Thermo Fisher Scientific Research Articles

Genomic alteration in sporadic adolescent and young adult-onset colorectal adenocarcinoma.

154 Background: Colorectal cancer (CRC) incidence is increasing in adults younger than 50 years. This study evaluated genomic alteration in adolescent and young adult (AYA)-onset sporadic CRC patients who aged between 15 and 39 years. Methods: DNA from 90 archived formalin-fixed, paraffin-embedded (FFPE) tissue samples that histologically confirmed adenocarcinoma with proficient mismatch repair tumors at Siriraj Hospital (Bangkok, Thailand) were extracted. Patients who clinically suspected familial adenomatous polyposis were excluded. Gene mutational analysis was performed by next-generation sequencing (NGS) using an Oncomine Comprehensive Assay Plus kit (Thermo Fisher Scientific, Inc., Waltham, MA, USA) and compared with previous reported molecular data in adult-onset CRC from our group. Results: The top 5 mutations frequency observed were TP53, KRAS, FBXW7, PIK3CA, and SMAD4 mutations which were comparable to what reported in adult-onset CRC. However, FBXW7, PIK3CA, NOTCH1, FGFR3, ERBB2, and PTEN were reported more frequent in AYA group. No difference in number of KRAS, NRAS, and BRAF mutations among 2 groups. Table below shows key cancer genes mutation frequencies. Conclusions: This study is the comprehensive report hotspot mutations using NGS in sporadic AYA-onset sporadic CRC patients. The most commonly identified gene mutation frequencies among AYA-onset were similar to those reported in adult-onset, except for FBXW7, PIK3CA, NOTCH1, FGFR3, ERBB2, and PTEN mutations that had a slightly higher frequency. Further studies on larger sample set for genetic and epigenetic landscape are required. [Table: see text]

Effects of three disinfection solutions on residual monomers released from resin nanoceramic CAD/CAM blocks

The aim of this study was to evaluate the effects of three disinfection solutions on the amount of monomers released from resin nanoceramic CAD/CAM blocks using high performance liquid chromatography (HPLC). Forty resin nanoceramic CAD/CAM (Cerasmart, GC, Japan) samples (12x14 × 2 mm) were divided into four groups; each group was disinfected using one of four solutions (Group 1: no disinfectant; Group 2: 70 % ethanol; Group 3: 2 % glutaraldehyde; and Group 4: 1 % sodium hypochlorite) for 5 min. Analysis of residual monomers (UDMA and Bis-EMA) amounts was performed using an HPLC instrument (Dionex Ultimate 3000, Thermo Fisher Scientific). After 30 days, the amounts of monomers found were as follows: 14.54 ppm for Group 1; 9.28 ppm for Group 2; 10.60 ppm for Group 3; and 2.76 ppm for Group 4 (the smallest monomer amount) (p < 0.001). Disinfection of indirect restorations prior to cementation can reduce the amount of residual monomers remaining from resin nanoceramic CAD/CAM blocks.

Diagnostic accuracy of MRI–CBCT fused images in assessment of clinically diagnosed internal derangement of the temporomandibular joint

ObjectivesTo evaluate the diagnostic accuracy of Fused (MRI)–CBCT images in the assessment of internal derangement of the temporomandibular joint.MethodsMRI and CBCT images of the TMJ were evaluated bilaterally in 10 patients with clinically diagnosed internal derangement. Image fusion was performed using Amira 3D Software (version 5.4.3, Thermo Fisher Scientific Inc.).ResultsThe AUC index for MRI–CBCT fused images was 0.83, which was significantly different from the null hypothesis value of 0.5. This was confirmed by inter-examiner reliability index of 0.87, which is statistically significant.ConclusionMRI–CBCT fused images can significantly improve the accuracy and inter-examiner reliability in the evaluation of clinically diagnosed cases with internal derangement.

Detection of SARS-CoV-2 B.1.1.529 (Omicron) variant by SYBR Green-based RT-qPCR.

The coronavirus disease 2019 (COVID-19) pandemic is unceasingly spreading across the globe, and recently a highly transmissible Omicron SARS-CoV-2 variant (B.1.1.529) has been discovered in South Africa and Botswana. Rapid identification of this variant is essential for pandemic assessment and containment. However, variant identification is mainly being performed using expensive and time-consuming genomic sequencing. In this study, we propose an alternative RT-qPCR approach for the detection of the Omicron BA.1 variant using a low-cost and rapid SYBR Green method. We have designed specific primers to confirm the deletion mutations in the spike (S Δ143-145) and the nucleocapsid (N Δ31-33) which are characteristics of this variant. For the evaluation, we used 120 clinical samples from patients with PCR-confirmed SARS-CoV-2 infections, and displaying an S-gene target failure (SGTF) when using TaqPath COVID-19 kit (Thermo Fisher Scientific, Waltham, USA) that included the ORF1ab, S, and N gene targets. Our results showed that all the 120 samples harbored S Δ143-145 and N Δ31-33, which was further confirmed by whole-genome sequencing of 10 samples, thereby validating our SYBR Green-based protocol. This protocol can be easily implemented to rapidly confirm the diagnosis of the Omicron BA.1 variant in COVID-19 patients and prevent its spread among populations, especially in countries with high prevalence of SGTF profile.

Assessing the Precision of Radon Measurements from Beta-Attenuation Monitors

Atmospheric radon measurements assist in many aspects of climate and meteorological research, notably as an airmass tracer and for modelling boundary layer development, mixing heights and stability. Daughter products from radon decay are sometimes incorporated into the particle pollution measurements of commercially available beta-attenuation monitors (BAM). BAMs incorporating radon measurements are used in air quality monitoring networks and can supplement traditional radon measurements. Here we compare in-situ radon measurements from Thermo Fisher Scientific (Franklin, MA, USA) BAM instruments (Thermo Scientific 5014i, Thermo Scientific 5030 SHARP, Thermo Anderson FH62C14) at two air quality monitoring stations in New South Wales, Australia. Between systems we find strong correlations for hourly measurements (r = 0.97–0.99); daily means (r = 0.97–0.99); hour of the day (r = 0.84–0.98); and month (r = 0.82–0.98). The regression analysis for radon measurements between systems showed strong linear responses, although there are some variations in the slopes of the regressions. This implies that with correction BAM measurements can be comparable to standard measurement techniques, for example, from the Australian Nuclear Science and Technology Organisation (ANSTO) dual flow loop monitors. Our findings imply that BAM derived radon measurements are precise, although their accuracy varies. BAM radon measurements can support studies on boundary layer development or where radon is used as an atmospheric transport tracer.

Genomics landscape of mitochondrial DNA variations in patients from South Italy affected by mitochondriopathies

Mitochondrial DNA (mtDNA) is a 16,569 base pairs, double-stranded, circular molecule that contains 37 genes coding for 13 subunits of the respiratory chain plus 2 rRNAs and 22 tRNAs. Mutations in these genes have been identified in patients with a variety of disorders affecting every system in the body. The advent of next generation sequencing technologies has provided the possibility to perform the whole mitochondrial DNA sequencing, allowing the identification of disease-causing pathogenic variants in a single platform. In this study, the whole mtDNA of 100 patients from South Italy affected by mitochondrial diseases was analyzed by using an amplicon-based approach and then the enriched libraries were deeply sequenced on the ION Torrent platform (Thermofisher Scientific Waltham, MA, USA). After bioinformatics analysis and filtering, we were able to find 26 nonsynonymous variants with a MAF <1% that were associated with different pathological phenotypes, expanding the mutational spectrum of these diseases. Moreover, among the new mutations found, we have also analyzed the 3D structure of the MT-ATP6 A200T gene variation in order to confirm suspected functional alterations. This work brings light on new variants possibly associated with several mitochondriopathies in patients from South Italy and confirms that deep sequencing approach, compared to the standard methods, is a reliable and time-cost reducing strategy to detect all the variants present in the mitogenome, making the possibility to create a genomics landscape of mitochondrial DNA variations in human diseases.

Differences in molecular sensitization profiles between a Spanish and Latin American mite allergic patients.

To analyze the sensitization pattern to Dermatophagoides pteronyssinus and to associate the diagnostic findings and clinical severity in 218 allergic patients from two different continents. Mite allergic patients were recruited by the Allergology departments from Latin America (n=88: Colombia, Costa Rica and Guatemala) and Spain (N=130). All patients had allergic rhinitis with or without asthma and positive skin prick test results to D. pteronyssinus. Specific IgE levels to D. pteronyssinus, D. farinae, Der p 1, Der p 2, and Der p 23 were quantified by ImmunoCAP system (ThermoFisher Scientific). Allergenic profile was also determined by western blot. Comparative Statistical analysis was performed by GraphPad software. Patients recognized most frequently Der p 2 (79%) followed by Der p 1 (73%), and Der p 23 (69%) allergens. The percentage of asthmatic patients increases with the number of sensitizations however none statistically significant differences were found. Interestingly, asthmatic patients presented the highest median levels of total IgE and specific IgE levels of D. pteronyssinus and molecular allergens, mainly Der p 2. Analysing the two different populations, Spanish patients were predominantly sensitized to Der p 2 (88.46%) and Der p 1 (83.84%), whereas Latin American population were more sensitized to Der p 23. Our data support the relevance of Der p 2 in mite allergy as the major allergen, with the high number of patients sensitized to it and its importance in the development of asthma. Sensitization to Der p 23 was more important in Latin America.

PREVALENCE OF CONTAGIOUS BOVINE MASTITIS PATHOGENS IN SAMPLES OF COLLECTED MILK

Dairy producers suffer significant economic losses due to various diseases of contagious and non-contagious etiology, among which inflammation of the mammary gland is one of the main ones. Mastitis is one of the most common diseases on dairy farms and causes significant economic losses associated with the cost of treating sick cows, reduced milk yield and quality of dairy products, and the risk of premature culling. The causative agents of mastitis are divided into two groups, among which the most important are contagious pathogens, including the five most important infectious agents: Staphylococcus aureus, Streptococcus agalactiae, Mycoplasma bovis, Streptococcus uberis, Streptococcus dysgalactiae, their habitat is sick animals and they are usually transmitted mainly from a healthy cow to a sick cow, especially through milking equipment, hands of machine milking operators, which in turn can lead to infection of the entire dairy herd. Therefore, establishing the prevalence of contagious mastitis pathogens is quite relevant today, it provides information on the possibility of creating rational and effective mastitis control programs on dairy farms, and also fills an important gap in veterinary medicine and farm management practice. The aim of this study was to identify and establish the prevalence of contagious mastitis pathogens in tank milk samples using bacteriological and molecular genetic (PCR-RFLP) studies. Molecular genetic studies were performed by real-time polymerase chain reaction (RT-PCR). Nucleic acids were isolated using the KingFisher Purification System automatic isolation system using the MagMAX™ CORE Nucleic Acid Purification Kit (Thermo Fisher Scientific, USA). For the amplification reaction of the isolated nucleic acids, commercial VetMAX MastiType Micro4 Kit (Applied Biosystems™ by Thermo Fisher Scientific, USA) was used. The reaction was performed on a real-time PCR product detection system using a QuantStudio5 Real Time PCR System (Thermo Fisher Scientific, USA). Bacteriological studies of tank samples of cow milk were carried out according to generally accepted methods, the identification of bacterial cultures obtained on blood agar was carried out by MALDI-TOF using a VITEK®MS device (BioMérieux, France). The VITEK MS KB V3.2.0 US Version database was used to analyze the mass spectra. The results of the study of contagious mastitis pathogens in tank milk samples by PCR-RF revealed that the most common are Streptococcus agalactiae 36% and Streptococcus uberis 35 %, Staphylococcus aureus occupies an intermediate position, accounting for 24 % of all isolates, Mycoplasma bovis, in turn, accounts for only 5% of isolates. When analyzing the data of the results of molecular genetic studies, it was found that the following associations of pathogens of contagious mastitis in tank milk samples were found Streptococcus uberis, Staphylococcus aureus, Streptococcus agalactiae – 29,8 %, Streptococcus agalactiae, Streptococcus uberis – 21,3 %, Streptococcus agalactiae, Staphylococcus aureus – 19,1 %, Streptococcus uberis, Staphylococcus aureus – 14,9 %, Streptococcus uberis, Staphylococcus aureus, Streptococcus agalactiae, Mycoplasma bovis and Streptococcus uberis, Mycoplasma bovis – 4,3 %, respectively. The associations of infectious agents Streptococcus uberis, Mycoplasma bovis, Streptococcus agalactiae; Mycoplasma bovis, Streptococcus agalactiae and Streptococcus uberis, Mycoplasma bovis, Staphylococcus aureus were found once each, which is 2,1 %. The bacteriological examination of tank milk samples showed that the most commonly detected contagious pathogens were Streptococcus agalactiae – 55 % and Streptococcus uberis – 28 %, Staphylococcus aureus accounted for 11 % of isolates, and Streptococcus dysgalactiae was identified in only 6 % of tank milk samples. The following associations of pathogens of contagious mastitis were most often found in tank milk samples: Streptococcus agalactiae and Staphylococcus aureus – 35,7 %, Streptococcus uberis and Staphylococcus aureus – 28,6 %, Streptococcus uberis and Streptococcus agalactiae – 21,4 %. The associations of pathogens Streptococcus uberis, Staphylococcus aureus, Streptococcus agalactiae and Streptococcus uberis, Streptococcus dysgalactiae were found in tank samples only once, which is 7,1 % each.

Comparison of zeolite adsorbent with N-permutite adsorbent

Zeolite is an aluminosilicate mineral with a three-dimensional framework structure with interconnected cavities and channels, which makes its surface area very large and effective as an adsorbent. There are currently natural and artificial types of zeolites. The high adsorption efficiency of zeolite is used as an adsorbent in all areas of the chemical industry. This scientific work obtained an alternative version of zeolite by synthesizing H-permutite based on natural raw materials. To study the characteristics of the structure of the molecular compounds of the studied samples, the Nicolet iS50 (Thermo Fisher Scientific, USA) IR-Fourier spectrometer was used. Measurements were made in the 4000-400 cm-1 spectral range, with spectral resolution not exceeding 0.1 cm. The test specimen was pressed against the mounting surface with a flat-tip probe. near, mid, and far IR attenuated total internal reflection (ATR) I S50 Analytical indicators were determined using a hole and a diamond crystal for measurement.

STEM SerialED: achieving high-resolution data for ab initio structure determination of beam-sensitive nanocrystalline materials.

Serial electron diffraction (SerialED), which applies a snapshot data acquisition strategy for each crystal, was introduced to tackle the problem of radiation damage in the structure determination of beam-sensitive materials by three-dimensional electron diffraction (3DED). The snapshot data acquisition in SerialED can be realized using both transmission and scanning transmission electron microscopes (TEM/STEM). However, the current SerialED workflow based on STEM setups requires special external devices and software, which limits broader adoption. Here, we present a simplified experimental implementation of STEM-based SerialED on Thermo Fisher Scientific STEMs using common proprietary software interfaced through Python scripts to automate data collection. Specifically, we utilize TEM Imaging and Analysis (TIA) scripting and TEM scripting to access the STEM functionalities of the microscope, and DigitalMicrograph scripting to control the camera for snapshot data acquisition. Data analysis adapts the existing workflow using the software CrystFEL, which was developed for serial X-ray crystallography. Our workflow for STEM SerialED can be used on any Gatan or Thermo Fisher Scientific camera. We apply this workflow to collect high-resolution STEM SerialED data from two aluminosilicate zeolites, zeolite Y and ZSM-25. We demonstrate, for the first time, ab initio structure determination through direct methods using STEM SerialED data. Zeolite Y is relatively stable under the electron beam, and STEM SerialED data extend to 0.60 Å. We show that the structural model obtained using STEM SerialED data merged from 358 crystals is nearly identical to that using continuous rotation electron diffraction data from one crystal. This demonstrates that accurate structures can be obtained from STEM SerialED. Zeolite ZSM-25 is very beam-sensitive and has a complex structure. We show that STEM SerialED greatly improves the data resolution of ZSM-25, compared with serial rotation electron diffraction (SerialRED), from 1.50 to 0.90 Å. This allows, for the first time, the use of standard phasing methods, such as direct methods, for the ab initio structure determination of ZSM-25.

Влияние ионизирующего излучения на активность антиоксидантных ферментов штамма Escherichia coli «ПЛ-6» при многократном облучении

The article presents the analysis of enzymatic activity of Escherichia coli “PL-6” cells following multiple gamma irradiation with increasing doses. For irradiation the stationary gamma ray unit “Researcher” (Co-60) with the dose rate of 36.7 Gy/min was used. Radiation doses to the bacterial cells increased from 3.5 to 10 kGy in 0.5 kGy increment, and from 10 to 30 kGy in 5 kGy increment. Unirradiated suspended bacterial cells were used as control. The irradiated cells enzymatic activity was estimated with the spectrophotometric method on the "MultiSkan Go" device (Thermo Fisher Scientific, Finland). The results of the study demonstrate the increase in the E. coli ”PL-6” cells radioresistance caused by gamma irradiation of the cells with gradually increased doses. Exposure of the bacteria to a higher gamma radiation dose caused decrease in the number of viable cells from 10Е+9 CFU/ml in the control culture sample to 10Е+2 CFU/ml after irradiation at a dose of 30 kGy. The most radiation-resistant cells were selected from cells undergone successive multiple irradiation with constantly increasing dose of ionizing radiation. The enzymes of the antioxidant system in irradiated cells exhibit increased specific activity as compared to the enzymes in control, unirradiated, cells. The activity of enzymes increases with an increase in the radiation dose. Catalase activity increased after multiple irradiation by 5 times and peroxidase activity increased by 4 times. To compare the results in the groups, the Student's t-test was applied at p&lt;0.05.

INVESTIGATION THE EXPRESSION LEVELS OF MIR-181 AND HOXA11 GENE IN EUTOPIC AND ECTOPIC ENDOMETRIAL TISSUE.

The exact pathogenesis of the endometriosis is not apparent. MicroRNAs (miRNAs/miRs) are non-coding RNAs that regulate gene expression at the post-transcriptional level. MicroRNAs can be used a diagnostic and therapeutic tools in different disorders such as endometriosis. MiR-181 has a function in embryo implantation. The main aim of this study is to evaluate the expression of miR-181 and its relationship with HOXA11 gene expression in ectopic and eutopic endometrium tissues in women with endometriosis. Thirty-four women participated in this study. Ectopic tissue samples (N=17) were collected via laparoscopic surgery, and eutopic tissue samples (N=17) were obtained from an endometrial biopsy. Endometrial tissue samples without endometriosis were considered the control group. Tissue samples were placed in RNase-free microtube with RNAlater™ Stabilization Solution (Thermo Fisher Scientific) and were kept at -80 °C. Quantitative real time-PCR for MiR-181 and HOXA11 genes were performed. MiR-181 expression level increased in eutopic tissue samples compared to the control group. This expression showed a significantly decrease in an ectopic group compared to the eutopic group. It was observed that HOXA11expression decreased remarkably in eutopic group compared to the control group and increased in ectopic group compared to the eutopic group. MiR-181 and HOXA11 are promising strategies in endometriosis disease. Understanding this relation and regulation roles contribute to realizing the etiology of endometriosis.

Simvastatin and amfenac act as immunomodulators during induced epithelial‐mesenchymal transition in ARPE‐19 cells

Purpose: Rhegmatogenous retinal detachment can develop into proliferative vitreoretinopathy (PVR) after surgery. Inflammation contributes to epithelial‐mesenchymal transition (EMT) in retinal pigment epithelial (RPE) cells, which takes place in PVR. The aim here was to explore the effects of simvastatin and amfenac on inflammation during the EMT.Methods: EMT in ARPE‐19 cells was stimulated by the commercial Stem X Vivo EMT supplement (R&amp;D Systems) either in non‐inflammatory or inflammatory conditions induced by 2.5 pg/mL IL‐1β (Gibco) or 10 pg/mL IL‐1α (R&amp;D Systems). Cells were treated concurrently with 5 μM simvastatin (Sigma Aldrich), 0.1 μM amfenac (Cayman Chemical) or both. The release of interleukin (IL)‐6 and IL‐8 (BD Biosciences), as well as monocytic chemoattractant protein‐1 (MCP‐1; Thermo Fisher Scientific) were measured using commercial enzyme‐linked immunosorbent assays. Lactate dehydrogenase (LDH) release and intracellular ATP levels were determined using commercial kits (Promega). Cells were photographed under an Axio Vert A1 Zeiss microscope.Results: Simvastatin alone or with amfenac reduced the release of IL‐6 in inflammatory and non‐inflammatory conditions upon experimental EMT, increased the IL‐8 release upon IL‐1α exposure and MCP‐1 in non‐inflammatory conditions and upon IL‐1β exposure. During EMT process, LDH release was increased by simvastatin alone and together with amfenac. They also increased ATP levels without inflammation and with IL‐1α. Amfenac had no effects on the cytokine levels (IL‐6, IL‐8, MCP‐1), LDH release, or ATP upon EMT process without inflammation. However, amfenac increased IL‐6 and MCP‐1 release and reduced LDH levels upon IL‐1β, and reduced IL‐6 upon the IL‐1a exposure. Under microscope, cells appeared alive and bigger but partly detached upon simvastatin.Conclusions: Simvastatin and amfenac showed immunomodulatory effects on ARPE‐19 cells during EMT. Further studies are needed concerning the effects of drugs on the EMT.

Испытания индустриального масла И-20А с нанододисперсными добавками шунгита и серпентина на машине трения

The paper presents the results of the experiments on the introduction of shungite and serpentine into industrial oil I-20A and the assessment of the friction coefficient in the load range of 50-200 N. The analysis of the results on the influence of the oil - shungite - serpentine ratios show that at the above loads the minimum friction coefficient is achieved when 0.9 % serpentine and 0.4 % shungite have been added to industrial oil I-20A. In this case, a reduction in the coefficient of friction by 25 % is obtained in comparison with the base industrial oil I-20A. The studies of the shungite powders and serpentine before grinding were carried out using a Thermo Fisher Scientific Quattro S scanning electron microscope. The microanalysis of the samples of shungite and serpentine studied in this work showed the presence of distinct particles with different compositions in them; this is often observed in minerals. The shungite sample contains a lot of iron, silicon, aluminum and potassium. To use shungite and serpentine obtained from other deposits as oil additives, additional studies of specific samples should be carried out.

Flow cytometric-based detection of CD80 is a useful diagnostic marker of acute myeloid leukemia in dogs.

CD80, a co-stimulatory molecule required for optimal T cell activation, is expressed on antigen-presenting cells, including monocytes and dendritic cells, in dogs and humans. We hypothesized that CD80 would be expressed on tumor cells in dogs from acute myeloid leukemia (AML) but not dogs with lymphoid neoplasms. We first evaluated the cellular staining pattern of a hamster anti-murine CD80 antibody (clone 16-10A1, ThermoFisher Scientific Cat# 17-0801-82, RRID: AB_469417) in blood and bone marrow aspirates from healthy dogs. Using flow cytometric analysis and examination of modified Wright's-stained cytologic smears of unsorted and flow cytometric or immunomagnetic bead-sorted leukocytes, we show that the antibody binds to mature and immature neutrophils and monocytes, but not lymphocytes or eosinophils, in blood and bone marrow. We then added the antibody to routine flow cytometric panels for immunophenotyping hematopoietic neoplasms in dogs. We found that the antibody labeled tumor cells in 72% of 39 dogs with AML and 36% of 11 dogs with acute leukemia expressing lymphoid and myeloid markers ("mixed lineage") but none of the dogs with B (n = 37) or T (n = 35) lymphoid neoplasms. A higher proportion of tumor cells in dogs with AML were labeled with the anti-CD80 antibody vs antibodies against other myeloid-associated antigens, including CD4 (36%, p = 0.003), CD11b (44%), CD11c (46%), CD14 (38%, p = 0.006) and CD18 (59%, clone YFC118). In contrast, antibodies against CD11b and CD11c bound to tumor cells in 8-32% of the lymphoid neoplasms. We show that CD80, as detected by antibody clone 16-10A1, is a sensitive and specific marker for AML and would be useful to include in flow cytometric immunophenotyping panels in dogs.

Cytomegalovirus Reactivation in Allogenic Haematopoietic Stem Cell Transplant Patients with Haploidentical Versus Fully Human Leukocyte Antigen Matched Donor

Objective: To determine CMV reactivation in hematopoietic stem cell transplant patients with haploidentical versus fully HLA matched donors Methodology: Retrospective Observational study was conducted at Armed Forces Bone Marrow Transplant Centre (AFBMTC) between March 2019 and October 2022. We enrolled a total of 60 consecutive patients.CMV DNA PCR analysis was carried out every week from day +14 to day +100 in order to monitor CMV reactivation during the 100-day post-transplantation period. The SaMag viral nucleic acid extraction kit (Sacace Biotechnologies, Como, Italy) was used to extract the DNA from two millilitres of blood collected in EDTA tubes. The kit's lower limit of detection for blood and serum was 5 copies/mL, while for other body fluids it was 500 copies/mL. An Applied Biosystems 7500 real-time PCR machine (Thermo Fisher Scientific, Waltham, MA) was used to conduct the PCR analysis. Result: The age range from 0.6 to 53 years ago was a median of 15±10.6. 39 patients had CMV reactivation (65%). With a range of 13 to 66 days, the median time to CMV DNAemia was 19.25 days.. Only two (3.3%) of the 39 patients received ganciclovir, while 26 (43.3%) received valganciclovir. Patients with aplastic anaemia had the highest rate of CMV infection (41%), followed by those with ALL (20.5%), AML (12.8%), and primary immunodeficiency (10.2%). A statistically significant connection between the underlying diagnosis and CMV infection. CMV infection was higher in haplo identical transplant recipients (66.6%) than in completely matched recipients (33.3%) and was statistically significantly. Sadly, 14 people passed away, yielding a mortality rate of 23.3%. Primary graft failure was the most frequent cause of mortality, resulting in neutropenic sepsis in 6 patients (42.85%), but 3 of these patients also had CMV reactivation. The rate of overall survival (OS) was found to be 76.7%. The rate of disease-free survival (DFS) was estimated at 71.7%. Notably, neither OS nor DFS in our analysis showed a statistically significant connection with CMV infection. Conclusion: The findings demonstrated a statistically significant association between haploidentical transplantation and CMV reactivation, with higher rates observed in the haploidentical group compared to fully matched recipients. Various factors, including underlying diagnosis, conditioning regimen, and GVHD prophylaxis, were also found to be correlated with CMV reactivation. However, while CMV reactivation was associated with specific factors, it did not impact overall survival or disease-free survival in the study population.

Exosomal microRNA following severe trauma: Role in bone marrow dysfunction.

Severe trauma disrupts bone marrow function and is associated with persistent anemia and altered hematopoiesis. Previously, plasma-derived exosomes isolated after trauma have been shown to suppress in vitro bone marrow function. However, the cargo contained in these vesicles has not been examined. We hypothesized that trauma plasma-derived exosomes exhibit microRNA (miRNA) changes that impact bone marrow function after severe injury. Plasma was collected from a prospective cohort study of trauma patients (n = 15; 7 males, 8 females) with hip and/or femur fractures and an Injury Severity Score of ≥15; elective total hip arthroplasty (THA) patients (n = 8; 4 males, 4 females) served as operative controls. Exosomes were isolated from plasma with the Invitrogen Total Exosome Isolation Kit (Thermo Fisher Scientific, Waltham, MA), and RNA was isolated using a miRNeasy Mini Kit (Qiagen, Hilden, Germany). Direct quantification of miRNA was performed by NanoString Technologies on a human miRNA gene panel and analyzed with nSolver with significance defined as p < 0.05. There were no differences in age or sex distribution between trauma and THA groups; the average Injury Severity Score was 23. Trauma plasma-derived exosomes had 60 miRNA identities that were significantly downregulated and 3 miRNAs that were upregulated when compared with THA ( p < 0.05). Twelve of the downregulated miRNAs have a direct role in hematopoiesis regulation. Furthermore, male trauma plasma-derived exosomes demonstrated downregulation of 150 miRNAs compared with male THA ( p < 0.05). Female trauma plasma-derived exosomes demonstrated downregulation of only four miRNAs and upregulation of two miRNAs compared with female THA ( p < 0.05). We observed downregulation of 12 miRNAs linked to hematopoiesis along with sexual dimorphism in miRNA expression from plasma-derived exosomes following severe trauma. Understanding sexually dimorphic miRNA expression provides new insight into sex-based changes in postinjury systemic inflammation, immune system dysregulation, and bone marrow dysfunction and will aid us in more precise future potential therapeutic strategies. Prognostic and Epidemiological; Level III.

Assessment of measurement accuracy of amplified DNA using a colorimetric loop-mediated isothermal amplification assay.

A colorimetric loop-mediated isothermal amplification assay detects changes in pH during amplification based on color changes at a constant temperature. Currently, various studies havefocused on developing and assessing molecular point-of-care testing instruments. In this study, we evaluated amplified DNA concentrations measured using the colorimetric LAMP assay of the 1POT™ Professional device(1drop Inc, Korea). Results of the 1POTanalysis of clinical samples were compared with measurements obtained from the Qubit™ 4 and NanoDrop™ 2000 devices (both from Thermo Fisher Scientific, MA, USA). These results showed a correlation of 0.98 (95%CI: 0.96-0.99) and 0.96 (95% CI: 0.92-0.98) between 1POTand the Qubitand NanoDrop. 1POTcan measure amplified DNA accurately and is suitable for on-site molecular diagnostics.

Morphological peculiarities of revascularization of perifocal areas of brain infarction

Aim. To study pathomorphological signs of revascularization of perifocal areas of brain infarction in the dynamics of the acute period of the disease. Material and methods. Perifocal areas of brain infarction in patients who died at the 1st, 3rd, 7th, and 14th days were studied. General histopathological study was followed by 10 % ammonium silver impregnation and immunohistochemical examination using monoclonal antibodies Mo a-Hu CD34, Clone QBEnd/10 (“Thermo Fisher Scientific Inc.” – USA) and Mo a-Hu CD105 Endoglin were used. clone: SN6h1 (“DAKO”, Denmark). Vascular density and total cross-sectional area of vessels were studied. For the morphometric study, Videotest – Morphology 5.2.0.158 software (VideoTest LLC) was used. Statistical processing of the obtained data was carried out using the Statistica® for Windows 13.0 software. Results. According to the data of CD34 expression, from the 3rd day in the perifocal areas of brain infarction, the angiogenesis processes were increased: the density of blood vessels increased by 18.48 %, and the total cross-sectional area of vessels increased by 1.93 times. On the 7th day, the maximal vessel density and total cross-sectional area of vessels were registered in these areas; there was a vascular budding, formation of endothelial proliferates, and increased expression of CD105 on the vascular endothelium and pericytes, which indicates the active participation of the latter in angiogenesis. On the 14th day, blood vessel density and indicators of the total cross-sectional area of vessels decreased. During this period, the density of the astrocytes increased in the infarct’s perifocal areas with hypertrophy of their bodies and processes. Conclusions. According to CD34 and CD105 expression data, in the perifocal areas of cerebral infarction, the processes of angiogenesis gradually increase from the 3rd day manifesting by increase in the blood vessel density and total vascular cross-sectional area, as well as in the vascular budding and the formation of endothelial proliferates. Vascular remodeling is accompanied by increased density and hypertrophy of astroglia in the perifocal areas of cerebral infarction.

Morphological Control of Functional TiO2 Films on Magnetic Nanoparticle’s Surface by Utilizing the Boric Acid-Containing Solutions

In recent years, various functional materials are used for electronic devices such as the smartphones and conductive materials. For the further expansion of these material’s usability and usage range, it is important to achieve the highly functionalized material’s manufacturing processes compatible with “green” processes. From this point of view, the development of a sustainable functional material synthesis method that achieves both high performance and reduced the environmental load has been required.As one of such functional materials, Kishimoto1)et al. reported the synthesis and application of colored magnetic toners powders by using the buffer solution of pH 7-12 as a reaction field. In their methods, metallic iron particles coated with a homogeneous nanometer ranged TiO2 films are synthesized, and their color can be freely controlled by controlling the thickness of their films because of the interference effect.However, it is also known that not only the homogeneous core-shell materials but also the plate-like or island-like structures can be sometimes formed on the core surface. Needless to say, contamination of these lowering the property of the functional materials. Conversely, if we can elucidate the formation mechanism of these films and can achieve the selectively formation of various structured films, new functional materials can be synthesized.Therefore, in this study, TiO2 film-coated particles with various morphologies such as plate-like and dot island-like films are tried to synthesized, through the elucidation of the interaction between the surface condition and reaction solution.Carbonyl iron powder (BASF EM grade, median diameter: 4-5 μm), and Clark& Lubs buffer solution (H3BO3-KCl-NaOHaq, pH 9, 0.4 M) were used. Tetra-isopropoxy titanium (C14H28O4Ti), hydrogen peroxide water, and ammonia water were used as the titanium oxide film raw materials in the subsequent film coating step. The temperature was optimized and film coating was performed so that the peroxotitanium bonds to the substrate surface.Boric acid buffer solution after suspended the carbonyl iron particles were analyzed by XPS (XPS K-Alpha, Thermo Fisher Scientific, USA), and it showed a tendency to be significantly oxidized as compared to the particles before suspension. Waveform separation of the XPS spectrum of the O1s peak indicated that these peaks consisted by the oxygen atoms in the OH- and Fe2O3 species. ESI-TOF-MS (Bruker Dltonics; microTOFII) analysis results of the solution obtained after contacting the buffer solution and metallic iron particles shows that borate ions tended to decrease after the reaction. Thus, by controlling the relationship between the iron surface, boric acid, and the buffer solution in the solution, surface oxidation condition of the iron substance particles affected the formation of the subsequent TiO2 coating reaction. Details of other results will be present in our presentation.1) WO/2003/031683, POWDER COATED WITH TITANIA FILM AND METHOD FOR PRODUCTION THEREOF.2）G. Bhargava et.al, Characterization of the “native”surface thin film on pure polycrystalline iron: A high resolution XPS and TEM study,” Applied Surface Science2007; 253, 4322-4329.