Nucleolar Research Articles

Nucleus-nucleolus features of certain species of the Pinaceae family

Aim: The aim of the study was to establish the interspecific features of the functioning of the nucleolus-forming organizer on the example of 6 species of the family Pinaceae. Methods. The study involved samples of different species of conifers that grow within the natural range and conditions of introduction. Silver nitric acid staining was used to visualize the nucleoli. Quantitative and dimensional features of the nucleus and nucleoli in the interphase cells of seedlings were studied, and the values of the nucleus-nucleolus ratio were calculated. Results. Interspecific polymorphism in the number and size of nucleoli, nuclei and nucleus-nucleolus ratio in six species and forms of the Pinaceae L. family was revealed. Conclusions. The studied species of conifers obviously have interspecific differentiation by nucleus-nucleolus parameters. The studied characteristics are influenced not only by the species, age and condition of the plant, but also by the natural and climatic conditions in which the population exists.

Porous Fly Ash/Aluminosilicate Microspheres-Based Composites Containing Lightweight Granules Using Liquid Glass as Binder

The modern energy-saving vector of development in building materials science is being implemented in a complex way through the development of new heat-insulating materials with the simultaneous exclusion of low-ecological cement from them. This article presents the results of the development of resource-saving technology for a heat-insulating composite material. The research is devoted to the development of scientific ideas about the technology and properties of effective cementless lightweight concretes. The aim of the work is to create a heat-insulating composite material based on porous granules and a matrix from mixtures of liquid glass and thermal energy waste. The novelty of the work lies in establishing the patterns of formation of a stable structure of a porous material during thermal curing of liquid glass with technogenic fillers. Studies of liquid glass mixtures with different contents of fly ash and aluminosilicate microspheres revealed the possibility of controlling the properties of molding masses in a wide range. To obtain a granular material, liquid glass mixtures of plastic consistency with a predominance of aluminosilicate microspheres are proposed. The matrix of composite materials is formed by a mobile mixture of liquid glass and a combined filler, in which fly ash predominates. The parameters of heat treatment of granular and composite materials are established to ensure the formation of a strong porous waterproof structure. The possibility of regulating the structure of composite materials due to different degrees of filling the liquid glass matrix with porous granules is shown. A heat-insulating concrete based on porous aggregate has been developed, characterized by the genetic commonality of the matrix and the granular component, density of 380–650 kg/m3, thermal conductivity of 0.095–0.100 W/(m °C) and strength of 3.5–9.0 MPa, resistance under conditions of variable values of humidity and temperature. A basic technological scheme for the joint production of granular and composite materials from liquid glass mixtures is proposed.

Functional imaging and targeted drug delivery in mice and patient tumors with a cell nucleolus-localizing and tumor-targeting peptide

Tumor-targeting peptides have profound clinical implications in early detection and delineation of microscopic lesions for surgical resection, and also delivery of therapeutics with reduced systemic toxicity. Here, we demonstrate that a peptide (RS), evolved from a previously reported hepatocellular carcinoma (HCC)-targeting peptide P47, enables improved HCC micrometastasis discrimination and delineation from noncancerous tissues in murine orthotopic mice and patient biopsies, with up to 21-fold contrast. Importantly, RS targets non-small cell lung (NSCLC) and colon cancers in mice and patient biopsies, with higher selectivity for highly proliferative tumor nodules. Moreover, RS localizes to cell nucleoli of HCC, NSCLC, breast, colon and cervical cancer cells and induces nucleolar stress when conjugated with chemotherapeutic Oxaliplatin (OXA) (RS-OXA), demonstrating both cellular and subcellular targeting. RS-delivered OXA elicits significant tumor retardation in orthotopic HCC mice with markedly reduced systemic toxicity compared to OXA alone. Injection of fluorescence-labeled RS enables dynamic visualization of tumor growth in RS-OXA-treated subcutaneous HCC mice. Our study demonstrates that RS targets a spectrum of tumors and localizes to cell nucleolus, thus enabling functional imaging and targeted delivery of OXA in HCC mice, and consequently provides a versatile tool for tumor imaging and targeted therapeutics.

CX-5461 causes nucleolar compaction, alteration of peri- and intranucleolar chromatin arrangement, an increase in both heterochromatin and DNA damage response

In this study, we characterize the changes in nucleolar morphology and its dynamics induced by the recently introduced compound CX-5461, an inhibitor of ribosome synthesis. Time-lapse imaging, immunofluorescence and ultrastructural analysis revealed that exposure of cells to CX-5461 has a profound impact on their nucleolar morphology and function: nucleoli acquired a compact, spherical shape and display enlarged, ring-like masses of perinucleolar condensed chromatin. Tunnels consisting of chromatin developed as transient structures running through nucleoli. Nucleolar components involved in rRNA transcription, fibrillar centres and dense fibrillar component with their major constituents ribosomal DNA, RNA polymerase I and fibrillarin maintain their topological arrangement but become reduced in number and move towards the nucleolar periphery. Nucleolar changes are paralleled by an increased amount of the DNA damage response indicator γH2AX and DNA unwinding enzyme topoisomerase I in nucleoli and the perinucleolar area suggesting that CX-5461 induces torsional stress and DNA damage in rDNA. This is corroborated by the irreversibility of the observed altered nucleolar phenotypes. We demonstrate that incubation with CX-5461, apart from leading to specific morphological alterations, increases senescence and decreases cell replication. We discuss that these alterations differ from those observed with other drugs interfering with nucleolar functions.

Box C/D snoRNA SNORD89 influences the occurrence and development of endometrial cancer through 2’-O-methylation modification of Bim

The small nucleolar RNA (snoRNA) is a type of small non-coding RNA widely distributed in the nucleoli of eukaryotic cells, promoting cancer development. The aim of this study was to assess box C/D snoRNA 89 (SNORD89) dysregulations in endometrial cancer. According to the TCGA database as well as the International Federation of Gynecology and Obstetrics (FIGO), higher SNORD89 expression is found in endometrial cancer tissues. In addition, the SNORD89 expression level was higher in endometrial carcinoma with lymph node metastasis than in endometrial carcinoma without lymph node metastasis. By interacting with the conservative chaperone protein methylase fibrillarin (Fbl), SNORD89 inhibits the translation process of the Bim gene, leading to a decrease in Bim protein. Cancer-promoting effect of SNORD89 can be reversed by Fbl knockdown or Bim overexpressing. What’s more, ASO-mediated silencing of SNORD89 could inhibit endometrial cancer cell proliferation and migration ability. Taken together, SNORD89 can modify Bim through 2′-O-methylation and affect downstream signaling pathways to promote endometrial cancer occurrence and development. The role of methylation modification in the prevention and treatment of endometrial cancer provides a new understanding and SNORD89 may be a new diagnostic and therapeutic target for endometrial cancer.

Small molecules enhancers of amyloid aggregation of C-terminal domain of Nucleophosmin 1 in acute myeloid leukemia

The “Acute Myeloid Leukemia with gene mutations'' group includes mutations in Nucleophosmin 1(NPM1) that is an abundant multifunctional protein with chaperon functions. This protein also takes part to rRNA maturation in ribosome biogenesis, tumor suppression and nucleolar stress response. Mutations of NPM1 associated to AML present in its C-terminal domain (CTD) unable its correct folding and confer it an aberrant cytoplasmatic localization (NPMc+). AML cells with NPM1 mutations retain a certain amount of wt NPM1 in the nucleolus and since NPM1 acts as a hub protein, the nucleolus of AML cells are more vulnerable with respect to cells expressing only wt NPM1. Thus, interfering with the levels or the oligomerization status of NPM1 may influence its capability to properly build up the nucleolus in AML cells. Our biophysical recent results demonstrated that AML-CTDs contain regions prone to amyloid aggregation and, herein, we present results oriented to exploit this amylodogenesis in a potential therapeutic way. We evaluated the different ability of two small molecules to enhance amyloid aggregation through complementary biophysical approaches as fluorescence and Circular Dichroism spectroscopies, Scanning Electron Microscopy and cell-viability assays, to evaluate the cytoxicity of these molecules in AML cells lines. These findings could pave the way into molecular mechanisms of NPM1c and in novel therapeutic routes toward AML progression.

CK2α as a prognostic factor in invasive ductal carcinomas of the breast: cancer recurrence prognosis by surgical sampling

Breast cancer is the most common cancer in Japanese and Asian women. Although the 5-year survival rate of these patients is relatively high, recurrences and fatalities do occur. The objective of this work was to develop a reliable prognostic indicator to guide therapeutic intervention without waiting for recurrence or metastasis. We have previously shown that positive immunohistochemical staining of a protein kinase, CK2α (CSNK2A1), in nucleoli of invasive ductal carcinoma cells is strongly associated with tumour recurrence and poor patient outcomes. For this study, we did an immunohistochemistry investigation of the location of CK2α in the nuclei and nucleoli of formalin-fixed, paraffin-embedded tissue block samples from patients with invasive ductal carcinoma who underwent surgery in 2008-14 but no neoadjuvant chemotherapy at Hoshi General Hospital, Fukushima, Japan. Furthermore, a new study with 59 patients with luminal type breast cancer was done. Patients were followed-up (clinicopathological information and outcomes) for more than 5 years. To characterise CK2α-associated proteins systematically, nuclear and nucleolar extracts were prepared from MCF-7 cells and immunoprecipitated with anti-CK2α antibodies followed by nano-flow liquid chromatography mass spectrometry analysis. We did a Kaplan-Meier analysis to evaluate whether nucleolar CK2 staining might be associated with poor recurrence-free survival. This retrospective study was approved by the Institutional Review Boards of Fukushima Medical University and Hoshi General Hospital. Samples from 112 patients were included. 105 (94%) of 112 patients with invasive ductal carcinoma had high CK2α expression in nuclei, and 41 (37%) of 112 displayed it predominantly in nucleoli. Clinical and pathological malignancy was strongly correlated with high nuclear and nucleolar CK2α expression. Recurrence-free survival was significantly reduced in patients with positive nucleolar CK2α staining (n=41; median 7·89 years [IQR 4·86-8·86]), comparing with patients with negative nucleolar staining (n=71; median 8·89 years [7·87-9·79]; p<0·0001). Kaplan-Meier analysis showed a decrease of approximately 50% in recurrence-free survival in patients with nucleolar CK2α-positive stain in the triple-negative breast cancer (p=0·0069) and post-operative stage 3 (p=0·0073) groups. By contrast, no patients who relapsed or died in the triple-negative breast cancer group had negative staining for nucleolar CK2α. Evaluation of nucleolar CK2α can be considered as an independent prognostic factor because it reached significance in multivariate analysis (hazard ratio [HR] 5·26 [95% CI 1·35-20·55]; p=0·017). In addition, the HR of nucleolar CK2α-positive staining was less affected by adjustments in Cox proportional multivariate analysis. Cluster analysis showed that nucleolar CK2α-positive staining contributed more than any other clinicopathological factor to predicting the variation in the future length of survival. Analysis with luminal type breast cancer (n=59), in which the risk of recurrence is generally low and which is treated postoperatively with hormone therapy alone, showed that cancer recurrence and poor patient outcomes strongly associate with nucleolar CK2α-positive stain (n=7; p=0·037) by Kaplan-Meier analysis. In this case, nucleolar CK2α is considered a sole independent prognostic factor (HR 2·11 [95% CI 0·27-3·69]; p=0·0087). Evaluation of this method in other solid tumours is ongoing. Our results suggest that the evaluation of nucleolar CK2α-positive staining might be a new and independent prognostic factor that can improve treatment efficacy and patient quality of life after surgical resection of the tumour. The application of this analysis could contribute to early treatment strategy decisions and to companion diagnostics. The protein interactome of nuclear CK2 complexes showed CK2α's role in protein synthesis and RNA post-transcriptional modification. Specifically, a more detailed evaluation of CK2α function in nucleolar events in malignant cells might lead to the development of novel, more accurately targeted anti-cancer therapeutics. In conclusion, evaluating nucleolar CK2α staining in combination with other clinicopathological indicators might allow for a more accurate prediction of future recurrence or metastasis, which cannot be accomplished by current methods. Japan Agency for Medical Research and Development (grant number 20lm0203006j0004).

Hybridisation chain reaction-based visualisation and screening for lncRNA profiles in clear-cell renal-cell carcinoma.

Analysis of long noncoding RNA (lncRNA) localisation at both the tissue and subcellular levels can provide important insights into the cell types that are important for their function. By applying new fluorescent in situ hybridisation technique called hybridisation chain reaction (HCR), we achieved a high-throughput lncRNA visualisation and evaluation of clinical samples. Assessing 1728 pairs of 16 lncRNAs and clear-cell renal-cell carcinoma (ccRCC) specimens, three lncRNAs (TUG1, HOTAIR and CDKN2B-AS1) were associated with ccRCC prognosis. Furthermore, we derived a new lncRNA risk group of ccRCC prognosis by combining the expression levels of these three lncRNAs. Examining genomic alterations underlying this classification revealed prominent features of tumours that could serve as potential biomarkers for targeting lncRNAs. We then derived combination of HCR with expansion microscopy and visualised nanoscale-resolution HCR signals in cell nuclei, uncovering intracellular colocalization of three lncRNA (TUG1, HOTAIR and CDKN2B-AS1) signals such as those located intra- or out of the nucleus or nucleolus in cancer cells. LncRNAs are expected to be desirable noncoding targets for cancer diagnosis or treatments. HCR involves plural probes consisting of small DNA oligonucleotides, clinically enabling us to detect cancerous lncRNA signals simply and rapidly at a lower cost.

Styryl Hemicyanine Dye (E)-3-Methyl-2-(4-thiomorpholinostyryl)benzo[d]thiazol-3-ium Iodide for Nucleic Acids and Cell Nucleoli Visualization

(E)-3-Methyl-2-(4-thiomorpholinostyryl)benzo[d]thiazol-3-ium iodide 1 was prepared by a convenient and reliable reaction procedure. The slight molar excess of the starting benzaldehyde and the mixture of ethanol: ethyl acetate in the ratio 3:1 as a solvent afforded a pure reaction product. The photophysical properties of the dye in a TE buffer in the absence and presence of double-stranded DNA (dsDNA) were elucidated. The low intrinsic fluorescence of 1 in TE buffer is followed by an increase in the fluorescence after dsDNA binding. The dye is nontoxic for stem cells from apical papilla and the most concentrated fluorescence is detected in the cell nucleoli.

Fluorescent carbon dots with real-time nucleolus-monitoring capability for gene delivery and biosensing of NO2– and pH

To achieve gene therapy, DNA, one of the most commonly used therapeutic nucleic acids, is typically employed to transfer into the nucleous. However, it remains extremely challenging to seek vectors with high transfection efficiency and low toxicity to introduce DNA into the nucleolus. Herein, we have designed and developed green fluorescent carbon dots (G-CDs) for nucleolus targeting, gene delivery, and biosensing of NO2– and pH. G-CDs were synthesized by a simple hydrothermal mean using 3,5-diaminobenzoic acid and citric acid as precursors. As-prepared G-CDs not only possess bright green fluorescence with respectable fluorescence quantum yield of 41%, but also exhibit low cytotoxicity and superior biocompatibility. Interestingly, G-CDs display accurate nucleolus targeting characteristic due to the response of G-CDs to DNA in the nucleolus. More unexpectedly, G-CDs could effectively deliver exogenous ROX-labelled DNA into the nucleolus of HeLa cells, which is the first time that fluorescent CDs are used as nanocarriers to transfer DNA to nucleolus. What’s more, G-CDs have been expanded to visually monitor NO2– and pH fluctuations in living cells.

Bovine Pancreatic RNase A: An Insight into the Mechanism of Antitumor Activity In Vitro and In Vivo.

In this investigation, we extensively studied the mechanism of antitumor activity of bovine pancreatic RNase A. Using confocal microscopy, we show that after RNase A penetration into HeLa and B16 cells, a part of the enzyme remains unbound with the ribonuclease inhibitor (RI), resulting in the decrease in cytosolic RNAs in both types of cells and rRNAs in the nucleoli of HeLa cells. Molecular docking indicates the ability of RNase A to form a complex with Ku70/Ku80 heterodimer, and microscopy data confirm its localization mostly inside the nucleus, which may underlie the mechanism of RNase A penetration into cells and its intracellular traffic. RNase A reduced migration and invasion of tumor cells in vitro. In vivo, in the metastatic model of melanoma, RNase A suppressed metastases in the lungs and changed the expression of EMT markers in the tissue adjacent to metastatic foci; this increased Cdh1 and decreased Tjp1, Fn and Vim, disrupting the favorable tumor microenvironment. A similar pattern was observed for all genes except for Fn in metastatic foci, indicating a decrease in the invasive potential of tumor cells. Bioinformatic analysis of RNase-A-susceptible miRNAs and their regulatory networks showed that the main processes modulated by RNase A in the tumor microenvironment are the regulation of cell adhesion and junction, cell cycle regulation and pathways associated with EMT and tumor progression.

The Highest Density of Phosphorylated Histone H1 Appeared in Prophase and Prometaphase in Parallel with Reduced H3K9me3, and HDAC1 Depletion Increased H1.2/H1.3 and H1.4 Serine 38 Phosphorylation.

Background: Variants of linker histone H1 are tissue-specific and are responsible for chromatin compaction accompanying cell differentiation, mitotic chromosome condensation, and apoptosis. Heterochromatinization, as the main feature of these processes, is also associated with pronounced trimethylation of histones H3 at the lysine 9 position (H3K9me3). Methods: By confocal microscopy, we analyzed cell cycle-dependent levels and distribution of phosphorylated histone H1 (H1ph) and H3K9me3. By mass spectrometry, we studied post-translational modifications of linker histones. Results: Phosphorylated histone H1, similarly to H3K9me3, has a comparable level in the G1, S, and G2 phases of the cell cycle. A high density of phosphorylated H1 was inside nucleoli of mouse embryonic stem cells (ESCs). H1ph was also abundant in prophase and prometaphase, while H1ph was absent in anaphase and telophase. H3K9me3 surrounded chromosomal DNA in telophase. This histone modification was barely detectable in the early phases of mitosis. Mass spectrometry revealed several ESC-specific phosphorylation sites of H1. HDAC1 depletion did not change H1 acetylation but potentiated phosphorylation of H1.2/H1.3 and H1.4 at serine 38 positions. Conclusions: Differences in the level and distribution of H1ph and H3K9me3 were revealed during mitotic phases. ESC-specific phosphorylation sites were identified in a linker histone.

EFFECT OFTHE COMPACTION PROCESS ON THE CONGLOMERATE STRUCTURE AT PRODUCTION OF CONSTRUCTION MATERIALS

Formulation of the problem. The process of densification of mixtures of construction products that are being formed or molded is an important stage in the formation of a macrostructure, since during this period, granular and other components of the composite that fill it are relatively firmly fixed in a viscous medium. A large number of publications are devoted to the study of this issue, but many aspects of the effect of compaction on the structure of a conglomerate are not yet known, and the results obtained have not been systematized. Introduction. Preliminary compaction of the mixture reduces the fragmentation of particles, transferring bonds from point to interfacial along the contact boundaries. With the approach of particles to very small distances, repulsive forces arise and increase. Ultimately, the resultant force acts, which, at a certain optimal distance of particles from one another, provides the initial cohesion of the raw product. Fixation occurs both directly by adjoining the components, including with the possibility of intergrowth (for example, crystals), and through interlayers of a completely hardened or gradually hardening binder. In conglomerate materials, contact through interlayers at the compaction stage is more typical than direct contact or coalescence of particles under the influence of surface energy, chemical bonds, or other, including complex, factors. Objective. The aim of the work was to establish the features in the formation of the structure of the conglomerate from the preliminary compaction of the mixture, as well as its influence on the functional and technological characteristics. Methods. Solving the problem of the optimal structure of a conglomerate requires identifying the relationship between the type, concentration, composition of raw materials, the method of compaction and the resulting functional characteristics of the product. Results. Optimization of the structure while maintaining the level of desired properties corresponds to achieving the highest quality in a material or product. Technology by this criterion becomes progressive. To reduce the water penetration of cement concretes, it is necessary to provide optimal conditions for cement hydration and the creation of a dense conglomerate with a minimum number of pores. In connection with the relevance of creating building materials and products from materials of the construction environment (soil blocks) and reducing energy consumption during molding and pressing, it is necessary to continue the development of technologies and equipment designs to obtain the optimal conglomerate structure with minimal water penetration. Conclusion. Studies of the effect of compaction of raw materials on the structure of conglomerates are ongoing, but already these results indicate that the rate and form of conglomerate formation change significantly at optimal values of pre-compacted raw materials.

Fingerprinting Small Molecule Modulators of Nucleolar Biophysics

Ribosomal RNA (rRNA) is the most abundant RNA in cells, reflecting its fundamental role in enabling protein synthesis in all living organisms. Nucleolus is a membraneless and multicompartmentalized nuclear body that governs rRNA biogenesis, a process that is misregulated in various diseases. Recent research has identified several small molecule therapeutics that alter rRNA biogenesis and subsequently nucleolar morphology, challenging the accepted mechanisms of action for these compounds and simultaneously establishing the nucleolus as an attractive therapeutic target. Nevertheless, there are currently no comprehensive and quantitative frameworks that capture and relate various drug‐induced changes in nucleolar morphology and its material properties to its function. We hypothesized that we can establish such a structure‐function framework by screening a panel of nucleolus‐perturbing small molecule drugs and measuring changes in biophysical properties of the nucleolus in cells from microscopy images. Here, we combine morphometric, thermodynamic, and kinetic measurements of drug‐treated nucleoli to reveal “fingerprints” for drugs that modulate distinct stages of rRNA biogenesis. We anticipate that this framework provides a starting point for robustly determining and predicting structure‐function relationships for nucleolar phenotype perturbations as well as for developing high‐content screening platforms to expedite the discovery of novel nucleolus‐targeting therapeutics.

CHARACTER OF THE ACTIVITY CHANGES IN THE LARGE CELL NUCLEI OF THE RAT HYPOTHALAMUS AT PHOTOPERIOD MODIFICATION

One of the important links of the hypothalamus neuroendocrine system is the largecell supraorbital nuclei, the neurons of which synthesize vasopressin and oxytocin,transport them to the neurohypophysis. Supraorbital nuclei also take part in theprovision of neuroendocrine response to various types of stress – light, immobilization,traumatic, emotional, painful, hypoxic and stresses of other etiologies.The mechanisms of the circadian pacemaker activity of the neural systems of thesupraorbital hypothalamus nuclei are currently under intensive investigation. Atthe same time, knowledge concerning the effects of photoperiod modifications (inparticular, constant lighting) on the activity of the most vulnerable hypothalamicstructures (supraorbital nuclei), which provide a sequence of neuroendocrine changesunder stress and stress-reactivity of the body, remains relatively limited.The purpose – to determine the influence of photoperiod modifications on the morphofunctional state of the supraorbital nuclei of the rat hypothalamus.Material and methods. The experiments were conducted on 40 nonlinear male whiterats, divided into 2 series of studies. Biomaterial sampling was carried out at 14.00and 02.00 using morphometry, densitometry and statistics research methods.Results. The circadian rhythm of the neurons’ morpho-functional activity of thehypothalamus supraorbital nuclei with maximum activity at a day time (up to 14.00) isregistered in rats under the standard light regime. In animals, subjected to a prolonged light exposure, pronounced changes in morphofunctional state of the neurons of the hypothalamus supraorbital nuclei wereestablished. In particular, at 02.00 h the area of the neuron nucleus was 94.08 ± 9.55μm2 and was probably larger than that in the intact animals. The nuclear-cytoplasmicratio of the neurons in the hypothalamus supraorbital nuclei at 02.00 h was lowerthan that in the intact animals due to a decrease in the specific volume of the nucleus.In comparison with the daytime period (14.00 h), the decrease in the neurons’ bodyarea of the hypothalamus supraorbital nuclei, caused by the probable decrease in thearea of the nucleus and nucleolus of cells, was detected by 02.00 h. This caused anincrease of the nuclear-cytoplasmic ratio in the neurons under study, which made 2.51± 0.023 units. Constant light regime does not cause inversion of the rhythm of morphofunctional activity of the neurons under study.Conclusions. 1. Photoperiod duration significantly influences upon the daily activityof the hypothalamus supraorbital nuclei, and constant light regime does not cause therhythm inversion of morpho-functional activity of the studied neurons.2. Light stress causes a probable increase in the area of the nucleus and nucleolusof neurons in the night interval of observation. At the same time, a decrease in thenuclear-cytoplasmic ratio, an increase in the concentration of RNA in the nucleolusand cytoplasm of the neuron of the supraorbital nuclei of the rat hypothalamus areobserved in the day time period.

Enzyme trafficking and coclustering precede and accurately predict human breast cancer recurrences: an interdisciplinary review.

Although great effort has been expended to understand cancer's origins, less attention has been given to the primary cause of cancer deaths-cancer recurrences and their sequelae. This interdisciplinary review addresses mechanistic features of aggressive cancer by studying metabolic enzyme patterns within ductal carcinoma in situ (DCIS) of the breast lesions. DCIS lesions from patients who did or did not experience a breast cancer recurrence were compared. Several proteins, including phospho-Ser226-glucose transporter type 1, phosphofructokinase type L and phosphofructokinase/fructose 2,6-bisphosphatase type 4 are found in nucleoli of ductal epithelial cells in samples from patients who will not subsequently recur, but traffic to the cell periphery in samples from patients who will experience a cancer recurrence. Large coclusters of enzymes near plasmalemmata will enhance product formation because enzyme concentrations in clusters are very high while solvent molecules and solutes diffuse through small channels. These structural changes will accelerate aerobic glycolysis. Agglomerations of pentose phosphate pathway and glutathione synthesis enzymes enhance GSH formation. As aggressive cancer lesions are incomplete at early stages, they may be unrecognizable. We have found that machine learning provides superior analyses of tissue images and may be used to identify biomarker patterns associated with recurrent and nonrecurrent patients with high accuracy. This suggests a new prognostic test to predict patients with DCIS who are likely to recur and those who are at low risk for recurrence. Mechanistic interpretations provide a deeper understanding of anticancer drug action and suggest that aggressive metastatic cancer cells are sensitive to reductive chemotherapy.

FKBP39 Controls the Larval Stage JH Activity and Development in Drosophila melanogaster.

Simple SummaryTwo endocrine hormones, ecdysone and juvenile hormone (JH), control insect development and reproduction. Some studies in the literature have suggested that FKBP39 functions as a transcriptional factor and regulates the JH pathway in Drosophila. However, the physiological roles of FKBP39 are still elusive. To determine the FKBP39 roles in vivo, we first developed an antibody to check the FKBP39 expression pattern and then detected JH activity-related phenotypes in fkbp39 mutants, such as pupariation, reproduction, and Kr-h1 expression. We found that FKBP39 expresses at a high level and controls JH activity at the larval stage. Moreover, we found that rp49, the most widely used reference gene for Real-time quantitative PCR (qRT-PCR), significantly decreased in the fkbp39 mutant. This work will provide valuable information for studies on JH activity and insect development.FK506-binding protein 39kD (FKBP39) localizes in the nucleus and contains multiple functional domains. Structural analysis suggests that FKBP39 might function as a transcriptional factor and control juvenile hormone (JH) activity. Here, we show that FKBP39 expresses at a high level and localizes in the nucleolus of fat body cells during the first two larval stages and early third larval stage. The fkbp39 mutant displays delayed larval-pupal transition and an increased expression of Kr-h1, the main mediator of the JH pathway, at the early third larval stage. Moreover, the fkbp39 mutant has a fertility defect that is independent of JH activity. Interestingly, the expression of rp49, the most widely used reference gene for qRT-PCR in Drosophila, significantly decreased in the fkbp39 mutant, suggesting that FKBP39 might regulate ribosome assembly. Taken together, our data demonstrate the expression pattern and physiological roles of FKBP39 in Drosophila.

Optical control of protein delivery and partitioning in the nucleolus.

The nucleolus is a subnuclear membraneless compartment intimately involved in ribosomal RNA synthesis, ribosome biogenesis and stress response. Multiple optogenetic devices have been developed to manipulate nuclear protein import and export, but molecular tools tailored for remote control over selective targeting or partitioning of cargo proteins into subnuclear compartments capable of phase separation are still limited. Here, we report a set of single-component photoinducible nucleolus-targeting tools, designated pNUTs, to enable rapid and reversible nucleoplasm-to-nucleolus shuttling, with the half-lives ranging from milliseconds to minutes. pNUTs allow both global protein infiltration into nucleoli and local delivery of cargoes into the outermost layer of the nucleolus, the granular component. When coupled with the amyotrophic lateral sclerosis (ALS)-associated C9ORF72 proline/arginine-rich dipeptide repeats, pNUTs allow us to photomanipulate poly-proline–arginine nucleolar localization, perturb nucleolar protein nucleophosmin 1 and suppress nascent protein synthesis. pNUTs thus expand the optogenetic toolbox by permitting light-controllable interrogation of nucleolar functions and precise induction of ALS-associated toxicity in cellular models.

Nucleolar localization of the ErbB3 receptor as a new target in glioblastoma

BackgroundThe nucleolus is a subnuclear, non-membrane bound domain that is the hub of ribosome biogenesis and a critical regulator of cell homeostasis. Rapid growth and division of cells in tumors are correlated with intensive nucleolar metabolism as a response to oncogenic factors overexpression. Several members of the Epidermal Growth Factor Receptor (EGFR) family, have been identified in the nucleus and nucleolus of many cancer cells, but their function in these compartments remains unexplored.ResultsWe focused our research on the nucleolar function that a specific member of EGFR family, the ErbB3 receptor, plays in glioblastoma, a tumor without effective therapies. Here, Neuregulin 1 mediated proliferative stimuli, promotes ErbB3 relocalization from the nucleolus to the cytoplasm and increases pre-rRNA synthesis. Instead ErbB3 silencing or nucleolar stress reduce cell proliferation and affect cell cycle progression.ConclusionsThese data point to the existence of an ErbB3-mediated non canonical pathway that glioblastoma cells use to control ribosomes synthesis and cell proliferation. These results highlight the potential role for the nucleolar ErbB3 receptor, as a new target in glioblastoma.

The Adsorption and Aging Process of Cadmium and Chromium in Soil Micro-aggregates.

The particle size and components of soil aggregates have played important roles in the migration and transformation of heavy metals. The present study focused on the adsorption behavior and aging characteristics of cadmium (Cd) and chromium (Cr) in various granular aggregates and soil components, which were studied by analyzing the adsorption isotherm, adsorption kinetics, and heavy metal speciation distribution. The results showed that, compared with other aggregates, clay aggregates (0-0.002 mm) had the strongest adsorption effect on Cd and Cr and that there was no significant positive correlation between the adsorption amount and the particle size of aggregates for Cd and Cr. In general, the influence of three components on Cd was organic matter > amorphous iron > free iron oxide, and the influence on Cr was free iron oxide > amorphous iron > organic matter. The adsorption isotherm showed that the correlation coefficient of the Langmuir model (R2 ) was higher than that of the Freundlich model (R2 ), indicating that the adsorption of Cd and Cr by soil aggregates can be well described by the Langmuir model with monolayer adsorption behavior. Kinetic adsorption studies showed that quasi-first-order kinetics and quasi-second-order kinetics were more consistent with the actual adsorption amounts of Cd and Cr in soil aggregates, respectively. At the same time, the forms of Cd and Cr gradually transformed from unstable to stable after entering the soil. After 60 days, Cd was mainly oxidized, and residual, and Cr was mainly reduced and residual. These results provide a theoretical basis for assessing the environmental risks of Cd and Cr and providing prevention and treatment methods. Environ Toxicol Chem 2022;41:975-990. © 2022 SETAC.