Search Results (651)

Preclinical investigations involving in vivo animal studies, are considered a crucial and mandatory step in pharmacological industry regulations. In recent years, as the ethical concerns associated with animal experimentation are becoming more relevant, substantial research efforts have been directed towards the development of novel in vitro methodological approaches aimed at minimizing the utilization of animals. Among those, three-dimensional (3D) cell culture systems have become one of the most promising substitutes for animal models. Skeletal muscle can be affected by genetic disorders and injuries. In this study, two types of 3D scaffolds (Coll and CollMA), prepared starting from a fibrillar collagen suspension extracted from sea urchin food waste, were used as platforms for the generation of 3D skeletal muscle tissue models. The scaffolds were cellularized with C2C12 myoblasts. As the culture period progressed, C2C12 cells gradually infiltrated and were viable throughout both scaffold types, establishing a multilayered cellular population. In the CollMA scaffold, the reciprocal Pax7/MyoD expression pattern suggests a progression of myoblasts towards myogenic commitment. Taken together, our results, although preliminary, suggest that sea urchin–derived collagen matrix represents a promising scaffold for skeletal muscle tissue engineering. Full article

Primary effusion-based lymphomas are uncommon and may pose significant diagnostic challenges. Fluid overload-associated large B-cell lymphoma is a recently recognized entity in the 5th edition of the World Health Organization Classification of Hematolymphoid Tumors and should be included in the differential diagnosis of effusion-based lymphomas, particularly in elderly immunocompetent patients with conditions that predispose to fluid overload. Background and Clinical Significance: We report a case of fluid overload-associated large B-cell lymphoma to add to the limited literature and highlight distinguishing features from other primary effusion lymphomas. Case Presentation: A 77-year-old male with end-stage renal disease on hemodialysis and heart failure with reduced ejection fraction was admitted for respiratory failure and found to have a right-sided pleural effusion. Two pleural fluid specimens examined several weeks apart revealed sheets of large atypical lymphoid cells positive for CD20, Pax-5, CD79a, CD45, MUM1, BCL2, BCL6 (weak) and negative for TTF1, CD68, MOC31, BER EP4, WT1, Calretinin, CD3, CD138, CD30, and cMYC. Human Herpesvirus-8 and Epstein–Barr virus were negative. Staging showed a few mildly fluorodeoxyglucose-avid mediastinal lymph nodes which were benign. Ultimately, the patient was diagnosed with fluid overload-associated large B-cell lymphoma and treated with rituximab, cyclophosphamide, vincristine sulfate, and prednisone, but passed away three months after diagnosis. Conclusions: Fluid overload-associated large B-cell lymphoma is a new and important diagnostic consideration in effusion-based lymphomas. It may be mistaken for other conditions such as primary effusion lymphoma or other diffuse large B-cell lymphomas. The presence of a Human Herpesvirus-8-negative effusion-based lymphoma in an elderly immunocompetent patient without nodal or tissue involvement should prompt consideration of fluid overload-associated large B-cell lymphoma. Full article

Background: Differentiating atypical endometrial hyperplasia, also known as endometrial intraepithelial neoplasia (EAH/EIN) from endometrial hyperplasia without atypia is crucial due to the higher risk of progression to endometrioid adenocarcinoma, associated with atypical lesions. Immunohistochemical markers such as PAX2 and PTEN have emerged as potential adjuncts to improve diagnostic accuracy in morphologically challenging cases. Objective: To evaluate the diagnostic value of PAX2 and PTEN expression in distinguishing atypical from endometrial hyperplasia without atypia in the Bulgarian population. Materials and Methods: A total of 96 endometrial hyperplasia cases (48 typical, 48 atypical) were included. Histopathological evaluation was performed on hematoxylin and eosin–stained sections, with two experienced pathologists confirming diagnoses according to the WHO criteria. Immunohistochemical analysis of PTEN and PAX2 was conducted on formalin-fixed, paraffin-embedded tissue sections. Results: PTEN expression loss was observed in 6% (3/48) of hyperplasia without atypia cases, compared with 81.3% (38/48) of EAH/EIN cases. For PAX2, strong nuclear staining was retained in 60% (29/48) of endometrial hyperplasia without atypia cases, with no complete loss of expression. In contrast, 64.6% (31/48) of EAH/EIN cases showed complete loss of PAX2 expression, while only 35.4% (17/48) preserved nuclear immunoreactivity. Together, these results highlight clear and consistent differences in PTEN and PAX2 expression between hyperplasia without atypia and EAH/EIN and may aid pathologists in distinguishing these two entities in routine diagnostic practice. Conclusions: Expression loss of PAX2 and PTEN is significantly associated with EAH/EIN. Immunohistochemical evaluation of these markers provides valuable adjunctive information for the diagnosis of morphologically ambiguous cases and may enhance diagnostic reproducibility and accuracy. Incorporating PAX2 and PTEN into routine assessment may guide appropriate clinical management and risk stratification of patients with endometrial hyperplasia. Full article

PAX6 haploinsufficiency-related congenital aniridia is frequently associated with ocular surface disease, including meibomian gland dysfunction (MGD), dry eye, limbal stem cell deficiency (LSCD), aniridia-associated keratopathy (AAK), and inflammation. This study measured ocular surface temperature (OST) at the corneal center and four paracentral points (2 mm from center) in patients with congenital aniridia and examined factors influencing OST. Forty-five eyes from 26 aniridia patients (55.6% female; 26.29 ± 17.78 years) with PAX6 pathogenic variants and 47 eyes from 25 controls (68.1% female; 24.81 ± 4.73 years; p = 0.1639) were included. Body temperature, OSDI, and OST (TG-1000) were recorded; clinical assessment evaluated MGD, LSCD, AAK, iris malformation, epithelial defects, Salzmann nodules, glaucoma and previous ocular surgery. Body temperature and OSDI did not differ in aniridia and controls (p ≥ 0.606). LSCD was mainly Grade 2 (46.7%) or Grade 4 (40.0%), and AAK Grade 1 (33.3%) or Grade 2 (31.1%). MGD affected 51.1%, Salzmann nodules 22.2%, epithelial defects 2.2%, glaucoma 60.0%, and previous ocular surgery 35.5%. Superior OST was higher in aniridia (34.98 ± 0.55 °C vs. 34.75 ± 0.47 °C; p = 0.012). Exploratory univariate analyses identified that higher AAK grade correlated with lower inferior OST (p = 0.030), iris malformation with reduced central/paracentral OST (p ≤ 0.029), and Salzmann nodules with lower OST overall (p ≤ 0.011). However, in a multivariate model, age, Salzmann nodular degeneration, and prior ocular surgery emerged as key determinants of OST. OST may serve as a noninvasive biomarker in congenital aniridia. Full article

The Asian black bear (Ursus thibetanus, UT), a Class II protected species in China, presents significant challenges for dermatological research due to limited availability of skin specimens and technical difficulties in fibroblast isolation. To address this, this study developed an immortalized fibroblast cell line through lentiviral-mediated introduction of human telomerase reverse transcriptase (hTERT). The hTERT gene fragment was amplified via PCR and cloned into a eukaryotic expression plasmid using homologous recombination. The recombinant plasmid was packaged into lentiviral particles using a three-plasmid system (psPAX2, pMD2.G) for stable transduction of primary Ursus thibetanus skin fibroblasts (UTSF). Puromycin selection enabled isolation of positive clones, which were subsequently expanded to establish an immortalized cell line. Comprehensive validation through Western blotting, PCR and immunofluorescence confirmed successful genomic integration and sustained expression of hTERT. This study successfully immortalized the skin fibroblasts of black bears and established a fibroblast cell line of Asian black bears. This immortalized UTSF cell line provides a critical in vitro model system for investigating cutaneous physiology and pathogen interactions in this protected species. Full article

This paper presents the results of research on an innovative, integrated IACMS (Intelligent Integrated Automation, Control, and Monitoring System), developed for energy-efficient operation of auxiliary machinery in ship engine rooms. The system, validated both in the laboratory and during full-scale operation on the MF Skania Ro-Pax ferry, integrates process monitoring, diagnostics, predictive maintenance, and intelligent energy optimization within a unified control architecture. This approach enables a significant reduction in electricity consumption while maintaining thermal safety and operational reliability. Laboratory tests focused on a pump cooling system with PLC and frequency converter control, achieving a 90.5% reduction in energy consumption compared to conventional constant-speed operation. During full-scale validation, the IACMS managed the seawater pump via adaptive frequency control (30–60 Hz). Two consecutive voyages demonstrated energy savings of 84.6% and 86.0%, with a daily energy reduction of 0.84 MWh, resulting in a decrease of approximately 0.5 tons of CO₂ emissions per day. Additionally, an observed reduction of about 6–7% in daily generator-set energy was recorded during the analyzed period; this vessel-level value is indicative, as the generator supplies multiple onboard consumers. All trials confirmed stable cooling system temperatures, and comprehensive diagnostics revealed no negative impact of inverter control on the technical condition of equipment. The findings indicate that IACMS is a universal and scalable tool for improving energy efficiency and enabling predictive maintenance in ship engine room auxiliary systems. The system was positively validated in commercial operation and certified by the Polish Register of Shipping, confirming its technological maturity and readiness for widespread adoption in the maritime industry. The results pave the way for further deployments of intelligent energy management solutions in shipping, supporting maritime decarbonization goals. Full article

Long-chain non-coding RNAs (lncRNAs) play important regulatory roles in the growth and development of skeletal muscle, but systematic identification and functional studies of lncRNAs related to porcine skeletal muscle development remain limited. Based on a previously constructed panoramic map of porcine skeletal muscle lncRNAs, lncRNA-ssc.37456 was identified as differentially expressed in porcine skeletal muscle before and after birth. Its function and potential mechanisms were investigated using a porcine skeletal muscle regeneration model, a primary skeletal muscle cell differentiation model, and knockdown and overexpression experiments in vitro. lncRNA-ssc.37456 was upregulated on day 7 of regeneration, with expression positively correlated with the muscle differentiation marker MYHC and negatively correlated with the proliferation marker PAX7. During differentiation of porcine primary myoblasts, expression continuously increased, peaking on day 4. Knockdown of lncRNA-ssc.37456 by small interfering RNA (siRNA) significantly increased cell proliferation, upregulated mRNA and protein levels of proliferation-related genes KI67 and PCNA, and increased the proportion of EdU-positive cells. Conversely, expression of differentiation-related genes MYOG and MYHC decreased, and immunofluorescence analysis revealed reduced myotube formation and differentiation index. Overexpression of lncRNA-ssc.37456 promoted differentiation and inhibited proliferation, showing effects opposite to those observed in knockdown experiments. Nucleocytoplasmic fractionation indicated predominant cytoplasmic localization, suggesting potential function through a ceRNA mechanism. An interaction network with miRNAs was constructed based on the miRDB database, indicating a potential miRNA “sponge” regulatory mechanism. These results indicate that lncRNA-ssc.37456 participates in porcine skeletal muscle development by regulating the transition of muscle cells from proliferation to differentiation, providing molecular insights and potential targets for muscle biology research and the molecular breeding of growth traits. Full article

Maritime transport is responsible for 3% of global greenhouse gas (GHG) emissions, making it a focus of decarbonization efforts. Ro-pax ferries, operating in the domestic shipping, are particularly emission-intensive due to their high operational frequency, while advances in battery-powered propulsion suggest that electrification is feasible on short to medium distance routes. This paper uses a systematic literature review of studies published between 2014 and 2024 to investigate the application of battery-powered ferries from a maritime transport system perspective. Using the PRISMA 2020 guidelines, the authors identified 15 case-study-based papers on battery-powered ferries, with a specific focus on the methodological approaches applied to domestic shipping routes. The goal of this review is to identify and systematize the methodologies used in case study research to analyze the implementation of battery-powered ferries on specific routes. The review contributes a structured synthesis of (I) methodological approaches, grouped into four clusters, and (II) route framing and selection practices using a three-level route classification, revealing an increasing methodological complexity, from single-route feasibility assessments to diversified, maritime network-integrated approaches. The paper systematically links existing methodologies to operational and conceptual case studies, providing practical insights for future decarbonization projects. Full article

Neonatal Diabetes Mellitus (NDM) is a rare, heterogeneous monogenic disorder typically presenting within the first six months of life. Unlike type 1 or type 2 diabetes, NDM is caused by single-gene mutations that disrupt pancreatic β-cell function or development. With the advent of next-generation sequencing, the genetic spectrum of NDM has expanded significantly, necessitating a shift from symptomatic management to precision medicine. This narrative review summarizes the genetic basis and pathogenic mechanisms of NDM, categorizing them into three major pathways: (1) ATP-sensitive potassium (K_ATP) channelopathies (e.g., ABCC8, KCNJ11), where gain-of-function mutations inhibit insulin secretion; (2) Transcription factor defects (e.g., GLIS3, PAX6, GATA6), which impair pancreatic development and often present with syndromic features; and (3) Endoplasmic reticulum (ER) stress-mediated β-cell apoptosis, exemplified by WFS1 mutations. Furthermore, we highlight the clinical complexity of these mutations, including the “biphasic phenotype” observed in ABCC8 and HNF1A variants. Understanding these molecular mechanisms is critical for clinical decision-making. We discuss the transformative impact of genetic diagnosis in treatment, particularly the successful transition from insulin to oral sulfonylureas in patients with K_ATP channel mutations, and emphasize the importance of early genetic testing to optimize glycemic control and prevent complications. Full article

This study examines the environmental and economic impacts of transitioning RORO Pax ships from diesel to green and blue hydrogen fuel, focusing on the Jazan case study vessel. It evaluates the environmental and economic effects for both retrofitted and new vessels. Findings reveal that hydrogen-powered PEMFC engines achieve a 99.13% reduction in NOx emissions and reduce both SO_x and CO₂ emissions to minimum values. The analysis indicates that retrofitting with blue hydrogen can achieve a lifetime emission reduction of approximately 134 kton, yielding a net benefit of USD 4.46 per ton of emissions reduced. Newbuilding options present a more favorable financial profile at USD 19.31 per ton, surpassing green hydrogen’s USD 16.61 per ton. The study highlights the economic infeasibility of retrofitting existing vessels due to insufficient operational life, while hydrogen fuel becomes viable for sustainable new builds after 6 to 10 years, potentially resulting in annual cost savings of USD 2 to USD 3 million and competitive hydrogen production costs of up to USD 0.30 per kWh. Full article

In recent years, heavy metal emissions in Lhasa have been increasing, which has an impact on the local water environment. The negative effects of copper (Cu²⁺) on aquatic ecosystems have attracted much attention, as even low concentrations of Cu²⁺ can exert toxic effects on aquatic organisms. However, the impact of Cu²⁺ on native fish species from the Lhasa River remains poorly understood. In this study, Schizopygopsis younghusbandi (S. younghusbandi) larvae were exposed to Cu²⁺ at concentrations of 0. 5, 5, 50, and 500 μg/L for 7 or 14 days to evaluate its toxic effects on thyroid function and the antioxidant system. The results indicate that whole-body total thyroxine (T4) and triiodothyronine (T3) levels were significantly decreased following Cu²⁺ exposure. This decrease was accompanied by a marked increase in dio1 and dio2 gene expression and decreased expression of thyroid hormone synthesis genes (nis, tg, ttf1 and pax8) after exposure to Cu²⁺. Furthermore, the activity of superoxide dismutase (SOD), catalase (CAT), and glutathione reductase (GR) and the content of lipid peroxidation were increased, while the content of glutathione (GSH) was decreased. In addition, the survival rates and body lengths of S. younghusbandi larvae were significantly reduced following 7- and 14-day Cu²⁺ exposure. The Integrated Assessment of Biomarker Response (IBR) analysis further revealed dose- and time-dependent effects of Cu²⁺ on the larvae. In conclusion, the findings demonstrate that Cu²⁺ exposure induced disruption of thyroid endocrine and antioxidant systems and caused developmental toxicity in S. younghusbandi larvae. Full article

Biphenotypic sinonasal sarcoma (BSNS) is a very rare, locally aggressive sarcoma arising in the sinonasal region, initially recognized as low-grade sinonasal sarcoma with neural and myogenic differentiation. Here, we report a case of BSNS extending into the intracranial and intraorbital regions, finally diagnosed by a break-apart fluorescence in situ hybridization (FISH) assay for rearrangements of PAX3. A 50-year-old woman presented with left diplopia and exophthalmos. CT and MRI revealed a large ethmoidal mass with intracranial and intraorbital extension. Since preoperative biopsy suggested a benign tumor, endoscopic endonasal resection was performed while preserving the anterior skull base and intraorbital structures. Postoperative histopathological diagnosis indicated synovial sarcoma, and proton beam therapy with adjuvant chemotherapy was subsequently administered. After treatment, FISH demonstrated rearrangements of PAX3 and MAML3 genes, leading to a revised diagnosis of BSNS, which typically does not require chemotherapy due to its non-metastatic behavior. Eleven years after treatment, the patient remains free of recurrence. Understanding BSNS is essential to avoid excessive intervention, and confirmation of PAX3 rearrangement by FISH or equivalent molecular testing is crucial for accurate diagnosis. Full article

Cervical cancer remains a major global public health concern, with persistent infection by high-risk human papillomavirus (hrHPV) types recognized as the primary etiological factor. This review explores the multifactorial nature of the disease, focusing on the complex interplay between host genetic susceptibility and epigenetic alterations that drive cervical carcinogenesis. Evidence from genome-wide association studies (GWAS) is discussed, highlighting the contribution of specific genetic loci, predominantly within the HLA region, to susceptibility to HPV infection and disease progression. In parallel, the review examines the molecular mechanisms by which the viral oncoproteins E6 and E7 promote genetic instability and epigenetic reprogramming, including histone modifications and dysregulation of non-coding RNAs. Particular emphasis is placed on DNA methylation, affecting both the viral genome and host genes such as FAM19A4, CADM1, PAX1, and MAL, as a promising biomarker for triage and detection of high-grade intraepithelial lesions (CIN2+). Finally, the review evaluates currently available methylation-based assays and self-sampling devices, highlighting their potential to enhance diagnostic accuracy and increase participation in cervical cancer screening programs. Full article

Electroplating wastewater poses a serious environmental threat due to its high concentrations of heavy metals and persistent organic pollutants. This study evaluated the efficiency of a combined coagulation and activated carbon filtration process for the treatment of real electroplating wastewater containing Ni²⁺, Zn²⁺, Cu²⁺, and Cr⁶⁺ ions. The research was conducted in two stages. In the first stage, laboratory-scale experiments were performed to determine the optimal coagulant type (Fe- and Al-based), dosage, and pH (5.0–10.0) for contaminant removal. In the second stage, the selected operating conditions were applied and validated under real industrial plant conditions at a pilot scale. The laboratory studies demonstrated that the highest Cr removal efficiency was achieved using an iron-based coagulant (PIX), while polyaluminum chloride (PAX) proved most effective for the removal of Ni and Zn. Subsequent filtration through activated carbon further enhanced heavy metal removal, increasing overall efficiencies to above 90%. The reported removal efficiencies represent the overall performance of the integrated treatment process. The results confirm that the integration of chemical coagulation and activated carbon filtration is an effective, environmentally friendly, and economically viable approach for treating real electroplating wastewater, enabling compliance with current environmental standards. Full article

Dicer is crucial for the generation of microRNAs (miRNAs), which are essential for regulating gene expression and keeping neuronal health. Dicer’s conditional deletion cuts all spiral ganglion neurons but spares a small fraction of vestibular ganglion neurons, innervating the utricle and part of the saccule. Hair cells develop in the utricle, saccule, posterior crista, and the cochlea in Pax2^Cre; Dicer^f/f. Cochlear hair cells develop at the base and expand the OHC and IHC in the middle, or split into a base/middle and the apex. In contrast, Foxg1^Cre; Dicer^f/f cuts all canal cristae and cochlea hair cells, leaving a reduced utricle and an exceedingly small saccule. Likewise, Foxg1^Cre; Gata3^f/f shows no cochlear hair cells and is absent in the horizontal and reduced in the posterior crista. In contrast, the utricle, saccule, and anterior crista are nearly normal, underscoring the intricate regulatory networks involved in hair cell and neuronal development. The central projections have been described as the topology of various null deletions. Still, without spiral ganglion neurons, fibers from Dicer null mice navigate to the cochlear nuclei and expand into the vestibular nuclei to innervate the caudal brainstem. Beyond a ramification around the CN, no fibers expand to reach the cerebellum, likely due to Pax2 and Foxg1 that cut these neurons. Genetic alterations, such as Dicer deletion, can lead to hearing loss and impairments in auditory signal processing, illustrating the critical role of microRNAs in the development and function of auditory and vestibular neurons. Further studies on this topic could help in understanding potential therapeutic targets for hearing loss associated with neuronal degradation of miRNA. Full article