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BioRxiv : the Preprint Server For... Jun 2024Cellular mechanical properties influence cellular functions across pathological and physiological systems. The observation of these mechanical properties is limited in...
Cellular mechanical properties influence cellular functions across pathological and physiological systems. The observation of these mechanical properties is limited in part by methods with a low throughput of acquisition or with low accessibility. To overcome these limitations, we have designed, developed, validated, and optimized a microfluidic cellular deformation system (MCDS) capable of mechanotyping suspended cells on a population level at a high throughput rate of ∼300 cells pers second. The MCDS provides researchers with a viable method for efficiently quantifying cellular mechanical properties towards defining prognostic implications of mechanical changes in pathology or screening drugs to modulate cytoskeletal integrity.
PubMed: 38948841
DOI: 10.1101/2024.06.17.599307 -
BioRxiv : the Preprint Server For... Jun 2024Both endogenous antibodies and a subset of antibody therapeutics engage Fc gamma receptor (FcγR)IIIa / CD16a to stimulate a protective immune response. Increasing the...
Both endogenous antibodies and a subset of antibody therapeutics engage Fc gamma receptor (FcγR)IIIa / CD16a to stimulate a protective immune response. Increasing the FcγRIIIa/IgG1 interaction improves the immune response and thus represents a strategy to improve therapeutic efficacy. FcγRIIIa is a heavily glycosylated receptor and glycan composition affects antibody-binding affinity. Though our laboratory previously demonstrated that natural killer (NK) cell N-glycan composition affected the potency of one key protective mechanism, antibody-dependent cell-mediated cytotoxicity (ADCC), it was unclear if this effect was due to FcγRIIIa glycosylation. Furthermore, the structural mechanism linking glycan composition to affinity and cellular activation remained undescribed. To define the role of individual amino acid and N-glycan residues we measured affinity using multiple FcγRIIIa glycoforms. We observed stepwise affinity increases with each glycan truncation step with the most severely truncated glycoform displaying the highest affinity. Removing the N162 glycan demonstrated its predominant role in regulating antibody-binding affinity, in contrast to four other FcγRIIIa N-glycans. We next evaluated the impact of the N162 glycan on NK cell ADCC. NK cells expressing the FcγRIIIa V158 allotype exhibited increased ADCC following kifunensine treatment to limit N-glycan processing. Notably, an increase was not observed with cells expressing the FcγRIIIa V158 S164A variant that lacks N162 glycosylation, indicating the N162 glycan is required for increased NK cell ADCC. To gain structural insight into the mechanisms of N162 regulation, we applied a novel protein isotope labeling approach in combination with solution NMR spectroscopy. FG loop residues proximal to the N162 glycosylation site showed large chemical shift perturbations following glycan truncation. These data support a model for the regulation of FcγRIIIa affinity and NK cell ADCC whereby composition of the N162 glycan stabilizes the FG loop and thus the antibody-binding site.
PubMed: 38948809
DOI: 10.1101/2024.06.17.599285 -
BioRxiv : the Preprint Server For... Jun 2024Drugs of abuse activate defined neuronal ensembles in brain reward structures such as the nucleus accumbens (NAc), which are thought to promote the enduring synaptic,...
Drugs of abuse activate defined neuronal ensembles in brain reward structures such as the nucleus accumbens (NAc), which are thought to promote the enduring synaptic, circuit, and behavioral consequences of drug exposure. While the molecular and cellular effects arising from experience with drugs like cocaine are increasingly well understood, the mechanisms that sculpt NAc ensemble participation are largely unknown. Here, we leveraged unbiased single-nucleus transcriptional profiling to identify expression of the secreted glycoprotein Reelin (encoded by the gene) as a marker of cocaine-activated neuronal ensembles within the rat NAc. Multiplexed in situ detection confirmed selective expression of the immediate early gene in neurons after cocaine experience, and also revealed enrichment of mRNA in + medium spiny neurons (MSNs) in both the rat and human brain. Using a novel CRISPR interference strategy enabling selective knockdown in the adult NAc, we observed altered expression of genes linked to calcium signaling, emergence of a transcriptional trajectory consistent with loss of cocaine sensitivity, and a striking decrease in MSN intrinsic excitability. At the behavioral level, loss of prevented cocaine locomotor sensitization, abolished cocaine place preference memory, and decreased cocaine self-administration behavior. Together, these results identify Reelin as a critical mechanistic link between ensemble participation and cocaine-induced behavioral adaptations.
PubMed: 38948801
DOI: 10.1101/2024.06.17.599348 -
BioRxiv : the Preprint Server For... Jun 2024We take a unique approach to understanding the causes of podocyte injury in collagen IV nephropathies, a crucial step in developing targeted therapies for conditions...
RATIONALE
We take a unique approach to understanding the causes of podocyte injury in collagen IV nephropathies, a crucial step in developing targeted therapies for conditions like Alport Syndrome.
OBJECTIVES
We characterize the structural, functional, and biophysical properties of glomerular capillaries and podocytes in mice and analyze kidney cortex transcriptional profiles at various disease stages. We investigate the effects of the ER stress mitigator TUDCA on these parameters. Furthermore, we used human FSGS associated podocyte enriched genes to identify molecular pathways rescued by TUDCA thereby offering potential therapeutic targets for Alport Syndrome.
FINDINGS
We find a clear disease progression timeline in mice. Podocyte injury develops by 3 months, with glomeruli reaching maximum deformability at 4 months, associated with a 40% loss of podocytes. This is followed by progressive stiffening of glomerular capillaries, increasing proteinuria, reduced renal function, inflammatory infiltrates, and fibrosis from months 4 to 8. Bulk RNA sequencing at 2, 4, and 7 months reveals a progressive increase in expression of genes related to cytokine and chemokine signaling, matrix and cell injury, and activation of the TNF pathway, similar to observations in a NEPTUNE FSGS cohort. Podocyte-enriched genes from FSGS patients mapped to mice found that TUDCA, which mitigated glomerular and renal injury suppressed molecular pathways associated with extracellular matrix and basement membrane synthesis, podocyte stress and hypertrophy.
CONCLUSIONS
We uncover two distinct phases of nephropathy progression. The first is characterized by podocytopathy, increased glomerular capillary deformability and accelerated podocyte loss, and the second by increased capillary wall stiffening and renal inflammatory and profibrotic pathway activation. The response of podocytes to TUDCA treatment provides novel insights into downstream signaling pathways, offering potential therapeutic targets for treating Alport and related nephropathies.
PubMed: 38948788
DOI: 10.1101/2024.02.26.582201 -
BioRxiv : the Preprint Server For... Jun 2024Primary hypertension in childhood tracks into adulthood and may be associated with increased cardiovascular risk. Studies conducted in children and adolescents provide...
BACKGROUND
Primary hypertension in childhood tracks into adulthood and may be associated with increased cardiovascular risk. Studies conducted in children and adolescents provide an opportunity to explore the early cardiovascular target organ injury (CV-TOI) in a population free from many of the co-morbid cardiovascular disease risk factors that confound studies in adults.
METHODS
Youths (n=132, mean age 15.8 years) were stratified by blood pressure (BP) as low, elevated, and high-BP and by left ventricular mass index (LVMI) as low- and high-LVMI. Systemic circulating RNA, miRNA, and methylation profiles in peripheral blood mononuclear cells and deep proteome profiles in serum were determined using high-throughput sequencing techniques.
RESULTS
gene expression was elevated in youths with high-BP with and without high-LVMI. expression levels positively correlated with systolic BP (r=0.3143, p=0.0034). The expression of hsa-miR-335-5p, one of the predicted miRNAs, was downregulated in high-BP with high-LVMI youths and was inversely correlated with systolic BP (r=-0.1891, p=0.0489). hypermethylation, circulating PROZ upregulation (log FC=0.61, p=0.0049 and log FC=0.62, p=0.0064), and SOD3 downregulation (log FC=-0.70, p=0.0042 and log FC=-0.64, p=0.010) were observed in youths with elevated BP and high-BP with high-LVMI. Comparing the transcriptomic and proteomic profiles revealed elevated levels in youths displaying high-BP and high-LVMI.
CONCLUSIONS
The findings are compatible with a novel blood pressure-associated mechanism that may occur through impaired angiogenesis and extracellular matrix degradation through dysregulation of Vasohibin-1 and Hyaluronidase1 was identified as a possible mediator of CV-TOI in youth with high-BP and suggests strategies for ameliorating TOI in adult-onset primary hypertension.
PubMed: 38948714
DOI: 10.1101/2024.06.17.599125 -
Frontiers in Endocrinology 2024Medullary thyroid cancer (MTC) is a challenging malignancy. The survival outcome of MTC based on AJCC staging system does not render a discriminant classifier among...
BACKGROUND
Medullary thyroid cancer (MTC) is a challenging malignancy. The survival outcome of MTC based on AJCC staging system does not render a discriminant classifier among early stages.
METHODS
3601 MTC patients from 2000 to 2018 were identified from the Surveillance, Epidemiology, and End Results (SEER) database. Smooth curve fitting, Cox proportional hazard regression and competing risk analysis were applied.
RESULTS
A linear correlation between age and log RR (relative risk of overall death) was detected. Overlaps were observed between K-M curves representing patients aged 45-50, 50-55, and 55-60. The study cohort was divided into 3 subgroups with 2 age cutoffs set at 45 and 60. Each further advanced age cutoff population resulted in a roughly "5%" increase in MTC-specific death risks and an approximately "3 times" increase in non-MTC-specific death risks.
CONCLUSIONS
The survival outcome disparity across age cutoffs at 45 and 60 for MTC has been well defined.
Topics: Humans; Thyroid Neoplasms; Middle Aged; Male; Female; Carcinoma, Neuroendocrine; Retrospective Studies; Age Factors; SEER Program; Survival Rate; Aged; Prognosis; Adult; Cohort Studies; Follow-Up Studies
PubMed: 38948527
DOI: 10.3389/fendo.2024.1393904 -
Frontiers in Pharmacology 2024Neurodevelopmental disorders (NDDs) include a broad spectrum of pathological conditions that affect >4% of children worldwide, share common features and present a... (Review)
Review
Neurodevelopmental disorders (NDDs) include a broad spectrum of pathological conditions that affect >4% of children worldwide, share common features and present a variegated genetic origin. They include clinically defined diseases, such as autism spectrum disorders (ASD), attention-deficit/hyperactivity disorder (ADHD), motor disorders such as Tics and Tourette's syndromes, but also much more heterogeneous conditions like intellectual disability (ID) and epilepsy. Schizophrenia (SCZ) has also recently been proposed to belong to NDDs. Relatively common causes of NDDs are copy number variations (CNVs), characterised by the gain or the loss of a portion of a chromosome. In this review, we focus on deletions and duplications at the 16p11.2 chromosomal region, associated with NDDs, ID, ASD but also epilepsy and SCZ. Some of the core phenotypes presented by human carriers could be recapitulated in animal and cellular models, which also highlighted prominent neurophysiological and signalling alterations underpinning 16p11.2 CNVs-associated phenotypes. In this review, we also provide an overview of the genes within the 16p11.2 locus, including those with partially known or unknown function as well as non-coding RNAs. A particularly interesting interplay was observed between MVP and MAPK3 in modulating some of the pathological phenotypes associated with the 16p11.2 deletion. Elucidating their role in intracellular signalling and their functional links will be a key step to devise novel therapeutic strategies for 16p11.2 CNVs-related syndromes.
PubMed: 38948459
DOI: 10.3389/fphar.2024.1407865 -
World Journal of Experimental Medicine Jun 2024Knowledge about refractive stabilization and the accuracy of postoperative refractive error measurements are crucial for improved patient outcomes after...
BACKGROUND
Knowledge about refractive stabilization and the accuracy of postoperative refractive error measurements are crucial for improved patient outcomes after phacoemulsification. Existing guidelines typically recommend waiting 4-6 wk before prescribing corrective lenses. Our research focused on identifying factors that influence refractive errors in the early stages of post-cataract surgery, thus contributing to the existing literature on this topic.
AIM
To investigate the time required for refraction stability after uneventful phacoemulsification surgery.
METHODS
We compared the variation and statistical significance of the difference in spherical, cylindrical components, and the spherical equivalent between the 1- and 6-wk follow-up period in a group of 257 eyes that underwent uneventful phacoemulsification with foldable intraocular lens implantation, all performed by a single experienced surgeon. The Wilcoxon-Signed Rank Test was utilized to assess the magnitude of the change and determine its statistical significance. The refractive stability was defined as the point at which the change in spherical equivalent was within ± 0.5 dioptres for two consecutive visits.
RESULTS
The average age of the patients was 64.9 ± 8.9 yr. The differences observed in both the visits in spherical power (0.1 ± 0.2), cylinder power (0.3 ± 0.4), and spherical equivalent (0.2 ± 0.2) were minimal and not statistically significant. The majority of eyes (93.4%) achieved refractive stability within 6 wk after the surgery. The cylindrical power differed between age groups at the 6 wk post-operative and the difference was statistically significant ( value 0.013). There were no significant differences in refractive stability when considering sex and axial length.
CONCLUSION
Phacoemulsification with foldable intraocular lens implantation results in no significant changes in refraction for the majority of cases during the 6-wk follow-up period. Therefore, a spectacle prescription can be given at the completion of 1 wk.
PubMed: 38948423
DOI: 10.5493/wjem.v14.i2.95016 -
Sichuan Da Xue Xue Bao. Yi Xue Ban =... May 2024Prior studies have established a connection between albuminuria and various inflammatory reactions, highlighting that an increase in C-reactive protein by 1 mg/L...
OBJECTIVE
Prior studies have established a connection between albuminuria and various inflammatory reactions, highlighting that an increase in C-reactive protein by 1 mg/L increases the likelihood of albuminuria by 2%. Recent investigations indicate a positive correlation between the systemic immune-inflammation index (SII) and increased urinary protein excretion. In addition, elevated levels of the systemic inflammatory response index (SIRI) also correlate with a higher prevalence of albuminuria. The aggregate index of systemic inflammation (AISI) offers a more comprehensive indicator of inflammation, providing an extensive assessment of systemic inflammatory status compared to SII and SIRI. Yet, the specific relationship between AISI and albuminuria remains unclear. This study aims to explore this association in U.S. adults.
METHODS
We analyzed data from the National Health and Nutrition Examination Survey (NHANES) for 2007-2018, excluding pregnant women and individuals under 18. Cases with missing data on AISI, urinary albumin concentration, and other covariates were also excluded. AISI was computed using the formula: AISI=(platelet count×neutrophil count×monocyte count)/lymphocyte count. Albuminuria was defined as the urinary albumin-to-creatinine ratio exceeding 30 mg/g. Continuous variables were presented in the form of the mean±standard error, and categorical variables in percentages. We utilized weighted -tests and chi-square tests for baseline comparisons. We applied weighted multivariable logistic regression and generalized additive models (GAM) to explore the association between AISI and albuminuria and to assess potential nonlinear relationships.
RESULTS
The study included 32273 participants, with an average age of (46.75±0.24) years old. The cohort comprised 48.73% males and 51.27% females. The prevalence of albuminuria was 9.64%. The average logarithmic value of logAISI was 7.95±0.01, and were categorized into tertiles as follows: Quartile 1 (Q1) (4.94 to 7.49), Q2 (7.49 to 8.29), and Q3 (8.29 to 10.85). As logAISI increased, so did the prevalence of hypertension, diabetes, congestive heart failure, and albuminuria, all showing statistically significant increases (<0.001). Similarly, the use of antihypertensive, lipid-lowering, and hypoglycemic drugs was also more prevalent (<0.001). Statistically significant differences were observed across the three groups concerning age, race and ethnicity, formal education, alcohol consumption, smoking status, systolic and diastolic blood pressures, body mass index, estimated glomerular filtration rate, HbA1c, alanine aminotransferase, aspartate aminotransferase, albumin, creatinine, uric acid, and high-density lipoprotein cholesterol (<0.05). However, no significant differences were noted in the total cholesterol or the sex ratios among the groups. The association between logAISI and albuminuria was assessed using weighted multivariable logistic regression, and the detailed results are presented in Table 2. In model 1, without adjusting for covariates, each unit increase in logAISI was associated with a 32% increase in the risk of albuminuria (odds ratio [OR]=1.32, 95% confidence interval [CI]: 1.27-1.38, <0.001). Model 2 was adjusted for age, gender, race, and education level, and showed a similar trend, with each unit increase in logAISI associated with a 31% increased risk (OR=1.31, 95% CI: 1.26-1.37, <0.001). Model 3, which was further adjusted for all covariates, revealed that each unit increase in logAISI was associated with a 20% increase in the risk of albuminuria (OR=1.20, 95% CI: 1.15-1.26, <0.001). The study also transformed logAISI from a continuous to a categorical variable for analysis. Compared with Q1, the risk of albuminuria in Q3, after adjusting for all covariates, significantly increased (OR=1.37, 95% CI: 1.22-1.55, <0.001). Q2 also demonstrated a higher risk compared with Q1 (OR=1.13, 95% CI: 1.06-1.36, =0.004). The trend test indicated a dose-effect relationship between increasing logAISI and the rising risk of albuminuria. GAM revealed a nonlinear relationship between logAISI and albuminuria, with distinct trends noted between sexes. Segmented regression based on turning points showed significant effects among women, although the slope difference between the segments was not significant. In men, a significant threshold effect was observed; below the logAISI of 7.25, increases in logAISI did not enhance the risk of albuminuria, but above this threshold, the risk significantly increased. As part of a sensitivity analysis, weighted multivariable logistic regression was performed by changing the outcome variable to macroalbuminuria and adjusting for all covariates. The analysis showed that for every unit increase in logAISI, the risk of developing macroalbuminuria increased by 31% (OR=1.31, 95% CI: 1.15-1.49, <0.001). Compared with Q1, the risk of albuminuria in Q3 increased by 69% (OR=1.69, 95% CI: 1.27-2.25, <0.001), and in Q2, it increased by 40% (OR=1.40, 95% CI: 1.03-1.92, =0.030). Subgroup analysis and interaction results showed that the positive association between AISI and proteinuria risk was stronger in men than in women. Similarly, the association was stronger in people with hypertension compared with those with normal blood pressure, and higher in overweight people compared with those of normal weight. Furthermore, smokers and drinkers showed a stronger positive association between AISI and the risk of proteinuria than non-smokers and non-drinkers do. These results suggest that sex, blood pressure, body mass index, smoking, and alcohol consumption interact with AISI to influence the risk of proteinuria.
CONCLUSION
There is a robust positive association between AISI and increased risks of albuminuria in US adults. As logAISI increases, so does the risk of albuminuria. However, further validation of this conclusion through large-scale prospective studies is warranted.
Topics: Humans; Albuminuria; Nutrition Surveys; Cross-Sectional Studies; Inflammation; Female; Male; Adult; Middle Aged; C-Reactive Protein; Platelet Count
PubMed: 38948283
DOI: 10.12182/20240560108 -
Imaging Science in Dentistry Jun 2024Ameloblastic fibrodentinoma (AFD) is a rare benign odontogenic tumor that resembles an ameloblastic fibroma with dysplastic dentin. This report presents a rare case of...
Ameloblastic fibrodentinoma (AFD) is a rare benign odontogenic tumor that resembles an ameloblastic fibroma with dysplastic dentin. This report presents a rare case of mandibular AFD with imaging features in a young patient. Panoramic radiography and computed tomography revealed a well-defined lesion with internal septa and calcified foci, causing inferior displacement of the adjacent molars as well as buccolingual cortical thinning and expansion of the posterior mandible. The lesion was surgically removed via mass excision, and the involved tooth was extracted under general anesthesia. During the 5-year follow-up period, no evidence of recurrence was observed. Radiologic features of AFD typically reveal a moderately to well-defined mixed lesion with varying degrees of radiopacity, reflecting the extent of dentin formation. Radiologists should consider AFD in the differential diagnosis when encountering a multilocular lesion with little dense radiopacity, particularly if it is associated with delayed eruption, impaction, or absence of involved teeth, on radiographic images of young patients.
PubMed: 38948190
DOI: 10.5624/isd.20230247