Differences In Sensitivity Research Articles

Identification of fossil juniper seeds from Rancho La Brea (California, USA): drought and extirpation in the Late Pleistocene.

Juniperus spp. are keystone shrubs in western North America and important climatic indicators in paleo-records. However, a lack of taxonomic resolution among fossil species limits our ability to track past environmental changes. Plant macrofossils at Rancho La Brea (RLB) allow for reconstructions of juniper occurrence to species across 60 000 yr. We use microscopy, image analysis, species distribution modeling (SDM), and radiocarbon dating to identify an unknown Juniperus species at RLB and put it into chronological context with fossil Juniperus californica at the site to infer past environmental conditions. We identify the unknown taxon as Juniperus scopulorum Sargent, 1897. The Pleistocene occurrence of this species in California expands its known distribution and documents its extirpation. Temporal ranges of the two fossil junipers alternate, revealing a pattern of differential climatic sensitivity throughout the end of the Pleistocene. Occurrence patterns suggest sensitivity to temperature, moisture availability, and the presence of two mega-droughts at c. 48-44.5 ka and c. 29.3-25.2 ka. Extirpation of both taxa by c. 13 ka is likely driven by climate, megafaunal extinction, and increasing fire. The extirpation of fossil junipers during these past climatic events demonstrates vulnerability of juniper species in the face of global change.

Living on the edge – physiological tolerance to frost and drought explains range limits of 35 European tree species

Species distribution models are key to evaluate how climate change threatens European forests and tree species distributions. However, current models struggle to integrate ecophysiological processes. Mechanistic models are complex and have high parameter requirements. Some correlative species distribution models have tried to include traits but so far have struggled to directly connect to ecophysiological processes. Here, we propose a new strategy in which species distributions are based on safety margins which represent species' proximity to their physiological thresholds. We derived frost and drought safety margins for 38 European tree species as the difference between physiological tolerance traits and local maximum stress. We used LT50 and Ψ50 as tolerance traits for frost and drought, respectively, and local minimum temperature and minimum soil water potential as maximum stress. We integrated these safety margins into a species distribution model, which tests if the probability of species presence declines rapidly when the safety margin reaches zero, when physiological stress exceeds the species' tolerance traits. Our results showed thaet 35 of the 38 studied species had their distribution explained by one or both safety margins. We demonstrated that safety‐margins‐based model can be efficiently transferred to species for which occurrence data are not available. The probability of presence dropped dramatically when the frost safety margin reached zero, whereas it was less sensitive to the drought safety margin. This differential sensitivity may be due to the more complex regulation of drought stress, especially as water is a shared resource, whereas frost is not. Our analysis provides a new approach to link species distributions to their physiological limits and shows that, in Europe, frost and drought safety margins are important determinants of species distributions.

Cascadable 2-D Vector Accelerometer Based on Differential Sensitivity Enhancement in Internal Seven-Core Fiber

Cascadable 2-D Vector Accelerometer Based on Differential Sensitivity Enhancement in Internal Seven-Core Fiber

Are boys more vulnerable to stunting? Examining risk factors, differential sensitivity, and measurement issues in Zambian infants and young children

BackgroundStunting remains a considerable public health problem globally, and sex differences in prevalence have been documented. While many risk factors for stunting have been identified, few studies examine how these factors may contribute to sex differences. We test whether: (1) boys and girls are differentially exposed to stunting risk factors, (2) boys and girls respond differently to similar exposures, and (3) these associations are sensitive to the growth measures used.MethodsData comes from 7486 Zambian infants, aged 0–23 months, participating in the SUN (Scaling Up Nutrition) 2.0 Program baseline survey. Surveys and dietary recalls were collected from primary caregivers, and anthropometry was collected for caregivers and children. Stunting was defined as height-for-age z-score (HAZ) <-2. Descriptive statistics and adjusted multilevel logistic regression models controlling for age and province were used to identify sex differences in exposures and risk factors for stunting, respectively. Interaction terms between exposure and infant sex were added to test for sex differences in response. Sensitivity testing with alternate measures of infant size, including height-for-age difference (HAD), was conducted.ResultsBoys were more likely to be stunted than girls (34.5% vs. 25.7%, respectively). Numerous maternal, care giving, diet, and household characteristics were associated with the risk of stunting, but few sex differences were seen in exposure to these factors. Only one risk factor—maternal marital status—showed evidence of moderation by sex at the p < 0.05 level, while a limited number of risk factors did so at the p < 0.10 level. At all ages, boys were longer than girls, and there were no sex-specific differences in the patterns of HAZ or HAD with age. Results were robust to alternate specifications.ConclusionsOur results show that Zambian boys have lower mean HAZ scores than girls and a greater prevalence of stunting throughout the first two years We do not find strong evidence that infant feeding practices, environmental exposures, or care giving differ consistently between boys and girls or that boys and girls respond differently to these exposures. Our results instead indicate that further investigation of prenatal factors and/or measurement issues is needed.

Identification of ETV5 as a prognostic marker related to epigenetic modification in pan-cancer and facilitates tumor progression in hepatocellular carcinoma

ETS variant transcription factor 5 (ETV5), a master transcription factor during development, exerts vital function on the occurrence and progression of various cancers. In order to systematically analyze and explore ETV5 potential specific regulatory mechanisms in pan-cancer, RNA sequencing data and clinicopathological features of patients with various tumors were obtained through the Cancer Genome Atlas (TCGA) and Genotype-Tissue Expression (GTEx) databases, and an integrated data mining analysis was carried out, including the association of ETV5 expression with patient prognosis, drug sensitivity and epigenetic modification. The results revealed that abnormally highly expressed ETV5 resulted in unfavorable prognosis and differential drug sensitivity in multiple malignancies, and its expression was associated with epigenetic modification modulators including EZH2. ETV5 related genes were enriched in tumorigenesis biological processes and signaling pathways. In hepatocellular carcinoma, ETV5 expression was correlated with patients’ tumor pathological stage and resulted in adverse outcome of patients. Our further experiments evidences indicated that ETV5 facilitated cell proliferation and reduced sensitivity to GSK126 via regulating EZH2. Collectively, this study comprehensively elucidates the carcinogenic effects and molecular mechanisms of ETV5 in tumorigenesis and development, and provides theoretical basis and guidance for tumor diagnosis, targeted therapy for ETV5 and clinical epigenetic drug research.

Genetic risk factors for Mesoamerican nephropathy

Mesoamerican nephropathy (MeN) is a progressive kidney disease found on the Pacific coast of Central America primarily in young male agricultural workers without typical kidney disease risk factors. While it is generally accepted that environmental exposures contribute to MeN, we hypothesized that there was also an important genetic component. We performed a genome-wide association study comparing individuals with MeN versus individuals with normal kidney function. We found that Native American ancestry was strongly associated with increased risk of MeN. We also identified candidate variants in the OPCML gene, which encodes a protein that binds opioid receptors, that were associated with ~sixfold reduced odds of MeN (allele frequency 0.029 in controls and 0.005 in cases, OR = 0.16; P = 4 × 10-8). Sugarcane workers with the protective OPCML variants had markedly increased urine osmolality suggesting greater ability to defend against hypovolemia. Experiments with Opcml knock-out mice revealed roles for OPCML in fluid balance and temperature regulation consistent with our findings in humans. Our data suggest that heritable differential sensitivity to heat stress and dehydration contributes to high rates of kidney disease in Central America.

Complex IIa formation and ABC transporters determine sensitivity of OSCC to Smac mimetics

Small molecule inhibitors of apoptosis proteins (IAPs) antagonists, known as Smac mimetics (SMs), activate non-canonical NF-κB and sensitize cancer cells to TNF-induced cell death. SMs are currently in phase III clinical trials for head and neck squamous cell carcinoma (HNSCC) after promising phase II trials. To explore the utility of SMs in oral squamous cell carcinoma (OSCC), we tested nine human OSCC cell lines and correlated SM sensitivity with both IAP mutation and expression levels. cIAP1 protein expression was shown to be higher in OSCC and a predictor of poor prognosis. However, our in vitro and in vivo testing demonstrated differential sensitivity to SMs, which did not correlate with cIAP1 and cIAP2 expression in these OSCC cell lines. Exogenous TNF failed to effectively increase the sensitivity of SM-resistant OSCC cells to SM-induced cell death. SM resistance was associated with a deficiency in Complex IIa formation, but activation of non-canonical NF-κB was not a determinant of SM efficacy. Finally, metabolic analysis revealed that the ABC transporter pathway was activated in SM-resistant OSSC cells, and SMs combined with ABC transporter inhibitors improved cell death sensitivity to overcome SM resistance. These studies highlight the therapeutic potential of SMs in OSCC and support patient stratification to improve efficacy with the addition of adjuvant therapy.

Psychometric Functions for α-Ionone in Young Adults

The mathematical relationship between the ability to detect an odorant and its concentration appears for some odorants to be non-monotonic, with reversals (“notches”) in performance appearing at points along the psychometric function. Like visual adaptation curves that reflect the differential sensitivities of cones and rods, such reversals may provide information about underlying olfactory receptor processes. However, the presence of such reversals is rarely acknowledged, few participants and odorants have been tested, and methodological concerns abound. In this study, we examined in detail the psychometric function for the odorant α-ionone using a sizable number of young participants and 10 log-based concentrations of alpha-ionone presented in a random fashion. A trial consisted of the counterbalanced presentation of an odorless mineral oil and a concentration of α-ionone in rapid succession using Snap & Sniff® wands. The participants reported which of the two seemed stronger and indicated their confidence on a 9-point scale. In Study 1, 24 participants completed a single 30-minute test session of 60 trials. In Study 2, 600 trials were obtained from each of nine participants over the course of ten 30-minute sessions. In both studies, notches were consistently found in the psychometric function near the 10−5 and 10−3.5 vol/vol concentrations. Participants’ trial-by-trial confidence judgments corresponded with their detection performance, but their self-rated sense of smell did not. This research definitively demonstrates the presence of reliable reversals in the human α-ionone psychometric function and begs the question as to whether such reversals reflect the recruitment of receptive elements with differing response profiles.

The auxin-like effect of substrate and auxin sensitivity of plant modulate the PGPR activity of Lysinibacillus pinottii sp. nov. PB211

BackgroundBacterial auxin production is a mechanism that promotes plant growth. However, it can also be linked to deleterious rhizobacteria and plant-pathogenic bacteria. High levels of auxins have the potential to impede plant growth. Additionally, certain environmental factors can enhance auxin activity and impact the activity of plant growth-promoting rhizobacteria (PGPR).ResultsIn this work, we boarded how the auxin-like effect in the soil–plant system modulates the plant growth-promoting activity of Lysinibacillus pinottii sp. nov. PB211. The effect of humified organic matter (HOM) soil was simulated with humic acids (HA) coming from vermicompost; the spectral configuration (13C-CP-TOSS-NMR and FTIR) and coleoptile elongation test indicated its bioactivity. Cucumber exhibits a PGP effect of PB211 and HA at a lower concentration compared to corn. This disparity agrees with the differential sensitivity to auxins of both plant models. Monocot plants, such as corn, generally exhibit greater resistance to exogenous auxin activity compared to eudicot plants, such as cucumber. The presence of HA in the growth substrate (sand) is found to modulate the PGPR activity of PB211, resulting in a loss of PGPR activity.ConclusionsThe findings highlight the importance of considering the cumulative auxin-like effects in the soil–plant system when utilizing bacterial inoculants for plant growth promotion in agriculture.Graphical

TMET-02. CELL-INTRINSIC METABOLIC PHENOTYPES IDENTIFIED IN GLIOBLASTOMA PATIENTS USING MASS SPECTROMETRY IMAGING OF 13C-LABELED GLUCOSE METABOLISM

Abstract Glioblastoma (GB) is an inherently heterogeneous and invasive primary brain cancer. Genomic and transcriptomic studies have attempted to classify GB into subtypes that predict survival and that have different therapeutic vulnerabilities. An outstanding question and technical challenge is to visualize the metabolism of a tumour within its native microenvironment and understand to what extent its metabolism is driven by the microenvironment. Patients with GB tumours were infused with [U-13C]glucose and intra-operative multi-regional tumour samples were rapidly snap frozen for analysis using mass spectrometry imaging (MSI). Tissue microenvironment was assessed using multiplexed antibodies in immunocytochemistry (IMC) of contiguous sections. Isotope tracking in human-derived neurospheres and patient-derived xenografts were used to confirm the presence of metabolic phenotypes detected in the human samples and to test the susceptibility of each phenotype to a panel of drugs targeting cellular metabolism. Three metabolic signatures were identified in patient tumours: glycolytic, oxidative and a mixed glycolytic/oxidative phenotype. The phenotypes did not correlate with microenvironmental features, including proliferation rate, immune cell infiltration, and vascularisation. The phenotypes were retained when patient-derived cells were grown in vitro, or as orthotopically implanted xenografts, and were robust to changes in oxygen concentration, demonstrating their cell intrinsic nature. The spatial extent of the regions occupied by cells displaying these distinct metabolic phenotypes are large enough to be detected using clinically applicable metabolic imaging techniques. The phenotypes also demonstrate differential drug sensitivities, suggesting imaging of these phenotypes may be used to stratify patients for personalised therapy trials. This is the first report to demonstrate high resolution imaging of metabolic fluxes in a human tumour in vivo and the presence of different metabolic phenotypes within GB that are tumour cell intrinsic and largely independent of the tumour microenvironment.

DDDR-05. SPATIALLY RESOLVED DRUG SENSITIVITY PROFILING IN GLIOBLASTOMA

Abstract BACKGROUND Glioblastoma is the most common malignant primary brain tumor in adults, characterized by poor overall survival and an urgent need for more effective treatment strategies. Macroscopic heterogeneity, as observed through MR imaging, divides the tumor into distinct regions: a tumor-cell dense contrast-enhancing tumor core and a non-contrast-enhancing infiltration zone that appears hyperintense on T2/FLAIR imaging. The infiltration zone contains both infiltrating tumor cells and non-malignant cells from the local brain microenvironment. Although genomic and transcriptomic differences between the tumor core and the infiltration zone have been reported, functional differences, such as differential drug sensitivities, remain largely unknown. METHODS To investigate this, we collected region-specific glioblastoma patient tissue samples from both the contrast-enhancing tumor core and the non-contrast-enhancing but 5-aminolevulinic acid (5-ALA) fluorescent infiltration zone during surgery. Using pharmacoscopy, a microscopy-based single-cell drug screening platform, we assessed drug responses across regions and patients. RESULTS To determine whether drug responses within a single tumor region were more similar compared to responses across different regions we collected multiple tissue samples from each tumor region in seven patients. We identified region-specific drug responses for temozolomide and lomustine, among other drugs, in some of the patients. However, drug sensitivities were not consistent within the same region across different patients. Thus, across patients, the region-specific drug responses were obscured by patient heterogeneity. To address intra- and inter-tumor drug response heterogeneity, we developed a complementarity score based on region-specific drug response data of 60 drugs across 23 paired samples. The complementarity score identified promising drug combinations that demonstrated strong anti-tumor activity across tumor regions and patients. CONCLUSIONS Monotherapy approaches have largely failed in glioblastoma, despite extensive molecular profiling efforts aimed at identifying tumor-specific vulnerabilities. Identifying drug combinations that synergistically target the locoregional heterogeneity of glioblastoma may open up new avenues of personalized therapy.

TMOD-13. IDENTIFYING MOLECULAR SIGNATURES OF PPM1D-MUTANT DIPGS

Abstract Diffuse Intrinsic Pontine Glioma (DIPGs) are universally fatal pediatric tumors with a median survival of only 9-12 months. PPM1D-truncating mutations (PPM1Dtr), which promote its phosphatase activity and suppress TP53 function, are found in approximately 15% of DIPG tumors. PPM1Dtr are mutually exclusive to TP53 mutations, and present unique therapeutic opportunities to reactivate wild-type TP53. Another distinct feature of PPM1Dtr DIPGs is the co-occurrence of PI3K/AKT/mTOR pathway alterations, such as PIK3CA. There is an unmet need to elucidate the underlying molecular mechanisms of DIPG and identify novel therapeutic vulnerabilities that will improve outcomes of this devastating tumor. We hypothesize that PPM1D-mutant DIPGs can be therapeutically targeted by reactivating the p53 signaling pathway and other complementary pathways which cooperate with PPM1Dtr. To investigate this, we have generated Trp53- and Ppm1d-mutant isogenic DIPG murine models to determine how specific oncogenic alterations function in DIPG formation, growth and treatment sensitivity. Current studies aim to examine differences in sensitivity to standard of care treatment modalities and leverage next generation sequencing techniques to uncover additional vulnerabilities. We found notable differences upon addition of PIK3CA mutations in the context of PPM1Dtr DIPG tumor formation, growth, survival, and lineage identity. Ongoing studies using whole genome transcriptomics to compare tumor models will provide insight into the molecular signals and cell state changes driven by individual genetic alterations. Furthermore, by comparing isogenic TP53 and PPM1Dtr mutant DIPG models, we aim to further delineate the molecular mechanisms that drive differential sensitivity of TP53 wild-type tumors. The immune-competent nature of our murine models also provides us with a unique opportunity to delineate tumor immune microenvironment changes. Leveraging this information and our novel murine models, our long-term goal is to develop combinatorial therapeutic strategies to improve outcomes for this deadly disease.

TMOD-22. MULTIREGION BIOPSIES AND CORRESPONDING NEUROSPHERE CULTURES REVEAL SPATIAL DIVERGENCE IN GLIOBLASTOMA EVOLUTION

Abstract Spatial organization of normal tissues is key for their function, yet the role of distribution of distinct clones of cancer cells in tumor is not usually studied as a potential vulnerability in cancer. This is of particular interest in glioblastoma (GBM), in which extensive intratumor heterogeneity hampers effectiveness of treatment. To better understand tumor evolution and the complexity of cancer ecosystem in GBM, we performed profiling of biopsies collected from MRI-defined regions of the tumors, spanning the surface, tumor core and deep tumor margins, which drive local recurrence after surgery. To test how spatially distinct biopsies differ in their cancer cell composition and to assess the tumor microenvironment heterogeneity, we took advantage of 5-ALA metabolite fluorescence, which accumulates in GBM cells and allows the surgeon to visualize cancer under blue light. We used it to separate cancer cells (predominantly 5-ALAhigh) from tumor microenvironment (5-ALAlow) and subjected the fractions to bulk and single cell transcriptomics, respectively. We found a gradient of cellular states associated with location in the tumor, with the deep margin samples having more 5-ALAlow cancer cells and generally less defined phenotype. We have also developed neurosphere cultures from our collection of spatially distant biopsies. Despite the selection pressure of the in vitro culture conditions, cells from different regions of the tumor adopted different morphologies, suggesting retention of some of their differential features. The differences between cultures were reflected in metabolism, proliferation, and differential sensitivity to a drug panel. Whole exome sequencing and transcriptomic analysis were also performed to identify the selection bottleneck of culture conditions, establish evolutionary trajectory of the distinct regions and neurosphere cultures, and test how strongly our cultures represent both the 5-ALAhigh and 5-ALAlow subpopulations of GBM cells. Our study identified properties associated with different spatial locations within GBM tumors, which will help uncover the cellular and microenvironmental dependencies that could lead to novel therapeutic targets for this highly heterogeneous tumor.

Differential sensitivity and specificity of Aedes aegypti and Anopheles gambiae to adenine nucleotide phagostimulants—an all-or-none response?

BackgroundThe decision to imbibe a blood meal is predominantly dependent on the sensitivity and specificity of haematophagous arthropods to blood-derived adenine nucleotides, in particular adenosine triphosphate (ATP). Despite previous efforts to identify and characterise the specificity and sensitivity to ATP and other adenine nucleotides, as well as the role of other blood-derived phagostimulants across the Culicidae, comparisons across species remain difficult.MethodsThe feeding response of the yellow fever mosquito Aedes aegypti and the African malaria vector Anopheles gambiae to adenine nucleotides in the presence of a carbonate buffer was assessed using a membrane feeding assay. The proportion of mosquitoes engorged and the volume imbibed by all mosquitoes was scored visually and spectrophotometrically. In addition, the proportion of prediuresing An. gambiae, as well as the volume engorged and prediuresed, was examined.ResultsAedes aegypti was more sensitive to adenine nucleotides than An. gambiae, but both species maintained specificity to these phagostimulants, demonstrating a dose-dependent bimodal feeding pattern, thereby expanding our understanding of the all-or-none blood-feeding hypothesis. Feeding on the bicarbonate buffer by An. gambiae—but not that of Ae. aegypti—demonstrated a species-specific variation in how blood phagostimulants are encoded. Adenine nucleotides, with and without bovine serum albumin, were observed to dose-dependently regulate the proportion of An. gambiae prediuresing and the volumes prediuresed but not volumes engorged.ConclusionsTaken together, the results of this study expand our understanding of how mosquitoes differentially assess and respond to blood meal constituents, and provide a basis for further physiological and molecular studies.Graphical

Mild focal cooling selectively impacts computations in dendritic trees.

Focal cooling is a powerful technique to temporally scale neural dynamics. However, the underlying cellular mechanisms causing this scaling remain unresolved. Here, using targeted focal cooling (with a spatial resolution of 100 micrometers), dual somato-dendritic patch clamp recordings, two-photon calcium imaging, transmitter uncaging, and modeling we reveal that a 5°C drop can enhance synaptic transmission, plasticity, and input-output transformations in the distal apical tuft, but not in the basal dendrites of intrinsically bursting L5 pyramidal neurons. This enhancement depends on N-methyl-D-aspartate (NMDA) and Kv4.2, suggesting electrical structure modulation. Paradoxically, and despite the increase in tuft excitability, we observe a reduced rate of recovery from inactivation for apical Na+ channels, thereby regulating back-propagating action potential invasion, coincidence detection, and overall burst probability, resulting in an "apparent" slowing of somatic spike output. Our findings reveal a differential temperature sensitivity along the basal-tuft axis of L5 neurons analog modulates cortical output.

Infant preference for specific phonetic cue relations in the contrast between voiced and voiceless stops.

Acoustic variability in the speech input has been shown, in certain contexts, to be beneficial during infants' acquisition of sound contrasts. One approach attributes this result to the potential of variability to make the stability of individual cues visible. Another approach suggests that, instead of highlighting individual cues, variability uncovers stable relations between cues that signal a sound contrast. Here, we investigate the relation between Voice Onset Time and the onset of F1 formant frequency, two cues that subserve the voicing contrast in German. First, we verified that German-speaking adults' use of VOT to categorize voiced and voiceless stops is dependent on the value of the F1 onset frequency, in the specific form of a so-called trading relation. Next, we tested whether 6-month-old German learning infants exhibit differential sensitivity to stimulus continua in which the cues varied to an equal extent, but either adhered to the trading relation established in the adult experiment or adhered to a reversed relation. Our results present evidence that infants prefer listening to speech in which phonetic cues conform to certain cue trading relations over cue relations that are reversed.

Differential redox sensitivity of tRNA dihydrouridylation.

Various transfer RNA (tRNA) modifications have recently been shown to regulate stress-dependent gene expression by modulating messenger RNA translation. Among these modifications, dihydrouridine stands out for its increase of tRNA structural flexibility. However, whether and how dihydrouridine synthesis reacts to environmental stimuli is largely unknown. In this study, we manipulated the intracellular redox state of Escherichia coli using paraquat, revealing differential sensitivities of the three tRNA-dihydrouridine synthases towards oxidative stress. Using liquid chromatography-mass spectrometryquantification of dihydrouridine in various knockout strains, we validated the use of a specific RNA sequencing method, namely AlkAnilineSeq, for the precise mapping of dihydrouridines throughout E. coli tRNAs. We found DusA showing high activity, followed by DusB and DusC, whose activity was decreased under paraquat treatment. The relative sensitivity is most plausibly explained by a paraquat-dependent drop of NADPH availability. These findings are substantiated by in vitro kinetics, revealing DusA as the most active enzyme, followed by DusB, while DusC showed little activity, likely related to the efficacy of the redox reaction of the flavin coenzyme with NADPH. Overall, our study underscores the intricate interplay between redox dynamics and tRNA modification processes, revealing a new facet of the regulatory mechanisms influencing cellular responses to oxidative stress.

Differential Sensitivity of Midline Patterning to Mitosis during and after Primitive Streak Extension.

Midline establishment is a fundamental process during early embryogenesis for Bilaterians . Midline patterning in nonamniotes can occur without mitosis, through Planar Cell Polarity (PCP) signaling. By contrast, amniotes utilize both cell proliferation and PCP signaling for patterning early midline landmark, the primitive streak (PS). This study examined their roles for midline patterning at post PS-extension. In contrast to PS extension stages, embryos under mitotic arrest during the post PS-extension preserved notochord (NC) extension and Hensen's node (HN)/PS regression judged by both morphology and marker genes, although they became shorter, and laterality was lost. Remarkably, no or background level of expression was detected for the majority of PCP core components in the NC-HN-PS area at post PS-extension stages, except for robustly detected prickle-1 . Morpholino knockdown of Prickle-1 showed little influence on midline patterning, except for suppressed embryonic growth. Lastly, associated with mitotic arrest-induced size reduction, midline tissue cells displayed hypertrophy. Thus, the study has identified at least two distinct mitosis sensitivity phases during early midline pattering: One is PS extension that requires both mitosis and PCP, and the other is mitotic arrest-resistant midline patterning with little influence by PCP at post PS-extension stages.

Differential sensitivity of hemocyte subpopulations (Mytilus edulis) to aged polyethylene terephthalate micro- and nanoplastic particles

Bivalve hemocytes, particularly granulocytes and hyalinocytes, play a crucial role in cell-mediated immunity. However, their interactions with aged plastic particles, exhibiting altered properties that more closely resemble those in natural environments, remain largely underexplored. This study assesses the differential responses of hemocyte subpopulations (Mytilus edulis) to chemically aged polyethylene terephthalate (PET) microplastic (MPs) and nanoplastic (NPs) particles across multiple cellular effect endpoints. Particle characteristics were analyzed using Single Particle Extinction and Scattering, Raman Spectroscopy, Scanning Electron Microscopy, and Dynamic Light Scattering. In vitro experiments with aged PET MPs (1.9 µm) and NPs (0.68 µm) were conducted at three internally relevant concentrations: 10 (C1), 10³ (C2), and 10⁵ particles/mL (C3). Cellular responses were assessed by measuring lysosomal content stability, reactive oxygen species (ROS) production, cellular mortality, and morphological parameters using flow cytometry at 6, 12, 24, and 48 hours. Our findings provide mechanistic insights into the differential sensitivities of granulocytes and hyalinocytes to aged PET, influenced by particle size and concentration. Specifically, aged PET MPs and NPs induce distinct size and concentration-dependent patterns of lysosomal destabilization, coinciding with the loss of functional integrity. Elevated ROS levels were observed only in granulocytes and hyalinocytes exposed to high concentrations of aged PET NPs, underscoring the effects on oxidative stress. Both aged PET MPs and NPs induce significant increases in cellular mortality, particularly after 24 h of exposure at high concentrations. These findings reveal the complex cellular mechanisms underlying hemocyte functional impairment following exposure to aged PET particles under environmentally and biologically relevant conditions.

Mechanisms of Differential Sensitivity to Ethanol-Induced Apoptosis in Mouse Spinal Cord at Different Developmental Stages-Akt/GSK Signaling and BAX.

The current study investigated differences in ethanol-induced apoptosis of spinal cord dorsal horn neurons at different developmental stages and the molecular mechanisms involved. A mouse ethanol intervention model was established on postnatal days 4, 7, and 12. Primary cells were derived from the spinal cord at postnatal day 4. Western blotting, immunofluorescence, and flow cytometry were used to detect apoptosis-related proteins in the spinal cord and primary cells. Kyoto Encyclopedia of Genes and Genomes enrichment analysis of differentially expressed genes originating from the Gene Expression Omnibus dataset GSE184615 was conducted. Effects on Akt/GSK3β pathway proteins were investigated using the GSK3β inhibitor AR-A014418, and the Akt inhibitor DHA. Lentiviral knockdown and overexpression of intervening GSK3β were used in HT22 cell lines to investigate the effects of alcohol on GSK 3β and caspase proteins. J-aggregates, reactive oxygen species assays, and calcein-AM assays were used to investigate mitochondrial function and cell viability. Ethanol caused downregulation of Akt activity and upregulation of GSK3β activity and apoptosis. DHA, AR-A014418, and knockdown of GSK3β effectively counteracted ethanol-induced apoptosis, whereas overexpression of GSK3β enhanced the injury process. PI3K activity was unchanged during these processes. Fluorescence colocalization analysis indicated that BAX was translocated to mitochondria during the apoptotic process. BAX was downregulated as the spinal cord developed, consistent with a reduced susceptibility to ethanol-induced apoptosis. Akt/GSK3β signaling and BAX together determine the direction of alcohol-induced apoptosis and its susceptibility to change during developmental stages in the spinal cord.