Strong Expression Research Articles

Spatial Silhouette: A Study on the Creation Strategy of Strong Bamboo Architecture with “Negative Space” as the Main Feature—A Case Study of Phu Quoc Island Visitor Centre, Vietnam

The Gestalt theory of mental completeness in architecture gave rise to the ideas of “positive space” and “negative space”. This research digs into the sturdy structural building process of bamboo architecture, which is essentially distinguished by “negative space”. It examines how bamboo is articulated in architectural space, while attempting to establish a balance between form and structure, with the goal of discovering the current value and spiritual position that bamboo in architecture represents. Using the Phu Quoc Island Visitor Center in Vietnam as an example, we introduce the strong structure concept and examine its design process in terms of spatial operation technique and strong structural expression logic. The fundamental strategy for creating bamboo architecture under this concept is to take the lead in negative space design and use the material capabilities of bamboo to build structural space prototypes. This further encourages the use of green building materials and offers architects working with bamboo a reference.

Nerve injury triggers time-dependent activation of the locus coeruleus, influencing spontaneous pain-like behavior in rats.

Dynamic changes in neuronal activity and in noradrenergic locus coeruleus (LC) projections have been proposed during the transition from acute to chronic pain. Thus, we explored the cellular cFos activity of the LC and its projections, in conjunction with spontaneous pain-like behavior in neuropathic rats. Tyrosine hydroxylase:Cre and wild-type Long-Evans rats, males and females, were subjected to chronic constriction injury (CCI) for 2 (short-term, CCI-ST) or 30 days (long-term, CCI-LT), evaluating cFos and Fluoro-Gold expression in the LC, and its projections to the spinal cord (SC) and rostral anterior cingulate cortex (rACC). These tests were carried out under basal conditions (unstimulated) and following noxious mechanical stimulation. LC activity was evaluated through chemogenetic and pharmacological approaches, as were its projections, in association with spontaneous pain-like behaviors. CCI-ST enhanced basal cFos expression in the LC and in its projection to the SC, which increased further after noxious stimulation. Similar basal activation was found in the neurons projecting to the rACC, although this was not modified by stimulation. Strong basal cFos expression was found in CCI-LT, specifically in the projection to the rACC, which was again not modified by stimulation. No cFos expression was found in the CCI-LT LCipsi/contra→SC. Chemogenetics showed that CCI-ST is associated with greater spontaneous pain-like behavior when the LCipsi is blocked, or by selectively blocking the LCipsi→SC projection. Activation of the LCipsi or LCipsi/contra→SC dampened pain-like behavior. Moreover, DREADDs-mediated inactivation of the CCI-ST LCipsi→rACC or CCI-LT LCipsi/contra→rACC pathway, or intra-rACC antagonism of alpha-adrenoreceptors, also dampens pain-like behavior. In the short term, activation of the LC following CCI attenuates spontaneous pain-like behaviors via projections to the SC while increases nociception via projections to the rACC. In the long term, only the projections from the LC to the rACC contribute to modulate pain-like behaviors in this model.

Factors influencing Agrobacterium-mediated genetic transformation efficiency in bread wheat (Triticum aestivum L.)

Assessment of the efficiency of Agrobacterium tumefaciens mediated transformation in four bread wheat varieties was focused and upon optimization of experiments done to obtain tangible results. The four varieties, Fielder, Navojoa, Baj and Kachu/saul, were transformed with optimized protocols and randomly selected from a selection medium amalgamated with phosphinothricin (Glufosinate). Initially, the variety Fielder has proceeded with 36 different concentrations of synthetic auxins 2,4-D and picloram as 36 treatments. The treatment 30 (48 μg of 2,4-D and 64 μg of picloram) has shown higher efficiency and is comparable in terms of the three traits studied. Additionally, antioxidants and growth regulators were adjusted to obtain better results towards reasonable transformation frequency. Additional varieties were also tested and selected against the selectable marker PPT based on the optimization for the variety Fielder. Selected genotypes were subjected for gusA or Bar gene PCR amplification. The result revealed the frequency of transgenesis in the range of 60 to 70% with the varieties studied. However, the whole experiment needs further repetitions for confirmatory results. Transgenic genotypes of the Fielder variety showed moderate to strong GUS expression in leaves, anthers, seeds, seed coats, and roots compared to non-transgenic control plant tissues. PPT leaf painting assay showed the lack of necrosis on the painted area of leaves in transgenic genotypes compared to the non-transgenic ones.

Bioluminescence imaging of Cyp1a1-luciferase reporter mice demonstrates prolonged activation of the aryl hydrocarbon receptor in the lung

Aryl hydrocarbon receptor (AHR) signalling integrates biological processes that sense and respond to environmental, dietary, and metabolic challenges to ensure tissue homeostasis. AHR is a transcription factor that is inactive in the cytosol but upon encounter with ligand translocates to the nucleus and drives the expression of AHR targets, including genes of the cytochrome P4501 family of enzymes such as Cyp1a1. To dynamically visualise AHR activity in vivo, we generated reporter mice in which firefly luciferase (Fluc) was non-disruptively targeted into the endogenous Cyp1a1 locus. Exposure of these animals to FICZ, 3-MC or to dietary I3C induced strong bioluminescence signal and Cyp1a1 expression in many organs including liver, lung and intestine. Longitudinal studies revealed that AHR activity was surprisingly long-lived in the lung, with sustained Cyp1a1 expression evident in discrete populations of cells including columnar epithelia around bronchioles. Our data link diet to lung physiology and also reveal the power of bespoke Cyp1a1-Fluc reporters to longitudinally monitor AHR activity in vivo.

Self-amplifying RNAs generated with the modified nucleotides 5-methylcytidine and 5-methyluridine mediate strong expression and immunogenicity in vivo

Abstract When utilized in therapeutic applications, synthetic self-amplifying RNA (saRNA) can lead to higher and more sustained expression than standard messenger RNA (mRNA). This feature is particularly important for gene replacement therapy applications where prolonged expression could reduce the dose and frequency of treatments. The inclusion of modified nucleotides in synthetic non-amplifying mRNA has been shown to increase RNA stability, reduce immune activation and enhance gene expression. Preclinical and clinical studies with saRNA have so far exclusively relied on RNA containing the canonical nucleotides adenosine, cytidine, guanosine and uridine. For the first time, we show that noncanonical nucleotides, such as 5-methylcytidine (m5C) and 5-methyluridine (m5U), are sufficiently compatible with a replicon derived from Venezuelan equine encephalitis alphavirus mediating protein translation in vitro, while those containing N1-methylpseudouridine in place of uridine show no detectable expression. When administered in vivo, saRNA generated with m5C or m5U mediates sustained gene expression of the luciferase reporter gene, with those incorporating m5U appearing to lead to more prolonged expression. Finally, distinct antigen-specific humoral and cellular immune responses were induced by modified saRNA encoding the model antigen ovalbumin. The use of modified nucleotides with saRNA-based platforms could enhance their potential to be used effectively in a variety of applications.

Expression of netrin-1 in uterine serous carcinoma and its association with prognosis.

At present, there are few biomarkers used to predict the prognosis of uterine serous carcinoma (USC). Netrin-1 may be a promising biomarker candidate. We investigated netrin-1 expression in USC tissues and healthy endometrial tissues to determine its relevance to disease prognosis. Netrin-1 expression was examined in the tissues of 48 patients with USC and 30 patients with healthy benign endometrial tissues via immunohistochemistry. None of the healthy tissues were stained with netrin-1. In tumor tissues, the overall positivity rate of netrin-1 was 75%, detected as high expression in 17 patients (35%) and low in 19 (40%). Patients who had tumors with no netrin-1 expression (n = 12) had a median overall survival (OS) of 60.0 months (95% confidence interval [CI], 47-98), whereas patients who had tumors with low to strong netrin-1 expression (n = 33) had a lower median OS of 50 months, but the difference was not statistically significant (95% CI, 58-108; P = 0.531). Disease-free survival (DFS) was not statistically significant between the groups (95% CI, 67.7-115.9; P = 0.566). Patients with a tumor diameter ≥2 cm had higher netrin-1 expression than those with a tumor diameter of 2 cm (P = 0.027). We did not find any difference in overall and DFS when age, tumor stage, histology, tumor diameter, p53 status, lymphovascular space invasion, myometrial invasion, and lymph node metastasis were compared according to netrin-1 expression (P > 0.05). Netrin-1 was expressed in USC but not in healthy tissues. Its expression was not associated with OS or DFS.

Super-enhancers: drivers of cells' identities and cells' debacles.

Precise spatiotemporal regulations of gene expression are essential for determining cells' fates and functions. Enhancers are cis-acting DNA elements that act as periodic transcriptional thrusters and their activities are cell type specific. Clusters of enhancers, called super-enhancers, are more densely occupied by transcriptional activators than enhancers, driving stronger expression of their target genes, which have prominent roles in establishing and maintaining cellular identities. Here we review the current knowledge on the composition and structure of super-enhancers to understand how they robustly stimulate the expression of cellular identity genes. We also review their involvement in the development of various cell types and both noncancerous and cancerous disorders, implying the therapeutic interest of targeting them to fight against various diseases.

Biosynthesis of C4-C8 3-Hydroxycarboxylic Acids from Glucose through the Inverted Fatty Acid β-Oxidation by Metabolically Engineered Escherichia coli.

Inverted fatty acid β-oxidation represents a versatile biochemical platform for biosynthesis by the engineered microbial strains of numerous value-added chemicals from convenient and abundant renewable carbon sources, including biomass-derived sugars. Although, in recent years, significant progress has been made in the production through this pathway of n-alcohols, 1,3-diols, and carboxylic acids and its 2,3-unsaturated derivatives, the potential of the pathway for the biosynthesis of 3-hydroxycarboxylic acids remained almost undisclosed. In this study, we demonstrate the microaerobic production of even-chain-length C4-C8 3-hydroxycarboxylic acids from glucose through the inverted fatty acid β-oxidation by engineered E. coli strains. The notable accumulation of target compounds was achieved upon the strong constitutive expression of the genes atoB, fadA, fadB, fadE/fabI, and tesB, which code for the key enzymes catalysing reactions of aerobic fatty acid β-oxidation and thioesterase II, in strains devoid of mixed-acid fermentation pathways and lacking nonspecific thioesterase YciA. The best performing recombinants were able to synthesise up to 14.5 mM of 3-hydroxycarboxylic acids from glucose with a total yield of 0.34 mol/mol and a C4/C6/C8 ratio averaging approximately 63/28/9. The results provide a framework for the development of highly efficient strains and processes for the bio-based production of valuable 3-hydroxycarboxylates from renewable raw materials.

Antennal Transcriptome Evaluation and Analysis for Odorant-Binding Proteins, Chemosensory Proteins, and Suitable Reference Genes in the Leaf Beetle Pest Diorhabda rybakowi Weise (Coleoptera: Chrysomelidae).

Diorhabda rybakowi Weise is one of the dominant pests feeding on Nitraria spp., a pioneer plant used for windbreaking and sand fixation purposes, and poses a threat to local livestock and ecosystems. To clarify the key olfactory genes of D. rybakowi and provide a theoretical basis for attractant and repellent development, the optimal reference genes under two different conditions (tissue and sex) were identified, and the bioinformatics and characterization of the tissue expression profiles of two categories of soluble olfactory proteins (OBPs and CSPs) were investigated. The results showed that the best reference genes were RPL13a and RPS18 for comparison among tissues, and RPL19 and RPS18 for comparison between sexes. Strong expressions of DrybOBP3, DrybOBP6, DrybOBP7, DrybOBP10, DrybOBP11, DrybCSP2, and DrybCSP5 were found in antennae, the most important olfactory organ for D. rybakowi. These findings not only provide a basis for further in-depth research on the olfactory molecular mechanisms of host-specialized pests but also provide a theoretical basis for the future development of new chemical attractants or repellents using volatiles to control D. rybakowi.

Clinicopathological characteristics and tumor infiltrating immune cells associations of PD-L1 tumor expression in non-small cell lung cancer patients.

Our study aimed to perform on Moroccan patients' non-small cell lung carcinoma (NSCLC) concerning the relationship between PD-L1 tumor expression, clinicopathological features and tumor infiltrating immune cells (ICs). This is a retrospective study (2019 to 2021) conducted on samples from Moroccan patients with NSCLC at the Pathological Anatomy Laboratory of Ibn Rochd University Hospital in Casablanca. Eligible participants for our study had to meet the following predefined criteria: age ≥18 years, histologically confirmed NSCLC, no prior therapeutic interventions, availability of clinical and pathological data, and a usable tumor sample for determining PD-L1 status. Exclusion criteria applied to patients with other types of lung cancer and unusable tumor samples. The evaluation of tumor and immune expression of PD-L1 was performed using immunohistochemistry (IHC), with the 22C3 clone on the Dako Autostainer Link 48 platform. Tumor PD-L1 expression was categorized into 3 levels: TPS <1% (negative expression), TPS 1-49% (low expression), and TPS ≥50% (high expression). ICs infiltrating the tumor expressing PD-L1 were considered positive when more than 1% of positive ICs were present. Among the 316 analyzed samples, 56.6% showed a negative expression of PD-L1, 16.8% displayed a low expression of PD-L1, and 26.6% exhibited a strong expression. Regarding the histological type, among patients with TPS ≥ 50%, 25.8% had adenocarcinoma. Among patients with TPS ≥ 50%, 24.81% were smokers. PD-L1 was also strongly expressed in the lung (28.2%) and bronchi (26.5%). PD-L1 expression (TPS ≥ 50%) was observed in 35.29% of early-stage patients. Concerning tumor cells (TCs), 27.5% of tumors infiltrated by ICs had TPS ≥ 50%. Furthermore, coexpression of PD-L1 on both TCs and ICs infiltrating the tumor was found in 27.8% of tumors. Statistical analysis demonstrated a significant association between tumor PD-L1 expression and smoking status (P=0.019). However, no significant difference was observed between PD-L1 expression and the presence of ICs infiltrating the tumor (P=0.652), as well as the IHC expression of PD-L1 on ICs (P=0.259). Our results demonstrate a significant association between PD-L1 expression and smoking status. However, no significant association was observed between PD-L1 expression and the presence of infiltrating ICs, nor with the IHC expression of PD-L1 on ICs. Our data underscore the importance of participating in the study of specific factors influencing PD-L1 expression in patients with NSCLC.

Abstract LB207: C5ar1 is a novel biomarker and potential therapeutic target in advanced cutaneous squamous cell carcinoma

Abstract Cutaneous squamous cell carcinoma (cSCC) is the most common metastatic skin cancer and the metastatic disease is associated with poor prognosis. At present, there are no established molecular markers to predict the metastasis risk and prognosis of patients with primary cSCC. Recently, the role of specific complement components in the progression of cSCC has been shown. Here, we have examined the role of complement C5a receptor C5aR1 in the progression and metastasis of cSCC. The results show that the expression of C5aR1 is increased in cSCC cells in 3D spheroid coculture model in the presence of fibroblasts and that treatment with recombinant C5a increases invasion of cSCC cells in this model. Staining for C5aR1 was detected on the surface of tumor cells in the invasive edge of human cSCC xenograft tumors in vivo. Staining of metastatic and non-metastatic primary human cSCCs, premalignant epidermal lesions and normal skin for C5aR1 with multiplexed fluorescence and chromogenic immunohistochemistry revealed stronger expression of C5aR1 on surface of cSCC tumor cells compared to epidermal cells in cSCC in situ, actinic keratosis and normal skin. Increased expression of C5aR1 on tumor cell surface was associated with the metastasis risk of primary cSCCs and independently predicted poor disease-specific survival of patients with primary cSCC. These results reveal the role of C5aR1 in cSCC progression and identify C5aR1 as a novel biomarker for metastasis risk and poor prognosis of patients with cSCC. The results also suggest C5aR1 as a novel therapeutic target for treatment of locally advanced and metastatic cSCC. Citation Format: Lauri Heiskanen, Liisa Nissinen, Elina Siljamäki, Jaakko Knuutila, Teijo Pellinen, Markku Kallajoki, Jyrki Heino, Pilvi Riihilä, Veli-Matti Kähäri. C5ar1 is a novel biomarker and potential therapeutic target in advanced cutaneous squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB207.

Exploring p53 protein expression and its link to TP53 mutation in myelodysplasia-related malignancies-Interpretive challenges and potential field of applications.

TP53 alterations have a significant prognostic effect in myeloid neoplasms. Our objective was to investigate the TP53 gene mutation status, p53 protein expression and their relationship in dysplasia-related myeloid neoplasms with varying levels of myeloblast counts. A total of 76 bone marrow biopsy samples with different blast counts were analysed. Total and strong (3+) p53 expression was determined. Dual immunohistochemical staining was performed to determine the cell population associated with p53 expression. NGS analysis was performed using the Accel-Amplicon Comprehensive TP53 panel. Both p53 expression and TP53 VAF showed a significant correlation with the myeloblast ratio (P < 0.0001); however, p53 expression was also present in other cell lineages. The VAF value exhibited a significant correlation with p53 expression. A high specificity (0.9800) was observed for TP53 mutation using the ≥ 10% strong (3+) p53 cut-off value, although the sensitivity (0.4231) was low. Strong (3+) p53 expression using a ≥ 10% cut-off value accurately predicts TP53 mutation but does not reveal the allelic state. The p53 expression is significantly influenced by myeloblast count, and histological interpretation should consider the presence of intermixed non-neoplastic marrow cells with varying physiological p53 expression.

ALK-positive large B-cell lymphoma (ALK + LBCL) with aberrant CD3 expression

ALK-positive ( +) large B cell lymphoma (ALK + LBCL) is a rare distinct subtype of diffuse large B cell lymphoma presenting with high stage and aggressive behavior. Although B cell markers such as CD20, CD19, and CD22 are generally negative, plasmacytic markers including CD138, CD38, and MUM1 are positive. T cell markers are negative with rare exceptions. We report an unusual case of ALK1 + LBCL in a 58-year-old man with partial expression of CD3 without other T cell antigen expression. The tissue was evaluated with flow cytometry, immunohistochemistry, fluorescent in situ hybridization, and gene rearrangement studies. Gene rearrangement studies for IGH and TCR gamma were performed. Flow cytometry did not demonstrate any abnormal lymphoid populations. Tissue sectioning shows a malignant plasmacytic large cell neoplasm which expresses CD45 but is negative for CD20, CD79a, and PAX5. Plasmacytic markers CD138 and MUM1 are positive with kappa light chain restriction. Strong granular cytoplasmic expression of ALK is present. FISH showing disrupted ALK supports the diagnosis while MYC, BCL6, and BCL2 are intact. Gene rearrangement studies show coexisting IGH and TCR gamma clones; however, the TCR peak was present within a polyclonal background suggesting the disputed cells are likely only a subset of the T cell population. ALK + LBCL can present with an ambiguous immunophenotype, which warrants the use of multiple B cell, T cell, and plasmacytic antibodies. CD3 expression in this entity is rare and of uncertain clinical significance, but warrants further study.

Assessment of the protective effect of saffron against Khat induced testicular dysfunction in male rats

ABSTRACT Catha edulis is a widespread herb used by many people for its euphoric effects, but it has an adverse effect on some vital organs. Saffron is abundant in secondary metabolites which have antioxidant and anti-inflammatory properties. So, this study was established to examine the impact of saffron in minimizing khat-induced testicular toxicity in rats. In this study, 40 male albino rats were used, the rats were divided into four groups (n = 10). Control, Saffron group, Khat group, and Khat + Saffron group. Rats treated with Khat showed marked changes in the testicular histological architecture. Histochemically, the testicular issues displayed low protein and polysaccharide contents. The immunohistochemical results showed low testicular expression for vimentin, E-cadherin proteins, and Bcl-2 while strong positive expression for BAX protein in compared with control. The antioxidant enzymes superoxide dismutase and catalase activity showed a marked decrease while malondialdehyde levels were elevated. However, rats given both khat followed by saffron showed an improvement in testicular histology, histochemistry, immunohistochemistry, and biochemical results; the results were close to the control ones. In conclusion, saffron proved its efficiency in improving testicular toxicity induced by khat in the testis of albino rats due to its richness with secondary metabolites at the histological, and biochemical levels and can be used as a safe daily supplementary.

Effect of Oxidative Stress on Histological and Immunohistochemical Changes in Testes of Albino Mice

Background: Fertility in male animals is an important issue in veterinary medicine. Exposure to severe crowding and fattening may affect the fertility of these animals. Objectives: This study compares the effect of oxidative stress caused by overcrowding or obesity on testicular tissue in male albino mice. Methods: A total of 30 male mice were divided into 3 groups. The first was a control group. The second group was subjected to crowding by cramming 20 mice in a small cage. The third group was fattened by eating a high-fat diet. Morphometric cells and histological measurements of the seminiferous tubule diameter, epithelium thickness, and Sertoli and Leydig cell counts were taken. Results: The crowding group testes showed the presence of necrosis and degeneration of spermatocytes, Sertoli, and Leydig cells. The obese group’s spermatocytes, Sertoli, and Leydig cells had shown severe necrosis, degeneration, and giant cell invasion. Histomorphometric measurements of seminiferous tubule diameter, epithelium-thickness, and Sertoli and Leydig cell counts revealed a considerable reduction in the obese group compared to the other groups. Furthermore, in the crowding and fattening groups, measurements of oxidative stress indicators revealed a decrease in glutathione value in the obese group compared to the control, whereas malondialdehyde rose in the overcrowded and obese groups compared to the control. The results of the immunohistochemical expressions for tumor necrosis factor-α (TNF-α) in the testes of mice revealed weak TNF-α positive expression in the control group, moderate expression in the crowding group, and strong expression in the obesity group. Conclusion: The study suggests that both obesity and crowding can impair testicular function, with obesity having the most significant impact. More research into the mechanisms underlying these effects, as well as potential interventions to improve testicular function in obese or overcrowded animals, is required.

Prognostic Model for High-Grade Neuroendocrine Carcinoma of the Lung Incorporating Genomic Profiling and Poly (ADP-ribose) Polymerase-1 Expression.

High-grade neuroendocrine carcinoma (HGNEC) of the lung is an aggressive cancer with a complex biology. We aimed to explore the prognostic value of genetic aberrations and poly(ADP-ribose) polymerase-1 (PARP1) expression in HGNEC and to establish a novel prognostic model. We retrospectively enrolled 191 patients with histologically confirmed HGNEC of the lung. Tumor tissues were analyzed using PARP1 immunohistochemistry (IHC; N = 191) and comprehensive cancer panel sequencing (n = 102). Clinical and genetic data were used to develop an integrated Cox hazards model. Strong PARP1 IHC expression (intensity 3) was observed in 153 of 191 (80.1%) patients, and the mean PARP1 H-score was 285 (range, 5-300). To develop an integrated Cox hazard model, our data set included information from 357 gene mutations and 19 clinical profiles. When the targeted mutation profiles were combined with clinical profiles, 12 genes (ATRX, CCND2, EXT2, FGFR2, FOXO1, IL21R, MAF, TGM7, TNFAIP3, TP53, TSHR, and DDR2) were identified as prognostic factors for survival. The integrated Cox hazard model, which combines mutation profiles with a baseline model, outperformed the baseline model (incremental area under the curve 0.84 v 0.78; P = 8.79e-12). The integrated model stratified patients into high- and low-risk groups with significantly different disease-free and overall survival (integrated model: hazard ratio, 7.14 [95% CI, 4.07 to 12.54]; P < .01; baseline model: 4.38 [2.56 to 7.51]; P < .01). We introduced a new prognostic model for HGNEC that combines genetic and clinical data. The integrated Cox hazard model outperformed the baseline model in predicting the survival of patients with HGNEC.

Output-Oriented Teaching Method of College English Speaking Based on Second Language Acquisition Theory

In recent years, English speaking has become one of the required courses for most college students in China. College students should have strong spoken English expression ability, and it has become an arduous task for them to improve their oral English level. This paper first describes the current situation of college oral English teaching, including some unsolved problems in class and a few teaching methods already in use. Then, the study synthesizes and sorts out previous studies on the Output-oriented teaching method and Second Language Acquisition (SLA) theory. The following parts analyzes and summarizes existing hypothesis on both SLA and Output-oriented teaching method and seek effective ways to advance college professor’s oral English teaching, so as to improve the oral capability of college English learners. This study also implements the teaching objectives and tasks of college students' English teaching, such as cultural accomplishment, comprehensive application ability, and intercultural language communication ability.

Correlation of prognosis of oral squamous cell carcinoma with CD44 expression: A retrospective immunohistochemical analysis

ABSTRACT Background: Cluster of differentiation (CD44) is a cancer stem cell marker responsible for angiogenesis, invasion, and migration in oral squamous cell carcinoma (OSCC). Objectives: The primary objective of the study was to analyze the difference in expression of CD44 among different grades of OSCC. The secondary objective was to correlate the prognosis with the expression. Materials and Methods: Based on histopathological diagnosis, we retrieved OSCC paraffin-embedded blocks from the archives of the Department of Oral Pathology, Sri Ramaswamy Memorial (SRM) Dental College, Chennai, India, reported from January 2012 to May 2022. We evaluated sections from 15 tissues of normal oral mucosa, 15 paraffin blocks of well-differentiated OSCC, 15 moderately differentiated OSCC, and 15 poorly differentiated OSCC for CD44 immunostaining. Immunohistochemical expression of CD44 was studied in these tissues and graded based on the intensity of staining. The difference in immune expression of CD44 between different grades was quantitatively analyzed using image analysis by Fiji image analysis software. Analysis of variance (ANOVA) was used to compare the mean difference in expression between different groups by using the Statistical Package for Social Sciences (SPSS) software version 16. Statistical significance was set at P &lt; 0.05. Results: We identified that 10% (n = 6) of the normal mucosa showed strong expression of CD44, whereas 13.3% (n = 8) of well-differentiated OSCC, 10% (n = 6) of moderately differentiated OSCC, and 0% (n = 0) of poorly-differentiated OSCC were strongly positive for CD44 expression. The difference in comparison was statistically significant (P, 0.003). By quantitative analysis, the mean expression of CD44-positive cells in well-differentiated OSCC was 38258 ± 4762, moderately-differentiated OSCC was 27353 ± 1357, and poorly-differentiated OSCC was 10265 ± 1685. Upon comparison, the difference in mean was found to be significant (P, 0.002). Conclusion: The expression of CD44 was noted to decrease from well-differentiated to poorly-differentiated OSCC.

За някои трайно очертаващи се тенденции в българската разговорна реч и в речта на медиите през последните десет години

The article describes three continuous trends in Bulgarian colloquial speech. The first one is related to changes in the use of verbs, consisting of: a) swarming of verbs derived from nouns, incl. from foreign words, and b) condensing of the expression by omission of: the verb to drive/cause, the reflexive particle se and some prepositions. A survey was conducted on the use of verbs registered as having undergone a process of dereflexivity and/or transitivisation. The answers of respondents point to an over-whelming disapproval of the described verb use strategies. Thus, a more appropriate and accurate way to define this process is as an effort to condense the expression by omitting certain elements, which is in accordance with the principle of economy in colloquial speech. The second trend presented is the progressive reduction of the use of forms expressing politeness and their increasingly frequent replacement with forms for informal ad-dressing. There is a relatively high degree of tolerance towards this speech practice from both researchers and native speakers. The third trend concerns the use of two types of short fixed expressions/clichés, rep-resenting constructions with one or two fixed components and one or two variables. They condense the maximum content into a short form and “leave” freedom for interpretation by the interlocutor/listener, being additionally marked either by strong expressiveness or originality (usually purposefully sought), which adds another element for “interpreting and decoding” by the interlocutor/listener.

Molecular identification of chalcone synthase-encoding genes EfCHSs and EfGATA8 as a negative regulator for flavonoid biosynthesis in Euryale ferox

Chalcone synthase is a crucial rate-limiting enzyme for flavonoid biosynthesis in plants. Despite the abundance of flavonoid resources in Euryale ferox, resolving their biosynthesis and regulatory pathways is remained scarce. Five putative CHS genes were obtained in E. ferox through genome-wide characterization. Phylogenetic, structural analysis and sequence alignment were performed to determine EfCHS1 and EfCHS2 as candidate genes for flavonoid biosynthesis. Enzyme activity assays and transient overexpression demonstrated that EfCHS1 and EfCHS2 exhibited potent activities. Interestingly, it was found that there was strong expression of EfCHS1 in the seed shells and EfCHS2 in the leaves. Moreover, the transcription factor EfGATA8 was screened in the E. ferox yeast library using a yeast one-hybrid system. Subcellular localization analysis displayed that EfGATA8 only existed fluorescent signals in the nucleus. It was revealed that EfGATA8 binds directly to the promoter of EfCHS2 and represses its expression according to further Y1H assay and luciferase (LUC) activity analysis. EfGATA8 transient transformation into tobacco leaves resulted in significantly reduced accumulation of flavonoids. Taken together, these findings clarify the position of EfCHS1 and EfCHS2 as key genes for flavonoid biosynthesis in E. ferox, while perfecting the regulatory network for flavonoid biosynthesis in plants.