Related Folds Research Articles

Development of infectious clones of mungbean yellow mosaic India virus (MYMIV, Begomovirus vignaradiataindiaense) infecting mungbean [Vigna radiata (L.) R. Wilczek] and evaluation of a RIL population for MYMIV resistance.

Yellow mosaic disease (YMD) is a major constraint for the low productivity of mungbean, mainly in South Asia. Addressing this issue requires a comprehensive approach, integrating field and challenge inoculation evaluations to identify effective solutions. In this study, an infectious clone of Begomovirus vignaradiataindiaense (MYMIV) was developed to obtain a pure culture of the virus and to confirm resistance in mungbean plants exhibiting resistance under natural field conditions. The infectivity and efficiency of three Agrobacterium tumefaciens strains (EHA105, LBA4404, and GV3101) were evaluated using the susceptible mungbean genotype PS16. Additionally, a recombinant inbred line (RIL) population comprising 175 lines derived from Pusa Baisakhi (MYMIV susceptible) and PMR-1 (MYMIV resistant) cross was developed and assessed for YMD response. Among the tested Agrobacterium tumefaciens strains, EHA105 exhibited the highest infectivity (84.7%), followed by LBA4404 (54.7%) and GV3101 (9.80%). Field resistance was evaluated using the coefficient of infection (CI) and area under disease progress curve (AUDPC), identifying seven RILs with consistent resistant reactions (CI≤9) and low AUDPC (≤190). Upon challenge inoculation, six RILs exhibited resistance, while RIL92 displayed a resistance response, with infection occurring in less than 10% of plants after 24 to 29 days post inoculation (dpi). Despite some plants remaining asymptomatic, MYMIV presence was confirmed through specific PCR amplification of the MYMIV coat protein (AV1) gene. Quantitative PCR revealed a very low relative viral load (0.1-5.1% relative fold change) in asymptomatic RILs and the MYMIV resistant parent (PMR1) compared to the susceptible parent (Pusa Baisakhi). These findings highlight the potential utility of the developed infectious clone and the identified MYMIV-resistant RILs in future mungbean breeding programs aimed at cultivating MYMIV-resistant varieties.

Establishing capsaicin's anti-cancer intricacies in chronic myelogenous leukemia care: insights from human K562 cells

Objective: We aimed to establish capsaicin as a potent natural chemotherapeutic agent in chronic myeloid leukemia by unveiling anti-cancer mechanisms, concentrating at pro-apoptotic and anti-proliferative effects on K562 cells. Methodology: After REC approval, this research proceeded with culturing K562 cells and treatment with serial concentrations of capsaicin for calculation of subsequent 50% Inhibitory Concentration (IC50) values via MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay, expression analysis of gene, comparative analysis of relative gene fold (intrinsic biomarkers caspase-3, caspase-9) as apoptosis mediator biomarkers followed by Quantitative Reverse Transcription Polymerase Chain Reaction (RT-qPCR). Anti-proliferative activity was confirmed via Nitric Oxide (NO) releasing Assay. Results: For K562 cells, the IC50 of capsaicin (12.1 µM) showed high gene expression of intrinsic pro-apoptotic biomarkers caspase-3 and caspase-9 with the relative gene fold of 12.1 and 13.9 respectively. Persistent peaks of NO levels confirmed the anti-proliferative potential of the compounds. Strongest growth inhibitory activity was seen at the peak time of 100-120 min. Conclusion: Capsaicin proved to be a strong pro-apoptotic and anti-proliferative phytochemical agent leading to therapeutic effects against K562 cells. Further studies would help in identifying key molecular intricacies by which capsaicin exhibits diversified anti-cancer potential. Abbreviations: CML - chronic myelogenous leukemia; NO - Nitric oxide; ROS - Reactive Oxygen Species; ROS - Reactive Oxygen Species Keywords: Capsaicin, Pro-apoptotic Effects, Anti-Cancer Mechanisms, K562 cells, Natural Chemotherapeutics Citation: Devi D, Nangdev P, Khaliq HMH, Anique M, Moqaddas A, Aziz O, Javed W. Establishing capsaicin's anti-cancer intricacies in chronic myelogenous leukemia care: insights from human K562 cells. Anaesth. pain intensive care 2024;28(5):830−835; DOI: 10.35975/apic.v28i5.2494 Received: June 26, 2024; Reviewed: August 10, 2024; Accepted: August 16, 2024

Exploring the Mechanisms of Neuronal Protection by Glial Cell Line-Derived Neurotrophic Factor in Autism Spectrum Disorder.

A complicated neurological disease known as autism spectrum disorder (ASD) is typified by issues with social interaction, communication, and repetitive behavior. The neural protective mechanisms in ASD are thought to be influenced by genetic variables, including the expression of neurotrophic genes such as glial cell line-derived neurotrophic factor (GDNF). The aim was to examine the relationship between neuronal protection and cognitive functioning by crosslinking GDNF gene expression and serum levels in individuals with relation to Mini-Mental State Examination (MMSE) scores in ASD patients. After getting study approval and informed consent of patients, this case-control study experimental study was conducted for six months between July 2023 and December 2023. The blood samples (5 ml each) were drawn from the study population (n = 140), including 100 ASD patients with a disease course of30 months based on patients' reports data and 40 healthy controls from four major clinical and hospital settings in Lahore, Karachi, and Bahawalpur from Pakistan. The analytical procedures included nucleic acid extraction, primer design and optimization, and GDNF-targeted real-time quantitative polymerase chain reaction expression analysis. To measure cognitive and behavioral deficits, enzyme-linked immunosorbent assay-based serum GDNF levels (pg/ml) and MMSE scores were compared, concluding the neuronal protection potential of GDNF. In patients with ASD, lower serum levels of GDNF (9.371 ± 2.388 pg/ml) were linked to more severe behavioral and cognitive deficits confirmed by MMSE scores (13.6 ± 3.5) of ASD patients in comparison with the control group (27.1 ± 2.1). Healthy individuals showed higher relative gene fold expression (11.71) compared to the ASD patients (5.51). There is a notable decrease in GDNF gene expression in people with ASD, which raises the possibility that GDNF is important for both cognitive performance and neuronal protection in these people. GDNF may be a useful biomarker for identifying ASD and comprehending its molecular causes, opening the door for focused treatment approaches.

Leveraging neural networks to correct FoldX free energy estimates.

Proteins play a pivotal role in many biological processes, and changes in their amino acid sequences can lead to dysfunction and disease. These changes can affect protein folding or interaction with other biomolecules, such as preventing antibodies from inhibiting a viral infection or causing proteins to misfold. The ability to predict the effects of mutations in proteins is crucial. Although experimental techniques can accurately quantify the effect of mutations on protein folding free energies and protein-protein binding free energies, they are often time-consuming and costly. By contrast, computational techniques offer fast and cost-effective alternatives for estimating free energies, but they typically suffer from lower accuracy. Enhancing the accuracy of computational predictions is therefore of high importance, with the potential to greatly impact fields ranging from drug design to understanding disease mechanisms. One such widely used computational method, FoldX, is capable of rapidly predicting the relative folding stability ( ) for a protein as well as the relative binding affinity ( ) between proteins using a single protein structure as input. However, it can suffer from low accuracy, especially for antibody-antigen systems. In this work, we trained a neural network on FoldX output to enhance its prediction accuracy. We first performed FoldX calculations on the largest datasets available for mutations that affect binding (SKEMPIv2) and folding (ProTherm4) with experimentally measured . Features were then extracted from the FoldX output files including its prediction for . We then developed and optimized a neural network framework to predict the difference between FoldX's estimated and the experimental data, creating a model capable of producing a correction factor. Our approach showed significant improvements in Pearson correlation performance. For single mutations affecting folding, the correlation improved from a baseline of 0.3 to 0.66. In terms of binding, performance increased from 0.37 to 0.61 for single mutations and from 0.52 to 0.81 for double mutations. For epistasis, the correlation for binding affinity (both singles and doubles) improved from 0.19 to 0.59. Our results also indicated that models trained on double mutations enhanced accuracy when predicting higher-order mutations (such as triple or quadruple mutations), whereas models trained on singles did not. This suggests that interaction energy and epistasis effects present in the FoldX output are not fully utilized by FoldX itself. Once trained, these models add minimal computational time but provide a substantial increase in performance, especially for higher-order mutations and epistasis. This makes them a valuable addition to any free energy prediction pipeline using FoldX. Furthermore, we believe this technique can be further optimized and tested for predicting antibody escape, aiding in the efficient development of watch lists.

Evidence for, and significance of, the Neoproterozoic Xuefeng Orogeny, South China

The recognition of unconformities is important for determining the long-term evolution of orogenic belts. We document an unconformity between Tonian and Cryogenian strata of the Nanhua Basin of South China. This stratigraphic break has previously been termed the Xuefeng Movement and is developed within the Yangtze Block and western Cathaysia Block. Deformation characteristics and detrital zircon U-Pb geochronology from rock units underlying and overlying the unconformity (i.e., the Niuguping and Gucheng formations, respectively) from the Anhua region of the northern Hunan Province in the southeastern Yangtze Block are documented. The Niuguping Formation displays a set of top-to-northwest imbricate thrust faults and related fault-propagation folds, NEE-plunging subparallel crenulation lineations, a foliation, and shows widespread greenschist facies metamorphism. In contrast, the overlying Gucheng Formation exhibits no obvious stratal deformation (apart from a mild inclination of bedding) and is unmetamorphosed. Detrital zircon age spectra of the samples from the Niuguping Formation are characterized by unimodal age peaks of ca. 750–730 Ma, while those of the samples from the Gucheng Formation yield a multimodal age distribution, with a few syn-sedimentary detrital zircons (<700 Ma), and much older zircons with age peaks of ca. 800 Ma, 2.1–1.9 Ga, and 2.6–2.3 Ga. Combining this data with previous geochronology indicates deposition of the Niuguping and Gucheng formations in the late Tonian (725–720 Ma) and Cryogenian (690–660 Ma), respectively. The time gap across the unconformity is some 30 Ma (720–690 Ma). The unconformity and associated structural and metamorphic break extend across the Yangtze Block and western Cathaysia Block, defining a major regional orogenic event. The significantly different detrital zircon spectra of strata across the unconformity suggest a dramatic change in the source due to the Xuefeng Orogeny in the Yangtze Block. Zircon U-Pb-Hf isotopic evidence indicates that detritus of the Niuguping Formation below the unconformity was derived from syn-depositional magmatic activity in the northern Yangtze Block, while a significant increase of Archean to Paleoproterozoic ages for the Gucheng Formation, suggests that the Xuefeng Orogeny resulted in crustal thickening and denudation in the northern Yangtze Block. These changes in provenance, along with changing deformational and metamorphic patterns, indicate evolution of the Nanhua Basin from a compressional to an extensional back-arc basin, which we infer is related to tectonic switching from an advancing to a retreating subduction system in the northern Yangtze Block at the time of the unconformity.

Mice microglia cytokine profile changes under the influence of HSV-1

Introduction. Today, prevalence of neurodegenerative diseases increases. In recent years, more studies have revealed a new knowledge about the role of microglia in the development of these diseases. Animal experiments showed that peripheral inflammation causes activation of microglia in brain. All this points to the essential role of the cells in the development of neurodegeneration. Under the influence of various factors, microglia can change the phenotype and participate in both repair and damage to brain cells. Chronic herpesvirus infection caused by HSV-1 is another known factor in the development of neurodegenerative pathology. However, the exact pathogenetic mechanisms are still unknown, nevertheless, studying the virus effect on microglia has great potential. The goal of our study in this connection was to assess the effect of HSV-1 on microglia polarization in mouse strain with normal susceptibility to this virus and in strain which is more resistant to the action of HSV-1. For this purpose, changes in the cytokine profile were detected. A comparison of interstrain differences in the expression of cytokine genes was also compared in control groups. Materials and methods. The study involved infecting C57BL/6 and BALB/c mice with herpes simplex virus type 1, an isolation of microglia was based on separation steps using a discontinuous gradient density, the cytokine profile was assessed by gene expression levels using a real-time reverse transcription PCR. To calculate the relative fold gene expression of samples the 2–ΔΔCt method was used. Statistical significance was determined using the Mann–Whitney U-test. Results. There were found no interstrain differences in cytokine gene expression in control groups of different mouse strains. At the same time, gene expression differed in the experimental groups: in BALB/c mice, the expression of genes for both pro-inflammatory and anti-inflammatory cytokines increased; in C57BL/6 mice, a slight increase in the expression of IL-1β genes was observed. Conclusion. The data indicate the formation of different microglial phenotypes after HSV-1 infection in different mouse strains. Apparently, in BALB/c mice there is a switch from the pro-inflammatory M1 phenotype of microglia to the anti-inflammatory M2 phenotype, while in C57BL/6 mice the attenuation of the infectious process occurs through a return to the original M0 phenotype.

Imprinting of IgA responses in previously infected individuals receiving bivalent mRNA vaccines (WT and BA.4/BA.5 or WT and BA.1)

ObjectivesThe emergence of new SARS-CoV-2 variants has led to the development of Omicron-targeting bivalent mRNA vaccines. It is crucial to understand how bivalent vaccines may improve antibody responses against new variants. MethodsA total of 107 participants, who had three COVID-19 WT mRNA vaccine doses, were recruited, and given either a monovalent (WT) or a bivalent mRNA vaccination (Pfizer/BioNTech Bivalent (WT and BA.4/BA.5) or Moderna Bivalent (WT and BA.1). Blood samples were taken before booster and at 28 days post-booster. ResultsWe found significantly lower fold change in serum binding IgA responses against BA.1, BA.5 and EG.5.1 spike in the bivalent booster group, compared with the monovalent (WT) booster group, following vaccination. However, this was only observed in individuals with prior infection. The relative fold change in serum binding IgA response was more skewed towards WT over variant (BA.1, BA.5 or EG.5.1) spike in previously infected bivalent-booster-vaccinees, as compared with previously infected monovalent-(WT)-booster-vaccinees. ConclusionThe findings suggest imprinting of antibody responses that is shaped by the first vaccination (WT spike). Previous infection also affects the boosting effect of follow-up vaccination. Studies are needed to understand how to induce a robust and long-lasting IgA immunity for protection against COVID-19 infection.

Cyclical heat stress during lactation influences the microstructure of the bovine mammary gland

This study aimed to evaluate the impact of heat stress on mammary epithelial cell (MEC) losses into milk, secretory mammary tissue structure, and mammary epithelial cell activity. Sixteen multiparous Holstein cows (632 ± 12 kg BW) approximately 100 d in milk housed in climate-controlled rooms were paired by body weight and randomly allocated to one of 2 treatments, heat stress (HS) or pair feeding thermoneutral (PFTN) using 2 cohorts. Each cohort was subjected to 2 periods of 4 d each. In period 1, both treatments had ad libitum access to a common total mixed ration and were exposed to a controlled daily temperature-humidity index (THI) of 64. In period 2, HS cows were exposed to controlled cyclical heat stress (THI: 74 to 80), while PFTN cows remained at 64 THI and daily dry matter intake was matched to HS. Cows were milked twice daily, and milk yield was recorded at each milking. Individual milk samples on the last day of each period were used to quantify MEC losses by flow cytometry using butyrophilin as a cell surface marker. On the final day of period 2, individual bovine mammary tissue samples were obtained for histomorphology analysis, assessment of protein abundance, and evaluation of gene expression of targets associated with cellular capacity for milk and milk component synthesis, heat response, cellular proliferation, and autophagy. Statistical analysis was performed using the GLIMMIX procedure of SAS. Milk yield was reduced by 4.3 kg by HS (n = 7) compared with PFTN (n = 8). Independent of treatment, MEC in milk averaged 174 cells/mL (2.9% of total cells). There was no difference between HS vs. PFTN cows for MEC shed or concentration in milk. Alveolar area was reduced 25% by HS, and HS had 4.1 more alveoli than PFTN. Total number of nucleated MEC per area were greater in HS (389 ± 1.05) compared with PFTN (321 ± 1.05); however, cell number per alveolus was similar between groups (25 ± 1.5 vs. 26 ± 1.4). There were no differences in relative fold expression for GLUT1, GLUT8, CSN2, CSN3, LALBA, FASN, HSPA5, and HSPA8 in HS compared with PFTN. Immunoblotting analyses showed a decrease abundance for phosphorylated STAT5 and S6K1, and an increase in LC3 II in HS compared with PFTN. These results suggest that even if milk yield differences and histological changes occur in the bovine mammary gland after 4 d of heat exposure, MEC loss into milk, nucleated MEC number per alveolus, and gene expression of nutrient transport, milk component synthesis, and heat stress related targets are unaffected. In contrast, the abundance of proteins related to protein synthesis and cell survival decreased significantly, while an upregulation of proteins associated with autophagy in HS compared with PFTN.

The nature of chronic rejection after lung transplantation: a murine orthotopic lung transplant study.

Chronic rejection is a major complication post-transplantation. Within lung transplantation, chronic rejection was considered as airway centred. Chronic Lung Allograft Dysfunction (CLAD), defined to cover all late chronic complications, makes it more difficult to understand chronic rejection from an immunological perspective. This study investigated the true nature, timing and location of chronic rejection as a whole, within mouse lung transplantation. 40 mice underwent an orthotopic left lung transplantation, were sacrificed at day 70 and evaluated by histology and in vivo µCT. For timing and location of rejection, extra grafts were sacrificed at day 7, 35, 56 and investigated by ex vivo µCT or single cell RNA (scRNA) profiling. Chronic rejection originated as innate inflammation around small arteries evolving toward adaptive organization with subsequent end-arterial fibrosis and obliterans. Subsequently, venous and pleural infiltration appeared, followed by airway related bronchiolar folding and rarely bronchiolitis obliterans was observed. Ex vivo µCT and scRNA profiling validated the time, location and sequence of events with endothelial destruction and activation as primary onset. Against the current belief, chronic rejection in lung transplantation may start as an arterial response, followed by responses in venules, pleura, and, only in the late stage, bronchioles, as may be seen in some but not all patients with CLAD.

Kinematic reconstruction of active tectonic and halokinetic structures in the 2021 NW Palagruža earthquake area (Central Adriatic)

In March 2021, a compressional earthquake sequence (mainshock Mw 5.2) occurred in the Central Adriatic Sea, offshore Croatia. The struck area is characterized by a complex tectonic settings due to due interaction between tectonic and halokinetic structures. Former studies in this region, mostly based on geophysical and seismological data, do not provide a comprehensive description of the geological complexities of the area, caused by the presence of different types of active structures. Following the interpretation and depth conversion of a set of seismic reflection profiles, we present a kinematic restoration to obtain the shortening rates for the last 6 Myr. We highlight the presence of three main types of active structures: i) shallow thrusts and related folds deforming the seafloor, ii) deep thrusts promoting large-scale deformation, iii) halokinetic structures deforming, at least, the Messinian. We highlight how the structural setting of the area is more complex than previously interpreted, with a possible decoupling of shallow and deep structures and an interaction between tectonic and halokinetic structures. This work opens new perspectives about the role of halokinetic processes in active seismogenic areas.

Tectonic Evolution of the Eastern Southern Alps (Italy): A Reappraisal From New Structural Data and Geochronological Constraints

AbstractThe Eastern Southern Alps fold‐and‐thrust belt (ESA) is part of the seismically active S‐verging retro‐wedge of the European Alps. Its temporal tectonic evolution during continental shortening has so far been constrained by few and low‐resolution indirect time constraints. Aiming at better elucidating the ESA spatiotemporal evolution, we gathered new structural and geochronological data from two regional thrust systems: the innermost south verging Valsugana Thrust (VT) and the more external Belluno Thrust System (BTS). Field work allowed us to constrain the geometry and kinematics of those thrusts and related folds and informed our sampling strategy to carry out fault gouge K‐Ar and tectonic carbonate U‐Pb dating from representative samples structurally associated with the VT and BTS. Our results suggest that the VT was active already in the Late Cretaceous (between ∼78 and 76 Ma) in response to far‐field stresses, with repeated reactivation continuing to the Late Miocene (∼6 Ma). The BTS recorded two distinct deformation events during the Oligocene (∼30 Ma) and at the Oligocene‐Miocene boundary (∼23 Ma). The VT was active for ∼72 Myr and partly acted during out‐of‐sequence thrusting. Based on regional correlations, we propose that the ESA share a similar spatiotemporal deformation history with the central Southern Alps farther to the west. We suggest a conceptual regional tectonic model wherein multiple, broadly coeval deformation events occurred in the entire Southern Alps during their long‐lived orogenic deformation in response to generally continuous NW‐SE shortening.

Investigation of neuroprotective and therapeutic effects of cannabidiol in an acute coronary syndrome model

PurposeThe ischemia–reperfusion (I/R) injury seen in the heart can cause severe damage to essential organs such as the brain. Cannabidiol (CBD) obtained from Cannabis sativa is used today to treat various diseases. This study aimed to demonstrate CBD's neuroprotective and therapeutic properties in rats with brain damage caused by I/R in the heart. MaterialsRats were divided into four groups; sham, I/R, I/R + Prophylactic CBD, and I/R + Therapeutic CBD. End of the experiment, brain tissues were collected for biochemical, histopathological, and genetic examinations. ResultsI/R damage increased the number of degenerative neurons, caspase-3 and TNF-α immunoexpression, total oxidant status levels, and oxidative stress index. Both prophylactic and therapeutic CBD administration reduced these increased values. In addition, the relative fold changes of AMPK, PGC-1α, SIRT1, and Bcl 2 decreased in the I/R group, and the relative fold change of Bax increased, which are indicators of ER stress and apoptosis. Both administrations of CBD reversed these genes' relative fold changes. ConclusionCBD can be protective against brain injury caused by cardiac I/R damage through antioxidant, anti-inflammatory, and anti-apoptotic mechanisms.

Potts Hamiltonian Models and Molecular Dynamics Free Energy Simulations for Predicting the Impact of Mutations on Protein Kinase Stability.

Single-point mutations in kinase proteins can affect their stability and fitness, and computational analysis of these effects can provide insights into the relationships among protein sequence, structure, and function for this enzyme family. To assess the impact of mutations on protein stability, we used a sequence-based Potts Hamiltonian model trained on a kinase family multiple-sequence alignment (MSA) to calculate the statistical energy (fitness) effects of mutations and compared these against relative folding free energies (ΔΔGs) calculated from all-atom molecular dynamics free energy perturbation (FEP) simulations in explicit solvent. The fitness effects of mutations in the Potts model (ΔEs) showed good agreement with experimental thermostability data (Pearson r = 0.68), similar to the correlation we observed with ΔΔGs predicted from structure-based relative FEP simulations. Recognizing the possible advantages of using Potts models to rapidly estimate protein stability effects of kinase mutations seen in cancer genomics data, we used the Potts statistical energy model to estimate the stability effects of 65 conservative and nonconservative mutations across three distinct kinases (Wee1, Abl1, and Cdc7) with somatic mutations reported in the Genomic Data Commons (GDC) database. The ΔEs of these mutations calculated from the Potts model are consistent with the corresponding ΔΔGs from FEP simulations (Pearson ratio of 0.72). The agreement between these methods suggests that the Potts model may be used as a sequence-based tool for high-throughput screening of mutational effects as part of a computational pipeline for predicting the stability effects of mutations. We also demonstrate how the scalability of the fitness-based Potts model calculations permits analyses that are not easily accessed using FEP simulations. To this end, we employed site-saturation mutagenesis in the Potts model in order to investigate the relative stability effects of mutations seen in different cancer evolutionary scenarios. We used this approach to analyze the effects of drug pressure in Abl kinase by contrasting the relative fitness penalties of somatic mutations seen in miscellaneous cancer types with those calculated for mutations associated with cancer drug resistance. We observed that, in contrast to somatic mutations of Abl seen in various tumors that appear to have evolved neutrally, cancer mutations that evolved under drug pressure in Abl-targeted therapies tend to preserve enzyme stability.

Genome-Wide Bioinformatics Analysis of SWEET Gene Family and Expression Verification of Candidate PaSWEET Genes in Potentilla anserina

Sugars act as the main energy sources in many fruit and vegetable crops. The biosynthesis and transportation of sugars are crucial and especially contribute to growth and development. SWEET is an important gene family that plays a vital role in plants’ growth, development, and adaptation to various types of stresses (biotic and abiotic). Although SWEET genes have been identified in numerous plant species, there is no information on SWEETs in Potentilla anserina. In the present study, we performed a comprehensive genome-wide bioinformatics analysis and identified a total of 23 candidate PaSWEETs genes in the Potentilla anserina genome, which were randomly distributed on ten different chromosomes. The phylogenetic analysis, chromosomal location, gene structure, specific cis-elements, protein interaction network, and physiological characteristics of these genes were systematically examined. The identified results of the phylogenetic relationship with Arabidopsis thaliana revealed that these PaSWEET genes were divided into four clades (I, II, III, and IV). Moreover, tissue-specific gene expression through quantitative real-time polymerase chain reaction (qRT-PCR) validation exposed that the identified PaSWEETs were differentially expressed in various tissues (roots, stems, leaves, and flowers). Mainly, the relative fold gene expression in swollen and unswollen tubers effectively revealed that PaSWEETs (7, 9, and 12) were highly expressed (300-, 120-, and 100-fold) in swollen tubers. To further elucidate the function of PaSWEETs (7, 9, and 12), their subcellular location was confirmed by inserting them into tobacco leaves, and it was noted that these genes were present on the cell membrane. On the basis of the overall results, it is suggested that PaSWEETs (7, 9, and 12) are the candidate genes involved in swollen tuber formation in P. anserina. In crux, we speculated that our study provides a valuable theoretical base for further in-depth function analysis of the PaSWEET gene family and their role in tuber development and further enhancing the molecular breeding of Potentilla anserina.

Effects of supplementation of live and heat-treated Bifidobacterium animalis subspecies lactis CECT 8145 on glycemic and insulinemic response, fecal microbiota, systemic biomarkers of inflammation, and white blood cell gene expression of adult dogs.

The popularity of functional ingredients such as probiotics and postbiotics has increased as pet owners seek ways to improve the health quality and longevity of their pets. Limited research has been conducted regarding the use of probiotics and postbiotics and their effects on canine health. The objective of this study was to evaluate the effects of daily supplementation of Bifidobacterium animalis subsp. lactis CECT 8145, in both live probiotic (PRO) and heat-treated postbiotic (POST) forms, on fecal fermentative end-products and microbiome, insulin sensitivity, serum gut hormones, oxidative stress, inflammatory biomarkers, and white blood cell gene expression of adult dogs. Eighteen adult beagles and 18 adult English pointers were used in a double-blinded placebo-controlled parallel group design, with 12 animals per group (6 English pointers and 6 beagles). The study began with a 60 d adaptation period followed by a 90 d period of daily supplementation with either PRO, POST, or placebo (maltodextrin; CON). Longitudinal assessment of body weight, body condition score, and pelvic circumference did not differ among dietary supplements (P > 0.05). Throughout the experimental period, fecal scores did not differ (P > 0.05); however, fecal pH was lower (P = 0.0049) in the dogs fed POST compared with CON. A higher fecal concentration of propionate (P = 0.043) was observed in dogs fed PRO and POST when compared with CON. While PRO and POST supplementation were associated with changes in bacterial composition at the family and genus level, the overall richness and diversity of the microbiome were not significantly affected. Functional analysis of the metagenome also suggests that PRO and POST supplementation induced potentially beneficial changes in the abundance of pathways involved in pathogenicity, amino acid biosynthesis, and DNA repair. No differences in glycemic or insulinemic responses were observed among the groups (P > 0.05). Dogs supplemented with PRO had a higher (P < 0.05) mean white blood cell leptin relative fold gene expression compared with groups POST and CON. Serum metabolites and complete blood cell counts were within normal ranges and all dogs remained healthy throughout the study. Together, these data suggest that the PRO and POST can safely be supplemented for dogs. Moreover, the results of this study support further investigation of the role of PRO and POST in supporting parameters related to gut health and hormonal regulation.

Pre and Postoperative Serum miR-21 Expression Levels in Oral Cancer and it’s Association with Clinical and Histological Parameters: An Observational Study

Introduction: The patterns of microRNA (miRNA) expression have opened up new avenues in the search for prognostic biomarkers and potential therapeutic targets for various tumours. In oral carcinogenesis research, analysing microRNAs expressed in Oral Squamous Cell Carcinoma (OSCC) unveils a complex network of interest. miR-21 is known to be overexpressed in numerous solid tumours and is linked to the progression of malignancies in hepatocellular carcinomas, breast cancer, and colon carcinomas. Aim: To determine the impact of surgical excision on serum miR-21 expression levels in OSCC cases and to establish correlations with clinicopathological parameters. Materials and Methods: The study was an exploratory prospective observational study conducted with proper institutional ethical approval at the Cancer Research Institute (CRI), Himalayan Institute of Medical Sciences (HIMS), Jolly Grant, Dehradun, Uttarakhand, India. Fifty-six histologically confirmed OSCC cases were enrolled along with 25 healthy subjects as controls from May 2021 to June 2023. The relative fold expression change was calculated using the Livak method (2-∆∆ct). Data analysis was performed using software such as Statistical Package for Social Sciences (SPSS) version 20.0 and MS Excel. Non parametric statistical tests such as Wilcoxon signed-rank test, MannWhitney test, and Kruskal-Wallis test were utilised. Results: Out of 56 sample included, 28 exhibited high expression of miR-21 (p-value&lt;0.001), while 27 cases showed downregulation postsurgery (p-value&lt;0.001), with one sample showing the same level of expression. In cases where the time difference between pre and postsurgery samples was over 25 days, no significant change was observed (p-value=0.06), and similarly, in cases with a time difference below 25 days, no significant difference was noted (p-value=0.14). A significant negative correlation was found between presurgery serum Albumin to Globulin ratio (A:G ratio) and miR-21 expression (p-value &lt;0.05), while no other parameters showed a significant correlation with miR-21. Conclusion: miR-21 expression decreased significantly in some OSCC cases; however, in the majority of cases, it increased postsurgery. A significant correlation was observed between miR-21 expression and the serum A:G ratio.

Lithospheric underthrusting beneath the northern Eastern Cordillera plateau, Colombian Andes: Insights from analyzing tomographic inversion using local earthquake travel time and gravity data

The northern Eastern Cordillera in the Colombian Andes is a wide (~200 km) high-elevation (>2.5 km) subduction-related asymmetric plateau. This plateau is currently subjected to shortening and magmatic addition, and yet the nature of the lowermost crust and its transition to the underlying mantle remains poorly constrained. To improve our knowledge about the structure of the deep crust beneath the EC plateau, we conducted a joint inversion of travel times of local earthquakes and gravity data. Gravity contributes to up to 0.5% in dVp and dVs with no affectation of the shape or location of the discussed anomalies. Along the latitudinal profile with the highest resolution (~5.7°N), two anomalies are identified at depths of 40–60 km beneath the plateau. A western low-velocity anomaly is interpreted as crustal material underthrust eastward beneath the northwestern EC. This process is triggering the abrupt change in topography between the adjacent low-elevation basin and the orogenic plateau. Additionally, a high-velocity anomaly beneath the eastern flank of the EC is likely related to mantle metasomatism and westward underthrusting of the foreland lithosphere. This metasomatism is either related to the interaction between mafic magmas and the uppermost mantle or silica enrichment that occurred during a past episode of flat subduction. Evidence for foreland underthrusting includes relocated earthquakes at 33–42 km depth within the thrust system between the EC and the foreland, along with increased shortening in the eastern part of the plateau, an increase in exhumation rates, the eastward migration of deformation towards the foreland, and dominated thick-skinned deformation in the upper crust of the foreland with an eastward vergence of thrusts and related folds. Furthermore, within the central part of the plateau, our results show low velocities between 30 and 40 km depth, consistent with previous constraints, suggesting magmatic underplating beneath the Paipa-Iza volcanic complex.

Changes in gene expression in the intestinal mucus of broilers with woody breast myopathy

Previous work has shown that dietary treatments affect woody breast (WB) incidence differently, which indicates that gut conditions such as gut barrier function, inflammation, and oxidative stress are likely related to WB. In this study, dietary supplementation with antibiotics (bacitracin) or probiotics (Bacillus subtilis) was investigated for their effects on the expression of transcripts related to gut barrier function, inflammation, and oxidative stress in the mucus lining of the jejunum from broilers with or without WB. A split-plot experimental design was used in this study. The dietary treatments served as the main plot factor and the breast muscle condition was the subplot factor. On d 41, jejunum mucus was collected from 1 bird from each of 3 replicate pens in each 3 dietary treatment groups that exhibited WB and an additional bird that contained a normal breast (3 biological replicates/treatment/phenotype; 3 × 3 × 2, total N = 18). Total RNA was extracted using a commercial RNA extraction kit. The expression levels of CLDN1, MUC6, TLR2A, TLR2B, TLR4, IFN-γ, IL-1β, IL-8L1, IL-10, NOS2, and SOD were determined using 2-step RT-qPCR analysis. The gene expression difference in ΔCt values was determined after normalizing with the chicken 18S rRNA gene. When the significant differences occurred between treatments, the relative fold change was calculated using the ΔΔCt method and the significance level was calculated. The PROC GLM procedure of SAS 9.4 was used, and the level of significance was set at P ≤ 0.05. There were no significant interactive effects between diet and the breast muscle condition on the expression of any of the genes tested. However, birds with WB exhibited higher MUC6 (P < 0.0001) gene expression levels than birds with normal breast muscles. In addition, the expression of SOD decreased in birds that were fed the antibiotic diet when compared to birds that were fed the probiotic diet (P = 0.014). In conclusion, WB identified in broilers tested in the current study is attributed to increased expression of mucin, indicating a correlation between WB incidence and gel-forming mucin secretion and pathogen signaling.

Structural and functional characteristics and expression profile of the 20S proteasome gene family in Sorghum under abiotic stress

The 26S proteasome is a molecular machine that catalyzes and degrades protein intracellularly with the help of its core complex called 20S proteasome. The 20S proteasomes degrade and cleave denatured, cytotoxic, damaged, and unwanted proteins via proteolysis and impart biotic and abiotic stress tolerance in model plants. This study identified 20 genes, namely, 10 SbPA and 10 SbPB that encode for α- and β-subunits of the 20S proteasome in Sorghum bicolor (L.) Moench (2n= 20). These genes have been found distributed on the 1st, 2nd, 3rd, 4th, 5th, 7th, and 10th chromosomes. These sorghum genes were orthologous to corresponding rice. Phylogenetic analysis clustered these genes into seven clades, each with one of the seven α-subunits (1 to 7) and one of the seven β-subunits (1 to 7). In silico gene expression analysis suggested that nine genes were involved in abiotic stress response (cold, drought, and abscisic acid hormone). The expression of these proteasomal genes was studied in shoots and roots exposed to different abiotic stresses (cold, drought, and abscisic acid) by quantitative real-time polymerase chain reaction. A significant increase in the relative fold expression of SbPBA1, SbPAA1, SbPBG1, SbPBE1, and SbPAG1 genes under ABA and drought stress provides an insight into its involvement in abiotic stress. No expression was observed for cold stress of these genes indicating their non-involvement. It is believed that additional investigation into the SbPA/SbPB genes would aid in the creation of S. bicolor cultivars that are resistant to climate change.

The Pleistocene tectono-stratigraphic evolution of the northern Po Plain (Italy) around the Castenedolo and Ciliverghe hillocks

AbstractWe studied the Pleistocene subsurface stratigraphy of an area in the northern Po Plain around the isolated tectonic hillocks of Castenedolo and Ciliverghe (Brescia, Italy) in order to estimate their long-term rates of tectonic deformation. Integrated stratigraphy of a new 100-m-long core (RL13) allowed better definition of the regional Y (0.45 Ma) and R (0.87 Ma) surfaces and the related magnetostratigraphically calibrated PS1, PS2, and PS3 depositional sequences. The Y surface in the RL13 core was placed at the base of the PS3 proximal braided river system that was deposited during middle Pleistocene within the Brunhes chron. The R surface is considered to be eroded within the PS2 braid-plain deposits at ca. 0.87 Ma between the top of Jaramillo subchron and the Bruhnes chron during the late Early Pleistocene.Based on different datasets, we evaluated the sedimentation rate, which has decreased from 0.09 mm/yr with deposition of PS2, to 0.06 mm/yr with deposition of PS3. The tectonic uplift, with an average rate of ~0.1 mm/yr in the last ca. 0.87 Ma, is interpreted to be associated with a fault and related fault-propagation folding. The Castenedolo and Ciliverghe hillocks then formed due to tectonic uplift during a change in the sedimentation regime since 0.45 Ma.