Mesenchymal Stromal Cell Senescence Induced by Dnmt3a -Mutant Hematopoietic Cells is a Targetable Mechanism Driving Clonal Hematopoiesis and Initiation of Hematologic Malignancy.

Clonal hematopoiesis (CH) can predispose to blood cancers due to enhanced fitness of mutant hematopoietic stem and progenitor cells (HSPCs), but the mechanisms driving this progression are not understood. We hypothesized that malignant progression is related to microenvironment-remodelling properties of CH-mutant HSPCs. Single-cell transcriptomic profiling of the bone marrow microenvironment in Dnmt3a R878H/+ mice revealed signatures of cellular senescence in mesenchymal stromal cells (MSCs). Dnmt3a R878H/+ HSPCs caused MSCs to upregulate the senescence markers SA-ß-gal, BCL-2, BCL-xL, Cdkn1a (p21) and Cdkn2a (p16), ex vivo and in vivo . This effect was cell contact-independent and can be replicated by IL-6 or TNFα, which are produced by Dnmt3a R878H/+ HSPCs. Depletion of senescent MSCs in vivo reduced the fitness of Dnmt3a R878H/+ hematopoietic cells and the progression of CH to myeloid neoplasms using a sequentially inducible Dnmt3a ; Npm1 -mutant model. Thus, Dnmt3a -mutant HSPCs reprogram their microenvironment via senescence induction, creating a self-reinforcing niche favoring fitness and malignant progression. Statement of Significance: Mesenchymal stromal cell senescence induced by Dnmt3a -mutant hematopoietic stem and progenitor cells drives clonal hematopoiesis and initiation of hematologic malignancy.

Cassava starch esterification with formic acid for fabrication of electrospun fibers.

Formic acid is utilized to induce esterification and chemical gelatinization in starch, particularly in the fabrication of electrospun fibers for nanomaterial production. This study investigated the impact of different concentrations (15, 20, 25, and 30 %) of cassava starch and formic acid as a solvent on the characteristics of the resultant polymeric solutions and electrospun fibers. Morphology, size distribution, thermogravimetric properties, diffraction patterns, and relative crystallinity were evaluated for the electrospun fibers. The amylose content of starch varied from 16.5 to 23.7 %, decreasing with esterification, achieving a degree of substitution of approximately 0.93. The solution-rheology exhibited elastic behavior, with viscosity increasing as starch concentration increased, hindering the fabrication of fibers at 25 and 30 % starch. Successful electrospun fibers were formed using 15 % and 20 % starch, displaying homogeneous morphologies with mean diameters of 165 nm and 301 nm, respectively. Esterification influenced thermogravimetric properties, leading to fibers with reduced degradation temperatures and mass loss compared to native starches. The electrospun fibers presented an amorphous structure, indicating a drastic reduction in relative crystallinity from 35.2 % in native starch to 8.5 % for esterified starches. This study highlights the intricate relationship between starch concentration, esterification, and solution viscosity, affecting the electrospinnability and properties of starch-polymeric solutions.

Evaluation of microbiome in primary and permanent dentition in grade C periodontitis in young individuals.

BACKGROUND: The aim of the present study was to evaluate the subgingival microbiome in patients with grade C molar-incisor pattern periodontitis (C-MIP) affecting the primary or permanent dentitions. METHODS: DNA was isolated from subgingival biofilm samples from diseased and healthy sites from 45 C-MIP patients and subjected to phylogenetic microarray analysis. C-MIP sites were compared between children affected in the primary to those affected in the permanent dentitions. Within-subject differences between C-MIP-affected sites and dentition-matched healthy sites were also evaluated. RESULTS: C-MIP sites of subjects affected in the primary dentition showed partially overlapping but distinct microbial communities from C-MIP permanent dentition sites (p < 0.05). Differences were due to increased levels in primary C-MIP sites of certain species of the genera Capnocytophaga and Leptotrichia, while C-MIP permanent dentition sites showed higher prevalence of Filifactor alocis. Aggregatibacter actinomycetemcomitans (Aa) was among species seen in high prevalence and levels in both primary and permanent C-MIP sites. Moreover, both permanent and primary C-MIP sites showed distinct microbial communities when compared to dentition-matched healthy sites in the same subject (p < 0.01). CONCLUSIONS: Primary and permanent teeth with C-MIP showed a dysbiotic microbiome, with children affected in the primary dentition showing a distinct profile from those affected in the permanent dentition. However, Aa was enriched in both primary and permanent diseased sites, confirming that this microorganism is implicated in C-MIP in both dentitions.

Defining Porphyromonas gingivalis strains associated with periodontal disease.

Porphyromonas gingivalis, a Gram-negative anaerobic bacterium commonly found in human subgingival plaque, is a major etiologic agent for periodontitis and has been associated with multiple systemic pathologies. Many P. gingivalis strains have been identified and different strains possess different virulence factors. Current oral microbiome approaches (16S or shotgun) have been unable to differentiate P. gingivalis strains. This study presents a new approach that aims to improve the accuracy of strain identification, using a detection method based on sequencing of the intergenic spacer region (ISR) which is variable between P. gingivalis strains. Our approach uses two-step PCR to amplify only the P. gingivalis ISR region. Samples are then sequenced with an Illumina sequencer and mapped to specific strains. Our approach was validated by examining subgingival plaque from 153 participants with and without periodontal disease. We identified the avirulent strain ATCC33277/381 as the most abundant strain across all sample types. The W83/W50 strain was significantly enriched in periodontitis, with 13% of participants harboring that strain. Overall, this approach can have significant implications not only for the diagnosis and treatment of periodontal disease but also for other diseases where P. gingivalis or its toxins have been implicated, such as Alzheimer's disease.

Epithelial-derived interleukin-23 promotes oral mucosal immunopathology.

At mucosal surfaces, epithelial cells provide a structural barrier and an immune defense system. However, dysregulated epithelial responses can contribute to disease states. Here, we demonstrated that epithelial cell-intrinsic production of interleukin-23 (IL-23) triggers an inflammatory loop in the prevalent oral disease periodontitis. Epithelial IL-23 expression localized to areas proximal to the disease-associated microbiome and was evident in experimental models and patients with common and genetic forms of disease. Mechanistically, flagellated microbial species of the periodontitis microbiome triggered epithelial IL-23 induction in a TLR5 receptor-dependent manner. Therefore, unlike other Th17-driven diseases, non-hematopoietic-cell-derived IL-23 served as an initiator of pathogenic inflammation in periodontitis. Beyond periodontitis, analysis of publicly available datasets revealed the expression of epithelial IL-23 in settings of infection, malignancy, and autoimmunity, suggesting a broader role for epithelial-intrinsic IL-23 in human disease. Collectively, this work highlights an important role for the barrier epithelium in the induction of IL-23-mediated inflammation.

Natural fermentation of potato (Solanum tuberosum L.) starch: Effect of cultivar, amylose content, and drying method on expansion, chemical and morphological properties.

Natural fermentation with sun-drying is a modification that promotes the expansion capacity of starch, and its effects on potato starch have not been reported so far. The aim of this study was to evaluate the effects of the amylose content of potato (Solanum tuberosum L.) starches and natural fermentation followed by oven or sun drying on its properties. Cassava starch was also used a control. Native and fermented starches were evaluated based on their chemical composition, amylose, carboxyl and carbonyl content as well as their thermal, pasty, and morphological properties. The fermentation water was evaluated by pH and titratable acidity to control the process. Puffed balls were prepared to evaluate expandability, mass loss, porosity and texture. The fermentation intensity was greater for potato and cassava starch with low-amylose content than for potato starch with higher amylose content. In addition, the acidity of the fermentation water increased faster with cassava starch than with potato starches. The fermented potato starches with the highest amylose content had low acidity and low expansion capacity compared to the fermented potato and cassava starches with low-amylose content. Fermentation and sun-drying of low-amylose potato and cassava starches increased the expansion and reduced the hardness of the puffed balls.

Seventeen-year study reveals fluctuations in key ecological indicators on two reef crests in Cuba.

Reef crests in the Caribbean have lost approximately 80% of the foundational habitat-forming coral Acropora palmata (Lamarck, 1816), with declines registered as early as the 1950s mainly from anthropogenic causes. We studied two reef crests in the northwestern region of Cuba over 17 years (2005 to 2021) to evaluate temporal changes in coral cover, dominated by A. palmata, and their potential drivers. The density of A. palmata generally showed a negative trend at both reefs, with the lowest density recorded in 2021 at 0.2 ± 0.05 col. m-2 at Playa Baracoa and 1.0 ± 0.1 col. m-2 at Rincon de Guanabo. The mean size of the colonies in the two reefs also decreased over time. In Playa Baracoa, the mean diameter of A. palmata colonies decreased from 2012 at 67 ± 5.9 cm to 2013 at 34 ± 2.2 cm, whereas in Rincon de Guanabo, a change in diameter was evident from 2015 at 44.3 ± 2.3 to 2021 at 21.6 ± 0.9 cm. Adult colonies (10 cm-50 cm diameter) predominated in most years on both reefs. The populations of A. palmata on both reefs were healthy, with an average of 70% colonies in good condition during the study period. However, A. palmata cover decreased by almost half by 2021, to 8.6% in Playa Baracoa and 16.8% in Rincon de Guanabo. By contrast, macroalgal cover increased two-fold to 87.1% in Playa Baracoa and four-fold to 77.2% in Rincon de Guanabo. The density of the sea urchin Diadema antillarum was higher in Playa Baracoa than in Rincon de Guanabo. The highest densities were 2.8 ± 0.2 ind. m-2 in Playa Baracoa in 2005 and 0.1 ± 0.03 ind. m-2 in Rincon de Guanabo in 2008. Although our results show an overall decline of A. palmata (density and percent cover) and an increase in macroalgae, these two reef crests are in better condition than most reefs in the Caribbean in terms of the density and health of A. palmata populations, and the density of D. antillarum at Playa Baracoa. Our results are important in establishing a management plan to ensure the condition of these reef crests does not degrade further.

Enhancing restoration success of rare plants in an arid-tropical climate through water-saving technologies: a case study of Scalesia affinis ssp. brachyloba in the Galapagos Islands.

Arid tropical archipelagos, such as the Galapagos Islands, host a high concentration of endemic plant species, many of which require restoration intervention to recover from past environmental degradation. Water-saving technologies (WSTs) have potential for hastening restoration by providing plants with additional water during the early stages of growth. However, it remains unclear whether such technologies provide an advantage for plant species of arid-tropical regions. This study examined the effect of the water-saving technology Groasis Waterboxx® (Groasis) on the rare endemic plant species Scalesia affinis ssp. brachyloba during early stages of restoration. Survival was monitored for 374 individuals planted across six sites on Santa Cruz Island, Galapagos (326 with technology and 48 as controls). Kaplan-Meier survival analysis showed that the use of Groasis reduced mortality during the first two years of the seedling survival. A mixed-effect logistic regression that modelled plant survival as a function of total precipitation, maximum temperature, and WST treatment (Groasis and no-technology control) found that despite low overall survival rates, plants grown with Groasis exhibited a three-fold higher predicted survival by the end of the 3.7 year duration of the study. Finally, through a resampling method, we demonstrate that the effect of the WST treatment is not dependent on the unbalanced design typical of a restoration project framework. We conclude that water-saving technologies such as the Groasis Waterboxx® can enhance survival of rare plant species such as S. affinis ssp. brachyloba in restoration programs in arid-tropical regions.

Does depth divide? Variable genetic connectivity patterns among shallow and mesophotic Montastraea cavernosa coral populations across the Gulf of Mexico and western Caribbean.

Despite general declines in coral reef ecosystems in the tropical western Atlantic, some reefs, including mesophotic reefs (30-150 m), are hypothesized to function as coral refugia due to their relative isolation from anthropogenic stressors. Understanding the connectivity dynamics among these putative refugia and more degraded reefs is critical to develop effective management strategies that promote coral metapopulation persistence and recovery. This study presents a geographically broad assessment of shallow (<30 m) and mesophotic (>30 m) connectivity dynamics of the depth-generalist coral species Montastraea cavernosa. Over 750 coral genets were collected across the Northwest and Southern Gulf of Mexico, Florida, Cuba, and Belize, and ~5000 SNP loci were generated to quantify high-resolution genetic structure and connectivity among these populations. Generally, shallow and mesophotic populations demonstrated higher connectivity to distant populations within the same depth zone than to adjacent populations across depth zones. However, exceptions to this pattern include the Northwest Gulf of Mexico and the Florida Keys which exhibited relatively high vertical genetic connectivity. Furthermore, estimates of recent gene flow emphasize that mesophotic M. cavernosa populations are not significant sources for their local shallow counterparts, except for the Northwest Gulf of Mexico populations. Location-based differences in vertical connectivity are likely a result of diverse oceanographic and environmental conditions that may drive variation in gene flow and depth-dependent selection. These results highlight the need to evaluate connectivity dynamics and refugia potential of mesophotic coral species on a population-by-population basis and to identify stepping-stone populations that warrant incorporation in future international management approaches.

Influence of gelatin type on physicochemical properties of electrospun nanofibers.

This study explores the fabrication of nanofibers using different types of gelatins, including bovine, porcine, and fish gelatins. The gelatins exhibited distinct molecular weights and apparent viscosity values, leading to different entanglement behavior and nanofiber production. The electrospinning technique produced nanofibers with diameters from 47 to 274 nm. The electrospinning process induced conformational changes, reducing the overall crystallinity of the gelatin samples. However, porcine gelatin nanofibers exhibited enhanced molecular ordering. These findings highlight the potential of different gelatin types to produce nanofibers with distinct physicochemical properties. Overall, this study sheds light on the relationship between gelatin properties, electrospinning process conditions, and the resulting nanofiber characteristics, providing insights for tailored applications in various fields.

Morphometric responses of two zooxanthellate octocorals along a water quality gradient in the Cuban northwestern coast.

Octocoral abundance is increasing on Caribbean reefs, and one of the possible causes is their vertical morphological plasticity that allows them to grow above the substrate to reduce the effect of processes that occur in it (e.g., scour by sediments) as well as adapt to environmental gradients. The aim of this study was to determine the morphometric response of two octocorals species (Eunicea flexuosa and Plexaura kükenthali) with different life strategies in a water quality gradient. The research was carried out between 2008 and 2016 on eight forereefs of northwest Cuba. Different morphometric indicators were measured in the colonies of both species found within a belt transect (100 x 2 m) randomly located at each site. The lowest means in height, diameter, number of terminal branches/colony, cover index, and least arborescent colonies of E. flexuosa were detected at the sites with the greatest anthropogenic pollution. The water quality gradient did not explain the variability of the five morphometric indicators of P. kükenthali. However, hydrodynamic stress was the factor that most negatively affected the morphometry of this species. The chronic effect of poor water quality over time resulted in more small sized colonies of E. flexuosa at the polluted site, probably due to higher mortality. The size distribution of P. kükenthali also showed the same trend but at the sites with greater hydrodynamic stress. These results show that the morphometric response of octocorals along a water quality gradient is species-specific. This study suggests that poor water quality decreases the size and thus availability of habitat provided by octocorals sensitive to that factor (e.g., E. flexuosa) while other tolerant species (e.g., P. kükenthali) could provide the habitat of several organisms in a scenario of increasing anthropogenic pollution.

Zeolite Nanoparticles Loaded with 2-Methoxystradiol as a Novel Drug Delivery System for the Prostate Cancer Therapy.

The estrogen metabolite 2-methoxyestradiol (2ME) is a promissory anticancer drug mainly because of its pro-apoptotic properties in cancer cells. However, the therapeutic use of 2ME has been hampered due to its low solubility and bioavailability. Thus, it is necessary to find new ways of administration for 2ME. Zeolites are inorganic aluminosilicates with a porous structure and are considered good adsorbents and sieves in the pharmaceutical field. Here, mordenite-type zeolite nanoparticles were loaded with 2ME to assess its efficiency as a delivery system for prostate cancer treatment. The 2ME-loaded zeolite nanoparticles showed an irregular morphology with a mean hydrodynamic diameter of 250.9 ± 11.4 nm, polydispersity index of 0.36 ± 0.04, and a net negative surface charge of -34 ± 1.73 meV. Spectroscopy with UV-vis and Attenuated Total Reflectance Infrared Fourier-Transform was used to elucidate the interaction between the 2ME molecules and the zeolite framework showing the formation of a 2ME-zeolite conjugate in the nanocomposite. The studies of adsorption and liberation determined that zeolite nanoparticles incorporated 40% of 2ME while the liberation of 2ME reached 90% at pH 7.4 after 7 days. The 2ME-loaded zeolite nanoparticles also decreased the viability and increased the mRNA of the 2ME-target gene F-spondin, encoded by SPON1, in the human prostate cancer cell line LNCaP. Finally, the 2ME-loaded nanoparticles also decreased the viability of primary cultures from mouse prostate cancer. These results show the development of 2ME-loaded zeolite nanoparticles with physicochemical and biological properties compatible with anticancer activity on the human prostate and highlight that zeolite nanoparticles can be a good carrier system for 2ME.

Neutrophil extracellular traps and extracellular histones potentiate IL-17 inflammation in periodontitis.

Neutrophil infiltration is a hallmark of periodontitis, a prevalent oral inflammatory condition in which Th17-driven mucosal inflammation leads to destruction of tooth-supporting bone. Herein, we document that neutrophil extracellular traps (NETs) are early triggers of pathogenic inflammation in periodontitis. In an established animal model, we demonstrate that neutrophils infiltrate the gingival oral mucosa at early time points after disease induction and expel NETs to trigger mucosal inflammation and bone destruction in vivo. Investigating mechanisms by which NETs drive inflammatory bone loss, we find that extracellular histones, a major component of NETs, trigger upregulation of IL-17/Th17 responses, and bone destruction. Importantly, human findings corroborate our experimental work. We document significantly increased levels of NET complexes and extracellular histones bearing classic NET-associated posttranslational modifications, in blood and local lesions of severe periodontitis patients, in the absence of confounding disease. Our findings suggest a feed-forward loop in which NETs trigger IL-17 immunity to promote immunopathology in a prevalent human inflammatory disease.

A randomized, double-blind, placebo-controlled clinical trial of the use of hydroxycitric acid adjuvant to shock wave lithotripsy therapy in patients with calcium stones. Stone fragmentation results.

Our objective was to improve the results of extracorporeal shock waves lithotripsy using hydroxycitric acid (HCA) like adjuvant therapy. Double blind randomized clinical trial using hydroxycitric acid versus placebo (ID NCT05525130). Multicenter study of adjuvant exposure to a food supplement with hydroxycitric acid (HCA), vs. placebo in patients with calcium oxalate and calcium phosphate lithiasis with indication for extracorporeal shock wave lithotripsy (ESWL). 81 patients were included in the study to compare the effect of HCA versus placebo. Stone fragmentation, the main efficacy variable. Other variables analyzed were stone size, Hounsfield Unit Stone and tolerability. Statistical study with SPSS, statistical significance p ≤ 0.05. Eighty-one patients were included, 40 in the intervention group with HCA and 41 in the control group with placebo. The average stone area was 174,36 mm2 (SD: 32,83 mm2) and the average hardness was 1128,11 (SD: 257,65), with no statistically significant differences between groups. Significant statistical differences were obtained in the analysis of the population by intention to treat and by protocol of the main variable, no fragmentation vs. fragmentation where 100% of the patients, who were given ESWL and took HCA, presented fragmentation while 17% of the patients with placebo did not reach fragmentation (p = 0.03). The adjuvant use of HCA in patients for whom ESWL has been indicated, facilitates stone fragmentation in all cases, which is not achieved in up to 17% of the patients who did not use HCA. We recommend the use of HCA in patients prior to shock wave treatment to improve their fragmentation in calcium stones.

Understanding the role of emotion and expertise in psychotherapy: An application of dynamical systems mathematical modeling to an entire course of therapy.

Introduction: The therapeutic relationship continues to be one of the most important factors in therapeutic outcomes. Given the place of emotion in the definition of the therapeutic relationship, as well as the demonstrated positive impact that emotional expression has on therapeutic process and outcome, it stands to reason that studying the emotional exchange between the therapist and client further would be warranted. Methods: This study used a validated observational coding system--the Specific Affect Coding System (SPAFF) and a theoretical mathematical model to analyze behaviors which make up the therapeutic relationship. Specifically, the researchers used to codify relationship-building behaviors between an expert therapist and his client over the course of six sessions. Dynamical systems mathematical modeling was also employed to create "phase space portraits" depicting the relational dynamics between the master therapist and his client over six sessions. Results: Statistical analysis was used to compare SPAFF codes and model parameters between the expert therapist and his client. The expert therapist showed stability in affect codes over six sessions while the client's affect codes appeared to be more flexible over time, though model parameters remained stable across the six sessions. Finally, phase space portraits depicted the evolution of the affective dynamics between the master therapist and his client as the relationship matured. Discussion: The clinician's ability to stay emotionally positive and relatively stable across the six sessions (relative to the client) was noteworthy. It formed the basis for a stable base from which she could explore alternative methods to relate to others that she had allowed to dictate her actions, which is in keeping with previous research on the role of therapist facilitation of the therapeutic relationship, emotional expression within the therapeutic relationship, and influence of these on client outcomes. These results provide a valuable foundation for future research on emotional expression as a key component of the therapeutic relationship in psychotherapy.

Evolutionary genomics of oceanic island radiations.

A recurring feature of oceanic archipelagos is the presence of adaptive radiations that generate endemic, species-rich clades that can offer outstanding insight into the links between ecology and evolution. Recent developments in evolutionary genomics have contributed towards solving long-standing questions at this interface. Using a comprehensive literature search, we identify studies spanning 19 oceanic archipelagos and 110 putative adaptive radiations, but find that most of these radiations have not yet been investigated from an evolutionary genomics perspective. Our review reveals different gaps in knowledge related to the lack of implementation of genomic approaches, as well as undersampled taxonomic and geographic areas. Filling those gaps with the required data will help to deepen our understanding of adaptation, speciation, and other evolutionary processes.

Periodontal disease is associated with increased gut colonization of pathogenic Haemophilus parainfluenzae in patients with Crohn's disease.

Intestinal colonization of the oral bacterium Haemophilus parainfluenzae has been associated with Crohn's disease (CD) severity and progression. This study examines the role of periodontal disease (PD) as a modifier for colonization of H. parainfluenzae in patients with CD and explores the mechanisms behind H. parainfluenzae-mediated intestinal inflammation. Fifty subjects with and without CD were evaluated for the presence of PD, and their oral and fecal microbiomes were characterized. PD is associated with increased levels of H. parainfluenzae strains in subjects with CD. Oral inoculation of H. parainfluenzae elicits strain-dependent intestinal inflammation in murine models of inflammatory bowel disease, which is associated with increased intestinal interferon-γ (IFN-γ)+ CD4+ T cells and disruption of the host hypusination pathway. In summary, this study establishes a strain-specific pathogenic role of H. parainfluenzae in intestinal inflammation and highlights the potential effect of PD on intestinal colonization by pathogenic H. parainfluenzae strains in patients with CD.

Hematopoietic stem cell aging and leukemia transformation.

With aging, hematopoietic stem cells (HSCs) have an impaired ability to regenerate, differentiate, and produce an entire repertoire of mature blood and immune cells. Owing to dysfunctional hematopoiesis, the incidence of hematologic malignancies increases among elderly individuals. Here, we provide an update on HSC-intrinsic and -extrinsic factors and processes that were recently discovered to contribute to the functional decline of HSCs during aging. In addition, we discuss the targets and timing of intervention approaches to maintain HSC function during aging and the extent to which these same targets may prevent or delay transformation to hematologic malignancies.