Zeitschriftenartikel zum Thema „Spectroscopic digital markers“

Breast cancer affects the female population worldwide. Radiotherapy (RT) is part of the therapeutic modality in the management of breast cancer, after radical mastectomy or conserving surgery. The FTIR spectroscopic “marker bands” will lead us to approach the mechanism of skin damage due to the interaction of ionizing radiation and skin, on a molecular level at the very early stages. FT-IR spectroscopy, breast digital pictures, and ImageJ software were used in the study. Healthy breast skin was irradiated <em>ex-vivo</em> with a 4 Gy dose of a γ-⁶⁰Co course Gammachamber 4000A. The FT-IR spectra showed that the low-dose irradiation induces skin dehydration, collagen secondary structure changes and advanced glycation end products (AGEs) as a result of free radicals as mediated products. The infrared “marker bands” at about 1743, 1160, and 870 cm^-1 are characteristic, indicating the development of inflammation, glycations, and peroxidations respectively, due to ionizing radiation-induced oxidative stress. ImageJ analysis provided the sharp surface of the skin after RT irradiation in contrast to the smooth surface of the non-irradiated healthy skin. The most important damages, induced by radiotherapy, were connective tissue lesions, glycosylation, and phosphorylation processes in the skin. The reactive oxygen species (ROS) free radicals prefer to abstract H atoms from lipids, sugar rings of glycoproteins, and base ribose of DNA. The produced intermediate free radicals, as a result of ROS reactions, led to the formation of AGEs and peroxides.

Plasmalemma vesicle-associated protein (PLVAP) is the main component of endothelial diaphragms in fenestrae, caveolae, and transendothelial channels. PLVAP is expressed in the adult kidney glomerulus upon injury. Glomerular endothelial injury is associated with progressive loss of kidney function in diabetic kidney disease (DKD). This study aimed to investigate whether PLVAP could serve as a marker for glomerular endothelial damage in DKD. Glomerular PLVAP expression was analyzed in different mouse models of DKD and their respective healthy control animals using automatic digital quantification of histological whole kidney sections. Transgenic mice expressing a dominant-negative GIP receptor (GIPRdn) in pancreatic beta-cells as a model for diabetes mellitus (DM) type 1 and black and tan brachyuric (BTBR) ob/ob mice, as a model for DM type 2, were used. Distinct PLVAP induction was observed in all diabetic models studied. Traces of glomerular PLVAP expression could be identified in the healthy control kidneys using automated quantification. Stainings for other endothelial injury markers such as CD31 or the erythroblast transformation-specific related gene (ERG) displayed no differences between diabetic and healthy groups at the time points when PLVAP was induced. The same was also true for the mesangial cells marker α8Integrin, while the podocyte marker nephrin appeared to be diminished only in BTBR ob/ob mice. Glomerular hypertrophy, which is one of the initial morphological signs of diabetic kidney damage, was observed in both diabetic models. These findings suggest that PLVAP is an early marker of glomerular endothelial injury in diabetes-induced kidney damage in mice.

Scientists today are pursuing the development of non-destructive and non-invasive methods for rapid and reliable diagnosis of diseases in digital form and reduction in the need for biopsies. In this paper we review the most recent studies supporting the application of Fourier Transform Infrared (FT-IR) spectroscopy and infrared thermography or medical thermography. Both are non-destructive digital techniques, which are promising to record and discriminate the local biochemical changes that are induced by the diseases, while the examined samples do not need any special preparation. The reflected infrared radiation from the affected areas of the body strongly depends on the metabolic steps of the cancer/or any other disease, which is also related to the structural changes at a molecular level of the biological molecules during enzymatic or non-enzymatic steps of the disease. The detection of the FT-IR spectral digital “marker bands” of the obtained changes of cell, liquids or tissue components are derived from the disease in the check point. Furthermore, ImageJ analysis of the thermal imaging in cancerous area showed aggregate formation upon cancer development as it was also indicated from the FT-IR spectra.

Glioblastoma, a highly aggressive brain tumor, poses significant treatment challenges. A deeper investigation into its molecular complexity is essential for the identification of novel prognostic biomarkers and therapeutic strategies, potentially improving patient outcomes in terms of survival and quality of life. While nuclear DNA mutations have been extensively studied, the role of mitochondrial DNA (mtDNA) mutations, specifically in the D-loop region, remains poorly understood. This prospective case-control study aimed to assess the prognostic significance of the mtDNA D-loop m.16126T>C variant in glioblastoma patients. Immunohistochemistry and droplet digital PCR (ddPCR) were employed for mutation analysis, complemented by statistical analyses and a literature review. The study cohort comprised 22 glioblastoma patients (mean age 59.36 ± 14.17, 12 (54.55%) females), and 25 controls (59.48 ± 13.22, 12 (80%) females). The D-loop m.16126T>C variant was observed in four (18%) of the glioblastoma samples and was associated with shorter median survival (9.5 vs. 18 months; p = 0.016, log-rank test). This study underscores the importance of investigating mtDNA, especially D-loop variants, in glioblastoma, suggesting its potential as a prognostic biomarker and, therefore, its possible therapeutic targets, warranting further exploration.

Neurotrophins constitute a group of growth factor that exerts important functions in the nervous system of vertebrates. They act through two classes of transmembrane receptors: tyrosine-kinase receptors and the p75 neurotrophin receptor (p75NTR). The activation of p75NTR can favor cell survival or apoptosis depending on diverse factors. Several studies evidenced a link between p75NTR and the pathogenesis of prion diseases. In this study, we investigated the distribution of several neurotrophins and their receptors, including p75NTR, in the brain of naturally scrapie-affected sheep and experimentally infected ovinized transgenic mice and its correlation with other markers of prion disease. No evident changes in infected mice or sheep were observed regarding neurotrophins and their receptors except for the immunohistochemistry against p75NTR. Infected mice showed higher abundance of p75NTR immunostained cells than their non-infected counterparts. The astrocytic labeling correlated with other neuropathological alterations of prion disease. Confocal microscopy demonstrated the co-localization of p75NTR and the astrocytic marker GFAP, suggesting an involvement of astrocytes in p75NTR-mediated neurodegeneration. In contrast, p75NTR staining in sheep lacked astrocytic labeling. However, digital image analyses revealed increased labeling intensities in preclinical sheep compared with non-infected and terminal sheep in several brain nuclei. This suggests that this receptor is overexpressed in early stages of prion-related neurodegeneration in sheep. Our results confirm a role of p75NTR in the pathogenesis of classical ovine scrapie in both the natural host and in an experimental transgenic mouse model.

Bipolar disorder (BD) is a complex neurobiological disorder characterized by a pathologic mood swing. Digital phenotyping, defined as the ‘moment-by-moment quantification of the individual-level human phenotype in its own environment’, represents a new approach aimed at measuring the human behavior and may theoretically enhance clinicians’ capability in early identification, diagnosis, and management of any mental health conditions, including BD. Moreover, a digital phenotyping approach may easily introduce and allow clinicians to perform a more personalized and patient-tailored diagnostic and therapeutic approach, in line with the framework of precision psychiatry. The aim of the present paper is to investigate the role of digital phenotyping in BD. Despite scarce literature published so far, extremely heterogeneous methodological strategies, and limitations, digital phenotyping may represent a grounding research and clinical field in BD, by owning the potentialities to quickly identify, diagnose, longitudinally monitor, and evaluating clinical response and remission to psychotropic drugs. Finally, digital phenotyping might potentially constitute a possible predictive marker for mood disorders.

Analysis of circulating tumor DNA (ctDNA) is a potential minimally invasive molecular tool to guide treatment decision-making and disease monitoring. A suitable diagnostic-grade platform is required for the detection of tumor-specific mutations with high sensitivity in the circulating cell-free DNA (ccfDNA) of cancer patients. In this multicenter study, the ccfDNA of 72 patients treated for advanced-stage non-small cell lung cancer (NSCLC) was evaluated using the UltraSEEK® Lung Panel on the MassARRAY® System, covering 73 hotspot mutations in EGFR, KRAS, BRAF, ERBB2, and PIK3CA against mutation-specific droplet digital PCR (ddPCR) and routine tumor tissue NGS. Variant detection accuracy at primary diagnosis and during disease progression, and ctDNA dynamics as a marker of treatment efficacy, were analyzed. A multicenter evaluation using reference material demonstrated an overall detection rate of over 90% for variant allele frequencies (VAFs) > 0.5%, irrespective of ccfDNA input. A comparison of UltraSEEK® and ddPCR analyses revealed a 90% concordance. An 80% concordance between therapeutically targetable mutations detected in tumor tissue NGS and ccfDNA UltraSEEK® analysis at baseline was observed. Nine of 84 (11%) tumor tissue mutations were not covered by UltraSEEK®. A decrease in ctDNA levels at 4–6 weeks after treatment initiation detected with UltraSEEK® correlated with prolonged median PFS (46 vs. 6 weeks; p < 0.05) and OS (145 vs. 30 weeks; p < 0.01). Using plasma-derived ccfDNA, the UltraSEEK® Lung Panel with a mid-density set of the most common predictive markers for NSCLC is an alternative tool to detect mutations both at diagnosis and during disease progression and to monitor treatment response.

DNA microarrays and RNA-based sequencing approaches are considered important discovery tools in clinical medicine. However, cross-platform reproducibility studies undertaken so far have highlighted that microarrays are not able to accurately measure gene expression, particularly when they are expressed at low levels. Here, we consider the employment of a digital PCR assay (ddPCR) to validate a gene signature previously identified by gene expression profile. This signature included ten Hedgehog (HH) pathways’ genes able to stratify multiple myeloma (MM) patients according to their self-renewal status. Results show that the designed assay is able to validate gene expression data, both in a retrospective as well as in a prospective cohort. In addition, the plasma cells’ differentiation status determined by ddPCR was further confirmed by other techniques, such as flow cytometry, allowing the identification of patients with immature plasma cells’ phenotype (i.e., expressing CD19+/CD81+ markers) upregulating HH genes, as compared to others, whose plasma cells lose the expression of these markers and were more differentiated. To our knowledge, this is the first technical report of gene expression data validation by ddPCR instead of classical qPCR. This approach permitted the identification of a Maturation Index through the integration of molecular and phenotypic data, able to possibly define upfront the differentiation status of MM patients that would be clinically relevant in the future.

Natural extracts are broadly utilized as remedies, nutrition additives, cosmetics or flavors as well as natural pesticides, fungicides or herbicides. Green manufacturing technologies are of added market value and are sustainable towards the climate neutrality politically demanded for 2045. The concept of digital twins involves experimentally distinct validated process models combined with process analytical technology that is to be adapted to the existing operations. This is a key technology for the autonomous operations in industry 4.0. This paper exemplifies this approach and evaluates the results of the application and implementation efforts of regulated industries. A conductivity sensor for the measurement of the dry residue content and/or Fourier-transformed infrared spectroscopy for marker/lead or reference substance concentration determination are the most feasible and straight forward solutions. Different process control concepts from simple PID controllers (proportional, integral and differential) to advanced process control using digital twin models are evaluated and discussed in terms of industrialization efforts and benefits. The global warming potential CO2 equivalent per kg of natural product could be decreased by a factor of 5–10 as well as the cost of goods, which makes the pay-out time for the industrialization investment less than 1 year and the approach highly competitive. The success rate of the extraction process under regulatory constraints can be raised to 100%, reducing waste, overall solvent consumption, personnel efforts and energy requirements to a minimum.

Lung cancer still has one of the highest morbidity and mortality rates among all types of cancer. Its incidence continues to increase, especially in developing countries. Although the medical field has witnessed the development of targeted therapies, new treatment options need to be developed urgently. For the discovery of new drugs, human cancer models are required to study drug efficiency in a relevant setting. Here, we report the generation of a non-small cell lung cancer model with a perfusion system. The bioprinted model was produced by digital light processing (DLP). This technique has the advantage of including simulated human blood vessels, and its simple assembly and maintenance allow for easy testing of drug candidates. In a proof-of-concept study, we applied gemcitabine and determined the IC50 values in the 3D models and 2D monolayer cultures and compared the response of the model under static and dynamic cultivation by perfusion. As the drug must penetrate the hydrogel to reach the cells, the IC50 value was three orders of magnitude higher for bioprinted constructs than for 2D cell cultures. Compared to static cultivation, the viability of cells in the bioprinted 3D model was significantly increased by approximately 60% in the perfusion system. Dynamic cultivation also enhanced the cytotoxicity of the tested drug, and the drug-mediated apoptosis was increased with a fourfold higher fraction of cells with a signal for the apoptosis marker caspase-3 and a sixfold higher fraction of cells positive for PARP-1. Altogether, this easily reproducible cancer model can be used for initial testing of the cytotoxicity of new anticancer substances. For subsequent in-depth characterization of candidate drugs, further improvements will be necessary, such as the generation of a multi-cell type lung cancer model and the lining of vascular structures with endothelial cells.

Background: Recently, there has been much debate in the US concerning drug importation from Canadian Internet pharmacies. The Food and Drug Administration and US drug manufacturers assert that drugs obtained from international markets via the Internet present a health risk to consumers from substandard products. The public's perception is that drugs from Canada are as safe as those from the US. Objective: To determine whether simvastatin tablets obtained via the Internet from Canadian generic manufacturers are comparable in blend uniformity, a major attribute of tablet quality, with the US innovator product. Methods: Generic simvastatin tablets from 4 Canadian Internet pharmacy Web sites and the US innovator product were obtained for pharmaceutical analysis, Tablet samples were analyzed using near-infrared spectroscopic imaging techniques, which are designed to detect formulation defects of drug products during the manufacturing process. Digital images were created, revealing the tablets’ internal structures. Results: The blend uniformity of the active pharmaceutical ingredient in the tablet samples from Canada was determined and compared with that of the US innovator product. Results indicated that there is little significant difference in blend uniformity among US innovator and Canadian generic tablets. Conclusions: Results of this study suggest comparable quality assurance manufacturing standards for the US innovator product and the Canadian generic drug products tested. These findings have clinical, legal, and economic implications that should be addressed by policy makers to safeguard consumers who choose to purchase Canadian-manufactured drugs via the Internet.

We report on oxygenation changes noninvasively recorded by multichannel continuous-wave near infrared spectroscopy (CW-NIRS) during endovascular neuroradiologic interventions requiring temporary balloon occlusion of arteries supplying the cerebral circulation. Digital subtraction angiography (DSA) provides reference data on the site, timing, and effectiveness of the flow stagnation as well as on the amount and direction of collateral circulation. This setting allows us to relate CW-NIRS findings to brain specific perfusion changes. We focused our analysis on the transition from normal perfusion to vessel occlusion, i.e., before hypoxia becomes clinically apparent. The localization of the maximal response correlated either with the core (occlusion of the middle cerebral artery) or with the watershed areas (occlusion of the internal carotid artery) of the respective vascular territories. In one patient with clinically and angiographically confirmed insufficient collateral flow during carotid artery occlusion, the total hemoglobin concentration became significantly asymmetric, with decreased values in the ipsilateral watershed area and contralaterally increased values. Multichannel CW-NIRS monitoring might serve as an objective and early predictive marker of critical perfusion changes during interventions—to prevent hypoxic damage of the brain. It also might provide valuable human reference data on oxygenation changes as they typically occur during acute stroke.

Uterine serous carcinoma (USC) and carcinosarcomas (CSs) are rare, highly aggressive variants of endometrial cancer. No reliable tumor biomarkers are currently available to guide response to treatment or detection of early recurrence in USC/CS patients. Circulating tumor DNA (ctDNA) identified using ultrasensitive technology such as droplet digital polymerase chain reaction (ddPCR) may represent a novel platform for the identification of occult disease. We explored the use of personalized ctDNA markers for monitoring USC and CS patients. Tumor and plasma samples from USC/CS patients were collected at the time of surgery and/or during the treatment course for assessment of tumor-specific somatic structural variants (SSVs) by a clinical-grade next-generation sequencing (NGS) platform (i.e., Foundation Medicine) and a droplet digital PCR instrument (Raindance, ddPCR). The level of ctDNA was quantified by droplet digital PCR in plasma samples and correlated to clinical findings, including CA-125 serum and/or computed tomography (CT) scanning results. The genomic-profiling-based assay identified mutated “driver” target genes for ctDNA analysis in all USC/CS patients. In multiple patients, longitudinal ctDNA testing was able to detect the presence of cancer cells before the recurrent tumor was clinically detectable by either CA-125 or CT scanning. Persistent undetectable levels of ctDNA following initial treatment were associated with prolonged progression-free and overall survival. In a USC patient, CA-125 and TP53 mutations but not PIK3CA mutations become undetectable in the plasma at the time of recurrence, suggesting that more than one customized probe should be used for monitoring ctDNA. Longitudinal ctDNA testing using tumor-informed assays may identify the presence of residual tumors, predict responses to treatment, and identify early recurrences in USC/CS patients. Recognition of disease persistence and/or recurrence through ctDNA surveillance may allow earlier treatment of recurrent disease and has the potential to change clinical practice in the management of USC and CS patients. CtDNA validation studies in USC/CS patients prospectively enrolled in treatment trials are warranted.

Adults do not engage in enough physical activity. Investigating cognitive and physiological factors related to improving this behavior—and reducing health risks—remains a public health priority. Our objective was to assess whether cognitive flexibility influenced perceptions and choice of exercise programs and whether flexibility was associated with cardiovascular disease (CVD) risk factors. Independent sample groups of college-aged adults (18–24 yrs) participated in two studies. Data were collected on individuals’ degree of cognitive flexibility (both self-reported and objectively measured), perceptions and choice of exercise programs, and health status markers known to be associated with CVD (vascular function, muscular strength, and body composition). Vascular function was assessed with a near-infrared spectroscopy device, strength was defined as handgrip, and body composition was estimated via digital circumferences. Self-reported flexibility reliably predicted individuals’ choice of exercise program and perceptions of effort required for success on an exercise program. The relationships among CVD risk factors and objectively measured cognitive flexibility were not significant, demonstrating that identifying a healthy individual’s degree of performance-based cognitive flexibility does not predict health status. Furthermore, although greater self-reported trait flexibility (rigidity) is known to predict higher (lower) likelihood of physical activity, this finding should not be extrapolated to also assume that flexibility (rigidity), as measured by objective cognitive tests, is associated with reduced CVD risk in healthy adults. We posit a rationale for how understanding cognitive flexibility and rigidity can play an impactful role in improving adherence to exercise prescriptions targeted to reducing risks.

In the setting of pronounced inflammation, changes in the epithelium may overlap with neoplasia, often rendering it impossible to establish a diagnosis with certainty in daily clinical practice. Here, we discuss the underlying molecular mechanisms driving tissue response during persistent inflammatory signaling along with the potential association with cancer in the gastrointestinal tract, pancreas, extrahepatic bile ducts, and liver. We highlight the histopathological challenges encountered in the diagnosis of chronic inflammation in routine practice and pinpoint tissue-based biomarkers that could complement morphology to differentiate reactive from dysplastic or cancerous lesions. We refer to the advantages and limitations of existing biomarkers employing immunohistochemistry and point to promising new markers, including the generation of novel antibodies targeting mutant proteins, miRNAs, and array assays. Advancements in experimental models, including mouse and 3D models, have improved our understanding of tissue response. The integration of digital pathology along with artificial intelligence may also complement routine visual inspections. Navigating through tissue responses in various chronic inflammatory contexts will help us develop novel and reliable biomarkers that will improve diagnostic decisions and ultimately patient treatment.

We introduce here a novel machine learning (ML) framework to address the issue of the quantitative assessment of the immune content in neuroblastoma (NB) specimens. First, the EUNet, a U-Net with an EfficientNet encoder, is trained to detect lymphocytes on tissue digital slides stained with the CD3 T-cell marker. The training set consists of 3782 images extracted from an original collection of 54 whole slide images (WSIs), manually annotated for a total of 73,751 lymphocytes. Resampling strategies, data augmentation, and transfer learning approaches are adopted to warrant reproducibility and to reduce the risk of overfitting and selection bias. Topological data analysis (TDA) is then used to define activation maps from different layers of the neural network at different stages of the training process, described by persistence diagrams (PD) and Betti curves. TDA is further integrated with the uniform manifold approximation and projection (UMAP) dimensionality reduction and the hierarchical density-based spatial clustering of applications with noise (HDBSCAN) algorithm for clustering, by the deep features, the relevant subgroups and structures, across different levels of the neural network. Finally, the recent TwoNN approach is leveraged to study the variation of the intrinsic dimensionality of the U-Net model. As the main task, the proposed pipeline is employed to evaluate the density of lymphocytes over the whole tissue area of the WSIs. The model achieves good results with mean absolute error 3.1 on test set, showing significant agreement between densities estimated by our EUNet model and by trained pathologists, thus indicating the potentialities of a promising new strategy in the quantification of the immune content in NB specimens. Moreover, the UMAP algorithm unveiled interesting patterns compatible with pathological characteristics, also highlighting novel insights into the dynamics of the intrinsic dataset dimensionality at different stages of the training process. All the experiments were run on the Microsoft Azure cloud platform.

There is a debate regarding the prediction of lymph node metastasis (LNM) in pedunculated T1 colorectal cancer (CRC). In this study with four cases of pedunculated T1 CRCs, we aimed to investigate gene expression variations based on the distance from the Haggitt line (HL) and identify potential molecular risk factors for LNM. By leveraging the Cancer Transcriptome Atlas and digital spatial profiling technology, we meticulously analyzed discrete regions, including the head, HL, proximal stalk region (300–1000 μm from HL), and distal stalk region (1500–2000 μm from HL) to identify spatially sequential molecular changes. Our findings showed significant overall gene expression variations among the head, proximal stalk, and distal stalk regions of pedunculated T1 CRCs compared to the control adenoma. Compared to LNM-negative T1 CRCs, LNM-positive T1 CRC showed that the expression of genes involved in immune-related pathways such as B2M, HLA-B, and HLA-E were significantly downregulated in the distal stalk region compared to the proximal stalk region. In summary, our results may tentatively suggest considering endoscopic resection of the stalk with a minimum 2000 μm margin from the HL, taking into account the gene expression alterations related to immune-related pathways. However, we acknowledge the limitations of this pilot study, notably the small case series, which may restrict the depth of interpretation. Further validation is imperative to substantiate these findings.

To the current data, there have been 6,955,141 COVID-19-related deaths worldwide, reported to WHO. Toll-like receptors (TLRs) implicated in bacterial and virus sensing could be a crosstalk between activation of persistent innate-immune inflammation, and macrophage’s sub-population alterations, implicated in cytokine storm, macrophage over-activation syndrome, unresolved Acute Respiratory Disease Syndrome (ARDS), and death. The aim of this study is to demonstrate the association between Toll-like-receptor-4 (TLR-4)-induced inflammation and macrophage imbalance in the lung inflammatory infiltrate of lethal COVID-19 disease. Twenty-five cases of autopsy lung tissues were studied by digital pathology-based immunohistochemistry to evaluate expression levels of TLR-4 (CD 284), pan-macrophage marker CD68 (clone KP1), sub-population marker related to alveolar macrophage Galectin-3 (GAL-3) (clone 9C4), and myeloid derived CD163 (clone MRQ-26), respectively. SARS-CoV-2 viral persistence has been evaluated by in situ hybridation (ISH) method. This study showed TLR-4 up-regulation in a subgroup of patients, increased macrophage infiltration in both Spike-1(+) and Spike-1(−) lungs (p < 0.0001), and a macrophage shift with important down-regulation of GAL-3(+) alveolar macrophages associated with Spike-1 persistence (p < 0.05), in favor of CD163(+) myeloid derived monocyte-macrophages. Data show that TLR-4 expression induces a persistent activation of the inflammation, with inefficient resolution, and pathological macrophage shift, thus explaining one of the mechanisms of lethal COVID-19.

Complex regional geologic structural controls have generated a lot of interest in the engineering, oil, and gas industries within the past few years. Digital elevation models (DEMs), multispectral remote sensing images using ArcGIS software, in combination with data cube and geomorphologic characterization, provide important markers that aid in spatial information analysis for the study area. We have validated the characterization and classification of DEMs using spatial statistics by mineral spectroscopy of multispectral remote sensing data. Our characterization was initiated by a joint interpretation of DEMs and multispectral remote sensing data in association with stratigraphic and geologic information. We have combined Landsat ETM+ images from visible (VIS), near-infrared (NIR), and mid-infrared (MID IR) to create red-green-blue (RGB) images, superimposed with high-spectral-resolution 15 m panchromatic band 8. Principal component analysis (PCA) further enhanced the image results. To characterize the geomorphology and near surface, specific bands used included RGB Landsat 742 and 321 data sets, whereas false-color Landsat RGB images (742 and 432) provided spatial data in delineating areas of lineations and fault systems. The tectonic lineaments extracted from the escarpments of the DEM and magnetic data provided structures related to tectonic forces to better understand the major faults, lineations, and geomorphology. Results of this study showed a strikingly reliable interpretative result of these faults that controlled the low-lying areas. These faults and lineations are high-permeability zones that can be saturated by water during active rainfall and flash-flood periods thereby disrupting the equilibrium of various fault zones in the area and raising tectonic activities within the active fault system. Such saturation presents a major environmental hazard for the study area. Generally, the use of Landsat data combined with PCA indicates promising evidence of possible plays within the huge sedimentary deposits and raised concerns about safety and hazard issues.

The management and screening of prostate cancer (PC) is still the main problem in clinical practice. In this study, we investigated the role of aggressiveness genetic markers for PC stratification. We analyzed 201 plasma samples from PC patients and controls by digital PCR. For selection and validation, 26 formalin-fixed paraffin-embedded tissues, 12 fresh tissues, and 24 plasma samples were characterized by RNA-Seq, immunochemistry, immunofluorescence, Western blot, and extracellular-vesicles analyses. We identified three novel non-invasive biomarkers; all with an increased expression pattern in patients (PCA3: p = 0.002, S100A4: p ≤ 0.0001 and MRC2: p = 0.005). S100A4 presents the most informative AUC (area under the curve) (0.735). Combination of S100A4, MRC2, and PCA3 increases the discriminatory power between patients and controls and between different more and less aggressive stages (AUC = 0.761, p ≤ 0.0001). However, although a sensitivity of 97.47% in PCA3 and a specificity of 90.32% in S100A4 was reached, the detection signal level could be variable in some analyses owing to tumor heterogeneity. This is the first time that the role of S100A4 and MRC2 has been described in PC aggressiveness. Moreover, the combination of S100A4, MRC2, and PCA3 has never been described as a non-invasive biomarker for PC screening and aggressiveness.

The identification of markers for early diagnosis, prognosis, and improvement of therapeutic options represents an unmet clinical need to increase survival in Non-Small Cell Lung Cancer (NSCLC), a neoplasm still characterized by very high incidence and mortality. Here, we investigated whether proline dehydrogenase (PRODH), a mitochondrial flavoenzyme catalyzing the key step in proline degradation, played a role in NSCLC tumorigenesis. PRODH expression was investigated by immunohistochemistry; digital PCR, quantitative PCR, immunoblotting, measurement of reactive oxygen species (ROS), and functional cellular assays were carried out. PRODH expression was found in the majority of lung adenocarcinomas (ADCs). Patients with PRODH-positive tumors had better cancer-free specific and overall survival compared to those with negative tumors. Ectopic modulation of PRODH expression in NCI-H1299 and the other tested lung ADC cell lines decreased cell survival. Moreover, cell proliferation curves showed delayed growth in NCI-H1299, Calu-6 and A549 cell lines when PRODH-expressing clones were compared to control clones. The 3D growth in soft agar was also impaired in the presence of PRODH. PRODH increased reactive oxygen species production and induced cellular senescence in the NCI-H1299 cell line. This study supports a role of PRODH in decreasing survival and growth of lung ADC cells by inducing cellular senescence.

Minimal residual disease (MRD) is of major importance in onco-hematology, particularly in acute myeloid leukemia (AML). MRD measures the amount of leukemia cells remaining in a patient after treatment, and is an essential tool for disease monitoring, relapse prognosis, and guiding treatment decisions. Patients with a negative MRD tend to have superior disease-free and overall survival rates. Considerable effort has been made to standardize MRD practices. A variety of techniques, including flow cytometry and molecular methods, are used to assess MRD, each with distinct strengths and weaknesses. MRD is recognized not only as a predictive biomarker, but also as a prognostic tool and marker of treatment efficacy. Expected advances in MRD assessment encompass molecular techniques such as NGS and digital PCR, as well as optimization strategies such as unsupervised flow cytometry analysis and leukemic stem cell monitoring. At present, there is no perfect method for measuring MRD, and significant advances are expected in the future to fully integrate MRD assessment into the management of AML patients.

Irritable bowel syndrome (IBS) involves low-grade mucosal inflammation. Among the various approaches capable of managing the symptoms, physical activity is still under investigation. Despite its benefits, it promotes oxidative stress and inflammation. Mitochondria impacts gut disorders by releasing damage-associated molecular patterns, such as cell-free mtDNA (cf-mtDNA), which support inflammation. This study evaluated the effects of a 12-week walking program on the cf-mtDNA and DNase in 26 IBS and 17 non-IBS subjects. Pro- and anti-inflammatory cytokines were evaluated by ELISA. Digital droplet PCR was used to quantify cf-mtDNA; DNase activity was assessed using a single radial enzyme diffusion assay. PCR-RFLP was used to genotype DNASE1 rs1053874 SNP. Significantly lower IL-10 levels were found in IBS than in non-IBS individuals. Exercise reduced cf-mtDNA in non-IBS subjects but not in IBS patients. DNase activity did not correlate with the cf-mtDNA levels in IBS patients post-exercise, indicating imbalanced cf-mtDNA clearance. Different rs1053874 SNP frequencies were not found between groups. The study confirms the positive effects of regular moderate-intensity physical activity in healthy subjects and its role in cf-mtDNA release and clearance. Walking alone might not sufficiently reduce subclinical inflammation in IBS, based on imbalanced pro- and anti-inflammatory molecules. Prolonged programs are necessary to investigate their effects on inflammatory markers in IBS.

Research on tumor angiogenesis has mainly focused on the vascular endothelial growth factor (VEGF) family and on methods to block its actions. However, reports on VEGF receptor (VEGFR) expression in tumor-associated endothelial cells (ECs) are limited. Thus, we evaluated VEGF, VEGFR-1 and VEGFR-2 expression in ECs of colorectal cancer (CRC) using immunohistochemistry. VEGF, VEGFR-1 and -2 expression in ECs was quantitatively evaluated by digital image analysis in a retrospective series of 204 tumor tissue samples and related to clinical variables. The data show that the VEGF, VEGFR-1 and VEGFR-2 expression in ECs is heterogeneous. Multivariate analysis including a set of clinicopathological variables reveals that high EC VEGFR-1 expression is an independent prognostic factor for overall survival (OS). The combination of low VEGFR-1 and high VEGFR-2 expression in ECs outperforms models integrating VEGFR-1 and VEGFR-2 as separate markers. Indeed, this VEGFR-1_VEGFR-2 combination is an independent negative prognostic factor for OS (p = 0.012) and metastasis-free survival (p = 0.007). In conclusion, this work illustrates the importance of studying the distribution of VEGF members in ECs of CRC. Interestingly, our preliminary data suggest that high VEGFR-1 and low VEGFR-2 expression in ECs appear to be involved in the progression of CRC, suggesting that targeting EC VEGFR-1 could offer novel opportunities for CRC treatment. However, a prospective validation study is needed.

Despite the success of current therapy concepts, patients with advanced non-small-cell lung cancer (NSCLC) still have a very poor prognosis. Therefore, biological markers are urgently needed, which allow the assessment of prognosis, or prediction of the success of therapy or resistance in this disease. Circulating microRNAs (miRs) have potential as biomarkers for the prognosis and prediction of response to therapy in cancer patients. Based on recent evidence that circulating miR-16, miR-29a, miR-144 and miR-150 can be regulated by ionizing radiation, the concentration of these four miRs was assessed in the plasma of NSCLC patients at different time points of radiotherapy by digital droplet PCR (ddPCR). Furthermore, their impact on patients’ prognosis was evaluated. The mean plasma levels of miR-16, miR-29a, miR-144 and miR-150 significantly differed intra- and inter-individually, and during therapy in NSCLC patients, but showed a strong positive correlation. The individual plasma levels of miR-16, miR-29a and miR-144 had prognostic value in NSCLC patients during or at the end of radiotherapy in Cox’s regression models. NSCLC patients with low levels of these three miRs at the end of radiotherapy had the worst prognosis. However, miR-150 plasma levels and treatment-dependent changes were not predictive. In conclusion, circulating miR-16, miR-29a and miR-144, but not miR-150, have a prognostic value in NSCLC patients undergoing radiotherapy.

The therapeutic potential of Musashi (MSI) RNA-binding proteins, important stemness-associated gene expression regulators, remains insufficiently understood in breast cancer. This study identifies the interplay between MSI protein expression, stem cell characteristics, radioresistance, cell invasiveness and migration. MSI-1, MSI-2 and Notch pathway elements were investigated via quantitative polymerase chain reaction (qPCR) in 19 triple-negative breast cancer samples. Measurements were repeated in MDA-MB-231 cells after MSI-1 and -2 siRNA-mediated double knockdown, with further experiments performed after MSI silencing. Flow cytometry helped quantify expression of CD44 and leukemia inhibitory factor receptor (LIFR), changes in apoptosis and cell cycle progression. Proliferation and irradiation-induced effects were assessed using colony formation assays. Radiation-related proteins were investigated via Western blots. Finally, cell invasion assays and digital holographic microscopy for cell migration were performed. MSI proteins showed strong correlations with Notch pathway elements. MSI knockdown resulted in reduction of stem cell marker expression, cell cycle progression and proliferation, while increasing apoptosis. Cells were radiosensitized as radioresistance-conferring proteins were downregulated. However, MSI-silencing-mediated LIFR downregulation resulted in enhanced cell invasion and migration. We conclude that, while MSI knockdown results in several therapeutically desirable consequences, enhanced invasion and migration need to be counteracted before knockdown advantages can be fully exploited.

CD8+ T cells are the main effector cells of anti-cancer immune response that can be regulated by various costimulatory and coinhibitory molecules, including members of the B7 family. B7 homolog 3 (B7-H3) appears as a promising marker for immunotherapy; however, its significance in gastric cancer (GC) is unclear yet. We evaluated the spatial distribution of CD8+ T cells in relation to the expression of B7-H3 by double immunohistochemical staining. The level of B7-H3 intensity was scored manually (0–3) and dichotomized into B7-H3-low and B7-H3-high groups. The distribution and density of CD8+ T cells was analysed using whole slide digital imaging. B7-H3 was expressed mainly in the stromal compartment of GC (n = 73, 76% of all cases). Tumours with high expression of B7-H3 showed larger spatial differences of CD8+ T cells (86.4/mm2 in tumour centre vs. 414.9/mm2 in invasive front) when compared to B7-H3-low group (157.7/mm2 vs. 218.7/mm2, respectively) (p < 0.001). This study provides insight into the expression pattern of B7-H3 in GC of Western origin. In GCs with higher level of B7-H3 expression, CD8+ T cells were spatially suppressed in the tumour centre suggesting that B7-H3 might be involved in tumour escape mechanisms from the immune response.

TRIM28 is a scaffold protein that interacts with DNA-binding proteins and recruits corepressor complexes to cause gene silencing. TRIM28 contributes to physiological functions such as cell growth and differentiation. In the chronic myeloid leukemia cell line K562, we edited TRIM28 using CRISPR/Cas9 technology, and the complete and partial knockout (KO) cell clones were obtained and confirmed using quantitative droplet digital PCR (ddPCR) technology. The amplicon sequencing demonstrated no off-target effects in our gene editing experiments. The TRIM28 KO cells grew slowly and appeared red, seeming to have a tendency towards erythroid differentiation. To understand how TRIM28 controls K562 cell proliferation and differentiation, transcriptome profiling analysis was performed in wild-type and KO cells to identify TRIM28-regulated genes. Some of the RNAs that encode the proteins regulating the cell cycle were increased (such as p21) or decreased (such as cyclin D2) in TRIM28 KO cell clones; a tumor marker, the MAGE (melanoma antigen) family, which is involved in cell proliferation was reduced. Moreover, we found that knockout of TRIM28 can induce miR-874 expression to downregulate MAGEC2 mRNA via post-transcriptional regulation. The embryonic epsilon-globin gene was significantly increased in TRIM28 KO cell clones through the downregulation of transcription repressor SOX6. Taken together, we provide evidence to demonstrate the regulatory network of TRIM28-mediated cell growth and erythroid differentiation in K562 leukemia cells.

Prostate cancer affects millions of men globally. The prostate cancer-associated gene ANO7 is downregulated in advanced prostate cancer, whereas benign tissue and low-grade cancer display varying expression levels. In this study, we assess the spatial correlation between ANO7 mRNA and protein using fluorescent in situ hybridization and immunohistochemistry for the detection of mRNA and protein in parallel sections of tissue microarrays prepared from radical prostatectomy samples. We show that ANO7 mRNA and protein expression correlate in prostate tissue. Furthermore, we show that ANO7 mRNA is enriched in the nuclei of the luminal cells at 89% in benign ducts and low-grade cancer, and at 78% in high-grade cancer. The nuclear enrichment of ANO7 mRNA was validated in prostate cancer cell lines 22Rv1 and MDA PCa 2b using droplet digital polymerase chain reaction (ddPCR) on RNA isolated from nuclear and cytoplasmic fractions of the cells. The nuclear enrichment of ANO7 mRNA was compared to the nuclearly-enriched lncRNA MALAT1, confirming the surprisingly high nuclear retention of ANO7 mRNA. ANO7 has been suggested to be used as a diagnostic marker and a target for immunotherapy, but a full comprehension of its role in prostate cancer progression is currently lacking. Our results contribute to a better understanding of the dynamics of ANO7 expression in prostatic tissue.

Background: This paper demonstrates the use of optical diagnostic methods to assess the dynamic skin changes observed in acute and chronic exposure to ultraviolet (UV) radiation in vivo. Methods: Firstly, in order to initiate photoaging (chronic UV exposure), animals (n = 40) were divided into two groups: chronic UV exposure (n = 30), and control (n = 10; without irradiation). Photoaging in animals was induced by chronic repeated exposure to UVA radiation three times per week, for 12 weeks continuously, while the UV dose increased stepwise over the course of the experiment (55 minimal erythema doses (MED) in total). Laser fluorescence spectroscopy (LFS), optical tissue oximetry (OTO), laser Doppler flowmetry (LDF), and optical coherence tomography (OCT) of the shaved dorsum skin were performed regularly, once per week until the conclusion of the study. At 0, 5, and 12 weeks of the experiment, histological examination of animal tissues using hematoxylin/eosin and Masson’s trichrome staining was performed. At the second stage, erythema was induced in mice (n = 15) by acute UV exposure at high doses. The colorimetric assay of the image from a digital RGB camera was used to evaluate the erythema index. Results: The tissue content index ηcollagen of collagen was appropriate for the characterization of skin photoaging. Significant differences (p < 0.05) in ηcollagen were found between the control and photoaging groups from the 5th to the 9th week of the experiment. In addition, the rate of collagen degradation in the control group was about half that of the photoaging group. This marker allows the differentiation of photo- and chronoaging. OCT revealed the main optical layers of the skin in compliance with the histological pattern. The analysis of the RGB camera images provided visualization of the acute skin reaction to UV radiation. Conclusions: This study demonstrates the applicability of optical methods for the quantitative assessment of acute and chronic skin effects of UV exposure in vivo.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is closely related to some metabolic disorders, such as central obesity and type 2 diabetes (T2D). Glucagon-like peptide 1 receptor agonists (GLP-1RAs), such as semaglutide, may have therapeutic roles in MASLD associated with T2D. This study aims to investigate the molecular mechanisms underlying the effectiveness of semaglutide on MASLD in terms of progression from liver steatosis to fibrosis. We characterized exosomes from ten patients with type 2 diabetes (T2D) before (T0) and after 12 months (T12) of treatment with once-weekly subcutaneous semaglutide. Six of ten patients were considered responders to therapy (R) based on MASLD severity downgrading by at least one class according to a validated ultrasonographic (US) score. Normal hepatocytes (HEPA-RG) and stellate (LX-2) cells were challenged with exosomes from R and NR patients, isolated before and after 12 months of therapy. Exosomes from both R and NR patients isolated at T0 significantly affected LX-2 viability. After 12 months of treatment, only those isolated from R patients restored cell viability, whereas those from NR patients did not. No effects were observed on HEPA-RG cells. Exosomes at T12 from R but not from NR patients significantly decreased the production of α-SMA, a marker of LX-2 activation, a liver stellate cell model, and ph-SMAD2 and CTGF, involved in fibrosis processes. TGF-β1 was not modulated by the exosomes of R and NR patients. As a downstream effect, Vimentin, Collagen 1A1, and Fibronectin extracellular matrix components were also downregulated, as measured by droplets digital PCR. In conclusion, these results shed light on the potential effectiveness of semaglutide in preventing liver fibrosis in MASLD.

ABSTRACTProstate cancer (PC) is the commonest malignancy in men that causes significant morbidity and mortality. The incidence has quadrupled in the last three decades. This is predominantly due to its increased detection by excellent newer techniques like Prostate-specific antigen (PSA) evaluation, Transrectal ultrasonography (TRUS), Transrectal ultrasound-guided biopsy, Contrast enhanced ultra sound studies, Multiparametric (Mp) MRI (MpMRI) and Nuclear medicine. Its incidence shows a rise in India. With the availability of PSA and trans-rectal biopsy, nowadays the majority of prostate cancers (PC) are diagnosed at an asymptomatic early stage (T1). Most PC are adenocarcinomas while a small percentage are ductal carcinomas, mucinous carcinomas, signet ring cell carcinomas and small cell carcinomas. These variants have poor prognosis.The anatomy of prostate will help us to further understand the basis of TRUS studies. The whole prostate can be divided into Transition Zone (TZ),Central Zone (CZ) and Peripheral Zone (PZ). This zonal anatomy of prostate is vital to understand the PC, since PC is predominantly seen as follows: TZ – 20%; CZ – 10%; and PZ – 70%.PSA is an extremely valuable tool in the evaluation of PC. It is exclusively produced by the prostate and to a lesser extent by the seminal vesicles. It is present in all post-pubertal men and absent in women and men following radical prostatectomy. Though the PSA is a vital parameter to detect PC, it can also be elevated in: i) Benign prostrate hypertrophy; ii) Prostate inflammation; iii) Prostatic infarct; iv) Post-digital rectal examination; and v) Sexual activity. The normal value of PSA is 0-4 ng/mL. The two techniques that are available to assess PSA levels are polyclonal assay or monoclonal assay. The monoclonal assay is the most commonly used method the world over. The accepted PSA values are: <4ng/mL (normal); 4.0-10.00ng/mL (borderline) and >10 ng/mL (abnormal). Other than normal PSA values, there are other PSA parameters which are often useful in confirming the diagnosis of PC. These are: i) PSA density; ii) PSA velocity; iii) PSA doubling time; iv) Other markers like PCA3; and v) PC is associated with more protein bound PSA (less free PSA) than in BPH. Free PSA (FPSA) can enhance the specificity of the total PSA value for detection of the PC while reducing the number of unnecessary biopsies.Another new finding is that of levels of insulin like growth factor binding protein-2 (IGFBP-2) appear to be directly associated with the presence of PC. Prostate Biopsy: Ultrasound guided biopsy of the prostate still remains the most important technique for the diagnosis of PC. Different biopsies which are used for diagnosis of PC are: Saturation Biopsy, MRI Guided Biopsy and Fusion Biopsy. Use of Gleason Score for grading the PC: Gleason score is the grading system used to determine the aggressiveness of PC. This grading system can be used to choose appropriate treatment options. The tumour grades provide important information regarding how fast the cancer is likely to be growing and the likelihood of the cancer spreading to other parts of the body such as lymph nodes or bones. The pathologist assigns the grade of the tumour when he or she looks at the malignant cells under the microscope. The higher the Gleason grade, the more aggressive is the tumour. Histopathology: Variants of usual acinar adenocarcinoma defined in 2004 by the WHO, include atrophic, pseudohyperplastic, foamy, colloid, signet ring, etc. Recently, variants not included in the 2004 WHO classification are microcysticadeno carcinoma, prostatic intraepithelial neoplasia – adenocarcinoma, large cell neuro endocrine carcinoma and pleomorphic giant cell carcinoma. Other diagnostic modalities for PC are Colour Flow Imaging, Elastography, Contrast Enhanced Ultrasound (CEUS), MR Imaging of Prostate Malignancies. The MR Imaging consisting of the following: Prostate Imaging Reporting and Data System (PIRADS) refers to a structured reporting scheme for evaluating the prostate for PC. T1-weighted images are not helpful in differentiating different zones or detecting the lesion. However, invasion of neurovascular bundle, haemorrhage within the gland and loco-regional lymphadenopathy is better visualized on this sequence. DWI imaging plays an important role in determining PIRADS score, predominantly in peripheral zone neoplasm. It utilizes proton diffusion properties in water to produce image contrast. Thus prostate malignancy appears bright (hyperintense) on DWI with corresponding low values on ADC map (darkhypo intense).Of all functional MR imaging techniques, DW imaging is the most practical and simple in its use. Dynamic Contrast Scan is considered positive if a suspected lesion/ nodule on T2-W or DWI image reveals earlier than normal or more than normal enhancement (hyper enhancement), as routinely seen in lesions with malignant etiology. MR Spectroscopy reflects resonance frequencies that are unique for protons in different metabolites present at the sampled location. A change in the ratios of concentrations of these metabolites suggests abnormality within the tissue. Normal PZ has high concentration of citrate and polyamines and low concentration of choline and creatinine. A reversal of these, i.e. decrease in citrate peak due to altered metabolism and increase in choline and creatinine peaks in a suspected nodule on T2-W image may suggest malignancy. Recent studies reveal that MR spectroscopy is more specific and less sensitive that anatomic T2-W scan. Current Modalities of Prostate Cancer Treatments There are a wide variety of treatments available for the management of prostate cancer. Radical prostatectomy, external beam radiation and radioactive prostate seed implant are potential cures for the prostate cancer. Hormone therapy may force the cancer into a prolonged remission but does not provide a cure unless it is combined with other treatments. The most commonly used treatments include the following:• Watchful Waiting (Active surveillance)• Radical Prostatectomy (Robot assisted radical prostatectomy; Laparoscopic prostatectomy)• Radiation Therapy (External beam radiation; Radioactive prostate seed implants)• Hormone Therapy• Combination of Therapies• High Intensity Focused Ultrasound (HIFU) or Magnetic Resonance-guided Focussed Ultrasound Surgery (MRgFUS)• Others (Cryotherapy; Photodynamic therapy)• Metastatic Disease1. Hormonal therapy (orchidectomy)2. Anti androgens3. Luteinizing Hormone-releasing Hormone (LHRH) Agonists (Leuprolide; Goserelin; Triptorelin; Histrelin) – monthly to annual depot injection S.C. implantsMetastatic PC responds to androgen- ablation/deprivation therapy, which heralded the beginning of a new era PC therapy.

AbstractFinding markers to detect and identify compounds and components related to an explosive is an important task that could reduce threats of unlawful uses of explosives. The capability of Raman spectroscopy to characterize chemical composition of explosive compounds and the analysis of stable isotope ratios could be useful to identify specific markers to identify the origin of the material used. Using Raman spectrometer directly on-site after the accident scene could be a useful tool for an immediate characterization of the components. In addition, carbon and nitrogen analysis on some fragments collected on the accident scene will then confirm the characterization of the components. The proposed configuration of double technique, coupled with a digital database and algorithms, could be useful to run analysis, with fast response time, in work environment related to emergency situations.

AbstractTeotihuacan, the main city of the Classic period in Central Mexico (ca. 150/200–650 CE), was among many things, a colorful city. Through its application on mural painting, ceramics, lapidary, bone, textiles, and the human body itself, coloring materials gave meaning to the Teotihuacan reality. This research presents color as a socio-technological phenomenon from the archaeology of color, the anthropology of technology, and the application of the concept of technological style.From this scope, pictorial palettes of mural painting fragments from three Teotihuacan architectural compounds were studied by digital microscopy, colorimetry, fiber optic reflectance spectroscopy (FORS), X-ray fluorescence (XRF), Raman spectroscopy, infrared spectroscopy (FTIR), and scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM-EDS) in order to identify the color technology of pigments. This approach led us to identify materials shared by the three studied compounds, red and yellow earths, cinnabar, malachite, pseudomalachite, azurite, and the Teotihuacan grayish blue, found to contain a mix of bone black with calcite. Our results contribute to the discussion on the use of raw materials and their mixtures. We observed a standardization of pigment technology in Teotihuacan mural painting through the use of shared and standard formulations with four indicators that we propose as markers of a technological style for pigments, along with variations in the color materials that evidence the use of different strategies for the procurement and distribution of raw materials among the diverse architectural assemblages of the city.

Background: Alzheimer’s disease (AD) is the most prevalent neurodegenerative disease. The detection of early-stage AD is particularly desirable because it would allow early intervention. However, a minimally invasive, low-cost, and accurate discrimination or diagnostic method for AD is especially difficult in the earliest stage of AD. Objective: The aim of this research is to discover blood plasma spectral digital biomarkers of AD, develop a novel intelligent method for the discrimination of AD and accelerate the translation of Fourier transform infrared (FTIR) spectral-based disease discrimination methods from the laboratory to clinical practice. Methods: Since vibration spectroscopy can provide the structure and chemical composition information of biological samples at the molecular level, we investigated the potential of FTIR spectral biomarkers of blood plasma to differentiate between AD patients and healthy controls. Combined with machine learning technology, we designed a hierarchical discrimination system that provides reagent-free and accurate AD discrimination based on blood plasma spectral digital biomarkers of AD. Results: Accurate segregation between AD patients and healthy controls was achieved with 89.3% sensitivity and 85.7% specificity for early-stage AD patients, 92.8% sensitivity and 87.5% specificity for middle-stage AD patients, and 100% sensitivity and 100% specificity for late-stage AD patients. Conclusions: Our results show that blood plasma spectral digital biomarkers hold great promise as discrimination markers of AD, indicating the potential for the development of an inexpensive, reagent-free, and less laborious clinical test. As a result, our research outcome will accelerate the clinical application of spectral digital biomarkers and machine learning.

Abstract Background Operating an aircraft is associated with a large mental workload; however, knowledge of the mental workload of ROV operators is limited. The purpose of this study was to establish a digital system for assessing the mental workload of remotely operated vehicle (ROV) operators using hemodynamic parameters, and compare results of different groups with different experience levels. Method Forty-one trainee pilots performed flight tasks once daily for 5 consecutive days in a flight simulation. Forty-five pilots experienced pilots and 68 experienced drivers were also included. Hemodynamic responses were measured by functional near-infrared spectroscopy (fNIRS). Results The median duration of peak oxyhemoglobin was 147.13 s (interquartile range [IQR] 21.97, 401.70 s) in the left brain and 180.74 s (IQR 34.37, 432.01 s) in the right brain in the experienced pilot group, and 184.42 s (IQR 3.41, 451.81 s) on day 5 in the left brain and 160.30 s (IQR 2.62, 528.20 s) in the right brain in the trainee group. Conclusion Navigation training reduces peak oxyhemoglobin duration, and may potentially be used as a surrogate marker for mental workload of ROV operators. Peak oxyhemoglobin concentration during s task may allow development of a simplified scheme for optimizing flight performance based on the mental workload of a pilot.

Abstract Background Skeletal muscle nicotinamide adenine dinucleotide (NAD) concentrations are low in people with sarcopenia. Increasing NAD concentrations may offer a novel therapy. This study tested if acipimox (a NAD precursor) improves skeletal muscle NAD concentration and function in people with probable sarcopenia. Participants aged 65 and over with low walk speed (< 0.8 m/s) and low muscle strength (by 2019 European Working Group criteria) were recruited to this before and after, proof-of-concept study. Participants received acipimox 250 mg orally (twice or thrice daily according to creatinine clearance) + aspirin 75 mg daily (to prevent facial flushing) for 4 weeks. Muscle biopsy of the vastus lateralis, 31P magnetic resonance spectroscopy and a 7-digital mobility assessment were performed before starting acipimox and after 3 weeks of treatment. The primary outcome was change in skeletal muscle NAD concentration. Secondary outcomes included change in phosphocreatine recovery rate and measures of physical performance. Eleven participants (8 women), mean age 78.9 years (SD 4.3), were recruited. Mean walk speed at baseline was 0.69 m/s (SD 0.07). All completed baseline and follow-up visits. Median medication adherence was 95% (range 91–104%). There was no statistically significant difference in the primary outcome of change in NAD concentrations in skeletal muscle between baseline and follow-up [median difference: − 0.003 umol/g (IQR − 0.058 to 0.210); P = 0.26] or secondary outcomes. Nineteen none-serious adverse events were reported. Although the study protocol was feasible and well tolerated, acipimox did not improve skeletal muscle NAD concentration, biochemical markers or physical function in people with probable sarcopenia. ClinicalTrials.gov Identifier: ISRCTN (ISRCTN87404878).

Abstract Background Circulating tumor DNA (ctDNA) has become one of the crucial components for cancer detection with the increase of precision medicine practice. ctDNA has great potential as a blood-based biomarker for the detection and treatment of cancer in its early stages. The purpose of this article was to discuss ctDNA and how it can be utilized to detect cancer. The benefits and drawbacks of this cancer detection technology, as well as the field’s future possibilities in various cancer management scenarios, are discussed. Main text ctDNA has clinical applications in disease diagnosis and monitoring. It can be used to identify mutations of interest and genetic heterogeneity. Another use of ctDNA is to monitor the effects of therapy by detecting mutation-driven resistance. Different technologies are being used for the detection of ctDNA. Next-generation sequencing, digital PCR, real-time PCR, and mass spectrometry are used. Using dPCR makes it possible to partition and analyze individual target sequences from a complex mixture. Mass-spectrometry technology enables accurate detection and quantification of ctDNA mutations at low frequency. Surface-enhanced Raman spectroscopy (SERS) and UltraSEEK are two systems based on this technology. There is no unified standard for detecting ctDNA as it exists in a low concentration in blood. As there is no defined approach, false positives occur in several methods due to inadequate sensitivities. Techniques used in ctDNA are costly and there is a limitation in clinical settings. Short conclusion A detailed investigation is urgently needed to increase the test's accuracy and sensitivity. To find a standard marker for all forms of cancer DNA, more study is needed. Low concentrations of ctDNA in a sample require improved technology to provide the precision that low concentrations of ctDNA in a sample afford.

Institute of Cluster Oncology named after L.L. Levshin of the First Moscow State Medical University (Institute) is the assignee of the first oncological Institute in Europe – Institute named after Morozov for the Treatment of Tumors of the Imperial Moscow University. Currently, the Institute is a university experimental - clinical cluster, which conducts multilateral research in the field of biology and medicine. The publication is an overview of the main results of the Institute as part of the implementation of work on the state assignment in 2020–2021. In the field of experimental and clinical development of new methods for the treatment of various diseases of the head and neck. So, within the state task, a study was conducted that demonstrated the high efficiency of replacing jaw defects using 3D printing and the feasibility of further clinical research in this direction. A study was made of cancer biomarkers derived from the tumor tissue microenvironment (TME), with particular attention to the extracellular matrix (ECM) and its products of activity and degradation. ECM-associated extracellular vesicles, ECM biomechanical characteristics, and ECM-derived biomarkers predictive of response to immunotherapy were studied. ECM has been shown to be a critical factor in malignant tumors. It is recommended that ECM-derived biomarkers be included in diagnostic and prognostic marker panels in the clinic. It is shown that the use of modern systems of THz spectroscopy and visualization opens up new possibilities in the label-free diagnosis of tumors, the treatment of cancer and inflammatory diseases. Models have been developed to fully describe the interaction of a THz wave with human brain tissues within the framework of classical electrodynamics, which is important for future research in the field of THz tumor neurodiagnosis. The need for a systematic study of various modes of THz irradiation of tissues and cells, including continuous and pulsed radiation, different powers, number and duration of exposure cycles, is emphasized. Liposomes containing MANα1-2MAN-PEG-DOPE were first tested as a nanocarrier DNA vaccine in cattle as a prophylactic against bovine herpes-1 (BoHV-1) infection. The proposed method increases immunogenicity and leads to long-term immunity. A dendritic cell (DC) targeting strategy has been proposed that uses a specific receptor known as DC-SIGN with its ability to bind α1,2-mannobiase, which is present at the ends of oligosaccharides in some viruses, bacteria and other pathogens. The Digital Display Precision Predictor (DDPP) algorithm is presented, aimed at identifying transcriptomic predictors of treatment outcome. Transcriptomic analysis, based on comparisons between tumor and normal tissue, compared with genomic analysis alone increased the number of patients eligible for targeted therapy by about a third. Work has been carried out to study the activation of neutrophils as an important therapeutic target in the treatment of cancer, and the use of neutrophils or neutrophil membrane vesicles as drug delivery and targeting vehicles. It has been shown that lipid-based nanocarriers can open up many opportunities to overcome the limitations of cell therapy and be used to deliver appropriate growth factors to the site of tissue damage and create conditions conducive to cell regeneration. Delivery of therapeutic agents to brain tissue is one of the most important problems in medicine. The use of transferrin, a protein, is promising for activating the surface of nanoparticles in order to direct them to the brain. An analysis was made of the use of polymer micelles in the targeted delivery of anticancer drugs, gene therapy, and diagnostic agents. The advantages of dendrimers, a unique drug delivery system, are analyzed: efficient loading of therapeutic and imaging materials, desired delivery, universal choice of the route of administration, monodisperse system, improved pharmacokinetic and pharmacodynamic profiling. Lipid nanoparticles, dynamic polyconjugates, GalNAc-siRNA conjugates, exosomes, and erythrocyte systems have been reviewed, demonstrating efficient siRNA delivery to cancer cells. The combination of salinomycin and paclitaxel delivered by a liposomal preparation modified with mAb 2C5 has been demonstrated to have high antitumor efficacy. The effectiveness of cell-penetrating peptides (CPPs) in delivering drugs and genes (siRNA, pDNA) to tumor sites to combat drug resistance was analyzed. An experimental study was made of the effectiveness of direct electrical stimulation of the posterior cricoarytenoid muscle for controlled opening of the glottis. Neurostimulation has a great potential for stimulating the human larynx. The efficiency of transplantation of parietal cells obtained from the olfactory lining of the nose of rats and humans on changing the size of post-traumatic spinal cord cysts has been established. The pathways and profiles of DNA expression in uveal melanoma have been determined, which will contribute to the creation of prognostic models that can lead to an improvement in the prognosis for patients with this disease. The effectiveness of cell-penetrating peptides (CPPs) in delivering drugs and genes (siRNA, pDNA) to tumor sites to combat drug resistance was analyzed. An experimental study was made of the effectiveness of direct electrical stimulation of the posterior cricoarytenoid muscle for controlled opening of the glottis. Neurostimulation has a great potential for stimulating the human larynx. The transplantation of ensheathing cells obtained from the olfactory lining of the nose of rats and humans was found to be effective in changing the size of post-traumatic spinal cord cysts. The pathways and profiles of DNA expression in uveal melanoma have been determined, which will contribute to the creation of prognostic models that can lead to an improvement in the prognosis for patients with this disease. The accumulation of methylene blue both in various tissues and in immune cells has been established, which indicates a potential correction of the immune response in patients with COVID-19 and a change in the macrophage phenotype, which can be achieved by deactivating inflammatory macrophages in tissues using red laser radiation. When modeling septoplasty, a change in heart rate variability, an increase in the concentration of corticosterone in the blood plasma in rats was established. As a result of a comparative analysis of the effect of septoplasty and modeling of sinus lift in rats on changes in the frequency domain of heart rate variability were established an increase in the activity of the sympathetic nervous system and a shift in metabolism under the influence of postoperative inflammation. Based on the studies, it was found that photobiomodulation therapy after septoplasty shows better results compared to the standard rehabilitation of patients after septoplasty: it helps to reduce the severity of pain and inflammatory response to surgical stress and leads to less pronounced changes in the autonomic nervous system in response to surgical stress. A separate staging system for oropharyngeal cancer associated with human papillomavirus is considered. The advantages of three-dimensional (3D) surface scanning over ultrasound during preoperative planning are shown, especially in "difficult" areas of the face. Data were obtained on the three-dimensional course of the angular artery within the nasolabial sulcus depending on age, sex and body mass index. With age, the depth and lateral distance between the arteries and sulci decreases significantly, and therefore there is no guaranteed safe place in the midface for minimally invasive procedures. The results of the study will help improve safety in minimally invasive treatments. Provided objective evidence of antagonistic skin movement between the medial and lateral midface. The functional border, identified by the 3D image, corresponds to the anatomical location of the ligament line. When studying the relationship of forehead fascial anatomy with the effectiveness of treatment with neuromodulators, it was found that the deep injection technique leads to a better result. The effectiveness of laser-induced fluorescent diagnostics and photodynamic therapy with sublingual administration of 5-ALA to patients in the treatment of precancerous lesions of the oral cavity and larynx has been demonstrated. Публикация представляет собой обзор основных результатов Института в рамках выполнения работ по государственному заданию в 2020–2021 гг. в области экспериментальной и клинической разработки новых методов лечения различных заболеваний головы и шеи. Так, в рамках государственного задания было проведено исследование, которое продемонстрировало высокую эффективность замещения дефектов челюстей с использованием 3D-печати. Проведено исследование биомаркеров рака, полученных из микроокружения опухолевой ткани (TME), с особым вниманием к внеклеточному матриксу (ECM) и продуктам его деятельности и деградации. Изучены внеклеточные везикулы, ассоциированные с ECM, биомеханические характеристики ECM и полученные из ECM биомаркеры, прогнозирующие ответ на иммунотерапию. Показано, что использование современных систем ТГц-спектроскопии и визуализации открывает новые возможности в безметочной диагностике опухолей, терапии рака и воспалительных заболеваний. Липосомы, содержащие MANα1-2MAN-PEG-DOPE, были впервые протестированы в качестве наноносителя ДНК-вакцины на крупном рогатом скоте в качестве профилактического средства против инфекции бычьего герпеса-1. Предложена стратегия нацеливания на дендритные клетки, которая использует преимущества специфического рецептора, известного как DC-SIGN, с его способностью связывать α1,2-маннобиозу, которая присутствует на концах олигосахаридов в некоторых вирусах, бактериях и других патогенах. Представлен алгоритм Digital Display Precision Predictor (DDPP), направленный на выявление транскриптомных предикторов исхода лечения. Проведена работа по изучению активации нейтрофилов как важной мишени терапии при лечении рака, и использованию нейтрофилов или везикул нейтрофильных мембран в качестве средств доставки и нацеливания лекарственных препаратов. Показано, что наноносители на основе липидов могут открыть множество возможностей для преодоления ограничений клеточной терапии, и быть использованы для доставки соответствующих факторов роста к месту повреждения тканей и создания условий, способствующих регенерации клеток. Доставка терапевтических средств в ткани головного мозга является одной из важнейших проблем в медицине. Отмечается перспективность использования трансферрина – белка для активации поверхности наночастиц с целью направления их в головной мозг. Проведен анализ использования полимерных мицелл для адресной доставки противоопухолевых препаратов, генной терапии и диагностических агентов. Проанализированы преимущества дендримеров – уникальной системы доставки лекарств с эффективной загрузкой терапевтических и визуализирующих материалов. Были рассмотрены липидные наночастицы, динамические поликонъюгаты, конъюгаты GalNAc-siRNA, экзосомы и системы эритроцитов, продемонстрировавшие эффективную доставку миРНК в раковые клетки. Продемонстрировали высокую противоопухолевую эффективность комбинации салиномицина и паклитаксела, доставляемой липосомальным препаратом, модифицированным mAb 2C5. Проанализирована эффективность проникающих в клетку пептидов (CPP) по доставке лекарств и генов (siRNA, pDNA) к участкам опухоли для борьбы с лекарственной устойчивостью. Проведено экспериментальное изучение эффективности прямой электрической стимуляции задней перстнечерпаловидной мышцы на контролируемое открытие голосовой щели. Отмечен большой потенциал нейростимуляции для активизации гортани человека. Установлена эффективность трансплантации обкладочных клеток, полученных из обонятельной выстилки носа крыс и человека, на изменение размеров посттравматических кист спинного мозга. Определены пути и профили экспрессии ДНК при увеальной меланоме, что будет способствовать созданию прогностических моделей, которые могут привести к улучшению прогноза для пациентов с данным заболеванием. Установлено накопление метиленового синего как в различных тканях, так и в иммунных клетках, что указывает на потенциальную коррекцию иммунного ответа у больных COVID-19 и изменение в фенотипе макрофагов, которое может быть достигнуто путем дезактивации воспалительных макрофагов в тканях с использованием лазерного излучения красного спектрального диапазона. При проведении моделирования септопластики установлено изменение вариабельности сердечного ритма (ВСР), повышение концентрации кортикостерона в плазме крови у крыс. В результате сравнительного анализа влияния септопластики и моделирования синус-лифтинга у крыс на изменения в частотной области ВСР установлено повышение активности симпатической нервной системы и сдвиг обмена веществ под влиянием послеоперационного воспаления. На основании проведенных исследований установлено, что фотобиомодуляционная терапия после септопластики показывает лучшие результаты по сравнению со стандартной реабилитацией пациентов после септопластики. Рассмотрена отдельная система стадирования рака ротоглотки, связанного с вирусом папилломы человека. Показаны преимущества трехмерного (3D) поверхностного сканирования перед ультразвуковым исследованием во время предоперационного планирования, особенно в «сложных» областях лица. Получены данные о трехмерном ходе угловой артерии в пределах носогубной борозды в зависимости от возраста, пола и индекса массы тела. Предоставлены объективные доказательства антагонистического движения кожи между медиальной и латеральной средней частью лица. Функциональная граница, идентифицированная с помощью 3D-изображения, соответствует анатомическому расположению линии связок. При изучении связи фасциальной анатомии лба с эффективностью лечения нейромодуляторами установили, что техника глубокой инъекции приводит к лучшему результату. Продемонстрирована эффективность лазерно-индуцированной флуоресцентной диагностики и фотодинамической терапии с сублингвальным введением 5-АЛК больным при лечении предраковых поражений полости рта и гортани.