Tissue Extracts Research Articles

Comparative analysis of in situ and ex situ postmortem brain MRI: Evaluating volumetry, DTI, and relaxometry

AbstractPurposeTo compare postmortem in situ with ex situ MRI parameters, including volumetry, diffusion tensor imaging (DTI), and relaxometry for assessing methodology‐induced alterations, which is a crucial prerequisite when performing MRI biomarker validation.MethodsMRI whole‐brain scans of five deceased patients with amyotrophic lateral sclerosis were performed at 3 T. In situ scans were conducted within 32 h after death (SD 18 h), and ex situ scans after brain extraction and 3 months of formalin fixation. The imaging protocol included MP2RAGE, DTI, and multi‐contrast spin‐echo and multi‐echo gradient‐echo sequences. Volumetry, fractional anisotropy, mean diffusivity, T1, T2, and have been assessed for specific brain regions.ResultsWhen comparing ex situ to in situ values, the following results were obtained. Deep gray matter as well as the thalamus and the hippocampus showed a reduced volume. Fractional anisotropy was reduced in the cortex and the whole brain. Mean diffusivity was decreased in white matter and deep gray matter. T1 and T2 were reduced in all investigated structures, whereas was increased in the cortex.ConclusionThe results of this study show that the volumes and MRI parameters of several brain regions are potentially affected by tissue extraction and subsequent formalin fixation, suggesting that methodological alterations are present in ex situ MRI. To avoid overlap of indistinguishable methodological and disease‐related changes, we recommend performing in situ postmortem MRI as an additional intermediate step for in vivo MRI biomarker validation.

Discovering Novel Bioherbicides: The Impact of Hemp-derived Phytocannabinoid Applications on Zea mays L. and Relevant Weeds

In addition to competition, phytotoxic plant metabolites contribute to the weed-suppressing properties of cover crops, which could be the basis for the development of novel bioherbicides. We investigated the impact of five Cannabis sativa L. -derived neutral phytocannabinoids and an aqueous C. sativa tissue extract (HE) at six concentrations on the germination rate (GR) and seedling root length (RL) of Zea mays L., two monocotyledonous and two dicotyledonous weed species in laboratory Petri dish bioassays. Additionally, the effect of pre-emergence applications of HE, cannabidiol (CBD), and cannabidivarin (CBDV) formulations on GR and shoot dry matter (SDM) were examined in greenhouse pot studies. The effects of phytocannabinoids and HE were analyzed in dose-response curves. For the highest rates, the effects on GR, RL and SDM were calculated by ANOVA and HSD test (p < 0.05). HE exhibited the greatest suppression on GR and RL for all plant species in the Petri dish bioassay, with RGR, RL exceeding −90%. Phytocannabinoids reduced mainly RL of all plants and decreased the GR of most weed species. Effects varied among plants and phytocannabinoids, with CBDV and CBD showing similar high inhibitory effects on RL as HE in the Petri dish bioassay. All pre-emergence applications resulted in a positive RGR across all studied plants and in a positive RSDM in Z. mays and Echinochloa crus-galli (L.) P. Beauv, whereas in the other weed species the RSDM was negative. In conclusion, phytocannabinoids play a major role in weed suppression of HEs. CBDV and CBD are the most promising candidates for bioherbicide development especially against annual dicotyledonous weed species.

Effect of salt stress on growth and phenolic compounds production in callus suspension culture of the dioecious species thyrse sorrel (Rumex thyrsiflorus Fingerh.)

The sex-dependent differences in the response to salt stress of the dioecious Rumex thyrsiflorus and the influence of different concentrations of sodium chloride (NaCl) on the biosynthesis of phenolic compounds in callus suspension cultures were evaluated. The cultures originated from callus obtained on hypocotyls isolated from male and female seedlings were cultured on Murashige and Skoog medium supplemented with 0.4 mg/L 6-benzylaminopurine and 1 mg/L 2,4-dichlorophenoxyacetic acid. The results showed that the response to salt stress depends on both the sex of the plant and the NaCl concentration in the culture medium. The analysis of the main morphometric parameters showed that callus tissue derived from the hypocotyls of female seedlings was less sensitive to salt stress than that from male seedlings, which was correlated with higher concentrations of phenolic compounds. The optimal NaCl concentration to increase the production of phenolic compounds was 129 mM for females and 43 mM for males. In the methanolic tissue extracts 22 compounds were determined using the HPLC–DAD method. In general, higher amounts of compounds were detected in the extracts from the female tissue. The following dominated: catechin (max. 213.31), cryptochlorogenic acid (max. 76.35) and epicatechin (max. 54.84) (mg/100 g DW). This comprehensive phytochemical analysis of the sex-related aspects of the response to salt stress was performed for the first time in this dioecious model species. The results revealed potential application of NaCl as the ecological friendly and inexpensive elicitor to increase the production of pharmaceutically valuable compounds and highlight the importance of dioecy in phytochemistry.

Understanding and controlling the variables for stromal vascular fraction therapy.

In regenerative medicine, the isolation of mesenchymal stromal cells (MSCs) from the adipose tissue's stromal vascular fraction (SVF) is a critical area of study. Our review meticulously examines the isolation process of MSCs, starting with the extraction of adipose tissue. The choice of liposuction technique, anatomical site, and immediate processing are essential to maintain cell functionality. We delve into the intricacies of enzymatic digestion, emphasizing the fine-tuning of enzyme concentrations to maximize cell yield while preventing harm. The review then outlines the filtration and centrifugation techniques necessary for isolating a purified SVF, alongside cell viability assessments like flow cytometry, which are vital for confirming the efficacy of the isolated MSCs. We discuss the advantages and drawbacks of using autologous vs allogeneic SVF sources, touching upon immunocompatibility and logistical considerations, as well as the variability inherent in donor-derived cells. Anesthesia choices, the selection between hypodermic needles vs liposuction cannulas, and the role of adipose tissue lysers in achieving cellular dissociation are evaluated for their impact on SVF isolation. Centrifugation protocols are also analyzed for their part in ensuring the integrity of the SVF. The necessity for standardized MSC isolation protocols is highlighted, promoting reproducibility and successful clinical application. We encourage ongoing research to deepen the understanding of MSC biology and therapeutic action, aiming to further the field of regenerative medicine. The review concludes with a call for rigorous research, interdisciplinary collaboration, and strict adherence to ethical and regulatory standards to safeguard patient safety and optimize treatment outcomes with MSCs.

Chlorophyll deficiency in Agave angustifolia Haw.: unveiling the impact on secondary metabolite production.

The albino phenotype of Agave angustifolia Haw. accumulates higher levels of phenylalanine and phenylpropanoids, while the green phenotype has a greater concentration of phenolic compounds. The metabolic consequences of chlorophyll deficiency in plants continue to be a captivating field of research, especially in relation to production of metabolic compounds. This study conducts a thorough analysis of the metabolome in green (G), variegated (V), and albino (A) phenotypes of Agave angustifolia Haw. Specifically, it examines the differences in the accumulation of compounds related to the phenylpropanoid and flavonoid biosynthesis pathways. Methanol extracts of leaf and meristem tissues from the three phenotypes grown in vitro were analyzed using liquid chromatography coupled with quadrupole time-of-flight high-resolution mass spectrometry (UPLC-MS-QTOF) for untargeted metabolomics and triple quadrupole (QqQ) mass spectrometry for targeted metabolomic analyses. By employing these methods, we discovered notable differences in the levels of important metabolites such as L-phenylalanine, 4-hydroxyphenylpyruvic acid, and various flavonoids among the different phenotypes. The results of our study indicate that the A phenotype shows a significant increase in the levels of phenylalanine and phenylpropanoids in both leaf and meristem tissues. This is in contrast to a decrease in flavonoids, suggesting a metabolic reprogramming to compensate for the lack of chlorophyll. Significantly, compounds such as kaempferol-3-O-glucoside and rutin exhibited significant quantitative reduction in the A leaves, suggesting a subtle modification in the production of flavonols and potentially a changed mechanism for antioxidant protection. This study emphasizes the complex metabolic changes in A. angustifolia´s chlorophyll-deficient phenotypes, providing insight into the complex interplay between primary and secondary metabolism in response to chlorophyll deficiency. Our research not only enhances the comprehension of plant metabolism in albino phenotypes but also opens new avenues for exploring the biochemical and genetic basis of such adaptations, with potential biotechnological applications of these distinct plant variants.

The Role of Adrenomedullin as a Predictive Marker of the Risk of Death and Adverse Clinical Events: A Review of the Literature.

Adrenomedullin (ADM) is a vasodilatory peptide that plays a crucial role in maintaining cardiovascular health through its various biological functions. ADM was discovered in the acidic extract of human pheochromocytoma tissue and has been recognized for its significant effects on the vascular system. The main functions of ADM include vasodilation, controlling blood pressure and maintaining vascular integrity, although its role on cardiovascular health is broader. Research has shown that elevated levels of adrenomedullin have been observed in a large number of severe diseases, with high risk of death. In this work, we examined the role of ADM as a predictive molecule of the risk of mortality and adverse clinical outcome through a narrative review of the scientific literature. The results were divided based on the pathologies and anatomical districts examined. This review demonstrates how ADM shows, in many diseases and different systems, a close correlation with the risk of mortality. These results prove the value of ADM as a prognostic marker in various clinical contexts and diseases, with utility in the stratification of the risk of clinical worsening and/or death and in the evaluation of therapeutic efficacy. The results open new perspectives with respect to the concrete possibility that ADM enters clinical practice as an effective diagnostic and prognostic marker of death as well as a molecular target for therapies aimed at patient survival.

Comparison of Peptidomes Extracted from Healthy Tissue and Tumor Tissue of the Parotid Glands and Saliva Samples.

Salivary gland tumors are highly variable in clinical presentation and histology. The World Health Organization (WHO) classifies 22 types of malignant and 11 types of benign tumors of the salivary glands. Diagnosis of salivary gland tumors is based on imaging (ultrasound, magnetic resonance imaging) and fine-needle aspiration biopsy, but the final diagnosis is based on histopathological examination of the removed tumor tissue. In this pilot study, we are testing a new approach to identifying peptide biomarkers in saliva that can be used to diagnose salivary gland tumors. The research material for the peptidomic studies was extracts from washings of neoplastic tissues and healthy tissues (control samples). At the same time, saliva samples from patients and healthy individuals were analyzed. The comparison of the peptidome composition of tissue extracts and saliva samples may allow the identification of potential peptide markers of salivary gland tumors in patients' saliva. The peptidome compositions extracted from 18 tumor and 18 healthy tissue samples, patients' saliva samples (11 samples), and healthy saliva samples (8 samples) were analyzed by LC-MS tandem mass spectrometry. A group of 109 peptides was identified that were present only in the tumor tissue extracts and in the patients' saliva samples. Some of the identified peptides were derived from proteins previously suggested as potential biomarkers of salivary gland tumors (ANXA1, BPIFA2, FGB, GAPDH, HSPB1, IGHG1, VIM) or tumors of other tissues or organs (SERPINA1, APOA2, CSTB, GSTP1, S100A8, S100A9, TPI1). Unfortunately, none of the identified peptides were present in all samples analyzed. This may be due to the high heterogeneity of this type of cancer. The surprising result was that extracts from tumor tissue did not contain peptides derived from salivary gland-specific proteins (STATH, SMR3B, HTN1, HTN3). These results could suggest that the developing tumor suppresses the production of proteins that are essential components of saliva.

Histological Changes in Dental Pulp During the Aging Process: Implications for Endodontic Treatment

on the results of endodontic treatment. Knowledge of these changes is essential in order to design treatment strategies that allow aging people a life with constant dental health. Objective: The objective of this study sought to perform an analysis of the histological changes that occur within dental pulp as a result of aging. Study Design: This is a cross-sectional study. Duration and Place of the Study: This study was conducted at Department Anatomy of General Hospital and Research Centre, Peshawar, from 3rd November 2022 to 2nd December 2023. Material and Methods: A total of 220 patients aged between 20-80 years, were admitted to this survey. According to age, they were classified into 20-29 years (n=41), 30-39 years (n=39),40-49 years (n=48), 50-59 years (n=45 ) and 60 -80 years( n=47). Extraction of dental pulp samples Pulp tissues were harvested from teeth that had been extracted for non-dental reasons. Histological analysis for cellularity, vascularity and fibrous content of the pulp tissue. Results: A total of 220 patients aged between 20 and 80 Years were included in this study. The biggest group of people was participants 40-49 years old (21.8%), and the smallest one is for 30-39-year-olds with (17.7%). The study found that Participants of the age groups 20–29 years showed a mean cellularity (1500 cells/mm²), with standard deviation of ±200, while those in the 60-80 years group had the lowest mean cellularity at 800 cells/mm² with a standard deviation of ±160. Conclusion: The histological changes in dental pulp with ageing have as consequences implications on endodontic treatment. Keywords: Dental Pulp, Histological Changes, Endodontic Treatment Implications

The role of Nuclear Magnetic Resonance (NMR) spectroscopy in cattle metabolism

Nuclear Magnetic Resonance (NMR) spectroscopy is a powerful analytical tool that has significantly advanced the understanding of cattle metabolism. Nuclear Magnetic Resonance (NMR) spectroscopy plays a pivotal role in the study of cattle metabolism, offering distinct advantages over other spectrometric methods. NMR spectroscopy is a powerful analytical tool that provides detailed molecular insights by exploiting the magnetic properties of atomic nuclei. Unlike mass spectrometry and infrared spectroscopy, NMR does not require extensive sample preparation or destruction, preserving the integrity of biological samples. This non-invasive nature is particularly beneficial for longitudinal studies in cattle, where metabolic changes over time are of interest. One of the key strengths of NMR spectroscopy is its ability to simultaneously detect and quantify a broad range of metabolites in complex biological matrices, such as blood, urine, and tissue extracts. This comprehensive metabolic profiling is crucial for understanding the biochemical pathways and physiological states in cattle. NMR's high reproducibility and quantitative accuracy further enhance its suitability for metabolic studies, enabling precise monitoring of metabolic fluctuations in response to dietary changes, environmental stressors, or disease conditions. NMR spectroscopy also offers unique advantages in elucidating structural information about metabolites. Through multidimensional NMR techniques, researchers can determine the molecular structure and conformation of metabolites, providing deeper insights into metabolic functions and interactions. This structural elucidation is often challenging with other spectrometric methods, which may lack the resolution or require derivatization of samples. Moreover, NMR spectroscopy's non-destructive nature allows for the analysis of living tissues and in vivo studies, facilitating real-time monitoring of metabolic processes. This capability is instrumental in studying dynamic metabolic responses and adaptations in cattle under different physiological states. Additionally, the development of advanced NMR techniques, such as high-resolution magic angle spinning (HR-MAS) and hyperpolarization, has further expanded the scope of NMR applications in metabolic research. NMR spectroscopy stands out as a superior method for studying cattle metabolism due to its non-destructive approach, comprehensive metabolic profiling, structural elucidation capabilities, and potential for in vivo analysis. These advantages make NMR an indispensable tool in advancing our understanding of cattle metabolism and improving livestock health and productivity. Keywords: Nuclear Magnetic Resonance (NMR), Cattles, Metabolomics.

Steroid sulfatase in mouse liver and testis: Characterization, ontogeny and localization

Steroid hormones often circulate in the plasma as inactive sulfated forms, such as estrone sulfate and dehydroepiandrosterone sulfate. The enzyme steroid sulfatase (STS) converts these steroids into active forms, mainly estrogens, in peripheral tissues. STS is present in most tissues, but it occurs at higher levels in certain organs, notably liver and placenta. In this study, we examined the tissue distribution of STS in a prominent laboratory model, the house mouse (Mus musculus). Tissues included were heart, liver, small intestine, skeletal muscle, and gonads of both sexes. An 3H-estrone-sulfate conversion assay was used to measure STS activity in tissue homogenates and extracts. STS activities were high for hepatic tissue homogenates of both genders. Testicular STS levels were similar to those of liver, while STS activities of ovary, small intestine, heart, and muscle were considerably lower. The specific STS inhibitors, EMATE and STX-64 virtually eliminated STS activity in hepatic microsomes and cytosols, verifying that the observed enzyme activity was due to STS. Enzyme kinetic assays showed Km values of 8.6 µM for liver and 9.1 µM for testis, using E1S as substrate. Hepatic and testicular STS activities, measured in CHAPS-extracted microsome, were low up to 5 weeks of age and were higher through 56 weeks. Western blotting, with a specific STS antibody, confirmed the presence of STS protein (65 Da) in both liver and testis. Immunofluorescence of tissue sections detected the presence of STS protein in hepatocytes, in testicular Leydig cells and in seminiferous tubules (Leydig cells and developing germ cells). These results suggest that STS may have a significant role in testicular function.

Understanding metal contamination in Anemonia sulcata: Effects of proximity to a submarine sewage pipe

The study conducted in June 2022 collected 120 Anemonia sulcata samples from Punta del Hidalgo, Tenerife, Spain, focusing on areas near a sewage pipe. The samples underwent meticulous preparation, including tissue extraction, drying, and filtration, followed by metal concentration (Al, Cd, Cu, Fe, Li, Ni, Pb, and Zn) analysis using ICP-OES. Results showed varying metal concentrations across zones, with higher concentrations nearer to the pipe. PERMANOVA analysis revealed significant differences in metal concentrations between zones, indicating spatial variations. Environmental factors such as pipe proximity, transport contaminant, and dispersion influence on metal accumulation in A. sulcata. Previous studies corroborate these findings, highlighting the pipe's significant impact on intertidal organisms. Effective monitoring strategies combining direct organism sampling with water and sediment analysis are crucial for assessing and mitigating the environmental and ecological impacts of metal contamination. These findings underscore the necessity of ongoing monitoring efforts and management interventions to safeguard marine ecosystems from detrimental effects of anthropogenic activities such as an underwater sewage pipe.

Analysis of the UV filter Benzophenone-3 assimilation in Crossostrea gigas oysters post-exposure in a controlled environment by LC-MS/MS

Benzophenone-3 (BP-3), utilized as a UV filter in cosmetic products, is an emerging contaminant that constitutes a threat to natural resources and environmental health. This study investigated the assimilation of the UV filter BP-3 in Crassostrea gigas oysters collected in Florianópolis, Santa Catarina, Brazil. Lyophilized oyster tissue extracts were prepared using the QuEChERS method, and LC-MS/MS was employed to determine the BP-3 concentration in the samples. The method was applied to specimens intentionally exposed to two concentrations of the contaminant, for different periods of exposure (1 and 7 days). Samples from treatment 1 (T1) were exposed to a concentration of 1 μg L−1 of the BP-3 standard, and samples from treatment 2 (T2) were exposed to a concentration of 100 μg L−1 of the BP-3 standard. The results revealed rapid absorption of BP-3, with an increase of 126% for lower concentrations, reaching 1.13 μg of BP-3 per gram of oyster tissue, and 17% for higher concentrations, reaching 34.6 μg of BP-3 per gram of oyster tissue after 7 days. The presence of BP-3 even in samples not directly exposed to the contaminant indicates its widespread environmental distribution. The rapid bioaccumulation suggests the need to consider seasonal variations, such as increased tourism in the summer. The validated analytical method demonstrated efficacy in quantifying BP-3, providing an integrated approach for long-term monitoring of pollution levels and their dynamic variations over time. In addition, variation in BP-3 levels in the samples may be related to transport patterns influenced by tides and discharges from septic system, highlighting the need to improve wastewater treatment. These findings underscore the necessity for continuous biomonitoring and effective environmental management to safeguard the health of marine ecosystems and humans.

Subject-Specific Probability Maps of Scalp, Skull and Cerebrospinal Fluid for Cranial Bones Segmentation in Neonatal Cerebral MRIs

ObjectivesSegmentation of cranial bones in magnetic resonance images (MRIs) is a challenging and indispensable task to study neonatal brain development and injury. This paper presents a new approach for creating subject-specific probability maps of the scalp, skull and cerebrospinal fluid (CSF) from retrospective bimodal (MR and CT) images acquired from neonates in the gestational age range of 39 to 42 weeks. These maps are subsequently employed for the segmentation of cranial bones in cerebral MRIs from neonates in the same age range. Material and methodsRetrospective MR and CT of neonates with normal head in the gestational age range of 39-42 weeks were preprocessed, segmented semi-automatically and employed as atlas data. For an input MR image acquired from a subject under study, a preprocessing stage and three main processing blocks were performed: First, subject-specific head and intracranial templates and CSF probability map were created using retrospective MR atlas data. Second, the CT atlas data were coregistered to MR templates and the resulted deformation matrices were fed to the next block to create subject-specific scalp and skull probability maps. Finally, some novel performance measures were presented to evaluate the performance of subject-specific CSF, scalp and skull probability maps for skull and intracranial segmentation in neonatal MRIs. ResultsThe subject-specific probability maps were employed for brain tissue extraction and compared with two public methods such as Brain Extraction Tool (BET) and Brain Surface Extractor (BSE). They were also applied for cranial bone extraction. Then, the similarity in shape between the frontal and occipital sutures (which had been reconstructed from segmented cranial bones) and the ground truth landmarks was evaluated. For this purpose, modified versions of the Dice similarity coefficient (DSC) were used. Finally, a retrospective bimodal (MR-CT) data acquired from a neonate within a short time interval was used for evaluation. After co-alignment of the two images, the DSC and modified Hausdorff distance (MHD) were used to compare the similarity of cranial bones in the MR and CT images. ConclusionSignificant improvements were achieved compared to conventional methods which rely solely on MR image intensities. These advancements hold promise for enhancing neurodevelopmental studies in neonates. The algorithm for creating subject-specific atlases is publicly accessible through a graphical user interface at medvispy.ee.kntu.ac.ir.

ATRT-03. PRIMARY DIFFUSE LEPTOMENINGEAL ATYPICAL TERATOID/RHABDOID TUMOR (ATRT) OF CHILDHOOD DEMONSTRATING UNEXPECTED IMPROVEMENT UPON CESSATION OF CONVENTIONAL THERAPY FOR PRESUMED PROGRESSION; A MOLECULARLY CHARACTERIZED CASE REPORT

Abstract BACKGROUND Atypical teratoid/rhabdoid tumors (ATRTs) are malignant tumors of the central nervous system, typically presenting in the posterior fossa of very young children. Prognosis remains poor despite current therapy, while tumorigenesis implicates both genomic and epigenetic dysregulation. Primary diffuse leptomeningeal disease (PDL ATRT), without a defined intraparenchymal lesion, is an extremely rare ATRT variant with only seven cases reported. None of these preceding cases had undergone molecular characterization. We compared our case, who demonstrated an atypical clinical and radiological trajectory, against these seven cases. CASE PRESENTATION We describe a PDL ATRT, with hitherto novel molecular and imaging characteristics, in a pediatric patient who subsequently demonstrated radiological and clinical recovery from radiologically diagnosed metastatic relapse in the absence of conventional ATRT salvage treatment. Our case is the first to utilize Magnetic Resonance Imaging Arterial Spin Labelling perfusion imaging to suggest an underlying malignancy as the responsible etiology for this patient’s complex presentation, and the first to use methylation profiling to establish a tumor molecular subgroup for a PDL ATRT patient (identifying a MYC-subgroup in this case). CONCLUSIONS For this reported case of pediatric MYC-subgroup PDL ATRT, MR perfusional imaging aided in establishing the diagnosis and predicting potential relapse as part of a multi-parametric imaging assessment. We administered upfront intensive multimodal therapy, including high-dose CSRT to the patient, resulting in the longest overall survival for any patient reported with this extremely challenging ATRT variant. His unanticipated clinical and radiological improvement following a subsequent radiological diagnosis of tumor relapse highlights the importance of confirming recurrence by tissue extraction where feasible, but suggests that further evaluation of histone deacetylase inhibition as a potential future therapy in ATRT is warranted.

Effect of accelerated extraction and sonication on the antioxidant capacity of wood and bark extracts of wet-hearted silver fir (Abies alba Mill.)

The aim of this study was to compare the antioxidant capacity of hydrophilic extracts of different tissues of wet-hearted silver fir (Abies alba Mill.) logs. Wet heart is a wood defect of silver fir and lowers the value of wood. The exploitation of extractives could increase utilization and financial returns. The bark, sapwood, heartwood and knotwood were investigated by using two green extraction methods: ultrasound assisted extraction (US) and accelerated solvent extraction (ASE), with aqueous ethanol as solvent. Total extractive content (TEC), total polyphenol content (TPC) and antioxidant capacities (FRAP, DPPH, ABTS) from the extracts were determined and compared. The highest contents of total hydrophilic extractives were measured in knotwood (23.07%, ASE) and bark (10.31%, ASE), and the lowest values were determined for sapwood (2.00%, ASE) and heartwood (3.56%, ASE). The ASE method resulted in significantly higher TPC (0.65–10.58%) than the US (0.46–9.19%) method. Nevertheless, the simplicity of instrumentation and costs can make US also a potential candidate for future extraction and utilization. The highest antioxidant capacities were measured in knotwood (FRAP: 159.75 mg AAE/g, ABTS: 316.15 mg TE/g, DPPH: 189.23 mg TE/g) and bark (FRAP: 159.75 mg AAE/g, ABTS: 126.81 mg TE/g, DPPH: 74.52 mg TE/ g) extracts, prepared with ASE, which complements well the existing literature data on silver fir extractives. The knotwood and bark of wet-hearted silver fir is an abundant source of antioxidant polyphenols, whereas sapwood and heartwood are poor in these extractives and potentially unsuitable for the valorization by the extraction of natural antioxidants.

Interaction of pRb and beta-catenin in cancer and normal tissue in the human prostate

Prostate cancer (PCa) is one of the most common oncological diseases, which goes through two stages in its development. The first stage, localized prostate cancer, can proceed indefinitely in a dormant form that does not require active medical intervention, or suddenly turn into an aggressive metastatic form with lethal outcome. The pathogenesis of the transition of the dormant form of PCa to the metastatic form remains not fully understood. The signaling pathways of the tumor suppressor pRb and the proto-oncogene β-catenin are probably the most involved in the pathogenesis of PCa but the role of their interaction in the pathogenesis of prostate cancer has not been studied. The publication on the pathogenesis of tumors in other tissues suggests that pRb may lose some properties of a tumor suppressor at the initial stage of PCa development due to its interaction with β-catenin that enables tumor cells to gain competitive advantages for reproduction. In this work, we have shown that the RB and β-catenin (CTNNB1) genes are well expressed in tumor and normal prostate tissue. Unlike β-catenin, pRb is not detected by immunoblotting in tumor and normal prostate tissue, but is easily determined in this way in extracts of control T98G cells. Co-immunoprecipitation with antibodies to pRb from extracts of tumor and normal prostate tissue makes it possible to detect this protein and β-catenin by subsequent immunoblotting, which indicates the physical interaction of these proteins in prostate tissue. On the other hand, immunoprecipitation of β-catenin with antibodies to its C-terminal fragment does not detect this protein in prostate extracts by subsequent immunoblotting using the same antibody. In contrast to prostate tissue, β-catenin is readily detected by immunoprecipitation combined with immunoblotting in T98G control cell extracts. The obtained data suggest that pRb and β-catenin physically interact with each other in cells of different tissue specificity. In T98G cells, this interaction probably occurs through the C-terminal fragment of β-catenin, but in prostate cells it occurs in a different way, since the C-fragment of β-catenin is shielded from such interaction, possibly due to its physical association with pRb.

Automated DeepLabV3+ based model for left ventricle segmentation on short-axis late gadolinium enhancement-magnetic cardiac resonance imaging images

Accurate segmentation of myocardial scar tissue on late gadolinium enhancement-magnetic cardiac resonance imaging (LGE-CMR) is exceptionally vital for clinical applications, enabling precise diagnosis and effective treatment of various cardiac diseases, such as myocardial infarction and cardiomyopathies. However, the ventricle (LV) variations in the size and shape, artifacts, and image resolution of LGE-CMR has made automatic segmentation of myocardial scar tissue more challenging. While many existing approaches delineate the LV myocardium region using multi-modal segmentation, these models may be computationally complex and suffer from misalignment. Therefore, this study proposed an automatic dual-stage DeepLabV3+ based approach tailored for myocardial scar segmentation on short-axis LGE-MRI exclusively. To segment myocardial scar tissue, the second stage employs the segmented LV chamber from the previous stage. The encoder part of the framework utilizes a MobileNetV2 and ResNet50 backbone for the first and second segmentation, respectively, aiming for optimal resolution of feature maps. Both stages tailor an improved Atrous Spatial Pyramid Pooling module in the DeepLabV3+ model with fine-tuned dilated atrous rates to effectively extract the LV chamber and myocardial scar from the complex LGE-MRI background. Based on the results, the proposed dual-stage network recorded an outstanding segmentation performance, with mean Dice score of 96.02% for LV chamber segmentation and 68.01% for scar tissue extraction.

Dome-type extracorporeal manual morcellation during laparoscopic uterine surgery: Two years’ experience in a teaching hospital

ObjectiveThis study aims to describe the dome-type manual morcellation technique, a modified form of C-type incision, its comparative advantages over existing morcellation methods, the perioperative outcomes of trainees with varying experience levels, and the variables influencing morcellation speed based on our two years of experience. MethodsThis retrospective cohort study included women who underwent laparoscopic myomectomy or hysterectomy using dome-type morcellation for tissue extraction at a tertiary teaching hospital between May 2020 and September 2022. Morcellation was performed by either a single surgeon or a trainee (resident). Basic patient characteristics, perioperative outcomes, and morcellation time and speed were compared between the surgeon and trainee group. Regression models were employed to analyze variables influencing morcellation speed. ResultsA total of 41 women were enrolled. Among them, 20 procedures were performed by a surgeon alone, while the remaining 21 procedures were completed by trainees under the surgeon's supervision. The median weight of the specimens was 378 g (range 91–1345 g), and the median time for morcellation was 10 min (range 1–55 min). The median morcellation speed of surgeon and trainees was 70.25 and 31.7 g/min, respectively. Trainees' level of experience was found to be associated with morcellation speed, particularly for soft specimens. Additionally, both incision size and specimen stiffness were significantly associated with morcellation speed. No morcellation-related complications or bag ruptures were observed. ConclusionDome-type manual morcellation is an intuitive, efficient and safe method for specimen removal and is easy to learn for beginners.

Quantification of multiple steroid hormones in serum and human breast cancer tissue by liquid chromatography-tandem mass spectrometry analysis.

Systemic and local steroid hormone levels may function as novel prognostic and predictive biomarkers in breast cancer patients. We aimed at developing a novel liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous measurement of multiple, biologically pivotal steroid hormones in human serum and breast cancer tissue. The quantitative method consisted of liquid-liquid extraction, Sephadex LH-20 chromatography for tissue extracts, and analysis of steroid hormones by liquid-chromatography-tandem mass spectrometry. We analyzed serum and tissue steroid hormone levels in 16 and 40 breast cancer patients, respectively, and assessed their correlations with clinical parameters. The method included quantification of nine steroid hormones in serum [including cortisol, cortisone, corticosterone, estrone (E1), 17β-estradiol (E2), 17α-hydroxyprogesterone, androstenedione (A4), testosterone and progesterone) and six (including cortisone, corticosterone, E1, E2, A4, and testosterone) in cancer tissue. The lower limits of quantification were between 0.003-10 ng/ml for serum (250 µl) and 0.038-125 pg/mg for tissue (20 mg), respectively. Accuracy was between 98%-126%, intra-assay coefficient of variations (CV) was below 15%, and inter-assay CV were below 11%. The analytical recoveries for tissue were between 76%-110%. Tissue levels of E1 were positively correlated with tissue E2 levels (p<0.001), and with serum levels of E1, E2 and A4 (p<0.01). Tissue E2 levels were positively associated with serum E1 levels (p=0.02), but not with serum E2 levels (p=0.12). The levels of tissue E2 and ratios of E1 to A4 levels (an index for aromatase activity) were significantly higher in patients with larger tumors (p=0.03 and p=0.02, respectively). The method was convenient and suitable for a specific and accurate profiling of clinically important steroid hormones in serum. However, the sensitivity of the profile method in steroid analysis in tissue samples is limited, but it can be used for the analysis of steroids in breast cancer tissues if the size of the sample or its steroid content is sufficient.

Proteomic reference map for sarcopenia research: mass spectrometric identification of key muscle proteins of organelles, cellular signaling, bioenergetic metabolism and molecular chaperoning.

During the natural aging process, frailty is often associated with abnormal muscular performance. Although inter-individual differences exit, in most elderly the tissue mass and physiological functionality of voluntary muscles drastically decreases. In order to study age-related contractile decline, animal model research is of central importance in the field of biogerontology. Here we have analyzed wild type mouse muscle to establish a proteomic map of crude tissue extracts. Proteomics is an advanced and large-scale biochemical method that attempts to identify all accessible proteins in a given biological sample. It is a technology-driven approach that uses mass spectrometry for the characterization of individual protein species. Total protein extracts were used in this study in order to minimize the potential introduction of artefacts due to excess subcellular fractionation procedures. In this report, the proteomic survey of aged muscles has focused on organellar marker proteins, as well as proteins that are involved in cellular signaling, the regulation of ion homeostasis, bioenergetic metabolism and molecular chaperoning. Hence, this study has establish a proteomic reference map of a highly suitable model system for future aging research.