Brain Perception of Different Oils on Appetite Regulation: An Anorectic Gene Expression Pattern in the Hypothalamus Dependent on the Vagus Nerve.

(1) Background: We examined the effect of the acute administration of olive oil (EVOO), linseed oil (GLO), soybean oil (SO), and palm oil (PO) on gastric motility and appetite in rats. (2) Methods: We assessed food intake, gastric retention (GR), and gene expression in all groups. (3) Results: Both EVOO and GLO were found to enhance the rate of stomach retention, leading to a decrease in hunger. On the other hand, the reduction in food intake caused by SO was accompanied by delayed effects on stomach retention. PO caused an alteration in the mRNA expression of NPY, POMC, and CART. Although PO increased stomach retention after 180 min, it did not affect food intake. It was subsequently verified that the absence of an autonomic reaction did not nullify the influence of EVOO in reducing food consumption. Moreover, in the absence of parasympathetic responses, animals that received PO exhibited a significant decrease in food consumption, probably mediated by lower NPY expression. (4) Conclusions: This study discovered that different oils induce various effects on parameters related to food consumption. Specifically, EVOO reduces food consumption primarily through its impact on the gastrointestinal tract, making it a recommended adjunct for weight loss. Conversely, the intake of PO limits food consumption in the absence of an autonomic reaction, but it is not advised due to its contribution to the development of cardiometabolic disorders.

Effectiveness of alcohol warning labels for at-risk groups and the general public: A policy-informing randomized experiment in Chile.

OBJECTIVE: The World Health Organization recommends using health-risk warnings on alcoholic beverages. This study examines the impact of separate or combined warning labels for at-risk groups and the general population on alcohol purchase decisions. METHODS: In 2022, 7758 adults who consumed alcohol or were pregnant/lactating women (54.0 % female, mean age = 40.6 years) were presented with an online store's beverage section and randomly assigned to one of six warning labels in a between-subjects experimental design: no-warning, pregnant/lactating, drinking-driving, general cancer risk, combined warnings, and assorted warnings across bottles. The main outcome, the intention to purchase an alcoholic vs. non-alcoholic beverage, was examined with adjusted risk differences using logistic regressions. RESULTS: Participants exposed to the general cancer risk warning decreased their alcoholic choices by 10.4 percentage points (pp.) (95 % CI [-0.139, -0.069], p < 0.001, OR = 0.561), while those in the pregnancy/lactation warning condition did it by 3.8 pp. (95 % CI [-0.071, -0.005], p = 0.025, OR = 0.806). The driving-drinking warning had no significant effect. Participants exposed to the combined warnings label, or the assorted warnings reduced alcohol purchase decisions by 6.1 pp. (95 % CI [-0.095, -0.028], p < 0.001, OR = 0.708) and 4.3 pp. (95 % CI [-0.076, -0.010], p = 0.011, OR = 0.782), respectively. Cancer warning outperformed other labels and was effective for subgroups such as pregnant/lactating women, young adults, and low-income individuals. CONCLUSIONS: General cancer risk warnings are more effective at reducing alcohol purchase decisions compared to warning labels for specific groups or labels using multiple warnings. In addition to warning labels, other policies should be considered for addressing well-known alcohol-related risks (e.g., drinking and driving).

A systematic review of mechanisms of PTEN gene down-regulation mediated by miRNA in prostate cancer.

Background: The Phosphatase and Tensin Homolog gene (PTEN) is pivotal in regulating diverse cellular processes, including growth, differentiation, proliferation, and cell survival, mainly by modulating the PI3K/AKT/mTOR pathway. Alterations in the expression of the PTEN gene have been associated with epigenetic mechanisms, particularly the regulation by small non-coding RNAs, such as miRNAs. Modifications in the expression levels of miRNAs that control PTEN have been shown to lead to its underexpression. This underexpression, in turn, impacts the PI3K/AKT/mTOR pathway, thereby influencing crucial mechanisms like proliferation and apoptosis, playing an important role in the initiation and progression of prostate cancer (PCa). Thus, we aimed to systematically reviewed available information concerning the regulation of PTEN mediated by miRNA in PCa. Methods: Electronic databases were searched to identify studies assessing PTEN regulation via PCa miRNAs, the search included combination of the words microRNAs, PTEN and prostatic neoplasms. The quality assessment of the articles included was carried out using an adapted version of SYRCLE and CASP tool. Results: We included 39 articles that measured the relative gene expression of miRNAs in PCa and their relationship with PTEN regulation. A total of 42 miRNAs were reported involved in the development and progression of PCa via PTEN dysregulation (34 miRNAs up-regulated and eight miRNAs down-regulated). Sixteen miRNAs were shown as the principal regulators for genetic interactions leading to carcinogenesis, being the miR-21 the most reported in PCa associated with PTEN down-regulation. We showed the silencing of PTEN could be promoted by a loop between miR-200b and DNMT1 or by direct targeting of PTEN by microRNAs, leading to the constitutive activation of PI3K/AKT/mTOR and interactions with intermediary genes support apoptosis inhibition, proliferation, invasion, and metastasis in PCa. Conclusion: According to our review, dysregulation of PTEN mediated mainly by miR-21, -20a, -20b, -93, -106a, and -106b up-regulation has a central role in PCa development and could be potential biomarkers for diagnosis, prognostic, and therapeutic targets.

Translational insights into the hormetic potential of carbon dioxide: from physiological mechanisms to innovative adjunct therapeutic potential for cancer.

Background: Carbon dioxide (CO2), traditionally viewed as a mere byproduct of cellular respiration, plays a multifaceted role in human physiology beyond simple elimination through respiration. CO2 may regulate the tumor microenvironment by significantly affecting the release of oxygen (O2) to tissues through the Bohr effect and by modulating blood pH and vasodilation. Previous studies suggest hypercapnia (elevated CO2 levels) might trigger optimized cellular mechanisms with potential therapeutic benefits. The role of CO2 in cellular stress conditions within tumor environments and its impact on O2 utilization offers a new investigative area in oncology. Objectives: This study aims to explore CO2's role in the tumor environment, particularly how its physiological properties and adaptive responses can influence therapeutic strategies. Methods: By applying a structured translational approach using the Work Breakdown Structure method, the study divided the analysis into six interconnected work packages to comprehensively analyze the interactions between carbon dioxide and the tumor microenvironment. Methods included systematic literature reviews, data analyses, data integration for identifying critical success factors and exploring extracellular environment modulation. The research used SMART criteria for assessing innovation and the applicability of results. Results: The research revealed that the human body's adaptability to hypercapnic conditions could potentially inform innovative strategies for manipulating the tumor microenvironment. This could enhance O2 utilization efficiency and manage adaptive responses to cellular stress. The study proposed that carbon dioxide's hormetic potential could induce beneficial responses in the tumor microenvironment, prompting clinical protocols for experimental validation. The research underscored the importance of pH regulation, emphasizing CO2 and carbonic acid's role in modulating metabolic and signaling pathways related to cancer. Conclusion: The study underscores CO2 as vital to our physiology and suggests potential therapeutic uses within the tumor microenvironment. pH modulation and cellular oxygenation optimization via CO2 manipulation could offer innovative strategies to enhance existing cancer therapies. These findings encourage further exploration of CO2's therapeutic potential. Future research should focus on experimental validation and exploration of clinical applications, emphasizing the need for interdisciplinary and collaborative approaches to tackle current challenges in cancer treatment.

"Opportunities and responsibilities": how do pharmacists assess their professionalism?

INTRODUCTION: Professionalism is fundamental to the existence of professions. In pharmacy, interest in this theme improved with events that examined the resocialization of pharmacists in care. With this, evaluating professionalism can help the operationalization of the theme and, consequently, the development of strategies for pharmacy consolidation before its challenges. Therefore, this study aimed to evaluate the professionalism of Brazilian pharmacists. METHODS: To meet the objective, a cross-sectional study was conducted between March 2022 and August 2023. Data were collected using the Brazilian version of the "Modification of Hall's Professionalism Scale for Use with Pharmacists". The scale has 39 items grouped into the domains: autonomy, vocation, professional council, self-regulation, continuing education, and altruism. Data were analyzed using descriptive statistics and an ANOVA analysis of variance with post-hoc Hochberg or Games-Howell tests with Bootstrapping was conducted to verify differences between groups. RESULTS: 600 pharmacists participated in this study. The majority (69%) was female and carried out their professional activities in community pharmacies (50%). Professionalism scores ranged between 14 and 29 points, with an average of 22.8 points. Pharmacists working in outpatient clinics had higher scores in most factors, namely, altruism, continuing education, professional council, vocation, and autonomy. This indicates that the inclination of pharmacists to occupy areas focused on care can be significant to assess professionalism. CONCLUSIONS: The data obtained indicate that pharmacists working in outpatient clinics had higher professionalism scores compared to others. This corroborates the worldwide trend experienced by pharmacy in recent decades, which is the execution of increasingly patient-centered practice models.

Peltier-based temperature regulation: A method for performance optimization in solid-state lasers.

This article presents a direct method for temperature control in solid-state lasers, where temperature stability is crucial for optimizing the performance and reliability of such lasers. The proposed method utilizes Peltier chips for both cooling and heating the laser crystal to achieve precise temperature regulation. The system design is based on the step response of the open-loop thermal system and employs a proportional-integral (PI) controller for closed-loop temperature control. Comprehensive testing on a femtosecond Titanium-Sapphire Laser (Ti:Sapphire laser) demonstrated that the system is capable of maintaining the desired operating temperature with remarkable stability and efficiency, highlighting its practicality for real-world applications. Method Outline:•Utilization of Peltier chips for precise temperature control.•Estimation of first-order transfer function based on step response.•Implementation of a proportional-integral (PI) controller for closed-loop temperature regulation.

Type 1 fimbria and P pili: regulatory mechanisms of the prototypical members of the chaperone-usher fimbrial family.

Adherence to both cellular and abiotic surfaces is a crucial step in the interaction of bacterial pathogens and commensals with their hosts. Bacterial surface structures known as fimbriae or pili play a fundamental role in the early colonization stages by providing specificity or tropism. Among the various fimbrial families, the chaperone-usher family has been extensively studied due to its ubiquity, diversity, and abundance. This family is named after the components that facilitate their biogenesis. Type 1 fimbria and P pilus, two chaperone-usher fimbriae associated with urinary tract infections, have been thoroughly investigated and serve as prototypes that have laid the foundations for understanding the biogenesis of this fimbrial family. Additionally, the study of the mechanisms regulating their expression has also been a subject of great interest, revealing that the regulation of the expression of the genes encoding these structures is a complex and diverse process, involving both common global regulators and those specific to each operon.

Use of animals for scientific and educational purposes by Mexican institutions.

In Mexico, it is not known which institutions use animals for scientific purposes. This work reports, based on data requested from the National Institute of Transparency, Access to Information and Protection of Personal Data (INAI), the types of institutions that use animals for research and how many of these have an ethics committee. Research centres, colleges, hospitals, national institutes, technical colleges, and public universities are the types of institutions that report using animals for experimentation. Only 54% of institutions have ethics committees. Mexican institutions from 2015 to 2021 used a total of 2,112,786 animals. Mammals are the most widely used animal group. The scientific purposes for using animals depend on the type of institution that uses them. In Mexico, it is necessary to update the regulations in order to regulate the use, protection and the care of laboratory animals.

En México se desconoce cuáles son las instituciones que utilizan animales con fines científicos. Se reporta, a partir de datos solicitados al Instituto Nacional de Transparencia, Acceso a la Información y Protección de Datos Personales (INAI), los tipos de instituciones que usan animales y cuántas poseen un comité interno para el cuidado y uso de los animales de laboratorio. Los centros de investigación, colegios, hospitales, institutos nacionales, tecnológicos y universidades públicas son los tipos de instituciones que reportaron usar animales. El 54% de las instituciones poseen comités de ética. Un total de 2,112,786 animales fueron usados por instituciones del 2015 al 2021. Los mamíferos es el grupo animal más utilizado. El uso de los animales se encuentra en función del tipo de institución que los utiliza. En México, es necesario actualizar la normatividad, con el fin de regular el uso, la protección y el cuidado de los animales de laboratorio.

Jumping with control: the interplay between psychological constructs and run-up variability in elite jumpers.

Objectives: The purpose of this study was to examine the relationship between psychological aspects (emotional regulation, self-control, mood states, and perceived stress) and components of run-up variability in horizontal jumps and to conduct comparisons based on sex, events (long jump and triple jump), and contextual situations (training versus competition). Methods: A total of 10 elite-level athletes (five males and five females) with a mean age of 27.14 (±4.25) years were recruited for the study. All participants had competed nationally or internationally and had 13.10 (±3.48) years of athletic experience. Data were collected during competitions and training sessions for 5 weeks. The participants completed the Brunel Mood Scale, Emotional Regulation Questionnaire, Brief Self-Control Scale, and Visual Analogical Scale of Perceived Stress before each session. The components of run-up variability of successful and failed attempts were measured using video analysis. Data were analyzed using a t-test, Pearson's correlation, and Cohen's d. Results: Athletes specializing in long jump and triple jump displayed similar psychological and run-up variability characteristics. However, females showed higher values for tension and depression, whereas males had higher run-up speeds and vigor. In competitions, athletes tended to have higher vigor, lower fatigue and confusion, an earlier beginning of the adjustment phase, fewer failed attempts, and higher run-up speed than during training. Emotional regulation is inversely related to depression in women during competitions, whereas higher self-control is associated with fewer failed jumps. Conclusion: Athletes competing in the long jump and the triple jump do not differ in psychological traits and run-up characteristics, which suggests that similar training strategies can be used in both events. However, different solutions should be used considering the sex of athletes, with a particular focus on utilizing emotion regulation tools to modulate depression in female jumpers. It is recommended to include training sessions that simulate competition demands, primarily to ensure the early onset of the run-up adjustment phase.

Differential Regulation of Hemichannels and Gap Junction Channels by RhoA GTPase and Actin Cytoskeleton: A Comparative Analysis of Cx43 and Cx26.

Connexins (Cxs) are transmembrane proteins that assemble into gap junction channels (GJCs) and hemichannels (HCs). Previous researches support the involvement of Rho GTPases and actin microfilaments in the trafficking of Cxs, formation of GJCs plaques, and regulation of channel activity. Nonetheless, it remains uncertain whether distinct types of Cxs HCs and GJCs respond differently to Rho GTPases or changes in actin polymerization/depolymerization dynamics. Our investigation revealed that inhibiting RhoA, a small GTPase that controls actin polymerization, or disrupting actin microfilaments with cytochalasin B (Cyto-B), resulted in reduced GJCs plaque size at appositional membranes and increased transport of HCs to non-appositional plasma membrane regions. Notably, these effects were consistent across different Cx types, since Cx26 and Cx43 exhibited similar responses, despite having distinct trafficking routes to the plasma membrane. Functional assessments showed that RhoA inhibition and actin depolymerization decreased the activity of Cx43 GJCs while significantly increasing HC activity. However, the functional status of GJCs and HCs composed of Cx26 remained unaffected. These results support the hypothesis that RhoA, through its control of the actin cytoskeleton, facilitates the transport of HCs to appositional cell membranes for GJCs formation while simultaneously limiting the positioning of free HCs at non-appositional cell membranes, independently of Cx type. This dynamic regulation promotes intercellular communications and reduces non-selective plasma membrane permeability through a Cx-type dependent mechanism, whereby the activity of Cx43 HCs and GJCs are differentially affected but Cx26 channels remain unchanged.

Effect of HDAC9 on the differentiation of chicken embryonic stem cells into male germ cells.

Histone deacetylase 9 (HDAC9) is a histone deacetylase (HDAC) subtype IIa protein that deacetylates histone 3 (H3), histone 4 (H4), and nonhistone proteins in vivo to alter chromosomal shape and regulate gene transcription. There have been few studies on the regulatory influence of the HDAC9 gene on the differentiation of chicken embryonic stem cells (cESCs) into male germ cells, and the significance of HDAC9 is still unknown. Therefore, we explored the specific role of HDAC9 during differentiation of the cESCs of Jilin Luhua chickens through inhibition or overexpression. In medium supplemented with 10-5 mol/L retinoic acid (RA), cESCs were stimulated to develop into germ cells. HDAC9 and germline marker gene mRNA and protein levels were measured using qRT‒PCR and western blotting. During the differentiation of cESCs into male germ cells, overexpression of the HDAC9 gene greatly increased the mRNA and protein expression levels of the germline marker genes Stra8, Dazl, c-kit, and integrin ɑ6. The HDAC9 inhibitor TMP195 significantly decreased the mRNA and protein expression levels of the above markers. In summary, HDAC9 positively regulates the differentiation of cESCs.

Emotional regulation in the classroom: detection of multiple cases from systematic observation.

Introduction: Emotional education is learned by living with others. This study analyzes how different actors participate in the classroom, influencing students' emotional regulation. Methods: Using a mixed methods approach [structured in macro-stages QUAL-QUAN-QUAL], 9 classes in three Chilean schools with children aged 10 to 12 are systematically observed. The audio-recorded sessions are transcribed (qualitative data) for coding from the observation instrument, and then, once the data quality is verified, they are analyzed quantitatively (quantitizing). A lag sequential analysis is performed to detect regularities and existing sequences, and a polar coordinate analysis is performed to observe the relationships among the categories in each class. Results: Once the results of the analyses have been obtained, similarities are sought to detect the presence of multiple cases based on the two analysis techniques. The multiple cases detected are presented, detailing how interactions between teachers and students contribute to the emotional regulation and climate in the daily life of the classroom. Discussion: The interpretation of the results makes it possible to revisit the qualitative aspect of the mixed methods and to discuss the need to problematize the role of teachers in students' development and autonomy.

Brain Networks, Neurotransmitters and Psychedelics: Towards a Neurochemistry of Self-Awareness.

PURPOSE OF REVIEW: Self-awareness can be defined as the capacity of becoming the object of one's own awareness and, increasingly, it has been the target of scientific inquiry. Self-awareness has important clinical implications, and a better understanding of the neurochemical basis of self-awareness may help clarifying causes and developing interventions for different psychopathological conditions. The current article explores the relationship between neurochemistry and self-awareness, with special attention to the effects of psychedelics. RECENT FINDINGS: The functioning of self-related networks, such as the default-mode network and the salience network, and how these are influenced by different neurotransmitters is discussed. The impact of psychedelics on self-awareness is reviewed in relation to specific processes, such as interoception, body ownership, agency, metacognition, emotional regulation and autobiographical memory, within a framework based on predictive coding. Improved outcomes in emotional regulation and autobiographical memory have been observed in association with the use of psychedelics, suggesting higher-order self-awareness changes, which can be modulated by relaxation of priors and improved coping mechanisms linked to cognitive flexibility. Alterations in bodily self-awareness are less consistent, being potentially impacted by doses employed, differences in acute/long-term effects and the presence of clinical conditions. Future studies investigating the effects of different molecules in rebalancing connectivity between resting-state networks may lead to novel therapeutic approaches and the refinement of existing treatments.

In Silico Identification of Dysregulated miRNAs Targeting KRAS Gene in Pancreatic Cancer.

Pancreatic cancer (PC) is highly lethal, with KRAS mutations in up to 95% of cases. miRNAs inversely correlate with KRAS expression, indicating potential as biomarkers. This study identified miRNAs targeting KRAS and their impact on PC characteristics using in silico methods. dbDEMC identified dysregulated miRNAs in PC; TargetScan, miRDB, and PolymiRTS 3.0 identified miRNAs specific for the KRAS gene; and OncomiR evaluated the association of miRNAs with clinical characteristics and survival in PC. The correlation between miRNAs and KRAS was analysed using ENCORI/starBase. A total of 210 deregulated miRNAs were identified in PC (116 overexpressed and 94 underexpressed). In total, 16 of them were involved in the regulation of KRAS expression and 9 of these (hsa-miR-222-3p, hsa-miR-30a-5p, hsa-miR-30b-5p, hsa-miR-30e-5p, hsa-miR-377-3p, hsa-miR-495-3p, hsa-miR-654-3p, hsa-miR-877-5p and hsa-miR-885-5p) were associated with the clinical characteristics of the PC. Specifically, the overexpression of hsa-miR-30a-5p was associated with PC mortality, and hsa-miR-30b-5p, hsa-miR-377-3p, hsa-miR-495-3p, and hsa-miR-885-5p were associated with survival. Correlation analysis revealed that the expression of 10 miRNAs is correlated with KRAS expression. The dysregulated miRNAs identified in PC may regulate KRAS and some are associated with clinically relevant features, highlighting their potential as biomarkers and therapeutic targets in PC treatment. However, experimental validation is required for confirmation.

MicroRNA Transcriptomes Reveal Prevalence of Rare and Species-Specific Arm Switching Events During Zebrafish Ontogenesis.

In metazoans, microRNAs (miRNAs) are essential regulators of gene expression, affecting critical cellular processes from differentiation and proliferation, to homeostasis. During miRNA biogenesis, the miRNA strand that loads onto the RNA-induced Silencing Complex (RISC) can vary, leading to changes in gene targeting and modulation of biological pathways. To investigate the impact of these "arm switching" events on gene regulation, we analyzed a diverse range of tissues and developmental stages in zebrafish by comparing 5p and 3p arms accumulation dynamics between embryonic developmental stages, adult tissues, and sexes. We also compared variable arm usage patterns observed in zebrafish to other vertebrates including arm switching data from fish, birds, and mammals. Our comprehensive analysis revealed that variable arm usage events predominantly take place during embryonic development. It is also noteworthy that isomiR occurrence correlates to changes in arm selection evidencing an important role of microRNA distinct isoforms in reinforcing and modifying gene regulation by promoting dynamics switches on miRNA 5p and 3p arms accumulation. Our results shed new light on the emergence and coordination of gene expression regulation and pave the way for future investigations in this field.

Building bio-innovation systems through advanced biotechnology education.

We discuss the role of advanced biotechnology education in fostering sustainable bio-innovation systems. As a case study, we focus on Paraguay's Graduate Diploma in Innovation Management and Biotechnological Projects, which emphasizes interdisciplinary collaboration, stakeholder integration, and professionals skilled in the interplay between biotechnology, society, and governance. We highlight the relevance of educational programs in addressing the gap between academic research and industrial needs, thereby contributing to sustainable growth in the biotechnology sector.

RNA interference of the clock gene period disrupts circadian rhythms in the expression of genes related to insecticide resistance in the chagas disease vector Triatoma infestans (Hemiptera: Reduviidae).

In Triatoma infestans it was observed pyrethroid resistance attributed in part to an elevated oxidative metabolism mediated by cytochromes P450. The nicotinamide adenine dinucleotide phosphate (NADPH) cytochrome P450 reductase (CPR) plays a crucial role in catalysing the electron transfer from NADPH to all cytochrome P450s. The daily variations in the expression of CPR gene and a P450 gene (CYP4EM7), both associated with insecticide resistance, suggested that their expressions would be under the endogenous clock control. To clarify the involvement of the clock in orchestration of the daily fluctuations in CPR and CYP4M7 genes expression, it was proposed to investigate the effect of silencing the clock gene period (per) by RNA interference (RNAi). The results obtained allowed to establish that the silencing of per gene was influenced by intake schemes used in the interference protocols. The silencing of per gene in T. infestans reduced its expression at all the time points analysed and abolished the characteristic rhythm in the transcriptional expression of per mRNA. The effect of the per gene silencing in the expression profiles at the transcriptional level of CPR and CYP4EM7 genes showed the loss of rhythmicity and demonstrated the biological clock involvement in the regulation of t heir expression.

Phaseolotoxin: Environmental Conditions and Regulatory Mechanisms Involved in Its Synthesis.

Phaseolotoxin is an antimetabolite toxin produced by diverse pathovars of Pseudomonas syringae which affects various plants, causing diseases of economic importance. Phaseolotoxin contributes to the systemic dissemination of the pathogen in the plant, therefore it is recognized as a major virulence factor. Genetic traits such as the Pht cluster, appear defining to the toxigenic strains phaseolotoxin producers. Extensive research has contributed to our knowledge concerning the regulation of phaseolotoxin revealing a complex regulatory network that involves processes at the transcriptional and posttranscriptional levels, in which specific and global regulators participate. Even more, significant advances in understanding how specific signals, including host metabolites, nutrient sources, and physical parameters such as the temperature, can affect phaseolotoxin production have been made. A general overview of the phaseolotoxin regulation, focusing on the chemical and physical cues, and regulatory pathways involved in the expression of this major virulence factor will be given in the present work.

Dynamic changes in mRNA nucleocytoplasmic localization in the nitrate response of Arabidopsis roots.

Nitrate is a nutrient and signal that regulates gene expression. The nitrate response has been extensively characterized at the organism, organ, and cell-type-specific levels, but intracellular mRNA dynamics remain unexplored. To characterize nuclear and cytoplasmic transcriptome dynamics in response to nitrate, we performed a time-course expression analysis after nitrate treatment in isolated nuclei, cytoplasm, and whole roots. We identified 402 differentially localized transcripts (DLTs) in response to nitrate treatment. Induced DLT genes showed rapid and transient recruitment of the RNA polymerase II, together with an increase in the mRNA turnover rates. DLTs code for genes involved in metabolic processes, localization, and response to stimulus indicating DLTs include genes with relevant functions for the nitrate response that have not been previously identified. Using single-molecule RNA FISH, we observed early nuclear accumulation of the NITRATE REDUCTASE 1 (NIA1) transcripts in their transcription sites. We found that transcription of NIA1, a gene showing delayed cytoplasmic accumulation, is rapidly and transiently activated; however, its transcripts become unstable when they reach the cytoplasm. Our study reveals the dynamic localization of mRNAs between the nucleus and cytoplasm as an emerging feature in the temporal control of gene expression in response to nitrate treatment in Arabidopsis roots.

Molecular insights into the regulatory landscape of PKC-related kinase-2 (PRK2/PKN2) using targeted small compounds.

The PKC-related kinases (PRKs, also termed PKNs) are important in cell migration, cancer, hepatitis C infection, and nutrient sensing. They belong to a group of protein kinases called AGC kinases that share common features like a C-terminal extension to the catalytic domain comprising a hydrophobic motif. PRKs are regulated by N-terminal domains, a pseudosubstrate sequence, Rho-binding domains, and a C2 domain involved in inhibition and dimerization, while Rho and lipids are activators. We investigated the allosteric regulation of PRK2 and its interaction with its upstream kinase PDK1 using a chemical biology approach. We confirmed the phosphoinositide-dependent protein kinase 1 (PDK1)-interacting fragment (PIF)-mediated docking interaction of PRK2 with PDK1 and showed that this interaction can be modulated allosterically. We showed that the polypeptide PIFtide and a small compound binding to the PIF-pocket of PRK2 were allosteric activators, by displacing the pseudosubstrate PKL region from the active site. In addition, a small compound binding to the PIF-pocket allosterically inhibited the catalytic activity of PRK2. Together, we confirmed the docking interaction and allostery between PRK2 and PDK1 and described an allosteric communication between the PIF-pocket and the active site of PRK2, both modulating the conformation of the ATP-binding site and the pseudosubstrate PKL-binding site. Our study highlights the allosteric modulation of the activity and the conformation of PRK2 in addition to the existence of at least two different complexes between PRK2 and its upstream kinase PDK1. Finally, the study highlights the potential for developing allosteric drugs to modulate PRK2 kinase conformations and catalytic activity.