ABSTRACT
Amyotrophic Lateral Sclerosis (ALS) is a poorly understood and fatal disease. It has a low prevalence and a 2-4 year survival period. Various theories and hypotheses relating to its development process have been proposed, albeit with no breakthrough in its treatment. Recently, the role of the adaptive immune system in ALS, particularly CD4+ T cells, has begun to be investigated. CD4+ T cells are a heterogeneous group of immune cells. They include highly pro-inflammatory types such as Th1 and Th17, as well as highly anti-inflammatory cells such as Tregs. However, the landscape of the role of CD4+ T cells in ALS is still not clearly understood. This review covers current hypotheses that elucidate how various CD4+ T cells can contribute to ALS development. These hypotheses include the SWITCH model, which suggests that, in the early stages of the disease, Tregs are highly capable of regulating the immune response. However, in the later stages of the disease, it seems that pro-inflammatory cells such as Th1 and Th17 are capable of overwhelming Treg function. The reason why this occurs is not known. Several research groups have proposed that CD4+ T cells as a whole might experience aging. Others have proposed that gamma delta T cells might directly target Tregs. Additionally, other research groups have argued that less well-known CD4+ T cells, such as Emoes+ CD4+ T cells, may be directly responsible for neuron death by producing granzyme B. We propose that the ALS landscape is highly complicated and that there is more than one feasible hypothesis. However, it is critical to take into consideration the differences in the ability of different populations of CD4+ T cells to infiltrate the blood-brain barrier, taking into account the brain region and the time of infiltration. Shedding more light on these still obscure factors can help to create a personalized therapy capable of regaining the balance of power in the battle between the anti-inflammatory and pro-inflammatory cells in the central nervous system of ALS patients.
ABSTRACT
Diabetes mellitus (DM) is a prevalent metabolic disease. The clinical impact of sustained hyperglycemia on ligament healing has not been well characterized. Diabetes is a known cause of macro-, microvascular, and diabetic ulcer healing difficulties among tissues. Therefore, we aimed to investigate the healing potential occurring in injured and healthy ligaments among diabetic and healthy individuals using a rat model. We hypothesize that DM may contribute to altering the knee medial collateral ligament (MCL), thus its morphology, biochemical fitness, and functionality. The study cohort consisted of 40 rats. The animals were randomized into four equal groups. Groups I and II (20 rats) received saline subcutaneously and served as controls. Groups III and IV (20 rats) were injected with a single dose of streptozotocin (STZ). All animals underwent surgery to cut the left tibial collateral ligament in the hind limb and suture it. The access site was sutured to create inflammation and study the regenerative capacities of animals with normal carbohydrate metabolism and pharmacologically induced diabetes. Each animal then underwent sham surgery to access and suture the right tibial collateral ligament in the hind limb without ligament intervention. After the animals had undergone surgeries, groups II and IV were given melatonin supplementation for 4 weeks. Rats with DM presented with more fibrosis and calcification of the MCL and decreased healing potential. Treatment with melatonin in diabetic rats mitigated alterations and improved the antioxidant status of ligaments from the diabetic group.
ABSTRACT
The chiroptical activity of various semiconductor inorganic nanocrystalline materials has typically been tested using circular dichroism or circularly polarized luminescence. Herein, we report on a high-throughput screening method for identifying and differentiating chiroptically active quantum-sized ZnO crystals using Raman spectroscopy combined with principal component analysis. ZnO quantum dots (QDs) coated by structurally diverse homo- and heterochiral aminoalcoholate ligands (cis- and trans-1-amino-2-indanolate, 2-amino-1-phenylethanolate, and diphenyl-2-pyrrolidinemethanolate) were prepared using the one-pot self-supporting organometallic procedure and then extensively studied toward the identification of specific Raman fingerprints and spectral variations. The direct comparison between the spectra demonstrates that it is very difficult to make definite recognition and identification between QDs coated with enantiomers based only on the differences in the respective Raman bands' position shifts and their intensities. However, the applied approach involving the principal component analysis performed on the Raman spectra allows the simultaneous differentiation and identification of the studied QDs. The first and second principal components explain 98, 97, 97, and 87% of the variability among the studied families of QDs and demonstrate the possibility of using the presented method as a qualitative assay. Thus, the reported multivariate approach paves the way for simultaneous differentiation and identification of chirotopically active semiconductor nanocrystals.
ABSTRACT
OBJECTIVES: Monitoring tobacco use is one of the most important tobacco control interventions recommended by the World Health Organization. This study aimed to assess the prevalence of cigarette smoking, e-cigarette use, and heated tobacco use in a nationwide sample of adults in Poland, as well as to identify factors associated with smoking, e-cigarette use, and heated tobacco use. MATERIAL AND METHODS: This nationwide crosssectional survey was carried out in February 2-4, 2024 using the computer-assisted web interviewing method. The study questionnaire included 17 closed questions on smoking, e-cigarette use, and heated tobacco use. A sample of 1080 adults was selected using a non-probability quota sampling method. RESULTS: Data were collected from 1080 adults, age M±SD 48.4±15.5 years, 53% females. Past 30-day smoking was declared by 30.4% of adults in Poland, including 24.5% of daily smokers. Ever e-cigarette use was declared by 33% of respondents, and 20.6% had ever tried heated tobacco. Past 30-day e-cigarette use was declared by 15.2% of adults, wherein 5.9% were daily e-cigarette users. Past 30-day heated tobacco use was declared by 10.9% of respondents, wherein 4.9% were daily heated tobacco users. One-tenth of adults were dual users (cigarette and e-cigarette or heated tobacco), and 6.5% were past 30-day triple users. Age 40-59 years, having less than higher education, and bad economic status were significantly associated with daily smoking (p < 0.05). Age 18-39 years, having less than higher education, and active occupational status (employment/ self-employment) were associated with daily e-cigarette use (p < 0.05). Age 18-39 years and active occupational status were associated with daily heated tobacco use (p < 0.05). CONCLUSIONS: This study provides comprehensive epidemiological data on the current prevalence of smoking, e-cigarette use, and heated tobacco use. The introduction of e-cigarettes and heated tobacco products evoked changes in social behaviors related to nicotine use. Int J Occup Med Environ Health. 2024;37(3).
ABSTRACT
In the human body, tumor cell occurrence can be indirectly monitored using the L-selectin concentration in the blood, since selectin ligands are present on the surface of tumor cells, and with tumor progression, a decrease in L-selectin levels can be expected and observed. In this study, we present a selective DNA-based surface-enhanced Raman spectroscopy (SERS) assay for the detection and determination of L-selectin in biological samples. Two calibration curves (linear in the 40-190 ng mL-1 region and exponential in the 40-500 ng mL-1 region) are fitted to the obtained SERS experimental data, i.e., the ratio of I732/I1334 band intensities (LOQ = 46 ng mL-1). Calculated determination coefficients are found to be R2 = 0.997 for the linear region of the calibration curve and R2 = 0.977 for the exponential region. Moreover, we demonstrate very good selectivity of the assay even in the presence of P- and E-selectin in a sample containing L-selectin. With our SERS assay, the L-selectin concentration in biological samples can be estimated directly from the calibration curves.
Subject(s)
Aptamers, Nucleotide , Metal Nanoparticles , Humans , Aptamers, Nucleotide/chemistry , Spectrum Analysis, Raman/methods , L-Selectin , Plasma , Metal Nanoparticles/chemistry , Gold/chemistryABSTRACT
The rapid, low cost, and efficient detection of SARS-CoV-2 virus infection, especially in clinical samples, remains a major challenge. A promising solution to this problem is the combination of a spectroscopic technique: surface-enhanced Raman spectroscopy (SERS) with advanced chemometrics based on machine learning (ML) algorithms. In the present study, we conducted SERS investigations of saliva and nasopharyngeal swabs taken from a cohort of patients (saliva: 175; nasopharyngeal swabs: 114). Obtained SERS spectra were analyzed using a range of classifiers in which random forest (RF) achieved the best results, e.g., for saliva, the precision and recall equals 94.0% and 88.9%, respectively. The results demonstrate that even with a relatively small number of clinical samples, the combination of SERS and shallow machine learning can be used to identify SARS-CoV-2 virus in clinical practice.
ABSTRACT
Tree mortality is an objective forest health criterion and is particularly suitable for long-term and large-scale studies of forest condition. However, it is impossible to determine actual tree mortality in Central European managed forests where trees are removed for various reasons. In this case, the only way to approximate tree mortality is to define the range in which it occurs. This can be carried out by including in the mortality calculations either dead trees that remain in the stand at the end of the assessment period or additionally trees that have been removed from the stand. We used data from the annual forest monitoring surveys in Poland from 2009 to 2022 for pine, spruce, oak and birch to perform a survival analysis in which we included all removals or sanitary cuttings either as censored or complete observations. The differences between the calculated mortality rates were significant, indicating the importance of how removals are treated in the analysis. To assess which method used for mortality calculation was more appropriate, we compared values for last recorded defoliation and severity of damage from live, dead and thinned or salvaged trees. For all species studied, significant differences were found between dead trees or trees removed by sanitation cuts and living trees or trees removed by thinning, suggesting that not only dead trees remaining in the forest, but also trees removed by sanitation cuts, should be considered when calculating mortality in managed stands. We also recommend the use of survival analysis in forest monitoring as a routine method for assessing the health of stands.
ABSTRACT
Urtica dioica is a species with well-established significance in folk medicine in many countries. It was utilized to support the treatment of arthritis, allergies, and urinary tract disorders; however, the substantial presence of antioxidants suggests that nettle extract could also have a positive impact on the skin. The objective of this study was to assess the impact of nettle extract on human skin fibroblasts subjected to oxidative stress. Various solvents were tested to prepare an extract rich in polyphenolic compounds with high antioxidant potential. The chemical composition was determined using ultra-high-performance liquid chromatography with mass spectrometry (UPLC-DAD-MS). H2O2 treatment was used to induce oxidative stress and cell viability, and the metabolism was evaluated through NR and MTT assays. Our study demonstrated that extraction with 80% ethanol, followed by the drying and re-dissolving of the extract in pure water, was more efficient than direct extraction with water. This yielded an extract rich in polyphenolic compounds, with chlorogenic acid and caffeoylmalic acid as the predominant compounds, averaging 64.9 and 114.4 µg/mL, respectively. The extract exhibited antioxidant properties in the DPPH and ABTS assays. Furthermore, it did not exhibit cytotoxicity and did not negatively affect cell metabolism. In addition, it effectively reduced ROS in the H2O2-stimulated cells, and at the highest concentration tested, the ROS levels returned to those of the untreated control. The extract also protected against H2O2-induced cytotoxicity. The cell viability was maintained at the level of the untreated control when the cells were pretreated with the extract before H2O2 exposure. These findings indicate that U. dioica extract is a valuable and safe additive in skincare products.
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Usnic acid (UA) is a compound with multiple biological activities that make it useful in various industries, e.g., pharmaceutical, cosmetic, dentistry, and agricultural sectors. Lichens are the primary source of UA, which is primarily extracted using acetone. This study aimed to investigate the solubility of UA in numerous natural deep eutectic solvents (NADESs) and use a mixture of thymol and camphor as a NADES in the optimization of the UA extraction process with the design of experiments method. For numerical optimization, the following parameters were employed in the experiment to confirm the model: a camphor-to-thymol ratio of 0.3, a liquid-to-solid ratio of 60, and a time of 30 min. The obtained experimental results aligned well with the predicted values, with the mean experimental value falling within the confidence interval, exhibiting deviations between 11.93 and 14.96. By employing this model, we were able to optimize the extraction procedure, facilitating the isolation of approximately 91% of the total UA content through a single extraction, whereas a single acetone extraction yielded only 78.4% of UA.
Subject(s)
Deep Eutectic Solvents , Thymol , Solvents , Acetone , Camphor , Plant ExtractsABSTRACT
INTRODUCTION AND OBJECTIVE: The eye examination is a simple and effective intervention that involves a series of tests to evaluate vision and check for eye disease. This study aimed to assess the frequency of eye examinations among adults in Poland and to identify factors associated with the frequency of such examinations. MATERIAL AND METHODS: The questionnaire-based cross-sectional survey was carried out in December 2022 on a non-probability quota sample of 1,076 adults in Poland. A computer-assisted web interview technique was used. The study questionnaire included a set of questions on eye health, eye examinations, and socio-demographic characteristics. RESULTS: Of the 1,076 respondents, 7.4% had an eye examination in the last 30 days, almost one-quarter (24.2%) had an eye examination more than 1 month but not more than 12 months ago, 13.9% had an eye examination in the last 1-2 years, and 24.1% had an eye examination more than 2 years ago but not more than 3 years ago. Among the respondents, 7.1% declared that they had never had an eye examination. Of the 12 different factors analyzed in this study, wearing spectacles or lenses and self-reported level of knowledge on eye diseases were the only factors significantly associated with higher odds of eye examinations in the last 12 months or 2 years. There was no significant impact of gender, educational level, place of residence, health status, or economic status (p>0.05) on the frequency of eye examinations, neither in the last 12 months nor in the last 2-3 years. CONCLUSIONS: The study showed that a significant percentage of adults in Poland do not have regular eye examinations. There was a lack of socio-economic differences (including place of residence and economic status) in the frequency of eye examinations. There is an urgent need for health education on preventive eye examinations and eye care among adults in Poland.
Subject(s)
Health Education , Humans , Adult , Poland/epidemiology , Cross-Sectional Studies , Socioeconomic Factors , Educational StatusABSTRACT
In this study, the intrinsic surface-enhanced Raman spectroscopy (SERS)-based approach coupled with chemometric analysis was adopted to establish the biochemical fingerprint of SARS-CoV-2 infected human fluids: saliva and nasopharyngeal swabs. The numerical methods, partial least squares discriminant analysis (PLS-DA) and support vector machine classification (SVMC), facilitated the spectroscopic identification of the viral-specific molecules, molecular changes, and distinct physiological signatures of pathetically altered fluids. Next, we developed the reliable classification model for fast identification and differentiation of negative CoV(-) and positive CoV(+) groups. The PLS-DA calibration model was described by a great statistical value-RMSEC and RMSECV below 0.3 and R2cal at the level of ~0.7 for both type of body fluids. The calculated diagnostic parameters for SVMC and PLS-DA at the stage of preparation of calibration model and classification of external samples simulating real diagnostic conditions evinced high accuracy, sensitivity, and specificity for saliva specimens. Here, we outlined the significant role of neopterin as the biomarker in the prediction of COVID-19 infection from nasopharyngeal swab. We also observed the increased content of nucleic acids of DNA/RNA and proteins such as ferritin as well as specific immunoglobulins. The developed SERS for SARS-CoV-2 approach allows: (i) fast, simple and non-invasive collection of analyzed specimens; (ii) fast response with the time of analysis below 15 min, and (iii) sensitive and reliable SERS-based screening of COVID-19 disease.
Subject(s)
COVID-19 , Humans , COVID-19/diagnosis , SARS-CoV-2/genetics , Saliva/chemistry , Nasopharynx , RNA, Viral/genetics , Spectrum Analysis, Raman , Specimen Handling/methods , COVID-19 TestingABSTRACT
BACKGROUND Vision health affects functioning in society, and the ability to learn and work. Ophthalmic symptoms may be caused by eye diseases, but also by environmental or lifestyle factors. This online questionnaire-based survey aimed to identify the prevalence of ophthalmic symptoms in 1076 individuals in Poland, as well as to identify factors associated with the presence of ophthalmic symptoms. MATERIAL AND METHODS An online questionnaire-based survey was carried out in December 2022 on a representative sample of 1076 adult Poles. Non-probability quota sampling was used. Respondents were asked about the presence of 16 different eye symptoms and vision problems in the last 30 days. The presence of ophthalmic symptoms was self-declared. Data were analyzed with IBM SPSS package version 28. RESULTS More than half of the respondents (57.8%) had at least 1 ophthalmic symptom in the last 30 days. Burning and stinging eyes (21.6%) and dry eyes (18.9%) were the most common ophthalmic symptoms declared by the respondents. Moreover, 21.3% of respondents reported vision deterioration in the last 30 days. Out of 10 different factors analyzed in this study, female gender, living in rural areas or small cities (below 100 000 inhabitants), living with at least 1 other person, having low economic status, having chronic diseases, and wearing spectacles/contact lenses were significantly associated (P.
Subject(s)
Contact Lenses , Dry Eye Syndromes , Adult , Humans , Female , Poland/epidemiology , Prevalence , Surveys and Questionnaires , Dry Eye Syndromes/epidemiologyABSTRACT
BACKGROUND: The aim of this study was to investigate the role of urine-derived extracellular vesicles (uEVs) in diabetic kidney disease (DKD) in patients diagnosed with type 2 diabetes mellitus (T2DM). METHODS: UEVs were characterized by size distribution and microRNA content by next-generation small RNA sequencing and quantitative reverse transcription PCR. RESULTS: A subset of sixteen miRNAs enriched in T2DM patients with DKD, including hsa-miR-514a-5p, hsa-miR451a, hsa-miR-126-3p, hsa-miR-214, or hsa-miR503 was identified. Eight miRNAs as hsa-miR-21-3p, hsa-miR-4792, hsa-miR375, hsa-miR-1268a, hsa-miR-501-5p, or hsa-miR-582 were downregulated. Prediction of potential target genes and pathway enrichment analysis of the Kyoto Encyclopedia of Genes and Genomes (KEGG) confirmed possible functions related to cellular processes such as apoptosis, inflammation, and tissue remodeling, that promote diabetic complications, such as DKD. Among them, hsa-miR-375, hsa-miR-503, and hsa-miR-451a make important contribution. Additionally, downregulated hsa-miR-582-5p has not been reported so far in any diabetes-related pathways. CONCLUSIONS: This study revealed the most significant miRNAs in uEVs of patients with T2DM. However, as this is a bioinformatic prediction that we performed based on the putative targets of the identified miRNAs. Thus, further in vitro functional studies are needed to confirm our findings. Knowing the fact that EVs are crucial in transferring miRNAs, there is a great need toto discover their involvement in the pathomechanism of T2DM-related kidney disease.
Subject(s)
Diabetes Mellitus, Type 2 , Diabetic Nephropathies , Extracellular Vesicles , MicroRNAs , Renal Insufficiency, Chronic , Humans , MicroRNAs/genetics , Extracellular Vesicles/metabolismABSTRACT
BACKGROUND The Nationwide General Hospital Morbidity Study Registry collects demographic, health, and medical data on patients hospitalized throughout Poland, and acts as a registry for epidemiological, public health, and hospital administrative studies. This epidemiological national registry-based study aimed to evaluate the characteristics of 395 646 patients hospitalized due to eye diseases in Poland in 2019. MATERIAL AND METHODS This study is a retrospective nationwide registry-based analysis. Data on all patients hospitalized due to eye diseases in 2019 were obtained from the Nationwide General Hospital Morbidity Study Registry. Demographic characteristics of patients hospitalized due to eye diseases as well as the hospitalization course were analyzed using descriptive epidemiology tools. RESULTS Disorders of the lens accounted for 68.6% of all hospital admissions, and disorders of the choroid and retina were the second most common (13.9%) cause of hospital admission due to eye diseases in 2019. Over three-quarters of hospital admissions (77.4%) lasted less than 24 h, and 86.8% were scheduled admissions. Most of the patients hospitalized due to eye diseases were females (59.9%) and over 70% lived in urban areas. There were regional differences in the hospital admissions rate per 100 000 inhabitants. According to the data presented in the registry, less than one-third of patients had comorbidities. CONCLUSIONS Hospital ophthalmological care in Poland is based mainly on planned hospitalizations lasting less than 24 h and related to ophthalmological surgery. Data on the comorbidity of ophthalmologic patients suggest that ophthalmologists do not pay enough attention to identifying and recording the patient's health status.
Subject(s)
Eye Diseases , Hospitalization , Female , Humans , Male , Poland/epidemiology , Retrospective Studies , Eye Diseases/epidemiology , RegistriesABSTRACT
Implementation of eye care behaviors may reduce the risk of eye symptoms and diseases. This study aimed to assess eye care behaviors and identify factors associated with eye care practices among adults in Poland. This cross-sectional survey was carried out between 9 and 12 December 2022 on a nationwide random quota sample of adults in Poland. The study questionnaire included a set of questions on 10 different eye care behaviors. The study population included 1076 participants with a mean age of 45.7 ± 16.2 years, and 54.2% of participants were females. The most common (30.2%) eye care behavior was the use of good lighting indoors, and 27.3% used sunglasses with a UV filter. More than one-fifth of participants declared taking regular screen breaks and limiting screen time. Less than one-tenth of participants used dietary supplements with lutein, beta-carotene, or zinc. Out of 12 factors assessed in this study, self-reported knowledge of eye diseases was the most important factor associated (p < 0.05) with eye care behaviors. There were no economic or educational gaps (p > 0.05) in the implementation of most of the eye care behaviors among adults in Poland. This study revealed a low level of implementation of eye care behaviors among adults in Poland.
Subject(s)
Lutein , beta Carotene , Female , Humans , Adult , Middle Aged , Male , Cross-Sectional Studies , Poland/epidemiology , Dietary SupplementsABSTRACT
Public knowledge and awareness of eye diseases may influence individuals' behaviors toward the use of eye care services and prevention methods. The objective of this study was to assess the awareness of common eye diseases and their risk factors among adults in Poland as well as to identify factors associated with knowledge of eye diseases. This nationwide cross-sectional web-based survey was carried out in December 2022 on a representative sample of 1076 adults in Poland. Most of the respondents had heard of cataracts (83.6%), glaucoma (80.7%), conjunctivitis (74.3%), and hordeolum (73.8%). Awareness of dry eye syndrome was declared by 50% of respondents, and 40% were aware of retinal detachment. Among the respondents, 32.3% had heard of AMD, and 16.4% had heard of diabetic retinopathy. A lack of awareness of glaucoma was declared by 38.1% of respondents, and 54.3% declared a lack of awareness of risk factors for AMD. Gender, age, and the presence of chronic diseases were the most important factors (p < 0.05) associated with awareness of common eye diseases and risk factors for glaucoma and AMD. This study demonstrated a low level of awareness of common eye diseases among adults in Poland. Personalized communication on eye diseases is needed.
Subject(s)
Eye Diseases , Glaucoma , Adult , Humans , Cross-Sectional Studies , Poland , Surveys and Questionnaires , Health Knowledge, Attitudes, Practice , Eye Diseases/complications , Glaucoma/complications , Risk FactorsABSTRACT
BACKGROUND: Manifestations of OAB can considerably diminish the quality of life. Botulinum toxin has emerged as a valuable treatment option in diseases whose symptoms cannot be controlled adequately with other available therapies. The aim of the present study was to compare the subjective quality of life of patients with OAB before the injection of botulinum toxin and three and six months after the intervention. METHODS: This study was based on a diagnostic survey with three validated questionnaires, ICIQ-OAB, ICIQ-OABqol, and ICIQ-LUTSqol, and an additional questionnaire developed by the authors to collect sociodemographic characteristics and selected medical data. RESULTS: This study demonstrated significant differences between pre-treatment scores and those at three and six months post injection. At three and six months after the intervention, mean scores for all three instruments (ICIQ-OAB, ICIQ-OABqol, ICIQ-LUTSqol) were significantly lower than the respective pre-treatment values, implying a significant attenuation of OAB symptoms and their lower impact on the quality of life. However, the severity of OAB symptoms and their impact on the quality of life at six months post intervention were significantly higher than at three months, except for the social interaction domain. CONCLUSIONS: Botulinum toxin is an effective treatment for OAB.
Subject(s)
Botulinum Toxins, Type A , Botulinum Toxins , Urinary Bladder, Overactive , Humans , Female , Urinary Bladder, Overactive/drug therapy , Urinary Bladder, Overactive/diagnosis , Botulinum Toxins/adverse effects , Quality of Life , Treatment Outcome , Surveys and Questionnaires , Botulinum Toxins, Type A/adverse effectsABSTRACT
The accurate identification of microorganisms belonging to vaginal microflora is crucial for establishing which microorganisms are responsible for microbial shifting from beneficial symbiotic to pathogenic bacteria and understanding pathogenesis leading to vaginosis and vaginal infections. In this study, we involved the surface-enhanced Raman spectroscopy (SERS) technique to compile the spectral signatures of the most significant microorganisms being part of the natural vaginal microbiota and some vaginal pathogens. Obtained data will supply our still developing spectral SERS database of microorganisms. The SERS results were assisted by Partial Least Squares Regression (PLSR), which visually discloses some dependencies between spectral images and hence their biochemical compositions of the outer structure. In our work, we focused on the most common and typical of the reproductive system microorganisms (Lactobacillus spp. and Bifidobacterium spp.) and vaginal pathogens: bacteria (e.g., Gardnerella vaginalis, Prevotella bivia, Atopobium vaginae), fungi (e.g., Candida albicans, Candida glabrata), and protozoa (Trichomonas vaginalis). The obtained results proved that each microorganism has its unique spectral fingerprint that differentiates it from the rest. Moreover, the discrimination was obtained at a high level of explained information by subsequent factors, e.g., in the inter-species distinction of Candida spp. the first three factors explain 98% of the variance in block Y with 95% of data within the X matrix, while in differentiation between Lactobacillus spp. and Bifidobacterium spp. (natural flora) and pathogen (e.g., Candida glabrata) the information is explained at the level of 45% of the Y matrix with 94% of original data. PLSR gave us insight into discriminating variables based on which the marker bands representing specific compounds in the outer structure of microorganisms were found: for Lactobacillus spp. 1400 cm-1, for fungi 905 and 1209 cm-1, and for protozoa 805, 890, 1062, 1185, 1300, 1555, and 1610 cm-1. Then, they can be used as significant marker bands in the analysis of clinical subjects, e.g., vaginal swabs.
Subject(s)
Microbiota , Vaginosis, Bacterial , Female , Humans , Least-Squares Analysis , Gardnerella vaginalis , Vagina/microbiology , Lactobacillus , Bacteria , BifidobacteriumABSTRACT
The detection of freely circulating cancer cells (CTCs) is one of the greatest challenges of modern medical diagnostics. For several years, there has been increased attention on the use of surface-enhanced Raman spectroscopy (SERS) for the detection of CTCs. SERS is a non-destructive, accurate and precise technique, and the use of special SERS platforms even enables the amplification of weak signals from biological objects. In the current study, we demonstrate the unique arrangement of the SERS technique combined with the deposition of CTCs cells on the surface of the SERS platform via a dielectrophoretic effect. The appropriate frequencies of an alternating electric field and a selected shape of the electric field can result in the efficient deposition of CTCs on the SERS platform. The geometry of the microfluidic chip, the type of the cancer cells and the positive dielectrophoretic phenomenon resulted in the trapping of CTCs on the surface of the SERS platform. We presented results for two type of breast cancer cells, MCF-7 and MDA-MB-231, deposited from the 0.1 PBS solution. The limit of detection (LOD) is 20 cells/mL, which reflects the clinical potential and usefulness of the developed approach. We also provide a proof-of-concept for these CTCs deposited on the SERS platform from blood plasma.
Subject(s)
Microfluidics , Neoplasms , Limit of Detection , Oligonucleotide Array Sequence Analysis , Spectrum Analysis, Raman/methodsABSTRACT
Nucleic acid extraction and purification are crucial steps in sample preparation for multiple diagnostic procedures. Routine methodologies of DNA isolation require benchtop equipment (e.g., centrifuges) and labor-intensive steps. Magnetic nanoparticles (MNPs) as solid-phase sorbents could simplify this procedure. A wide range of surface coatings employs various molecular interactions between dsDNA and magnetic nano-sorbents. However, a reliable, comparative evaluation of their performance is complex. In this work, selected Fe3O4 modifications, i.e., polyethyleneimine, gold, silica, and graphene derivatives, were comprehensively evaluated for applications in dsDNA extraction. A family of single batch nanoparticles was compared in terms of morphology (STEM), composition (ICP-MS/MS and elemental analysis), surface coating (UV-Vis, TGA, FTIR), and MNP charge (ζ-potential). ICP-MS/MS was also used to unify MNPs concentration allowing a reliable assessment of individual coatings on DNA extraction. Moreover, studies on adsorption medium (monovalent vs. divalent ions) and extraction buffer composition were carried out. As a result, essential relationships between nanoparticle coatings and DNA adsorption efficiencies have been noticed. Fe3O4@PEI MNPs turned out to be the most efficient nano sorbents. The optimized composition of the extraction buffer (medium containing 0.1 mM EDTA) helped avoid problems with Fe3+ stripping, which improved the validity of the spectroscopic determination of DNA recovery.