Abstract: Cytoskeletal structures shape and confer resistance to cells. The intermediate filament protein vimentin forms versatile structures that play key roles in cytoskeletal crosstalk, in the integration of cellular responses to a variety of external and internal cues, and in the defense against stress. Such multifaceted roles can be fulfilled thanks to the vast variety of vimentin proteoforms, which in turn arise from the combinations of a myriad of tightly regulated posttranslational modifications. Diverse vimentin proteoforms will differentially shape its polymeric assemblies, underlying vimentin ability to organize in filaments, bundles, squiggles, droplets, cell surface-bound and/or various secreted forms. Interestingly, vimentin dots or droplets have been lately categorized as biomolecular condensates. Biomolecular condensates are phase-separated membraneless structures that are critical for the organization of cellular components and play important roles in pathophysiology. Recent findings have unveiled the importance of low complexity sequence domains in vimentin filament assembly. Moreover, several oxidants trigger the transition of vimentin filaments into phase-separated biomolecular condensates, a reversible process that may provide clues on the role of condensates as seeds for filament formation. Revisiting previous results in the light of recent knowledge prompts the hypothesis that vimentin condensates could play a role in traffic of filament precursors, cytoskeletal crosstalk and cellular responses to stress. Deciphering the “vimentin PTM code”, that is, the structure-function relationships of vimentin proteoforms, constitutes a major challenge to understand the regulation of vimentin behavior and its multiple personalities. This will contribute to unveil essential cellular mechanisms and foster novel opportunities for drug discovery.

Abstract: Plants are confronted with a myriad of challenges that they must mitigate without being able to move to new environments. Systemic acquired resistance (SAR) is a secondary immune response in plants to pathogen infection. Little is known about SAR in red algae. The red alga Dasysiphonia japonica can survive for more than six years without lethal damage when infected with the pathogen Olpidiopsis heterosiphoniae but dies within a month when infected with another pathogen, Olpidiopsis dasysiphoniae. During this process, D. japonica infected with O. heterosiphoniae accumulated reactive oxygen species (ROS, DCFH-DA staining) in the host cell membrane, whereas this was not observed with O. dasysiphoniae infection. Transcriptome analysis identified four NADPH-oxidase genes (Djrboh) that generate ROS in the cell membrane of D. japonica. Real-time PCR of four NADPH oxidase genes (Djrboh) showed that two were upregulated (Djrboh1, 2) at the onset of Olpidiopsis heterosiphoniae infection, but not during infection with O. dasysiphoniae in early infection that does not induce SAR. The successful SAR of D. japonica was mediated by low concentrations of ROS signaling and regulation of NADPH-oxidase genes.

Abstract: Epidemiological studies have linked higher serum and dietary phosphorus to an in-creased risk of prostate cancer (PC) and its lethal state. These findings do not distinguish between the impact of inorganic phosphorus (Pi) per se or its homoeostatic regulators. Thus, this study aimed to determine the in vitro tumorigenic effects of elevated Pi concen-trations per se on androgen-dependent epithelial-like PLum-AD murine PC cells at molec-ular and cellular levels. Physiologically attainable elevated levels and supraphysiological levels of sodium (NaPi) and potassium phosphate (KPi) were used to assess PLum-AD cell proliferation, viability, migration, and epithelial-mesenchymal transition (EMT) marker expression, which were determined by the thiazolyl blue tetrazolium bromide cell assay, trypan blue exclusion assay, wound healing assay, and immunofluorescence staining, respectively. Treatment of Plum-AD cells with supraphysiological levels of NaPi (20 mM) significantly reduced cell proliferation, whereas KPi did not, suggesting a poten-tial sodium- dependent Pi uptake mechanism. Furthermore, physiologically relevant ele-vated concentrations of NaPi (3 mM) and KPi (1 and 3 mM) increased relative vimentin expression of PLum-AD PC cells, a biomarker of EMT. Our findings suggest that elevated Pi levels per se, in hyperphosphatemia range, can directly promote EMT in PC, highlight-ing its potential role in tumor progression.

Abstract:

Background: Antimicrobial resistance (AMR) is a pressing global health concern, leading to increased treatment costs, prolonged hospital stays, and higher mortality rates. This study analyzes the prevalence and trends of AMR in pathogenic bacteria isolated from various clinical specimens from Chattogram Medical College Hospital (CMCH) in Chittagong, Bangladesh. The objective is to track AMR over an extended period and provide comparative analytics for local and global surveillance efforts. Methods: Retrospective data from June 2017 to November 2019 were collected from a tertiary care hospital, en-compassing both inpatients and outpatients. Bacterial identification and antibiotic susceptibility testing followed standard methods. WHONET and Quick Analysis of AMR Patterns and Trends (QAAPT) software were utilized for data management and analysis. Results: The analysis included 6,896 records, with an average bacterial growth positivity rate of 39%. The most common specimen type was urine, accounting for 48.9% of all specimens. Among the bacterial isolates, variations in AMR prevalence were observed, particularly with E. coli displaying high resistance to commonly used antibiotics. Soft tissue and blood fluid samples exhibited a high positivity rate for bacterial growth. The study underscores the urgent need for AMR surveillance and evidence-based treatment guidelines tailored to local antibiotic susceptibility patterns. Conclusion: This study highlights the significance of monitoring AMR trends in Chittagong, Bangladesh. By understanding and addressing AMR patterns, policymakers, and stakeholders can develop informed national policies and strategies to combat AMR effectively. Sharing these findings with relevant parties is crucial for creating awareness and promoting evidence-based practices. The study emphasizes the importance of ongoing surveillance efforts and the development of targeted interventions to mitigate the impact of AMR and improve patient outcomes in the region.

Abstract:

The development of BPD in preterm neonates is increased by poor growth and nutritional deficits. The involvement of the fatty acid DHA in the development of BPD has been a focus for over a decade. However, recent clinical trials show that isolated DHA supplementation may increase BPD in subgroups of preterm neonates. One explanation for poor lung outcomes in DHA-supplemented neonates is a disruption of global fatty acid profiles and increased expression of a dominant-negative splice variant of a key driver of lung development, PPARγ. We previously developed rat model of postnatal growth restriction (PGR) in which pups have impaired lung function and altered PPARγ activity. Here we use our PGR rat model to assess the effects of DHA supplementation on lung outcomes. We hypothesize that the PPARγ splice variant, PPARγΔ5, will be expressed in the rat lung, and that DHA supplementation of PGR rat pups will alter circulating lipid profiles, lung mechanics, and PPARγ variant expression. Our findings demonstrate that PPARγΔ5 is expressed in the developing rat lung and that DHA supplementation of PGR rat pups alters global circulating fatty-acid profiles and does not restore PGR-induced impaired lung mechanics or PPARγ activity.

Abstract: Ubiquitination is dynamic and tightly regulated post-translational modifications essential for modulating protein stability, trafficking, and function to preserve cellular homeostasis. This process is orchestrated through a hierarchical enzymatic cascade involving three key enzymes: the E1 ubiquitin-activating enzyme, the E2 ubiquitin-conjugating enzyme, and the E3 ubiquitin ligase. The final step of ubiquitination is catalyzed by E3 ubiquitin ligase, which facilitates the transfer of ubiquitin from the E2 enzyme to the substrate, thereby dictating which proteins undergo ubiquitination. Emerging evidence underscores the critical roles of ubiquitin ligases in neurodevelopment, regulating fundamental processes such as neuronal polarization, axonal outgrowth, synaptogenesis, and synaptic function. Mutations in genes encoding ubiquitin ligases and the consequent dysregulation of these pathways have been increasingly implicated in a spectrum of neurodevelopmental disorders, including autism spectrum disorder, intellectual disability, and attention-deficit/hyperactivity disorder. This review synthesizes current knowledge on the molecular mechanisms underlying neurodevelopment regulated by Cullin-RING ubiquitin ligases—the largest subclass of ubiquitin ligases—and their involvement in the pathophysiology of neurodevelopmental disorders. A deeper understanding of these mechanisms holds significant promise for informing novel therapeutic strategies, ultimately advancing clinical outcomes for individuals affected by neurodevelopmental disorders.

Abstract:

Spermatogenesis is a complex and highly regulated process involving the proliferation, differ-entiation, and apoptosis of germ cells. This process is controlled by various hormonal, genetic, and environmental factors, including temperature. In Hormonal regulation, Follicle-Stimulating Hormone (FSH), Luteinizing Hormone (LH) and testosterone (T), are essential for a correct spermatogenesis development from early stages and spermatogonia proliferation to germ cells maturation. Other hormones, like inhibin and activin, finely participate tuning the process of spermatogenesis. Genetic regulation involves various transcription factors such as SOX9, SRY, and DMRT1, which are crucial for the development and maintenance of the testis and germ cells. MicroRNAs (miRNAs) play a significant role by regulating gene expression post-transcriptionally. Epigenetic modifications, including DNA methylation, histone modifica-tions, and chromatin remodelling, are also vital. Temperature regulation is another critical as-pect, with the testicular temperature maintained around 2-4°C below body temperature, essen-tial for efficient spermatogenesis. Heat shock proteins (HSPs) protect germ cells from heat-induced damage by acting as molecular chaperones, ensuring proper protein folding and preventing the aggregation of misfolded proteins during thermal stress. Elevated testicular temperature can impair spermatogenesis, increasing germ cell apoptosis, and inducing oxidative stress, DNA damage, and disruption of the blood-testis barrier, leading to germ cell death and impaired differentiation. Cellular mechanisms of germ cell proliferation, differentiation, and death include the mitotic divisions of spermatogonia to maintain the germ cell pool and pro-duce spermatocytes. Spermatocytes undergo meiosis to produce haploid spermatids, which then differentiate into mature spermatozoa. Apoptosis, or programmed cell death, ensures the re-moval of defective germ cells and regulates the germ cell population. Hormonal imbalance, ge-netic defects, and environmental stress can trigger apoptosis during spermatogenesis. Under-standing these mechanisms is crucial for addressing male infertility and developing therapeutic interventions. Advances in molecular biology and genetics continue to uncover the intricate de-tails of how spermatogenesis is regulated at multiple levels, providing new insights and poten-tial targets for treatment.

Abstract: In many vertebrates, mature ovulated eggs arrest at metaphase II prior to fertilization. The eggs exit meiotic arrest after fertilization-induced or parthenogenetic activation, followed by embryo development or egg degradation, respectively. Calcium-dependent activation of meiotically-arrested eggs has been thoroughly investigated in various species. In addition, several recent studies have detailed the excessive activation of ovulated frog eggs, so called overactivation. This overview highlights the major events of overactivation observed in mature ovulated eggs of the African clawed frog Xenopus laevis with a focus on similarities and differences between spontaneous, oxidative stress-induced, and mechanical stress-induced overactivation. The paper also underscores the dramatically different cell death scenarios that unfold in activated and overactivated eggs.

Abstract: By the mid-1960s hematopoietic stem cells (HSCs) were well described and they generate perhaps the most complex array of functionally mature cells in an adult organism. HSCs and their descendants have been studied extensively, and findings have provided principles that have been applied to the development of many cell systems. However, there are uncertainties about the process of HSC development. They center around when and how HSCs become affiliated to a single cell lineage. A longstanding view is that this occurs late in development and stepwise via a series of committed oligopotent progenitor cells which eventually give rise to unipotent progenitors. A very different view is that lineage affiliation can occur as early as within HSCs and the development of these cells to a mature end cell is then a continuous process. A key consideration is the extent to which lineage affiliated HSCs self-renew, to make a major contribution to hematopoiesis. This review examines the above aspects to our understanding of hematopoiesis.

Abstract: Glomerular podocytes act as a part of filtration barrier in the kidney. The activity of this filter is regulated by ionotropic and metabotropic glutamate receptors. Adjacent podocytes can potentially release glutamate into intercellular space; however, little is known about how podocytes release glutamate. Here, we demonstrated vesicular glutamate transporter 3 (VGLUT3)-dependent glutamate release from podocytes. Immunofluorescence analysis revealed that rat glomerular podocytes and an immortal mouse podocyte cell line (MPCs) express VGLUT1 and VGLUT3. Consistent with this finding, quantitative RT-PCR revealed the expression of VGLUT1 and VGLUT3 mRNA in undifferentiated and differentiated MPCs. In addition, the exocytotic proteins vesicle-associated membrane protein 2, synapsin 1, and synaptophysin 1 were present in punctate patterns, and colocalised with VGLUT3 in MPCs. Interestingly, approximately 30% of VGLUT3 colocalised with VGLUT1. By immunoelectron microscopy, VGLUT3 was often observed around clear vesicle-like structures in differentiated MPCs. Differentiated MPCs released glutamate following depolarization with high potassium levels and after stimulation with the muscarinic agonist pilocarpine. Depletion of VGLUT3 in MPCs by RNA interference reduced depolarization-dependent glutamate release. These results strongly suggest that VGLUT3 is involved in glutamatergic signaling in podocytes and maybe a new drug target for various kidney diseases.

Abstract:

The molecular mechanisms through which endometriosis-related ovarian neoplasms (ERONs) develop from benign endometrioma remain unclear. It is especially a long-standing mystery why ovarian endometrioma has the potential to develop into two representative histological subtypes: endometrioid ovarian carcinoma or clear cell ovarian carcinoma. The present study aimed to investigate the molecular carcinogenesis of ERONs using newly developed in vitro and in vivo carcinogenesis models. Epithelial cells were isolated and purified from surgically removed benign endometrioma samples, followed by immortalization by overexpressing cyclinD1/CDK4 in combination with the human TERT gene. Immortalized cells were subjected to various molecular manipulations by combining knockout or overexpression of several candidate drivers, including ARID1A, KRAS, PIK3CA, AKT, and MYC, based on previous comprehensive genome-wide studies of ERONs. These cells were then inoculated into immunocompromised mice and evaluated for malignant transformation. Inoculated cells harboring a combination of three genetic alterations successfully de-veloped tumors with malignant features in mice, whereas those with two genetic mutations failed to do so. Especially, ARID1A gene knockout combined with overexpressing the KRAS oncogenic mutant allele (or overexpressing AKT) and c-Myc overexpression led to efficient tumor formation. Of note, these three combinations of genetic alterations produced tumors that histologically represented typical clear cell carcinoma in SCID mice, while the same combination led to tumors with endometrioid histology in nude mice. A combination of ARID1A mutation, KRAS mutation or AKT activation, and c-Myc overexpression were confirmed to be the main candidate drivers for the development of ERONs, as suggested by comprehensive genetic analyses of ERONs. A tumor immune microenvironment involving B cell signaling may contribute to the diverse histological phenotypes. The present model may help to clarify the molecular mechanisms of ERON carcinogenesis and understand their histological diversity and novel molecular targets.

Abstract: Poor embryo quality is a major cause of poor clinical outcomes in assisted reproductive medicine, and there are no currently available interventions that can improve embryo quality. Mitochondria dysfunction is linked to low-quality female gametes and zygotes. Previously, microtubule integrity was also associated with mitochondrial function in oocytes. In the present study, we investigated the effects of the microtubule stabilizers (MTS) Taxol and Epothilone D and the microtubule disturber (MTD) vinorelbine on mouse preimplantation embryo quality and pregnancy outcome. We prepared young BDF1 mice (7~9 weeks old) and cultured preimplantation embryos with MTS or MTD. Mitochondrial functional activity and embryo development ratios including pregnancy ratios were then assessed. MTS-treated embryos showed significantly increased mitochondrial membrane potentials and motility. Blastocyst formation was significantly higher in MTS-treated embryos than in MTD-treated embryos. Especially, MTS-treated embryos exhibited higher hatched blastocyte formation than untreated embryos. The number of offspring was significantly higher in surrogate mice transplanted with MTS-treated embryos. These findings demonstrated that treatment of mouse preimplantation embryos with Taxol or EpD increased embryo development competence, which was associated with increased mitochondrial functional activity. Consistently, delivery ratios were significantly higher after transplantation with MTS-treated embryos than after transplantation with untreated embryos. These findings suggest that MTS could be used to supplement in vitro culture media to promote the recovery of poor-quality embryos.

Abstract: Chitin synthase 3 complex assembly begins at the endoplasmic reticulum where the formation of a Chs3/Chs7 complex facilitates its exit from the ER and its transport along the secretory route. In the present study, our work shows that orphan molecules of Chs7 can exit the ER and are later recycled from the early Golgi by coat protein I (COPI) machinery via the adaptor complex Erv41/Erv46. Moreover, an eventual excess of the protein in the Golgi is recognized by the GGA complex and targeted to the vacuole for degradation through the ESCRT machinery. Non-oligomerizable versions of Chs3, can also exit the ER individually and follow a similar route to that of Chs7. We therefore demonstrate the traffic of unassembled CS3 subunits and describe the cellular mechanisms that guarantee the correct assembly of this protein complex at the ER while providing a default traffic route to the vacuole in case of its failure. This traffic route is shared with canonical ER adaptors, such as Erv29 and Erv14, and other components of protein complexes. The comparative analysis of their traffic allows us to discern a cellular program that combines COPI recycling, proteasomal degradation, and vacuolar disposal for maintaining protein homeostasis at the ER.

Abstract:

Oxidative stress represents a significant contributing factor to male infertility and sperm dysfunction. In this condition, an increase in ROS production exceeds the body's antioxidant defences, resulting in a decline in spermatozoa quality and fertilising capacity. Furthermore, excessive ROS production has been linked to the promotion of genomic damage, lipid peroxidation, inflammation, altered enzyme activity, and ultimately, irreversible alterations, cell death, and a decline in seminal parameters associated with male infertility. It is established that physical activity (PA), acting on inflammatory parameters and improving antioxidant defence, can alleviate the negative effects caused by free radicals, offering numerous health benefits and positively influencing sperm quality. The objective of this review is to highlight the mechanisms of ROS production, the physiological and pathophysiological roles of ROS in relation to the male reproductive system, and recent knowledge on the impact of some protocols of PA on these systems and the molecular mechanisms involved.

Abstract: Amyotrophic lateral sclerosis (ALS) is a devastating neuromuscular disease characterized by progressive motor neuron degeneration and muscle atrophy, with no effective treatments available. Chronic inflammation, which impairs muscle regeneration and promotes proteolysis, is a key contributor to ALS-related muscle atrophy and a promising therapeutic target. Here, we applied extracellular vesicles (EVs) derived from regenerating skeletal muscles 14 days post-acute injury (CTXD14SkM-EVs), which possess a unique anti-inflammatory profile, to target muscle defects in ALS. We found that CTXD14SkM-EVs enhanced myoblast differentiation and fusion in a cellular muscle-wasting model induced by pro-inflammatory cytokine tumor necrosis factor alpha. Intramuscular administration of these EVs into an ALS mouse model mitigated muscle atrophy by promoting muscle regeneration, shifting macrophage polarization from pro-inflammatory M1 to anti-inflammatory M2 state, and suppressing the aberrant Nuclear Factor Kappa B (NF-κB) signaling, a key driver of muscle protein degradation. These results underscore the therapeutic potential of regenerating muscle-derived EVs for combating muscle atrophy in ALS.

Abstract:

Cardiovascular disease (CVD), a major global health concern, is characterized by cardiac complications that can lead to death. The commonly used treatments for this condition are synthetic drugs, but these often come with risky side effects. A potential alternative is the use of traditional medicinal plants, such as Amaranthus viridis, which is rich in bioactive compounds. This study aimed to determine the non-toxic concentration of A. viridis ethanolic extract, investigate its cardioprotective effects on zebrafish (Danio rerio) heart rate and cardiac phenotype, qualitatively assess the presence of phytochemicals, and assess its free radical scavenging activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH). Zebrafish larvae at 72 hours post-fertilization (hpf) were used to evaluate mortality and optimize dosing. Physio-morphological screening was conducted by pre-treating zebrafish larvae with the extract 4 hours prior to administering a heart failure inducer, verapamil. The maximum non-toxic concentration was found to be 25 µg/mL, as all zebrafish survived after 24 hours. Mortality began at 50 µg/mL, and concentrations from 100 µg/mL to 400 µg/mL resulted in 100% mortality. All tested concentrations of A. viridis leaf extract showed cardioprotective activity in the physio-morphological analysis. Phytochemical analysis detected the presence of alkaloids, flavonoids, and saponins. Furthermore, A. viridis exhibited free radical scavenging activity from all tested concentrations. Based on the results, A. viridis exhibited cardioprotective effects against verapamil-induced cardiotoxicity, as evidenced by the recovery of heart rate and cardiac phenotype in the zebrafish model.

Abstract: Many neurobehavioral tests are used for the assessment of human-like behaviors in animals. Most of them were developed in rodents and are used for the assessment of animal models that mimic human neurodevelopmental and neuropsychiatric disorders (NDD). We have described tests for assessing social behavior, social interaction and social communication; tests for restricted and repetitive behaviors; tests for cognitive impairment, for sensory stimuli, for anxiety like behavior and for motor coordination deviations. These tests are used to demonstrate autistic-like behavior as well as other NDD. We described possible general pitfalls in the performance of such studies, as well as probable individual errors for each group of tests assessing specific behavior. The mentioned pitfalls may induce crucial errors in the interpretation of the results, minimizing the reliability of specific models of defined human NDD. It is imperative to minimize these pitfalls and also use sufficient and reliable tests that are able to demonstrate as many of the traits of the human disorder, grade the severity of the specific deviations and the severity of the tested NDD by using a scoring system. Due to possible gender differences in the clinical presentations of NDD, it is important to carry out studies on males and females.

Abstract: Three-dimensional (3D) spheroids of mesenchymal stem cells (MSCs) have high biological significance due to more accurate imitation of physiological conditions, in vivo processes and greater therapeutic efficiency compared to monolayer MSCs. However, molecular processes and specific mechanisms regulating the effectiveness of spheroids remain unknown. Endometrial MSCs are considered as reliable and potent objects for stem cell-based therapy due to availability and minor ethical concerns. Thus, knowledge of the molecular mechanisms controlling eMSC processes is of clinical importance. Regulation of a number of important physiological processes in MSCs is associated with the functioning of Ca2+-permeable mechanosensitive Piezo1 channels. It has been shown that Piezo1 is a key determinant in the regulation of intracellular Ca2+, which controls the migratory activity of 2D eMSCs. We aimed to determine the presence and functions of Piezo1 in eMSC spheroids. We detected that Piezo1 mRNA expression was lower in spheroids compared to 2D culture. Also, we observed the decrease in spheroid spreading rate in the presence of a selective Piezo1 agonist, Yoda1, during spheroid spreading or formation. The selective activation of Piezo1 could be used for the modulation of spheroid spreading rates which could be favorable for some strategies in development of spheroid-based cell therapies.

Abstract:

Five major antimicrobial peptides (AMPs) in <i>Drosophila</i> are induced in <i>multiple sex combs</i> (<i>mxc</i>) mutant larvae harboring lymph gland (LG) tumors and exhibit anti-tumor effects. The effects of the other well-known AMPs, Cecropin A and Drosocin, remain unelucidated. We investigated the tumor-elimination mechanism of these AMPs. A half-dose reduction of either <i>Toll</i> or <i>Imd</i> gene reduced the AMPs’ induction in the fat body and enhanced tumor growth in <i>mxc^mbn1</i> mutant larvae, indicating that their anti-tumor effects depend on the innate immune pathway. Overexpression of these AMPs in the fat body suppressed tumor growth without affecting cell proliferation. Apoptosis was significantly promoted in the mutant LGs but not in normal tissues. Conversely, their knockdown inhibited apoptosis and enhanced tumor growth. Therefore, these AMPs inhibit LG tumor growth by inducing apoptosis. The AMPs from the fat body were incorporated into hemocytes of mutant but not normal larvae. Another AMP, Drosomycin, was taken up via phagocytosis factors. Enhanced phosphatidylserine signals were observed on the tumor surface. Inhibition of the cell-surface exposed signals impeded tumor growth suppression. AMPs may target phosphatidylserine in the tumors for apoptosis induction to execute tumor-specific effects. AMPs are potentially beneficial anti-cancer drugs with minimal side effects for clinical development.

Abstract:

We focused on two microenvironments within a 5-min jugular vein puncture wound thrombus. The goal was to infer from 3D platelet morphodynamics how microenvironments determine platelet activation state. Platelets at the collagen-rich adventitial interface were rod shaped and oriented vertical to flow vectors. The net effect was to maximize their interaction with collagen. At the wound hole interface, soluble signaling should predominant. Here, the typical platelet exhibited what appeared to be a “migratory”, granule-free, leading edge extending toward the wound hole from an organelle rich cell body. Morphologically, this structure resembled the leading edge of a wound associated fibroblast. Platelets were aligned parallel with one another and at ~45 degrees to blood flow. We submit the morphodynamics observed here indicate novel similarities between platelets and wound associated nucleated cells.