The main units of persimmon proanthocyanidins (PAs) are composed of flavan-3-ols including epigallocatechin gallate (EGCG) and gallocatechin gallate (GCG). Precise quantification of GCG is challenging due to its trace amounts in persimmon. In this study, to establish the optimal UHPLC-Q-Exactive Orbitrap/MS technique for the determination of PAs monomer composition in persimmon fruit flesh of different astringency types, mass spectrometry and chromatographic conditions were optimized. The results showed that when operating in negative ion mode, using a T3 chromatographic column (a type of C18 column with high-strength silica), acetonitrile as the organic phase, a 0.1% mobile phase acid content, and a mobile phase flow rate of 0.2 mL/min, the chromatographic peak shape and resolution of the PAs monomer composition improved. Additionally, there was no tailing phenomenon observed in the chromatographic peaks. At the same time, the intra-day and inter-day precision, stability, and recovery of the procedure were good. The relative standard deviation (RSD) of stability was less than 5%. The intra-day precision was in the range of 1.14% to 2.36%, and the inter-day precision ranged from 1.03% to 2.92%, both of which were less than 5%. The recovery rate ranged from 94.43% to 98.59% with an RSD less than 5%. The results showed that the UHPLC-Q-Exactive Orbitrap/MS technique established in this study can not only be used for the quantification of EGCG and GCG in persimmon fruit flesh but also be suitable for analyzing other PAs monomer compositions, providing robust support for the related research on persimmon PAs. Full article

In breast cancer, epithelial-mesenchymal transition (EMT) is positively associated with programmed death ligand 1 (PD-L1) expression and immune escape, and TWIST1 silences ERα expression and induces EMT and cancer metastasis. However, how TWIST1 regulates PD-L1 and immune evasion is unknown. This study analyzed TWIST1 and PD-L1 expression in breast cancers, investigated the mechanism for TWIST1 to regulate PD-L1 transcription, and assessed the effects of TWIST1 and PD-L1 in cancer cells on cytotoxic CD8+ T cells. Interestingly, TWIST1 expression is correlated with high-level PD-L1 expression in ERα-negative breast cancer cells. The overexpression and knockdown of TWIST1 robustly upregulate and downregulate PD-L1 expression, respectively. TWIST1 binds to the PD-L1 promoter and recruits the TIP60 acetyltransferase complex in a BRD8-dependent manner to transcriptionally activate PD-L1 expression, which significantly accelerates the exhaustion and death of the cytotoxic CD8+ T cells. Accordingly, knockdown of TWIST1 or BRD8 or inhibition of PD-L1 significantly enhances the tumor antigen-specific CD8+ T cells to suppress the growth of breast cancer cells. These results demonstrate that TWIST1 directly induces PD-L1 expression in ERα-negative breast cancer cells to promote immune evasion. Targeting TWIST1, BRD8, and/or PD-L1 in ERα-negative breast cancer cells with TWIST1 expression may sensitize CD8+ T-cell-mediated immunotherapy. Full article

This study investigates the impact of nitrogen (N), sulfur (S), and iron (Fe) fertilization on secondary metabolites, particularly quercetin and its forms, in onion bulbs (Allium cepa L.). Field experiments over two years examined four onion varieties with red, yellow, and white colors of bulbs: Kamal, Robin, Pueblo, and Mundo. The parameters investigated included the yield, dry matter content, and average onion weight. The phenolic and flavonoid contents were also analyzed. The free quercetin, bound quercetin, and total quercetin contents were determined using the HPLC method. The results demonstrated notable increases in yield following the application of nitrogen (NH⁴⁺) and sulfur (SO₄²⁻) fertilizers. Incorporating iron (Fe²⁺) alongside these fertilizers did not yield a significant impact compared to N+S variant. The phenolic and flavonoid content varied with fertilization, while the quercetin content did not yield statistically significant results. Overall, the study highlights the complex relationship between fertilization practices and secondary metabolite production in onions, emphasizing the need for sustainable intensification in modern agriculture. Full article

Background: Staphylococcus aureus is a leading cause of skin and bloodstream infections in humans. Antibiotic resistant strains of S. aureus continue to be a problem in treating patients that are infected, so treatment options are needed. A drug discovery project identified SK-03-92 as a novel anti-staphylococcal drug, but the SK-03-92 mechanism of action is unknown. We hypothesized that a lethal factor was being released by the bacteria that killed siblings. Methods: In this study, filtration through molecular weight cut-off filters as well as boiling, trypsin treatment, and proteinase K treatment were used to ascertain what the lethal factor was released by SK-03-92 treated S. aureus cells. Results: Filtration through molecular weight cut-off filters demonstrated the lethal factor released by SK-03-92 treated S. aureus cells had a molecular cut-off between 10,000 Da and 30,000 Da that killed fresh S. aureus cells but was not released by untreated cells. Through proteinase K digestion, trypsin digestion, and boiling experiments, the lethal factor was shown to be a protein. Further experiments are needed to identify what proteins released following SK-03-92 treatment cause the death of S. aureus cells. Conclusions: The data show that SK-03-92 treatment causes S. aureus to release a lethal factor protein that kills S. aureus cells, suggesting a new mechanism of action for an antibacterial drug. Full article

In tropical deciduous forests (TDFs), plants have developed various strategies to tolerate desiccation during the dry season. One strategy is osmotic adjustment, which includes the accumulation of secondary metabolites. Annona lutescens, a species that inhabits TDFs, increases and accumulates liriodenine alkaloid in its roots during the dry season. In this study, we evaluate the possible role of this molecule as an osmolyte and in pH homeostasis. We performed growth analyses and determined liriodenine concentrations during water stress in Annona lutescens seedlings grown under controlled temperature, water, and light conditions. We also calculated their osmotic adjustment based on pressure–volume curves and performed solubility tests along a pH gradient. Osmotic adjustment was compared between control plants (irrigated) and plants subjected to 15, 25, and 35 days of water stress. Osmotic adjustment was dramatically higher in plants subjected to 35 days of water stress compared to the control. The solubility of liriodenine was 54% at pH 4.5, and when liriodenine was in contact with malic acid solutions, the pH increased slightly. The highest concentration of liriodenine was in the roots, with a significant increase from 540.855 μg g⁻¹ after 15 days of water stress to 1239.897 μg g⁻¹ after 35 days. Our results suggest that liriodenine plays an important role in the response to water stress as an osmolyte and in pH homeostasis. Full article

Aquatic ecosystems can suffer inadvertent contamination from widely used herbicides. This study delves into the relative toxicity of 36 herbicides on green algae, exploring 11 distinct modes of action and 25 chemical structure classes. Through a 72-h algal growth inhibition test, it was found that herbicides targeting acetolactate synthase (ALS), photosystem II (PSII inhibitors), microtubule assembly, very-long-chain fatty acid (VLCFA) synthesis, and lipid synthesis exhibited high toxicity, with 72-h EC50 (half-maximal effective concentration) values ranging from 0.003 mg/L to 24.6 mg/L. Other pesticide types showed moderate to low toxicity, with EC50 values ranging from 0.59 mg/L to 143 mg/L. Interestingly, herbicides sharing the same mode of action but differing in chemical composition displayed significantly varied toxicity. For instance, penoxsulam and pyribenzoxim, both ALS inhibitors, demonstrated distinct toxicity levels. Similarly, terbuthylazine and bentazone, both PSII inhibitors, also exhibited differing toxicities. Notably, herbicides approved for rice cultivation showed lower toxicity to green algae compared to those intended for terrestrial plants. These data offer valuable insights for assessing the potential risks posed by these chemicals to aquatic organisms. Additionally, to prevent or minimize herbicide residual effects, modern management practices were reviewed to offer practical guidance. Full article

Complex atypical endometrial hyperplasia (CAH) carries a high probability of cancer. The intraoperative evaluation of endometrial cancer in cases of CAH has not been reliable. The safety and sensitivity of sentinel lymph node (SLN) sampling has been validated. In our study, we aimed to evaluate the efficacy and safety of SLN sampling in CAH managed by the da Vinci robotic platform. A total of 113 patients with a preoperative diagnosis of CAH were included in this retrospective cohort study. All of them underwent a robot-assisted total laparoscopic hysterectomy and bilateral salpingo-oophorectomy, with 69 patients undergoing SLN sampling. A statistical analysis calculated the probability of cancer, the SLN map rate, and surgical complications. The predictors of cancer were evaluated. Descriptive statistics were used to summarize the results; comparative statistics were used to compare the cohorts; and logistical regression analysis was used to predict the risk. Forty-seven percent of the entire cohort was diagnosed with endometrial cancer. The median age was 63 years in the SLN cohort (N = 69) and 61 in the No-SLN cohort (N = 44) (p = 0.363). The median BMI was 34 Kg/m² in the SLN cohort and 40 in the No-SLN cohort (p = 0.004). The bilateral SLN map was 92.8%, and the unilateral SLN map rate was 7.2%. There were no grade-3–4 complications in the SLN cohort, and four grade-3–4 complications in the No-SLN group (p = 0.021). A preoperative diagnosis of CAH bordering on or unable to rule out cancer was the only predictor of cancer. Sentinel lymph node sampling has a high map rate and low complications in CAH. We recommend a prospective study investigating the clinical benefit of the procedure. Full article

The COVID-19 pandemic, with increased home cooking and decreased restaurant dining, significantly altered seafood consumption patterns. By applying an ordered logit model to identify factors affecting seafood consumption during the pandemic, this study found that the shift in seafood consumption was driven by factors such as changes in meal preparation methods, more time spent at home, and shifts in financial situations. While take-out consumption boosted overall seafood intake, popular varieties saw a rise in home consumption, while high-end seafood suffered from decreased demand as consumers focused more on home dining. This study underscores the importance of supporting suppliers, restaurants, and retailers dealing with high-end seafood, as they face economic challenges due to reduced consumption. In summary, pandemic-induced restrictions on mobility led to a notable transition from restaurant-prepared seafood to home-cooked options, highlighting the need for targeted policies to aid affected sectors. Full article

The decentralized nature of blockchain enables data traceability, transparency, and immutability as complementary security features to the existing Internet of Things (IoT) systems. These Blockchain-based IoT (BIoT) systems aim to mitigate security risks such as malicious control, data leakage, and dishonesty often found in traditional cloud-based, vendor-specific IoT networks. As we steadily advance into the era of situation-aware IoT, the use of machine learning (ML) techniques has become essential for synthesizing situations based on sensory contexts. However, the challenge to integrate learning-based situation awareness with BIoT systems restricts the full potential of such integration. This is primarily due to the conflicts between the deterministic nature of smart contracts and the non-deterministic nature of machine learning, as well as the high costs of conducting machine learning on blockchain. To address the challenge, we propose a framework named Situ-Oracle. With the framework, a computation oracle of the blockchain ecosystem is leveraged to provide situation analysis as a service, based on Recurrent Neural Network (RNN)-based learning models tailored for the Situ model, and specifically designed smart contracts are deployed as intermediary communication channels between the IoT devices and the computation oracle. We used smart homes as a case study to demonstrate the framework design. Subsequently, system-wide evaluations were conducted over a physically constructed BIoT system. The results indicate that the proposed framework achieves better situation analysis accuracy (above 95%) and improves gas consumption as well as network throughput and latency when compared to baseline systems (on-chain learning or off-chain model verification). Overall, the paper presents a promising approach for improving situation analysis for BIoT systems, with potential applications in various domains such as smart homes, healthcare, and industrial automation. Full article

We studied the effect of succinimide derivatives on acetylcholinesterase activity due to the interest in compounds that influence this enzyme’s activity, which could help treat memory issues more effectively. The following parameters were established for this purpose based on kinetic investigations of the enzyme in the presence of succinimide derivatives: the half-maximal inhibitory concentration, the maximum rate, the inhibition constant, and the Michaelis–Menten constant. Furthermore, computational analyses were performed to determine the energy required for succinimide derivatives to dock with the enzyme’s active site. The outcomes acquired in this manner demonstrated that all compounds inhibited acetylcholinesterase in a competitive manner. The values of the docking energy parameters corroborated the kinetic parameter values, which indicated discernible, albeit slight, variations in the inhibitory intensity among the various derivatives. Full article

The biological activity of structural HIV-1 proteins is not limited to ensuring a productive viral infection but also interferes with cellular homeostasis through intra- and extracellular signaling activation. This interference induces genomic instability, increases the lifespan of the infected cell by inhibiting apoptosis, and subverts cell senescence, resulting in unrestricted cell proliferation. HIV structural proteins are present in a soluble form in the lymphoid tissues and blood of infected individuals, even without active viral replication. The HIV matrix protein p17, the envelope glycoprotein gp120, the transenvelope protein gp41, and the capsid protein p24 interact with immune cells and deregulate the biological activity of the immune system. The biological activity of HIV structural proteins is also demonstrated in endothelial cells and some tumor cell lines, confirming the ability of viral proteins to promote cell proliferation and cancer progression, even in the absence of active viral replication. This review corroborates the hypothesis that HIV structural proteins, by interacting with different cell types, contribute to creating a microenvironment that is favorable to the evolution of cancerous pathologies not classically related to AIDS. Full article

Chronic Lymphocytic Leukemia (CLL) is an indolent malignancy characterized by the accumulation of quiescent mature B cells. However, these cells are transcriptionally and translationally active, implicating an active metabolism. The recent literature suggests that CLL cells have an oxidative-type phenotype. Given the role of cell metabolism, which is able to influence the outcome of treatments, in other neoplasms, we aimed to assess its prognostic role in CLL patients by determining the ex vivo bioenergetic metabolic profile of CLL cells, evaluating the correlation with the patient clinical/biological characteristics and the in vivo response to BTK inhibitor treatment. Clustering analysis of primary samples identified two groups, characterized by low (CLL low) or high (CLL high) bioenergetic metabolic rates. Compared to the CLL high, CLL with lower bioenergetic metabolic rates belonged to patients characterized by a statistically significant higher white blood cell count and by unfavorable molecular genetics. More importantly, patients in the CLL low cluster displayed a better and more durable response to the BTK inhibitor ibrutinib, thus defining a bioenergetic metabolic subgroup that can benefit the most from this therapy. Full article

Alzheimer’s disease (AD) is the world’s most common neurodegenerative disease, expected to affect up to one-third of the elderly population in the near future. Among the major challenges in combating AD are the inability to reverse the damage caused by the disease, expensive diagnostic tools, and the lack of specific markers for the early detection of AD. This paper highlights promising research directions for molecular markers in AD diagnosis, including the diagnostic potential of microRNAs. The latest molecular methods for diagnosing AD are discussed, with particular emphasis on diagnostic techniques prior to the appearance of full AD symptoms and markers detectable in human body fluids. A collection of recent studies demonstrates the promising potential of molecular methods in AD diagnosis, using miRNAs as biomarkers. Up- or downregulation in neurodegenerative diseases may not only provide a new diagnostic tool but also serve as a marker for differentiating neurodegenerative diseases. However, further research in this direction is needed. Full article

A low-calcium microenvironment is imperative for spermatozoa maturation within the epididymis. Our previous work has shown that γ-glutamyl carboxylase (GGCX), the carboxylation enzyme of the matrix Gla protein (MGP), plays an essential role in epididymal calcium homeostasis and sperm maturation in rats and that the GGCX SNP mutation rs699664 was associated with asthenozoospermia (AZS) in humans. Here, we investigated the expression patterns of GGCX and MGP in the mouse epididymis and generated Ggcx^K325Q knock-in (KI) mice. We also tested the effects of this mutation on epididymal calcium homeostasis, sperm function, and male fertility in Ggcx^K325Q−/− mice. The results showed that both GGCX and MGP were enriched in all regions of the mouse epididymis, especially in the initial segment of the epididymis. Double immunofluorescence staining revealed that GGCX colocalized with MGP in the epithelial cells of the initial segment and caput regions as well as in the lumen of the corpus and cauda regions of the mouse epididymis. However, the Ggcx^K325Q−/− mice were fertile with normal epididymal morphology, sperm functions, and epididymal calcium concentration. Overall, our findings revealed that the Ggcx^K325Q mutation does not exert any discernible effect on male fertility in mice. Full article

The grid plays a prominent role in architecture, aiding in space organization and influencing all aspects of planning, ranging from urban design to intricate building details. This paper posits that the grid receives heightened emphasis in Brutalism, particularly in constructivist Brutalism, where materials and construction are intentionally exposed. A question arises regarding the grid’s characteristics—despite its subtle appearance, the grid can sometimes be deceptive, ambiguous, and manipulative. The paper analyzes the merits and drawbacks of employing the grid in architecture, shedding light on its contributions to both structural and perceptual comprehensibility, as well as its role in increasing usefulness. To illustrate the application and perception of the grid, the paper examines two primary planning levels: urban planning and building design. The case studies focus on examples from New York City housing developments, specifically those constructed between the 1950s and the 1970s, and projects by architect I. M. Pei, which offer valuable insights into practical implementation. The paper concludes that while the grid can establish order, it may also engender an “uncanny” feeling. Full article

Lactic acid bacteria (LAB) can produce peptides known as bacteriocins with antagonistic activity against foodborne pathogens. The potential of LAB isolated from the surface of jalapeno peppers to produce bacteriocins with antagonistic activity against Listeria monocytogenes, Staphylococcus aureus, Escherichia coli O157:H7, and Salmonella Typhimurium was evaluated. Previously isolated LAB strains were reactivated, and their cell-free supernatants (CFSs) were evaluated. Out of 390 reactivated strains, 60 produced bacteriocin-like inhibitory substances (BLIS) since their antagonistic activity was lost after proteases addition. Subsequently, 16 BLIS showed heat resistance (HR-BLIS), retaining their bioactivity after heat treatment (121 °C for 15 min). By 16S rRNA gene sequencing and antibiotic susceptibility tests, LAB strains producing HR-BLIS were identified as Enterococcus lactis. Four HR-BLIS exhibited a minimum inhibitory concentration (MIC) of 80 mg/mL against L. monocytogenes. MIC and minimum bactericidal concentration (MBC) of HR-BLIS-67 for S. aureus (MIC = 80 mg/mL; MBC = 320 mg/mL), S. Typhimurium (MIC = 150 mg/mL; MBC = 250 mg/mL), and E. coli O157:H7 (MIC = 250 mg/mL; MBC = 400 mg/mL) were determined. LAB isolated from the surface of jalapeno pepper produced HR-BLIS (possibly enterocin) that exhibited broad-spectrum antagonistic activity against foodborne pathogens; therefore, they are a promising source of natural antimicrobials to ensure food safety. Full article

Basal cell carcinoma (BCC) is the most frequent malignancy in the general population. To date, dermoscopy is considered a key tool for the diagnosis of BCC; nevertheless, line-field confocal optical coherence tomography (LC-OCT), a new non-invasive optical technique, has become increasingly important in clinical practice, allowing for in vivo imaging at cellular resolution. The present study aimed to investigate the possible correlation between the dermoscopic features of BCC and their LC-OCT counterparts. In total, 100 histopathologically confirmed BCC cases were collected at the Dermatologic Clinic of the University of Siena, Italy. Predefined dermoscopic and LC-OCT criteria were retrospectively evaluated, and their frequencies were calculated. The mean (SD) age of our cohort was 65.46 (13.36) years. Overall, BCC lesions were mainly located on the head (49%), and they were predominantly dermoscopically pigmented (59%). Interestingly, all dermoscopic features considered had a statistically significant agreement with the LC-OCT criteria (all p < 0.05). In conclusion, our results showed that dermoscopic patterns may be associated with LC-OCT findings, potentially increasing accuracy in BCC diagnosis. However, further studies are needed in this field. Full article

Onychomycoses are nail infections that require prolonged therapy and have high recurrence rates. Dermatophytes are the main etiological agents of these infections, followed by yeasts and non-dermatophyte filamentous fungi. The limited antifungal arsenal used to treat onychomycosis and the change in the susceptibility profile of these agents contribute to the chronicity and recalcitrant profile of infections. The present study aimed to determine the antifungal activity of extracts obtained from pure and mixed cultures of Candida parapsilosis, Trichophyton mentagrophytes, and Trichophyton rubrum. Additionally, in vivo toxicity tests with Galleria mellonella and time-kill assays were carried out. The susceptibility profiles of dermatophytes were determined using a microdilution technique with minimum inhibitory concentrations (MICs) between 250 and 8000 µg/mL. The time-kill assay, compared to growth control, resulted in the death of dermatophytes within 48 h. No toxicity of the extracts was detected in experiments with Galleria mellonella larvae under the test conditions. The extracts of pure and mixed cultures of Candida parapsilosis and dermatophytes present antifungal activity against T. mentagrophytes and T. rubrum. Isolating and identifying compounds in the extracts may allow the development of new therapeutic approaches to control fungal infections. Full article

Background: This study aimed to examine the efficacy of methylene blue (MB) and toluidine blue O (TBO) photodynamic therapy (PDT) as adjuncts to root surface debridement (RSD). Methods: This split-mouth, randomized, controlled clinical trial included eighteen patients, and a total of 332 sites (control = 102, MB = 124 and TBO = 106) were examined. Two sessions of PDT were completed at baseline and two weeks after RSD. Clinical parameters of bleeding on probing (BOP), plaque index (PI), probing pocket depth (PPD), and clinical attachment level (CAL) were measured pre- and post-treatment. Results: PPD and BOP reductions in sites treated by RSD with adjunctive photosensitizers (MB and TBO) were significantly higher than in control sites. RSD with MB showed higher efficacy in improving moderately deep pockets (OR 3.350), while adjunctive TBO showed better results in treating deeper pockets (OR 4.643). Conclusions: Results suggested that adjunctive use of MB and TBO to RSD could significantly improve periodontal pocket closure and reduce signs of inflammation. In addition, TBO seems to be more efficient in treating deep periodontal pockets than MB, which is more effective in resolving shallower pockets. Full article

Archaeologists deploy a variety of models and theories, often tailored to specific questions or situations, in making sense of the material record we study. The concept of the community of practice, originally developed in the context of modern work and learning situations, describes among other things how participation in shared activities can create and shape social relationships. It therefore offers a powerful and flexible framework for the many archaeological research agendas in which group dynamics play a role. Some archaeologists have already begun to use the community of practice approach (CoP) as an interpretive framework, and this essay argues that a wider embrace would be a benefit to individual archaeologists and to the field as a whole. Full article

The use of remote testing to collect behavioral data has been on the rise, especially after the COVID-19 pandemic. Here we present psychometric functions for a commonly used speech corpus obtained in remote testing and laboratory testing conditions on young normal hearing listeners in the presence of different types of maskers. Headphone use for the remote testing group was checked by supplementing procedures from prior literature using a Huggins pitch task. Results revealed no significant differences in the measured thresholds using the remote testing and laboratory testing conditions for all the three masker types. Also, the thresholds measured obtained in these two conditions were strongly correlated for a different group of young normal hearing listeners. Based on the results, excellent outcomes on auditory threshold measurements where the stimuli are presented both at levels lower than and above an individual’s speech-recognition threshold can be obtained by remotely testing the listeners. Full article

Portland cement has a limited shelf life because of the prehydration that can occur during storage. One approach to mitigating strength losses observed for concrete is to pretreat cement with a protective coating to slow the advance of prehydration. This study compared cement pretreatments with alkyl ketene dimer (AKD) wax and a combination of AKD + paraffin wax to a more traditional pretreatment approach using oleic acid. After exposing the treated cements to elevated temperature and humidity conditions, paste and mortar calorimetry tests showed improved resistance to prehydration reactions. The cements aged up to 12 weeks under the accelerated regime showed strength improvement for the mortars made with AKD and AKD + paraffin-treated cements relative to the mortars made with oleic acid-treated cement and mortars made with untreated cement. The wax can be added during clinker milling and additionally functions as a grinding aid. Full article