Medium Samples Research Articles

Development of a High-Throughput qPCR Assay for Detecting Waterborne Protozoa and Helminths Across Different Environmental Media in China.

The establishment of a high-throughput quantification approach for waterborne pathogenic protozoa and helminths is crucial for rapid screening and health risk assessment. We developed a high-throughput quantitative polymerase chain reaction (HT-qPCR) assay targeting 19 waterborne protozoa and 3 waterborne helminths and validated its sensitivity, specificity, and repeatability. The assay was then applied to test various environmental media samples. The HT-qPCR assay's limit of detection (LOD) was 5×102 copies/μL DNA, and its specificity was confirmed using Giardia and Cryptosporidium standards. Repeatability, assessed through intra- and inter-group experiments, yielded a coefficient of variation (CV) of 1.0%-4.6% and 1.2%-6.4% at concentrations of 1×105 and 1×104 copies/μL, respectively. The R 2 values of the 22 standard curves ranged from 0.983 to 0.998, with amplification efficiencies between 80% and 107%. In drinking water sources, sludge from municipal wastewater treatment plants (MWTPs), and livestock manure samples, 17 of 22 targets were detected, with Acanthamoeba genus (50.0%), Acanthamoeba castellanii (11.8%), and Enterocytozoon bieneusi (11.8%) showing high prevalence. Cryptosporidium spp., Enterocytozoon bieneusi, and Cyclospora cayetanensis were simultaneously found in all three sample types. This study presents a useful tool for the rapid detection of waterborne protozoa and helminths in complex environmental microbiomes, providing scientific data for monitoring cross-media transmission and controlling microbial risk from a One Health perspective.

Dolomite dissolution, pH neutralization, and potentially toxic element immobilization in stormwater bioretention beds.

The importance of pH in stormwater bioretention beds cannot be overstated since it impacts plant and microbial populations and removal of potentially toxic elements (PTEs) from stormwater runoff. This study investigated the effects of dolomite amendment on pH neutralization and subsequent PTE immobilization in bioretention media. To assess dolomite dissolution, pH neutralization, and PTE immobilization, engineered bioretention media was amended with different dolomite ratios and samples of dolomite-amended media were collected from two bioretention beds, one and two months after installation. The effects of inflow conditions and operational time on dolomite dissolution and PTE immobilization were investigated through a column study. Laboratory batch experiments revealed that pH neutralization was fast and reached the recommended pH range of 5 to 8 within 5min. Among the 1-D column conditions, intermittent inflow had the highest porewater Mg and Ca concentrations, indicating greater dolomite dissolution. Batch experiments on field-collected media showed that pH neutralization was substantial within 1month, and continued during the second month, due to dolomite dissolution. Dissolved (batch experiments) and column porewater Mg and Ca concentrations supported instantaneous Ca and a relatively slow Mg dissociation during dolomite dissolution. Among the monitored PTEs, dissolved and porewater concentrations (μg/L) were found to decrease with time, in the order Mn>Fe>Zn>Cu>Pb>Cd, with Cr mainly undetected in all experimental conditions. However, as pH became neutralized to slightly basic (pH~8), dissolved and porewater As concentrations increased. The study suggests that pH neutralization and PTE immobilization depend on the soil-to-dolomite ratio and hydrological properties such as inflow rate, dry-wet cycle, and soil-water contact time. Dolomite amendment to engineered media can be an effective measure for intercepting contaminants, improving ecosystem health, and enhancing biochemical contaminant breakdown, though consideration should be given to bioretention bed design features and system hydrology.

Metabolic profiling of endophytic fungi acting as antagonists of the banana pathogen Colletotrichum musae.

Three endophytic strains, Phomopsis sp., Fusarium proliferatum, and Tinctoporellus epimiltinus, isolated from various plants in the rainforest of the Philippines, were investigated regarding their ability to repress growth of the pathogenic fungus Colletotrichum musae on banana fruits causing anthracnose disease. An in vitro plate-to-plate assay and an in vivo sealed box assay were conducted, using commercial versus natural potato dextrose medium (PDA). All tested endophytes were able to significantly reduce C. musae growth compared to the control. However, the type of medium had no significant effect on lesion size of C. musae on banana. An interaction effect between fungal strain and medium could be shown. On the commercial medium, no differences between the biocontrol ability of the fungi and control treatments could be found, while there were significant differences between the fungal strains on natural medium. Lesions on banana incubated with Phomopsis sp. on natural medium were significantly but only slightly larger than those on banana incubated with F. proliferatum. Volatiles released by these two strains and one pathogenic strain of F. graminearum were collected using polydimethylsiloxane tubes and analyzed via gas chromatography mass spectrometry (GC-MS). Twelve volatile metabolites were detected. Benzaldehyde was the most prominent volatile emitted from the commercial and plain medium. 2-Undecanone, 2-nonanone, and phenylethyl-alcohol were detected in individual samples in both media. 1-Decanol and acoradiene were exclusive to the commercial medium, with acoradiene also being unique to F. proliferatum. Five volatileorganic compounds (VOCs)were emitted from all tested fungal species: 2-heptanone, 2-nonanone, 2-undecanone, 2-tridecanone, and phenylethyl-alcohol. Beta-acorenol was detected in F. proliferatum grown on both media. To reveal whether the medium (commercial PDA versus potato extract) affected the metabolism of the fungi, metabolic footprints were assessed via high performance liquid chromatography with quadrupole time of flight mass spectrometry MS (HPLC-QTOF-MS). A total of 388 metabolic signals were recorded. The intensities of 80-90% of these signals differed significantly between the two types of media. Metabolic footprints varied in response to different potato dextrose medium preparations. The two promising fungal strains may be used to reduce postharvest decay and losses in fruits.

Development of a Quinoa‐Based Fermentation Medium for Propagation of Lactobacillus Plantarum and Weissella Confusa in Opaque Beer Production

Product inconsistency of opaque beer has for long been a tenacious problem in the brewing industry since the current process relies on spontaneous lactic acid fermentation. In order to impede this challenge, there is a need to add lactic acid bacteria (LAB) starter cultures in opaque beer brewing to improve its organoleptic qualities. This study sought to develop a quinoa‐based fermentation medium for propagation of Lactobacillus plantarum and Weissella confusa as potential starter cultures in opaque beer production. An evaluation of the stability and tolerance of the LAB under various stress conditions was also done. Fermentation wort from opaque beer brewing and different quinoa‐based synthetic media with varying nutritional components was prepared for propagation of LAB. Physiochemical analyses which included pH, Brix value and total titratable acidity (TTA) of monocultured and cocultured synthetic media were measured. The measurements were done at 24 h time intervals ranging from 0 to 96 h. Tolerance studies which included the effect of heat shock, cold shock, oxidative stress and osmotic pressure on the survival rate of LAB were conducted to determine the stability of LAB. MRS with L. plantarum monoculture (MRSp) had a notable change in pH from 4.5 to 3.6 after 24 h. The cocultured (M5p + w) synthetic media and cocultured MRS (MRSp + w) also exhibited change in pH from 4.3 to 3.2 and 4.3 to 3.3, respectively, after 72 h. Brix value in all media samples decreased after 24 h except for the uninoculated MRS sample (MRS C). The synthetic and coculture medium (M5p + w) exhibited an increase in TTA (0.79% (m/v) lactic acid) within the first 24 h. Exposure to heat shock had a significance effect (p < 0.05) on the survival rate of L. plantarum and W. confusa. The W. confusa in synthetic media recorded a higher survival rate (27 ± 0.03%) upon exposure to heat shock than L. plantarum (7 ± 0.01%). In contrast, L. plantarum in MRS recorded a higher survival rate (67 ± 0.02%) upon exposure to cold shock and oxidative stress (34 ± 0.01%). The starter cultures tested survived upon exposure to the stress conditions, indicating their potential use in opaque beer production.

Sonochemically Modified Lapindo Mud Using Sulfuric Acid for Efficient Adsorption of Phenol in Aqueous Media and Real Wastewater Samples

Pharmaceutical industrial wastewater frequently contains high amounts of phenolic substances, which pose severe threats to the ecosystem and human health. Therefore, efficient removal of these pollutants is urgently needed. In the present work, sulfated Lapindo mud (SLM) was prepared using the sonochemical method and applied as an adsorbent for phenol removal in aqueous media and actual wastewater samples from Code River, Yogyakarta. Modification of Lapindo mud (LM) using sulfuric acid enables it to remove its impurities, resulting in a material containing 78.4% silica (SiO2) and 15.3% alumina (Al2O3). The SLM adsorbent demonstrated sufficient adsorption performance of 49.8% with an optimal initial phenol concentration of 120 mg/L with a contact time of 100 min at pH of 10. The maximum adsorption capacity (qmax) obtained by the Langmuir isotherm model was 27.2 mg/g. The adsorption process follows pseudo-second-order because it has two active sites, Brønsted acid sites (–SiOH and –SO3H) and Lewis acid sites (Si4+). Phenol in base condition undergoes a deprotonation reaction that is stabilized by the acid-active sites of the SLM adsorbent through intermolecular forces. Considering the large adsorption capacity and quick kinetic, the SLM adsorbent can be a promising cheap and green material to remove phenolic substances in wastewater, especially in the river near the medical facility. Copyright © 2024 by Authors, Published by BCREC Publishing Group. This is an open access article under the CC BY-SA License (https://creativecommons.org/licenses/by-sa/4.0).

Electrochemical and microstructural characterization of a precipitation hardened 17-4 steel in different environments

In this study, an in-depth electrochemical characterization of a precipitation hardened 17-4 stainless steel (SS17-4PH) was carried out in combined acidic, alkaline, and chloride environments. The characterizations were carried out through polarization-EIS analysis, Mott-Schottky analysis to characterize the semiconducting properties of the thin oxide films, and a post-mortem scanning electron microscopy (SEM-EDX) analysis to reveal the morphologies after anodic polarization. The electrochemical measurements revealed that the corrosion rate of SS17-4PH samples in acidic-chloride solutions is higher than the sample polarized in only NaCl or NaOH solution. From the Mott Schottky results, the straight lines with a positive slope obtained for the steel samples in different test media show that there is no change in the semi-conducting properties of the passive film. For samples polarized in acidic-chloride medium, the microstructural examination revealed the formation of a foot-like pit and zig-zag degradation structure. This was attributed to the presence of acid in the test media, and the deposition of corrosion products on the sample surface.

Effects of storage temperature on throat swabs preserved in different transport media for detection of SARS-CoV-2 by reverse transcriptase polymerase chain reaction

Objectives: The coronavirus disease 2019 (COVID-19) pandemic resulted in an increased need for molecular diagnostic testing. Delay in the specimen processing and storage of samples in laboratories leads to degradation of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viral RNA. Inactivation transport medium (ITM) contains chaotropic agents that inactivate the virus and stabilize SARS-CoV-2 RNA for a longer duration, even at room temperature. The effect of different temperatures and duration of storage of samples in viral transport media (VTM) and ITM for detection of SARS-CoV-2 RNA was assessed. Materials and Methods: Samples from COVID-19 patients were aliquoted in ITM and VTM and kept at ambient temperature, 37°C and 45°C. SARS-CoV-2 viral RNA was extracted. Multiplex real-time polymerase chain reaction was done on days 0, 1, 3, and 5, and cycle threshold (Ct) values were noted. Statistical Analysis: Data were analyzed using the Statistical Package for the Social Sciences version 26.0. Linear variables were summarized as mean and standard deviations. One-way analysis of variance test with post hoc Tukey honestly significant difference was used to compare mean value between different loops and for pair-wise comparison. P < 0.05 was taken as significant. Results: The mean Ct values of both the Orf and E genes of the samples in VTM and ITM were stable across all temperature conditions on day 1. On day 5, the increase in Ct values for both E and Orf genes were significantly higher for VTM than ITM at ambient temperature, 37°C and 45°C. Ribonuclease P failure was significantly higher for VTM than ITM at ambient temperature and 37°C on day 3 and at all temperatures on day 5. Conclusions: ITM is a valuable transport media that can preserve SARS-CoV-2 for up to 5 days at ambient temperature and 37°C. As it renders the samples non-infectious, thus reducing the potential of biohazard events, this transport medium can be used effectively for the collection and transportation of SARS-CoV-2 samples, especially from remote or isolated healthcare facilities.

Characterization and Differentiation of Candida auris on Dixon's Agar Using Raman Spectroscopy.

Candida auris, an emerging multidrug-resistant fungal pathogen, poses significant challenges in healthcare settings due to its high misidentification rate and resilience to treatments. Despite advancements in diagnostic tools, a gap remains in rapid, cost-effective identification methods that can differentiate C. auris from other Candida species, particularly on non-standard culture media. We used Raman spectroscopy to characterize C. auris grown on modified Dixon's agar (mDixon) and differentiated it from Candida albicans and Candida parapsilosis. Key Raman spectral markers at 1171 cm-1 and 1452 cm-1, linked to mannan and β-glucan composition, differentiated C. auris into two subgroups, A and B. Despite the spectral similarities of groups A and B with C. albicans and C. parapsilosis, respectively, all Candida species were distinguishable through principal component analysis (PCA). Additionally, this study is the first to demonstrate the distinct spectral signature of mDixon agar, achieved through spatially offset Raman spectroscopy (SORS), which enables accurate discrimination between the culture medium and fungal samples. The observed inter-individual variability within C. auris, coupled with the spectral overlap between C. auris subgroups and other Candida species, highlights a major challenge in differentiating closely related fungi due to their similar molecular composition. Enhancements in spectral resolution and further fluorescence minimization from the culture medium are needed to reliably detect the subtle biochemical differences within these species. Despite these challenges, the results underscore the potential of Raman spectroscopy as a real-time, non-destructive, and complementary tool for fungal pathogen identification.

A novel healthcare 4.0 system for testing respiratory diseases based on nanostructured biosensors and fog networking

New digital healthcare models based on advanced bio-sensing technologies are regarded as a possible solution to improve the screening and prevention processes and the overall performance of healthcare supply chains. This is particularly relevant for respiratory diseases, which are currently among the first causes of death and medical expenditures in industrialized countries. This research proposes a new e-health model based on a fog architecture and a smart device, enabling remote diagnostics of respiratory diseases and allowing for decentralized patient testing and self-testing. According to such a model, the patients’ testing is executed through the analysis of the exhaled breath collected using a smart device based on a novel nanostructured sensor and transferring relevant information to the medical staff involved in the diagnosis process. This research proposes an original testing method and system, validated in the lab through a comparative analysis of culture media samples collected from healthy patients, and subsequently exposed to cigarette smoke extract (CSE, an inducer of oxidative stress). The preliminary results obtained demonstrate the validity of the approach proposed, encouraging further experimental analyses on human patients for the implementation into clinical practice.

Detection of oomycete pathogens in UK peat-free growing media and implications for plant health

ABSTRACT Peat-based growing media products are being phased out in the UK. Alternative constituents may be at risk of harbouring oomycete plant pathogens. Thirty-five peat-free growing media samples were tested for oomycete pathogens by baiting and DNA metabarcoding. No Phytophthora species was detected by baiting, but seven species of Pythium, Phytopythium and Elongisporangium, including four species not previously reported in the UK, were baited. DNA matching eleven Phytophthora species was detected by metabarcoding across a range of samples. A coir sample was also found to contain DNA of Peronosclerospora, a genus of tropical downy mildew pathogens. DNA matching six Phytophthora pathogens, including P. bishii, a pathogen of strawberry not previously reported in the UK, was detected in recycled coir that had been used for strawberry production in The Netherlands. We discuss the implications of the species detected in terms of risks to plant health and the need for further evidence-gathering.

Raman-Enabled Predictions of Protein Content and Metabolites in Biopharmaceutical Saccharomyces cerevisiae Fermentations.

Raman spectroscopy, a robust and non-invasive analytical method, has demonstrated significant potential for monitoring biopharmaceutical production processes. Its ability to provide detailed information about molecular vibrations makes it ideal for the detection and quantification of therapeutic proteins and critical control parameters in complex biopharmaceutical mixtures. However, its application in Saccharomyces cerevisiae fermentations has been hindered by the inherent strong fluorescence background from the cells. This fluorescence interferes with Raman signals, compromising spectral data accuracy. In this study, we present an approach that mitigates this issue by deploying Raman spectroscopy on cell-free media samples, combined with advanced chemometric modeling. This method enables accurate prediction of protein concentration and key process parameters, fundamental for the control and optimization of biopharmaceutical fermentation processes. Utilizing variable importance in projection (VIP) further enhances model robustness, leading to lower relative root mean squared error of prediction (RMSEP) values across the six targets studied. Our findings highlight the potential of Raman spectroscopy for real-time, on-line monitoring and control of complex microbial fermentations, thereby significantly enhancing the efficiency and quality of S. cerevisiae-based biopharmaceutical production.

Synthesis of a Hematite‐Geopolymer Composite: Optimization of Formulation Parameters and Application to the Detection of Pb2+ Ions in Aqueous Solution

AbstractThe Composite Centered Experimental Design (CCED) using the response surface methodology was used to optimize the synthesis of a hematite‐geopolymer composite, applied to the preparation of a Pb2+ electrochemical sensor. Two important experimental parameters in the formulation (curing time and hematite content) were considered in the optimization of the synthesis of the composite material. The electrochemical signal of Pb2+ at a carbon paste electrode modified by the synthesized composite was used as a response to identify the best formulation. After applying the generated experimental plan, the optimal composite material was obtained using 17.08% hematite and a curing time of 0.16 days. The characterization of the optimized composite material revealed the presence of a porous structure and a poly(phospho‐ferro‐siloxo) network, but with a fraction of well dispersed and unreacted hematite. Preliminary experiments confirmed that the sensor prepared with the composite showed high stability and reproducibility of the Pb (II) signal, justifying its application for accurate sensing of the metal cation in an aqueous solution. A Pb2+ calibration curve was obtained after optimizing the experimental detection parameters (composition of the carbon paste electrode, pH of the accumulation solution, accumulation time, and electrolysis potential). A detection limit of 0.2 nM (based on a signal‐to‐noise ratio of 3) was obtained for a studied concentration range between 0.04 and 0.48 nM. The designed sensor also showed remarkable selectivity and excellent performance in real media samples (spring, tap, rain, and well water).

Probing the effects of polysaccharide hydrogel composition on the viability and pro‐angiogenic function of human adipose‐derived stromal cells

AbstractCell therapies harnessing the pro‐vascular regenerative capacities of mesenchymal stromal cell (MSC) populations, including human adipose‐derived stromal cells (hASCs), have generated considerable interest as an emerging treatment strategy for peripheral arterial disease (PAD) and its progression to critical limb ischemia (CLI). There is evidence to support that polysaccharide hydrogels can enhance therapeutic efficacy when applied as minimally‐invasive delivery systems to support MSC survival and retention within ischemic tissues. However, there has been limited research to date on the effects of hydrogel composition on the phenotype and function of encapsulated cell populations. Recognizing this knowledge gap, this study compared the pro‐angiogenic function of hASCs encapsulated in distinct but similarly‐modified natural polysaccharide hydrogels composed of methacrylated glycol chitosan (MGC) and methacrylated hyaluronic acid (MHA). Initial in vitro studies confirmed high viability (>85%) of the hASCs following encapsulation and culture in the MGC and MHA hydrogels over 14 days, with a decrease in the cell density observed over time. Moreover, higher levels of a variety of secreted pro‐angiogenic and immunomodulatory factors were detected in conditioned media samples collected from the hASCs encapsulated in the MGC‐based hydrogels compared to the MHA hydrogels. Subsequent testing focused on comparing hASC delivery within the MGC and MHA hydrogels to saline controls in a femoral artery ligation‐induced CLI (FAL‐CLI) model in athymic nu/nu mice over 28 days. For the in vivo studies, the hASCs were engineered to express tdTomato and firefly luciferase to quantitatively compare the efficacy of the two platforms in supporting the localized retention of viable hASCs through longitudinal cell tracking with bioluminescence imaging (BLI). Interestingly, hASC retention was significantly enhanced when the cells were delivered in the MHA hydrogels as compared to the MGC hydrogels or saline. However, laser Doppler perfusion imaging (LDPI) indicated that the restoration of hindlimb perfusion was similar between the treatment groups and controls. These findings were corroborated by endpoint immunofluorescence (IF) staining showing similar levels of CD31+ cells in the ligated limbs at 28 days in all groups. Overall, this study demonstrates that enhanced MSC retention may be insufficient to augment vascular regeneration, emphasizing the complexity of designing biomaterials platforms for MSC delivery for therapeutic angiogenesis. In addition, the data points to a potential challenge in approaches that seek to harness the paracrine functionality of MSCs, as strategies that increase the secretion of immunomodulatory factors that can aid in regeneration may also lead to more rapid MSC clearance in vivo.

Plastic Debris in the Aquatic Environment: An Emerging Substratum for Antimicrobial Resistant (AMR) Biofilms.

Plastic pollution has quadrupled over the past years and has become a global concern due to its direct impact on life forms. The present study analysed whether the plastic debris in aquatic environments could act as the substratum for the antimicrobial-resistant (AMR) bacteria to form biofilm for survival. We have collected various plastic debris (n = 32) from six sites of the Periyar River, the drinking water source for the entire city and one of the most polluted rivers in Kerala (India). The chemical composition of plastics was screened via FTIR analysis and found that they comprised two types, viz., polyethylene and polypropylene. Bacteria isolated from the samples were screened for the AMR characteristics towards eight different classes of antibiotics. All isolates showed 100% resistance towards colistin and obtained the MAR index value of 0.1-0.4 range. Six representative bacterial isolates with high multiple antibiotic resistance (MAR) index were selected and identified by 16sRNA sequencing as Lysinibacillus mangiferihumi, Bacillus pumilus, Bacillus safensis, Bacillus cereus, Bacillus altitudins and Bacillus pumilus. In vitro biofilm formation was experimented on the purchased plastic samples in artificial media and river water using two selected strains, Bacillus pumilus and Bacillus cereus. Significant variations were observed in biofilm growth in different media (P < 0.05) regardless of plastic types (P > 0.05). The extracellular polymeric substances (EPS) and the characteristic holes on the surface morphology were visualized in SEM analysis, thus indicating the conditioning of the plastics by the isolates for biofilm formation.

Automatic characterization of capillary flow profile of liquid samples on μTADs based on capacitance measurement

Capillary flow profile of liquid samples in porous media is closely related to the important properties of liquid samples, including the viscosity and the surface energy. Therefore, capillary flow profile can be used as an index to differentiate liquid samples with different properties. Fast and automatic characterization of capillary flow profile of liquid samples is necessary. In this work, we develop a portable and economical capacitance acquisition system (CASY) to easily obtain the capillary flow profile of liquid samples on microfluidic thread-based analytical devices (μTADs) by measuring the capacitance during the capillary flow. At first, we validate the accuracy of this method by comparing with the traditional method by video analysis in obtaining the capillary flow profiles in μTADs of cotton threads or glass fiber threads. Then we use it to differentiate liquid samples with different viscosity (mixture of water and glycerol). In addition, capillary flow profile on μTADs with chemical valves (chitosan or sucrose) can also be obtained on this device. Lastly, we show the potential of this device in measurement of hematocrit (HCT) of whole blood samples. This device can be used to catalog liquid biological samples with different properties in point-of-care diagnostics in the near future.

Ultrasonic diagnostics of concrete destruction under the influence of corrosion environment

Introduction. Modern methods of studying the corrosion damage of concrete are quite lengthy and time-consuming, which makes their use difficult. This necessitates the search for new approaches to study the destruction of concrete under the influence of aggressive media. Currently, the ultrasound diagnostic method is effectively used to identify various structural defects, but the possibility of its use for concrete corrosion studies is poorly understood. This work is devoted to filling this gap.The methods of research. Concrete samples of 2.5x2.5x10 cm and 2.5x2.5x16 cm of various composition (cement: placeholder = 1:9, 1:7, 1:5). The control samples hardened under normal conditions, the test samples were stored in various aggressive environments. The samples of all storage media were subjected to periodic tests on the Pulsar-2.2 ultrasound diagnostic device. In parallel, the strength parameters obtained by the destructive method on the PGM-100MG4 hydraulic press, as well as the resistance coefficient, were determined.Results. The relationship between the growth periods, extremes and abrupt changes in the velocity of passage of ultrasonic waves and the stages of strength gain, the conditions of hardening of samples, and time factors has been established. It is shown that ultrasound diagnostics is quite sensitive to the processes caused by the corrosion destruction of concrete and allows us to obtain reliable data, while the strength and coefficient of resistance do not always adequately reflect the development of corrosion processes. It has been established that the use of ultrasound diagnostics makes it possible to increase the reliability of the results of concrete corrosion studies.Conclusion. The use of the ultrasound diagnostic method enables to obtain new data on the processes of concrete destruction under the influence of aggressive media, which makes it possible to increase the reliability of the results of concrete corrosion studies.

Influence of inflammation on the expression of microRNA-140 in extracellular vesicles from 2D and 3D culture models of synovial-membrane-derived stem cells.

In osteoarthritis (OA), articular homeostasis is regulated by microRNA-140 that inhibits ADAMTS-5, an enzyme that cleaves aggrecan and stimulates the synthesis of other inflammatory mediators. This study aims to evaluate the expression of microRNA-140 in extracellular vesicles (EVs) derived from equine synovial-membrane-derived mesenchymal stem cells (eqSMMSCs) cultured in monolayer (2D) and three-dimensional (3D) culture models under an in vitro inflammatory environment. Four experimental groups of eqSMMSC cultures were defined for isolation of the EVs. The 2D and 3D control groups were cultured in a conventional cell culture medium, while the 2D-OA and 3D-OA treatment groups were exposed to an OA-like medium containing IL-1β and TNFα. The culture media samples were collected at 24h, 72h, and 120h time points for EV isolation and characterization using nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM). Reverse transcription quantitative polymerase chain reaction was employed to assess the expressions of microRNA-140 in both the cells and EVs. All statistical analyses were conducted at the 5% significance level. Encapsulation of the eqSMMSCs protected the cells from the inflammatory media compared to the monolayer cultures. EVs were found in higher concentrations in the 3D-OA cultures. Additionally, higher expressions of microRNA-140 were observed in the cells of the 3D-OA group at 24 and 72h, whereas microRNA-140 expressions in the EVs were higher in the 3D group at 72h and in the 2D-OA group at 120h (p < 0.001). However, the 3D-OA culture showed higher expression of the mRNA Adamts5 in the EVs at 120h. The responses of the eqSMMSCs to inflammatory stimuli involve intracellular expression of microRNA-140 and its subsequent transportation via the EVs, with quicker responses observed in the 3D than 2D cultures. This study sheds light on the behaviors of stem cells in restoring homeostasis in osteoarthritic joints.

Development of a PCR-based method to identify fetal sex during IVF cycles.

One of the most recognizable cases of preimplantation genetic diagnosis (PGD) is X-linked diseases. Diagnosis of fetal sex is essential for couples who are known to be at risk of some X-linked disorders. The objective of this study was to discriminate between female (XX) and male (XY) embryos by detecting sex chromosomes-specific sequences in spent culture medium and comparing these results to PGD/CGH array results. It may open new window for the development of a non-invasive PGD method. 120 Embryo's spent media from Day 3 and Day 5 embryos were collected. Modified phenol-chloroform solution was used for DNA extraction from spent media. Sex determination was performed using SRY, TSPY and AMELOGENIN evaluation through quantitative polymerase chain reaction (q-PCR) method. IBM SPSS and MedCalc were used for statistical analyses to compare sex determination of embryos by spent medium with PGD/CGH array results. Culture time was demonstrated to increase the DNA amount among day 5 embryos culture medium samples. Non-invasive PGD by means of spent culture medium gave a sensitivity, specificity, positive predictive value and negative predictive value of 100% for sex determination. Results of sex determination using spent medium by q-PCR were consistent with the results of PGD/CGH array. Improvements in cell-free DNA extraction and PCR amplification procedures provide us an effective method to perform a PGD test without biopsy in the future, especially about X-linked diseases.

IDENTIFIKASI BAKTERI PADA PRODUK DARAH THROMBOCYTE CONCENTRATE

TThe Thrombocte Concentrate component is the blood component that is most easily contaminated with bacteria due to its processing and storage at room temperature. Blood or blood products that are ready to be transfused must be safe and free from the risk of infection by infectious diseases and contamination by microorganisms. Sources of bacterial contamination can come from donor skin that is less than aseptic, donor bacteremia and processing of blood products. This type of research is Pre-Experiment. The design of this research is One-Shot Case Study. In One-Shot Case Study design. The sample in this study used 1 Thrombocyte Concentrate blood product. Then it was replicated into 10 samples for Nutrient Agar media and 10 samples for Mac conkey Agar media. From the results of research conducted on 10 samples of Nutrient Agar (NA) media, it was found that there was bacterial growth in all petri dishes on Nutrient Agar (NA) media. ), while in petri dish number 10 it was found that 10% of gram-positive bacteria were in the form of bacilli (rods). From the results of research conducted on 10 samples of Mac Conkey Agar (MCA) media, no bacterial growth was found in all of the MCA media petri dishes. There was contamination of Gram-Positive bacteria in the form of Cocci (Rounds) and Bacillus (Sticks) in Thrombocyte Concentrate blood products using NA media in 10 samples out of the total samples examined were 10 samples of NA media. There was no Gram-Negative bacterial contamination in Thrombocyte Concentrate blood products using MCA media in 10 samples from the 10 samples examined.

Radiation survey of the territory of Teriberka village and Kildin Island of Murmansk Region

The Arctic zone is a strategically important territory of the Russian Federation from the point of view of socio-economic, scientific, and technological components, as well as in terms of planning the development of a system for ensuring national defense, state, and public security. Intensive exploration and development of Arctic territories, combined with a rapidly changing environment due to global climate change, carries a few risks for the environment. This determines the need to assess potential environmental damage and determine the significance of factors of potential negative impact on unique Arctic ecosystems now and in the future. The article presents current data from a radiation survey of the coastal zone of residential areas close to the site of the sinking of the nuclear submarine ‘K-159’ – Teriberka village and Kildin Island, Murmansk region, as well as the sea area to assess the primary levels of man-made and natural radionuclides before starting work on the potential lifting of the submerged object. This work was carried out as part of the implementation of the State Program of the Russian Federation ‘Health Development’ (approved by Decree of the Government of the Russian Federation dated December 26, 2017, No. 1640). The results of measuring the ambient dose equivalent rate of gamma radiation in open areas and determining the content of man-made and natural radionuclides in samples of environmental media (soil, aquatic vegetation, marine aquatic organisms) and food products of local origin and production (mushrooms, berries, fish, milk). The research results showed that the state of the study area is generally stable in terms of the radiation factor, the content of radionuclides is at the background level. As part of this work, electronic database modules have been developed based on the results of radiation monitoring in Teriberka village and on Kildin Island. Certificate No. 2022621890 was received on registration of a database with the results of the radiation situation of the above-mentioned territories.