contamination. We report the case of an athlete who contaminated herself by exchanging body fluids while kissing her boyfriend, who took 25 mg per day of MK-2866 for 9 days prior to the athlete's AAF (urinary concentration evaluated at 13 ng/mL) without her knowledge. Both subjects came to our lab for hair testing. The athlete's hair was black and slightly frizzy. Six segments of 2 cm then 7 × 3 cm (33 cm) were analysed and showed increasing concentrations, from 2 pg/mg on the first segment to 17.8 pg/mg on the last segment. The boyfriend's hair, light-brown, analyzed on 4 × 2 cm, also showed increasing values, from 65 to 143 pg/mg. These gradients of concentration in the hair's athlete and in her boyfriend were compatible with external contamination of the hair, confirmed by analysis of washing baths, pillowcases (150 pg on each), and the athlete's hairbrush (250 pg). Fingernails were also contaminated, with 21 pg/mg in the athlete and 1041 pg/mg in the boyfriend, with highly contaminated washing baths, and toenails were less contaminated, with 2 pg/mg in the athlete and 17.3 pg/mg in the boyfriend. Urine samples taken 35 days after the start of MK-2866 treatment showed a value of 3690 ng/mL in the boyfriend and 5.7 ng/mL in the athlete. After 6 days off, these concentrations were 3.3 ng/mL and 0.1 ng/mL, respectively. A controlled transfer study was carried out 12 days after discontinuation (urine concentrations returned to negative level). After administration of 17 mg (the 25 mg/mL vial having been controlled at 17 mg/mL), urine samples were taken from the boyfriend and the athlete (n = 10 for each) for more than 25 h after they had been living normally with each other (regular kissing in particular). The boyfriend's urine concentrations ranged from 681 ng/mL to 12822 ng/mL (Tmax = 8:30 hrs), and the athlete's from 0.3 ng/mL to 13 ng/mL with Tmax = 8:30 hrs, i.e. at 22:30 hrs, which corresponded exactly to the time of collection of the urine that showed AAF, with a similar concentration. The dose ingested by the athlete was estimated at 15 µg. These results demonstrate the transfer of ostarine via body fluids between two subjects, with a high risk of AAF in one athlete, as observed in our case.
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Anabolizantes , Líquidos Corporais , Dopagem Esportivo , Feminino , Humanos , Anabolizantes/urina , Anilidas , Líquidos Corporais/química , Detecção do Abuso de Substâncias/métodos , MasculinoRESUMO
The circadian clock plays important roles in several physiological processes. With aging, some of these circadian rhythms have been shown to be disrupted and suggested contributing to age-related diseases. The aim of this scoping review was to examine and map the existing evidence of circadian differences between young and older people in body fluid composition. Literature search was carried out on PubMed, Embase, Scopus and OpenGrey. The studies were screened based on inclusion and exclusion criteria by two independent reviewers and the results were summarized tabularly and narratively. The review process resulted in the identification of 1889 publications, of which 42 were eligible for inclusion. Forty-eight parameters or families of parameters were identified, including cortisol and melatonin, sex hormones, thyroid-related hormones, steroids and aldosterone. However, many were reported by only a single study. The results from the studies were heterogeneous. Even though the majority suggested the flattening of several circadian oscillations in the elderly population, this was not always observed for all the parameters analyzed, and some contradictory results were found. This review revealed a substantial number of publications that explored this research question, but further studies would be important to elucidate the clinical significance of these alterations.
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Ritmo Circadiano , Melatonina , Humanos , Idoso , Ritmo Circadiano/fisiologia , Envelhecimento/fisiologia , HidrocortisonaRESUMO
Introduction: Infectious diseases are major causes of morbidity and mortality worldwide, necessitating the rapid identification and accurate diagnosis of pathogens. While unbiased metagenomic next-generation sequencing (mNGS) has been extensively utilized in clinical pathogen identification and scientific microbiome detection, there is limited research about the application of nanopore platform-based mNGS in the diagnostic performance of various infectious fluid samples. Methods: In this study, we collected 297 suspected infectious fluids from 10 clinical centers and detected them with conventional microbiology culture and nanopore platform-based mNGS. The objective was to assess detective and diagnostic performance of nanopore-sequencing technology (NST) in real-world scenarios. Results: Combined with gold-standard culture and clinical adjudication, nanopore sequencing demonstrated nearly 100% positive predictive agreements in microbial-colonized sites, such as the respiratory and urinary tracts. For samples collected from initially sterile body sites, the detected microorganisms were highly suspected pathogens, and the negative predictive agreements were relatively higher than those in the microbial-colonized sites, particularly with 100% in abscess and 95.7% in cerebrospinal fluid. Furthermore, consistent performance was also observed in the identification of antimicrobial resistance genes and drug susceptibility testing of pathogenic strains of Escherichia coli, Staphylococcus aureus, and Acinetobacter baumannii. Discussion: Rapid NST is a promising clinical tool to supplement gold-standard culture, and it has the potential improve patient prognosis and facilitate clinical treatment of infectious diseases.
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Doenças Transmissíveis , Mycobacterium tuberculosis , Sequenciamento por Nanoporos , Infecções Estafilocócicas , Humanos , Testes de Sensibilidade Microbiana , Escherichia coli/genética , Sequenciamento de Nucleotídeos em Larga Escala , Metagenômica , Sensibilidade e EspecificidadeRESUMO
Circular DNA segments isolated from chromosomes are known as extrachromosomal circular DNA (eccDNA). Its distinct structure and characteristics, along with the variations observed in different disease states, makes it a promising biomarker. Recent studies have revealed the presence of eccDNAs in body fluids, indicating their involvement in various biological functions. This finding opens up avenues for utilizing eccDNAs as convenient and real-time biomarkers for disease diagnosis, treatment monitoring, and prognosis assessment through noninvasive analysis of body fluids. In this comprehensive review, we focused on elucidating the size profiles, potential mechanisms of formation and clearance, detection methods, and potential clinical applications of eccDNAs. We aimed to provide a valuable reference resource for future research in this field.
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Líquidos Corporais , DNA Circular , Humanos , DNA Circular/genética , Cromossomos , BiomarcadoresRESUMO
BACKGROUND: Body fluid traceability inferences can provide important clues to the investigation of forensic cases. Microbiome has been proven to be well applied in forensic body fluid traceability studies. Most of the specimens at crime scenes are often exposed to the external environment when collected, so it is extremely important to exploring the structure characteristics of microbial communities of body fluid samples under different exposure durations for tracing the origin of body fluids based on microorganisms. METHODS: Full-length 16S rRNA sequencing technology and multiple data analysis methods were used to explore the microbial changes in three types of body fluid samples at five different exposure time points. RESULTS: With increasing exposure time, the Proteobacteria abundance gradually increased in the negative control and body fluid samples, and the Bacteroidetes and Firmicutes abundance decreased gradually, but the relative abundance of dominant genera in each body fluid remained dynamically stable. The microbial community structures of those samples from the same individual at different exposure durations were similar, and there were no significant differences in the microbial community structures among the different exposure time points. LEfSe and random forest analyses were applied to screen stable and differential microbial markers among body fluids, such as Streptococcus thermophilus, Streptococcus pneumoniae and Haemophilus parainfluenzae in saliva; Lactobacillus iners and Streptococcus agalactiae in vaginal fluid. CONCLUSIONS: There were no significant differences in microbial community structures of the three types of body fluid samples exposed to the environment for various time periods, although the relative abundance of some microbes in these samples would change. The exposed samples could still be traced back to their source of the body fluid samples using the microbial community structures.
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Líquidos Corporais , Microbiota , Feminino , Humanos , RNA Ribossômico 16S/genética , Saliva/microbiologia , Microbiota/genéticaRESUMO
Lateral flow immunochromatographic (LFI) tests are widely used in both biomedical and forensic sciences for different applications. In forensic sciences, their main use is to detect body fluids at crime scenes. However, there are situations in which the amount of potential biological evidence is so low that DNA extraction is favored with respect to the identification of body fluids. Here, an efficient and quick protocol is presented to integrate the detection of body fluids through LFI with DNA extraction from a sample swab and buffer, providing a complete characterization of the biological evidence. This protocol is a modification of a general DNA extraction silica-based kit, whose main application is for blood and tissues. Thus, it could be carried out in different settings (forensic labs, hospitals, other testing labs) without the necessity of buying a specific kit for swabs. The validation of this protocol is supported by the results presented here and previous publications from our group, obtaining DNA in good quantity and with good quality. This proves the potential application of the protocol in both forensic scenarios, to fully characterize biological evidence, and biomedical settings, to molecularly confirm the results of LFI tests.
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BACKGROUND AND AIM: This case involves a 20-year-old man with prior hallucinogen-use experience, who sniffed an unknown amount of dipropyltryptamine in an apartment. Dipropyltryptamine, a hallucinogenic compound belonging to the tryptamine class is recognized for inducing effects similar to dimethyltryptamine (DMT) but with a longer duration. Ten to fifteen minutes later he experienced visual hallucinations, followed by increasing apathy. Two hours post consumption he developed abdominal pain, leading to collapse, seizure, and vomiting. Despite emergency medical resuscitation on site, transport to hospital 2.5 hours post consumption and extracorporeal life support he died 21 hours later. Relevant toxicological and morphological findings are presented. METHODS: A serum sample was collected four hours post consumption. Autopsy was performed six days after death. Antemortem serum, as well as postmortem cardiac blood and urine were analyzed for alcohol and psychoactive drugs by systematic toxicological analyses employing gas chromatography-mass spectrometry (Maurer/Pfleger/Weber library among others), liquid chromatography-ion trap mass spectrometry (LC-MSn, Toxtyper™), and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Dipropyltryptamine was quantified by LC-MS/MS after solid-phase extraction. RESULTS: Autopsy revealed a state after deep aspiration of gastric contents with consecutive brain edema due to oxygen deprivation. Dipropyltryptamine concentrations were approximately 210 ng/ml, 110 ng/ml and 180 ng/ml in antemortem serum, postmortem cardiac blood and urine, respectively. To the best of our knowledge, these are the first reported concentrations of dipropyltryptamine in a fatal case. CONCLUSION: Unlike typical tryptamine overdose reports, this case did not present with agitation, hyperthermia, or tachycardia. Despite the individual's prior experience with tryptamines and the generally low toxicity associated with this class of hallucinogens, death in this case was an indirect consequence of the nasal consumption of a high dose of dipropyltryptamine.
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Espectrometria de Massas em Tandem , Triptaminas , Masculino , Humanos , Adulto Jovem , Adulto , Cromatografia Líquida , Triptaminas/efeitos adversosRESUMO
OBJECTIVE: Whether alpha-synuclein in peripheral body fluids can be used for the diagnosis of Parkinson's disease (PD) remains in controversy. This study evaluates diagnostic potential of alpha-synuclein for PD in various peripheral body fluids using a meta-analysis approach. METHODS: Studies published before October 2022 were searched in Web of Science and PubMed databases. The results were computed using the STATA 12.0 statistical software. RESULTS: In plasma, PD patients exhibited elevated alpha-synuclein levels relative to healthy controls (HCs) [standard mean difference (SMD) = 0.78, 95% confidence interval (CI) = 0.42 to 1.15] with a sensitivity of 0.79 (95% CI: 0.64-0.89) and a specificity of 0.95 (95% CI: 0.90-0.98). Higher plasma alpha-synuclein levels were correlated with longer disease durations, higher Unified Parkinson's Disease Rating Scale motor scores, and higher Hoehn and Yahr stages in PD patients. Plasma neural-derived exosomal alpha-synuclein levels (SMD = 1.82, 95% CI = 0.30 to 3.35), ratio of plasma neural-derived exosomal alpha-synuclein to total alpha-synuclein (SMD = 1.26, 95% CI = 0.19 to 2.33), and erythrocytic alpha-synuclein levels were also increased in PD patients (SMD = 6.57, 95% CI = 3.55 to 9.58). In serum, there was no significant difference in alpha-synuclein levels between PD patients and HCs (SMD = 0.54, 95% CI = - 0.27 to 1.34). In saliva, reduced alpha-synuclein levels were observed in PD patients (SMD = - 0.85, 95% CI = - 1.67 to - 0.04). CONCLUSIONS: Alpha-synuclein levels in plasma, plasma neural-derived exosome, erythrocyte, and saliva may serve as potential biomarkers for the diagnosis of PD.