Waad Bajunaid, Kyle Tomaro, Anwer Hasil Kottarampatel
et al.
ABSTRACT Nε-lysine-protein fatty acid acyltransferase (k-FAT) is a new class of enzyme found in bacteria that express either a MARTX toxin with a Rho-interacting domain (RID) or a type III secretion system (T3SS). The prototype k-FAT is the T3SS effector IcsB, which enables enteropathogenic Shigella species to evade destruction by autophagy in host cells. We mined protein databases using the acyltransferase domain of IcsB to expand this family. In addition to the known BopA homolog from Burkholderia pseudomallei, we identified several additional IcsB-like k-FATs in β- and γ-proteobacteria. These new proteins are poorly characterized and are primarily found in bacteria harboring a T3SS. Despite high sequence diversity, structure predictions indicate a highly conserved spatial disposition of catalytic residues. Furthermore, yeast cytotoxicity and in-gel acyltransferase activity depend on the integrity of the catalytic site, which supports the idea that most of the IcsB homologs identified are also acyltransferases. Finally, we used mass spectrometry to identify the yeast proteins acylated by seven of these novel enzymes. These proteins share a variety of substrates with IcsB, including membrane- and vesicle-associated proteins, such as SNAREs and small GTPases of the Rho, Rab, and Ras families. Taken together, this study provides evidence of a family of k-FAT related to IcsB.IMPORTANCEIcsB-like k-FATs are found in the related Pseudomonadota and Thermodesulfobacteriota phyla, suggesting that they are a recent biochemical innovation. Like IcsB, new k-FATs are primarily found in proteobacterial species with a T3SS. This leaves open the possibility that they may play a role in the colonization of plants or animals. However, we characterized one k-FAT from an environmental bacterium that is unlikely to possess a T3SS. Additionally, measurable fatty acid acyltransferase activity was not detected in approximately 25% of the proteins tested. These results imply that the IcsB-like k-FAT family has undergone functional diversification and may have a more complex evolutionary origin than previously thought. In summary, this study describes the properties of the IcsB-like k-FAT family and presents yeast-based assays for characterizing new family members and unrelated proteins with similar fatty acid acyltransferase activity.
Ayesha Mudasser, Mubina Laghari, Aman Ullah Siddiqui
et al.
Background: Salivary biomarkers are non-invasive molecules that indicate neurodegenerative illnesses, especially Alzheimer disease (AD) and Parkinson disease (PD).this study was conducted to determine the diagnostic precision of salivary proteomic and genomic biomarkers in terms of early AD and PD detection.
Methods: A systematic literature search was conducted in PubMed, web of science and Google Scholar, and studies included from 2016 to 2025. Research that examined salivary biomarkers in AD and PD was eligible. The data were analyzed with a random-effects model and odds ratios (OR), standard mean differences (SMD), and 95% confidence interval (CI) was estimated. Also, subgroup and sensitivity analysis were performed. To assess the risk of bias, the Newcastle-Ottawa Scale (NOS) was applied for included observational studies.
Results: A total of 11 eligible studies concerning proteomic biomarkers, including amyloid-β (Aβ42, Aβ40) and alpha-synuclein total (α-synTotal) and alpha-synuclein Oligomer (α-synOligo), and genomic biomarkers like different salivary microRNAs were included. Meta-analysis indicated that Aβ42 (OR=0.70; 95% CI: 0.41 to 1.1) and Aβ40 (OR=1.01; 95% CI: 0.97 to 1.06) had significant discriminatory potential in AD patients; but α-synOligo (SMD = 2.90; 95% CI: -0.59–6.39) and α-synTotal (SMD = 0.44; 95% CI: -3.14 to 4.02) was higher in PD patients as compared with controls. Genomic biomarkers demonstrated inconsistent findings (SMD = -0.18; 95% CI: -1.79–1.42) because of difference in microRNA types. Heterogeneity was high (I2 > 90%), which is caused by alterations in study design and in the methods to measure biomarkers.
Discussion: Salivary biomarkers were found to be an insignificant yet exceptional method of early examination of AD and PD. Nonetheless, the inconsistency of different studies points to develop standardized protocols.
Negar Jafari, Ali Zolfi Gol, Venus Shahabi Rabori
et al.
Kawasaki disease (KD) is an acute vasculitis primarily affecting children, with a potential risk of developing coronary artery aneurysms (CAAs) and cardiovascular complications. The emergence of non-coding RNAs (ncRNAs), including microRNAs (miRNAs), long non-coding RNAs (lncRNAs), and circular RNAs (circRNAs), has provided insights into Kawasaki disease pathogenesis and opened new avenues for diagnosis and therapeutic intervention. Furthermore, polymorphism analysis of ncRNA genes offers significant insights into genetic predisposition to Kawasaki disease, facilitating tailored treatment approaches and risk assessment to improve patient outcomes. Exosomal ncRNAs, which are ncRNAs encapsulated within extracellular vesicles, have garnered significant attention as potential biomarkers for Kawasaki disease and CAA due to their stability and accessibility in biological fluids. This review comprehensively discusses the biogenesis, components, and potential of exosomal and non-exosomal ncRNAs in Kawasaki disease diagnosis and prognosis prediction. It also highlights the roles of non-exosomal ncRNAs, such as miRNAs, lncRNAs, and circRNAs, in Kawasaki disease pathogenesis and their implications as therapeutic targets. Additionally, the review explores the current diagnostic and therapeutic approaches for Kawasaki disease and emphasizes the need for further research to validate these ncRNA-based biomarkers in diverse populations and clinical settings.
Membrane contact sites (MCSs) between the endoplasmic reticulum and the plasma membrane enable the transport of lipids without membrane fusion. Extended Synaptotagmins (ESYTs) act at MCSs, functioning as tethers between two membrane compartments. In plants, ESYTs have been mainly investigated in <i>A. thaliana</i> and shown to maintain the integrity of the plasma membrane, especially during stress responses like cold acclimatization, mechanical trauma, and salt stress. ESYTs are present at the MCSs of plasmodesmata, where they regulate defense responses by modulating cell-to-cell transfer of pathogens. Here, the analysis of ESYTs was expanded to the bryophyte <i>Physcomitrium patens</i>, an extant representative of the earliest land plant lineages. <i>P. patens</i> was found to contain a large number of ESYTs, distributed over all previously established classes and an additional class not present in <i>A. thaliana</i>. Motif discovery identified regions in the Synaptotagmin-like mitochondrial (SMP) domain that may explain phylogenetic relationships as well as protein function. The adaptation mechanisms of <i>P. patens</i> necessary to conquer land and its simple tissue structure make it highly suitable as a model organism to study ESYT functions in tip growth, stress responses, and plasmodesmata-mediated transport, and open new directions of research regarding the function of MCSs in cellular processes and plant evolution.
Abstract Little is known about the regulation and function of phase separation in craniofacial developmental disorders. MSX1 mutations are associated with human cleft palate, the most common craniofacial birth defect. Here, we show that MSX1 phase separation is a vertebrate-conserved mechanism underlying embryonic palatal fusion. Notably, MSX1 phase separation is triggered by its intrinsically disordered protein region (IDR) and regulated by PRMT1-catalyzed methylation, specifically asymmetric dimethylation of arginine in the MSX1 IDR including R150 and R157. Hypomethylated MSX1 due to methylation site mutations and PRMT1 deficiency consistently leads to abnormal MSX1 phase separation to form less dynamic gel-like condensates, resulting in proliferation defects of embryonic palatal mesenchymal cells and cleft palate. Besides, high frequency mutations in the MSX1 IDR, especially R157S, have been identified in humans with cleft palate. Overall, we reveal the function and regulatory pathway of MSX1 phase separation as a conserved mechanism underlying cleft palate, providing a proof-of-concept example of a phenotype-associated phase separation mechanism associated with craniofacial developmental disorders.
Saleema Rehman, Humaira Bilqis, Farah Deeba
et al.
Background: Morbidly adherent placenta (MAP) is one of the most dreaded antenatal complications leading to massive hemorrhage, immense blood transfusion, hysterectomy, intensive care unit admission, mechanical ventilation, multi-organ failure and maternal mortality. The study aimed to determine the maternal outcome in terms of complications, interventions, and mortality in patients with MAP in a tertiary care hospital.
Methods: During the study period of one year (2020-2021) a total of 68 patients with MAP were studied. Demographic profiles including age, parity, gestational age and history of previous cesarean sections were recorded. Other complications and interventions were also noted. Data was analyzed by using SPSS version 23 and a t-test was applied for comparison between the two groups. p-value <0.05 was considered statistically significant.
Results: Total number of deliveries during the study period was 20971. Among these 7183 women had a cesarian section (CS). The total number of CS done due to placenta previa was 319, out of which a total of 68(21.3%) patients were diagnosed with MAP. It was further observed that 0.6% of patients had no history of a previous uterine scar, 39.0% of patients with previous 1 scar and 80% with 4 scars (p>0.05). Peripartum hysterectomy was performed in 48.5% of patients. The complications noted were bladder injury 17.6%, ICU admission 45.5% and maternal mortality 4.4%.
Conclusion: Morbidly adherent placenta (MAP) was found directly related to cesarean section (p>0.05). Appropriate measures should be taken to reduce the primary scars to reduce the incidence of repeat scars and ultimately MAP related severe maternal outcomes.
Keywords: Morbidly Adherent Placenta (MAP); Caesarean Section; Placenta Previa.
Azra Kulovic-Sissawo, Carolina Tocantins, Mariana S. Diniz
et al.
Endothelial dysfunction is associated with several lifestyle-related diseases, including cardiovascular and neurodegenerative diseases, and it contributes significantly to the global health burden. Recent research indicates a link between cardiovascular risk factors (CVRFs), excessive production of reactive oxygen species (ROS), mitochondrial impairment, and endothelial dysfunction. Circulating endothelial progenitor cells (EPCs) are recruited into the vessel wall to maintain appropriate endothelial function, repair, and angiogenesis. After attachment, EPCs differentiate into mature endothelial cells (ECs). Like ECs, EPCs are also susceptible to CVRFs, including metabolic dysfunction and chronic inflammation. Therefore, mitochondrial dysfunction of EPCs may have long-term effects on the function of the mature ECs into which EPCs differentiate, particularly in the presence of endothelial damage. However, a link between CVRFs and impaired mitochondrial function in EPCs has hardly been investigated. In this review, we aim to consolidate existing knowledge on the development of mitochondrial and endothelial dysfunction in the vascular endothelium, place it in the context of recent studies investigating the consequences of CVRFs on EPCs, and discuss the role of mitochondrial dysfunction. Thus, we aim to gain a comprehensive understanding of mechanisms involved in EPC deterioration in relation to CVRFs and address potential therapeutic interventions targeting mitochondrial health to promote endothelial function.
In vitro regeneration was studied to protect the rare Chinese medicinal orchid Liparis nervosa (Thunb.) Lindl. The mixtures of protocorm and seeding and the stem tip were used as explants. The results revealed that the best essential medium for L. nervosa growth was 1/3 MS medium with 25 g · L–1 sucrose, 50 g · L–1 banana puree, 40 g · L–1 mashed potato, and 1.0 g · L–1 AC (MS1); MS1 medium with 0.5 mg · L–1 BA, 0.05 mg · L–1 2,4-D, and 1.5 mg · L–1 NAA was optimal for proliferation. When stem tips were cultured in a proliferation medium, four types of proliferation occurred: basal stem cluster bud (occurring at the basal node), tiller bud (occurring at the root), protocorm-like body (occurring at the plant’s base incision), and high-position bud (occurring on plant stem nodes other than the basal nodes). Four methods produced 10.12 proliferation coefficients. In the MS1 medium with 0.5 mg · L−1 NAA, the plantlets rooted 100%, and the rooted plantlets survived 100% after domestication and transplantation.
Alfonso Varela-López, José M. Romero-Márquez, María D. Navarro-Hortal
et al.
The rise of life expectancy in current societies is not accompanied, to date, by a similar increase in healthspan, which represents a great socio-economic problem. It has been suggested that aging can be manipulated and then, the onset of all age-associated chronic disorders can be delayed because these pathologies share age as primary underlying risk factor. One of the most extended ideas is that aging is consequence of the accumulation of molecular damage. According to the oxidative damage theory, antioxidants should slow down aging, extending lifespan and healthspan. The present review analyzes studies evaluating the effect of dietary antioxidants on lifespan of different aging models and discusses the evidence on favor of their antioxidant activity as anti-aging mechanisms. Moreover, possible causes for differences between the reported results are evaluated.
Armando G. Salinas, Jeong Oen Lee, Shana M. Augustin
et al.
Abstract The development of genetically encoded dopamine sensors such as dLight has provided a new approach to measuring slow and fast dopamine dynamics both in brain slices and in vivo, possibly enabling dopamine measurements in areas like the dorsolateral striatum (DLS) where previously such recordings with fast-scan cyclic voltammetry (FSCV) were difficult. To test this, we first evaluated dLight photometry in mouse brain slices with simultaneous FSCV and found that both techniques yielded comparable results, but notable differences in responses to dopamine transporter inhibitors, including cocaine. We then used in vivo fiber photometry with dLight in mice to examine responses to cocaine in DLS. We also compared dopamine responses during Pavlovian conditioning across the striatum. We show that dopamine increases were readily detectable in DLS and describe transient dopamine kinetics, as well as slowly developing signals during conditioning. Overall, our findings indicate that dLight photometry is well suited to measuring dopamine dynamics in DLS.
Stanisław Malicki, Barbara Pucelik, Edyta Żyła
et al.
Immune checkpoint targeting immunotherapy has revolutionized the treatment of certain cancers in the recent years. Determination of the status of immune checkpoint expression in particular cancers may assist decision making. Here, we describe the development of a single-stranded aptamer-based molecular probe specifically recognizing human PD-L1. Target engaging aptamers are selected by iterative enrichment from a random ssDNA pool and the binding is characterized biochemically. Specificity and dose dependence is demonstrated in vitro in the cell culture using human kidney tumor cells (786-0), human melanoma cells (WM115 and WM266.4) and human glioblastoma LN18 cancer cells. The utility of the probe in vivo is demonstrated using two mouse tumor models, where we show that the probe exhibits excellent potential in imaging. We postulate that further development of the probe may allow universal imaging of different types of tumors depending on their PD-L1 status, which may find utility in cancer diagnosis.
Mohammed I. Sorour, Andrew H. Marcus, Spiridoula Matsika
Accurate modeling of optical spectra requires careful treatment of the molecular structures and vibronic, environmental, and thermal contributions. The accuracy of the computational methods used to simulate absorption spectra is limited by their ability to account for all the factors that affect the spectral shapes and energetics. The ensemble-based approaches are widely used to model the absorption spectra of molecules in the condensed-phase, and their performance is system dependent. The Franck–Condon approach is suitable for simulating high resolution spectra of rigid systems, and its accuracy is limited mainly by the harmonic approximation. In this work, the absorption spectrum of the widely used cyanine Cy3 is simulated using the ensemble approach via classical and quantum sampling, as well as, the Franck–Condon approach. The factors limiting the ensemble approaches, including the sampling and force field effects, are tested, while the vertical and adiabatic harmonic approximations of the Franck–Condon approach are also systematically examined. Our results show that all the vertical methods, including the ensemble approach, are not suitable to model the absorption spectrum of Cy3, and recommend the adiabatic methods as suitable approaches for the modeling of spectra with strong vibronic contributions. We find that the thermal effects, the low frequency modes, and the simultaneous vibrational excitations have prominent contributions to the Cy3 spectrum. The inclusion of the solvent stabilizes the energetics significantly, while its negligible effect on the spectral shapes aligns well with the experimental observations.
Eleonora Olivotto, Manuela Minguzzi, Stefania D’Adamo
et al.
Abstract IKKα and IKKβ are essential kinases for activating NF-κB transcription factors that regulate cellular differentiation and inflammation. By virtue of their small size, chemokines support the crosstalk between cartilage and other joint compartments and contribute to immune cell chemotaxis in osteoarthritis (OA). Here we employed shRNA retroviruses to stably and efficiently ablate the expression of each IKK in primary OA chondrocytes to determine their individual contributions for monocyte chemotaxis in response to chondrocyte conditioned media. Both IKKα and IKKβ KDs blunted both the monocyte chemotactic potential and the protein levels of CCL2/MCP-1, the chemokine with the highest concentration and the strongest association with monocyte chemotaxis. These findings were mirrored by gene expression analysis indicating that the lowest levels of CCL2/MCP-1 and other monocyte-active chemokines were in IKKαKD cells under both basal and IL-1β stimulated conditions. We find that in their response to IL-1β stimulation IKKαKD primary OA chondrocytes have reduced levels of phosphorylated NFkappaB p65pSer536 and H3pSer10. Confocal microscopy analysis revealed co-localized p65 and H3pSer10 nuclear signals in agreement with our findings that IKKαKD effectively blunts their basal level and IL-1β dependent increases. Our results suggest that IKKα could be a novel OA disease target.