Effect of In Vitro Enzyme Digestion and Bile Treatment on Milk Extracellular Vesicles Stability.

SCOPE: Milk extracellular vesicles (EVs) are nanosized particles with potential immune bioactivities. This study examines their fate during in vitro infant gastrointestinal digestion (GI). METHODS AND RESULTS: Bovine milk is digested using the in vitro INFOGEST method, adjusted for the infant. To unravel the contribution of digestive enzymes from bile, milk is treated with digestive enzymes, bile, or a combination of both. EVs are collected posttreatment using differential ultracentrifugation. EVs characterization includes electrophoresis, immunoblotting, nanoparticle tracking analysis, and atomic force microscopy. EVs protein markers programmed cell death 6-interacting protein (ALIX), tumor susceptibility gene 101 (TSG101), cluster of differentiation 9 (CD9), and xanthine dehydrogenase (XDH) are detected after gastric digestion (G60), but their signal intensity is significantly reduced by intestinal conditions (p < 0.05). Enzyme digestion, compared to bile treatment (I60 + bile), results in a significant reduction of signal intensities for TSG101 and CD9 (p < 0.05). Nanoparticle tracking analysis shows a significant reduction (p < 0.05) of EV numbers at the end of the intestinal phase. EVs are detected by atomic force microscopy at the end of the intestinal phase, showing that intact EVs can survive upper gut digestion. CONCLUSION: Intact EVs can be found at the end of the intestinal phase. However, digestive enzymes and bile reduce the quantity and characteristics of EVs, with digestive enzymes playing a larger role.

A network of transcriptomic signatures identifies novel comorbidity mechanisms between schizophrenia and somatic disorders.

The clinical burden of mental illness, in particular schizophrenia and bipolar disorder, are driven by frequent chronic courses and increased mortality, as well as the risk for comorbid conditions such as cardiovascular disease and type 2 diabetes. Evidence suggests an overlap of molecular pathways between psychotic disorders and somatic comorbidities. In this study, we developed a computational framework to perform comorbidity modeling via an improved integrative unsupervised machine learning approach based on multi-rank non-negative matrix factorization (mrNMF). Using this procedure, we extracted molecular signatures potentially explaining shared comorbidity mechanisms. For this, 27 case-control microarray transcriptomic datasets across multiple tissues were collected, covering three main categories of conditions including psychotic disorders, cardiovascular diseases and type II diabetes. We addressed the limitation of normal NMF for parameter selection by introducing multi-rank ensembled NMF to identify signatures under various hierarchical levels simultaneously. Analysis of comorbidity signature pairs was performed to identify several potential mechanisms involving activation of inflammatory response auxiliarily interconnecting angiogenesis, oxidative response and GABAergic neuro-action. Overall, we proposed a general cross-cohorts computing workflow for investigating the comorbid pattern across multiple symptoms, applied it to the real-data comorbidity study on schizophrenia, and further discussed the potential for future application of the approach.

From sensing interactions to controlling the interactions: a novel approach to obtain biological transistors for specific and label-free immunosensing.

Antibody-antigen interactions are shaped by the solution pH level, ionic strength, and electric fields, if present. In biological field-effect transistors (BioFETs), the interactions take place at the sensing area in which the pH level, ionic strength and electric fields are determined by the Poisson-Boltzmann equation and the boundary conditions at the solid-solution interface and the potential applied at the solution electrode. The present study demonstrates how a BioFET solution electrode potential affects the sensing area double layer pH level, ionic strength, and electric fields and in this way shapes the biological interactions at the sensing area. We refer to this as 'active sensing'. To this end, we employed the meta-nano-channel (MNC) BioFET and demonstrate how the solution electrode can determine the antibody-antigen equilibrium constant and allows the control and tuning of the sensing performance in terms of the dynamic range and limit-of-detection. In the current work, we employed this method to demonstrate the specific and label-free sensing of Alpha-Fetoprotein (AFP) molecules from 0.5 µL drops of 1 : 100 diluted serum. AFP was measured during pregnancy as part of the prenatal screening program for fetal anomalies, chromosomal abnormalities, and abnormal placentation. We demonstrate AFP sensing with a limit-of-detection of 10.5 aM and a dynamic range of 6 orders of magnitude in concentration. Extensive control measurements are reported.

Comparative Efficacy of Epley, Semont and Gans Maneuver in Treating Posterior Canal Benign Paroxysmal Positional Vertigo.

Pharmacological therapies are used to control Benign paroxysmal positional vertigo (BPPV) symptoms for a brief period, discontinuing them usually results in recurrence. Canalolith repositioning maneuvers, including Epley, Semont, and Gans maneuvers, have been recommended for treating posterior canal BPPV with a high rate of success. A prospective, quasi-randomized study was carried out to compare the efficacy of Epley, Semont, and Gans maneuvers in the treatment of posterior canal BPPV and their durability. All patients over the age of 20 who met the BPPV diagnostic criteria, regardless of gender, were included in the study. Diagnosis of BPPV was done by Dix Hallpike maneuver. Typical posterior canal BPPV, the most frequent form of BPPV, is characterized by paroxysmal nystagmus evoked through the Dix-Hallpike test; the nystagmus is torsional clockwise for the left side, counter-clockwise for the right side, with a vertical up-beating component. The patients were uniformly quasi-randomized in a 1:1:1 ratio to be treated with Epley, Semont, and Gans maneuvers. After performing the maneuver, the patients were again subjected to the Dix-Hallpike test. Based on the result of the Dix-Hallpike test's positivity, the maneuvers were repeated up to three times. All the patients were called for a reassessment 30 days after the last intervention to assess the durability of the maneuver. In the study, 54.44% (49) of the 90 patients were female, whereas 45.56% (41) were male. Overall, 83.33% (75) of patients required only one attempt, 15.56% (14) required two attempts, and 1.11% (1) required three attempts to improve. In the Epley maneuvers group, 86.66% (26) required only one attempt, 10% (3) required two attempts, and 3.33% (1) required three attempts. Similarly, 83.33% (25) required only one attempt in the Gans maneuvers group, and 16.67% (5) required two attempts. In Semont maneuver groups, 80% (24) required only one attempt, and 20% (6) required two attempts. The recurrence of the symptoms was seen in a total of 11 patients: 27.27% (3 patients) of the Epley maneuvers group, 36.36% (4 patients) of the Gans maneuvers group, and 36.36% (4 patients) of the Semont maneuvers group. All three maneuvers show equal efficacy in reducing vertigo. The Epley maneuver may be more relevant in the treatment of BPPV compared to others, considering the slightly higher improvement rate and the requirement for fewer attempts for the treatment.

Visible light mediated synthesis of 1,3-diarylated imidazo[1,5-a]pyridines via oxidative amination of C-H catalyzed by graphitic carbon nitride.

Graphitic carbon nitride (g-C3N4) as a novel heterogeneous catalyst is employed for the visible light-mediated synthesis of the imidazo[1,5-a]pyridines via the oxidative amination of C-H bond at room temperature without the need for any additional solvent. Extensive characterization of the catalyst was performed using techniques such as FT-IR, PXRD, TGA, SEM and EDX analysis. The optimized conditions enabled the successful and expeditious conversion of a wide range of substrates to imidazo[1,5-a]pyridines in good yields; a notable advantage of this catalyst being recyclability, as it can be reused for up to five cycles without significant loss of activity. This feature makes it suitable for gram-scale synthesis of imidazo[1,5-a]pyridines. Additionally, this approach offers several benefits from a green chemistry perspective as affirmed by its favorable green chemistry metrics (GCM), including low process mass intensity (PMI), low E-factor, high atom economy (AE), and good reaction mass efficiency (RME) relative to existing protocols. In addition, chemical yield (CY), mass intensity (MI), mass productivity (MP) and optimum efficiency were also calculated. This environmentally friendly method offers multiple advantages and represents a significant advancement in the synthesis of imidazo[1,5-a]pyridines.

Recurrent Angiolymphoid Hyperplasia with Eosinophilia of the Parotid.

Angiolymphoid hyperplasia with eosinophilia (ALHE) is a benign vasculoproliferative disorder. It usually affects young adults presenting as papules or nodules involving the skin of head and neck region and rarely involves extracutaneous sites. ALHE involving parotid is rare and can be a diagnostic dilemma as it mimics a parotid neoplasm. This is a case of a 23 year old male presenting with a recurrent swelling over the left parotid region post surgery. Ultrasonography revealed a vascular soft tissue lesion in the preauricular region suggestive of a benign lesion. Contrast enhanced magnetic resonance imaging showed a hyperintense lesion involving the superficial lobe of the left parotid gland. Patient underwent superficial parotidectomy and histopathologically was diagnosed to have ALHE. Very few cases have been reported and this case is highlighted as the facial nerve was enmeshed by the intraparotid lesion which was a surgical challenge.

PLAE Web App Enables Powerful Searching and Multiple Visualizations Across One Million Unified Single-Cell Ocular Transcriptomes.

Purpose: To create a high-performance reactive web application to query single-cell gene expression data across cell type, species, study, and other factors. Methods: We updated the content and structure of the underlying data (single cell Eye in a Disk [scEiaD]) and wrote the web application PLAE (https://plae.nei.nih.gov) to visualize and explore the data. Results: The new portal provides quick visualization of over a million individual cells from vertebrate eye and body transcriptomes encompassing four species, 60 cell types, six ocular tissues, and 23 body tissues across 35 publications. To demonstrate the value of this unified pan-eye dataset, we replicated known neurogenic and cone macula markers in addition to proposing six new cone human region markers. Conclusions: The PLAE web application offers the eye community a powerful and quick means to test hypotheses related to gene expression across a highly diverse, community-derived database. Translational Relevance: The PLAE resource enables any researcher or clinician to study and research gene expression patterning across a wide variety of curated ocular cell types with a responsive web app.

Distinct neural encoding of glimpsed and masked speech in multitalker situations.

Humans can easily tune in to one talker in a multitalker environment while still picking up bits of background speech; however, it remains unclear how we perceive speech that is masked and to what degree non-target speech is processed. Some models suggest that perception can be achieved through glimpses, which are spectrotemporal regions where a talker has more energy than the background. Other models, however, require the recovery of the masked regions. To clarify this issue, we directly recorded from primary and non-primary auditory cortex (AC) in neurosurgical patients as they attended to one talker in multitalker speech and trained temporal response function models to predict high-gamma neural activity from glimpsed and masked stimulus features. We found that glimpsed speech is encoded at the level of phonetic features for target and non-target talkers, with enhanced encoding of target speech in non-primary AC. In contrast, encoding of masked phonetic features was found only for the target, with a greater response latency and distinct anatomical organization compared to glimpsed phonetic features. These findings suggest separate mechanisms for encoding glimpsed and masked speech and provide neural evidence for the glimpsing model of speech perception.

Extrafollicular IgD-CD27-CXCR5-CD11c- DN3 B cells infiltrate inflamed tissues in autoimmune fibrosis and in severe COVID-19.

Although therapeutic B cell depletion dramatically resolves inflammation in many diseases in which antibodies appear not to play a central role, distinct extrafollicular pathogenic B cell subsets that accumulate in disease lesions have hitherto not been identified. The circulating immunoglobulin D (IgD)-CD27-CXCR5-CD11c+ DN2 B cell subset has been previously studied in some autoimmune diseases. A distinct IgD-CD27-CXCR5-CD11c- DN3 B cell subset accumulates in the blood both in IgG4-related disease, an autoimmune disease in which inflammation and fibrosis can be reversed by B cell depletion, and in severe COVID-19. These DN3 B cells prominently accumulate in the end organs of IgG4-related disease and in lung lesions in COVID-19, and double-negative B cells prominently cluster with CD4+ T cells in these lesions. Extrafollicular DN3 B cells may participate in tissue inflammation and fibrosis in autoimmune fibrotic diseases, as well as in COVID-19.

A New Conformationally Symmetrical Calix[4]pyrrole Based Supramolecular System for Liquid Crystalline and Window Layer Solar Cell Applications.

The newly symmetrical liquid crystalline compounds (CPB1 -CPB4 ) based on calix[4]pyrrole as central rigid core are synthesized via esterification reaction. All the four functionalized compounds exhibit columnar hexagonal phase (Colh ) over a higher mesophase temperature range and further stabilized mesophase upto room temperature. The thermal behavior and optical texture are identified by using differential scanning calorimetry (DSC), Polarizing optical microscopy (POM) while the molecular organization of compound in mesogenic state by X-ray diffraction technique. The molecular system based on calix[4]pyrrole core with symmetrical nature exhibited columnar type self-assembly at room temperature. All these four supramolecules with different side spacer show higher thermal stability. Based upon the optimization, compound CPB2 has been further tested to implicate as optical window layer in thin films solar cell devices. The calix[4]pyrrole functionalized supramolecular liquid crystalline compound based thin films showed suitable transmittance, optical energy band gap together with absorbance and extinction coefficient. The linear dependence of current on the voltage demonstrated Ohmic behavior of the CPB2 films. The surface morphology to the developed samples designated nearly uniform deposition of the CPB2 thin films together with grain growth. The findings warrant suitability of the films to implicate these as an eco-friendly optical window layer in thin films based solar cells.

Thiacalixarene Appended Azo-based Supramolecular Systems: Self-assembly and Photo Tuning Reversible Liquid Crystalline Properties.

Four new azo-based supramolecular materials containing thiacalixarene core substituted by variable alkoxy groups (TFA1 -TFA4 ) have been designed and synthesized for the mesomorphic and photoswitching properties. The liquid crystalline behavior were accomplished by using DSC, POM, and XRD studies. All azo-based thiacalixarene based materials with short and higher chain length display columnar hexagonal mesophase with broad temperature range. The thermal behavior of all the materials was investigated by DSC and TGA study. The structural and conformational study of the lower rim functionalized materials was confirmed by using different techniques. These thiacalixarene moulded liquid crystalline compounds shows columnar self-assembly type behavior and higher thermal stability. The introduction of bi-substituted azo-ester network towards the lower rim of thiacalixarene core has impact on the electron delocalization and liquid crystalline properties. The photoswitching properties suggested cis and trans azo-isomerization under radiation of UV light and higher thermal back relaxation time. The mesogenic behaviour of compound TFA2 and TFA4 were demolished by the influence of cis and trans isomerization. The structure-property correlation is studied to understand the variation in mesogenic properties with the substitution of variable alkoxy side chain.

Fructose-induced topographical changes in fructophilic, pseudofructophilic and non-fructophilic lactic acid bacterial strains with genomic comparison.

Fructophilic Lactic Acid Bacteria (FLAB), Fructobacillus fructosus DPC7238 and pseudofructophilic Leuconostoc mesenteroides DPC7261 and non-FLAB Limosilactobacillus reuteri DSM20016 strains were studied for their growth and morphological evolution as a function of increased fructose concentrations (0, 25, and 50% w/v) in the media. A comparison of the genomics of these strains was carried out to relate observed changes and understand fructose-rich adaptations. The viability of FLAB strains were reduced by approx. 50% at a 50% fructose concentration, while the Limosilactobacillus reuteri strain was reduced to approx. 98%. Electron microscopy demonstrated that FLAB strain, Fructobacillus. fructosus and pseudofructophilic Leuc. mesenteroides, were intact but expanded in the presence of high fructose in the medium. Limosilactobacillus reuteri, on the other hand, ruptured as a result of excessive elongation, resulting in the formation of cell debris when the medium contained more than 25% (w/v) fructose. This was entirely and quantitatively corroborated by three-dimensional data obtained by scanning several single cells using an atomic force microscope. The damage caused the bacterial envelope to elongate lengthwise, thus increasing width size and lower height. The cell surface became comparatively smoother at 25% fructose while rougher at 50% fructose, irrespective of the strains. Although Fructobacillus fructosus was highly fructose tolerant and maintained topological integrity, it had a comparatively smaller genome than pseudofructophilic Leuc. mesenteroides. Further, COG analysis identified lower but effective numbers of genes in fructose metabolism and transport of Fructobacillus fructosus, essentially needed for adaptability in fructose-rich niches.

Fluorescently Guided Optical Photothermal Infrared Microspectroscopy for Protein-Specific Bioimaging at Subcellular Level.

Infrared spectroscopic imaging is widely used for the visualization of biomolecule structures, and techniques such as optical photothermal infrared (OPTIR) microspectroscopy can achieve <500 nm spatial resolution. However, these approaches lack specificity for particular cell types and cell components and thus cannot be used as a stand-alone technique to assess their properties. Here, we have developed a novel tool, fluorescently guided optical photothermal infrared microspectroscopy, that simultaneously exploits epifluorescence imaging and OPTIR to perform fluorescently guided IR spectroscopic analysis. This novel approach exceeds the diffraction limit of infrared microscopy and allows structural analysis of specific proteins directly in tissue and single cells. Experiments described herein used epifluorescence to rapidly locate amyloid proteins in tissues or neuronal cultures, thus guiding OPTIR measurements to assess amyloid structures at the subcellular level. We believe that this new approach will be a valuable addition to infrared spectroscopy providing cellular specificity of measurements in complex systems for studies of structurally altered protein aggregates.

Institution of Metal-Organic Frameworks as a Highly Sensitive and Selective Layer In-Field Integrated Soil-Moisture Capacitive Sensor.

The ongoing global industrialization along with the notable world population growth is projected to challenge the global environment as well as pose greater pressure on water and food needs. Foreseeably, an improved irrigation management system is essential and the quest for refined chemical sensors for soil-moisture monitoring is of tremendous importance. Nevertheless, the persisting challenge is to design and construct stable materials with the requisite sensitivity, selectivity, and high performance. Here, we report the introduction of porous metal-organic frameworks (MOFs), as the receptor layer, in capacitive sensors to efficiently sense moisture in two types of soil. Namely, our study unveiled that Cr-soc-MOF-1 offers the best sensitivity (≈24,000 pF) among the other tested MOFs for any given range of soil-moisture content, outperforming several well-known oxide materials. The corresponding increase in the sensitivities for tested MOFs at 500 Hz are ≈450, ≈200, and ≈30% for Cr-soc-MOF-1, Al-ABTC-soc-MOF, and Zr-fum-fcu-MOF, respectively. Markedly, Cr-soc-MOF-1, with its well-known water capacity, manifests an excellent sensitivity of ≈450% in clayey soil, and the analogous response time was 500 s. The noted unique sensing properties of Cr-soc-MOF-1 unveils the great potential of MOFs for soil-moisture sensing application.

SHP1 loss augments DLBCL cellular response to ibrutinib: a candidate predictive biomarker.

SHP1, a tyrosine phosphatase, negatively regulates B-cell receptor (BCR) signaling. Ibrutinib selectively inhibits BTK and has been approved for the treatment of several types of B-cell lymphomas, but not yet in diffuse large B-cell lymphoma (DLBCL). A phase 3 clinical trial of ibrutinib-containing regimen has been completed to evaluate its activity in subtypes or subsets of DLBCL patients. Although the subtype of activated B-cell like (ABC) DLBCL is characterized by chronic active BCR signaling, only a fraction of ABC-DLBCL patients seem to benefit from ibrutinib-containing regimen. New alternative predictive biomarkers are needed to identify patients who better respond. We investigated if SHP1 plays a role in defining the level of the BCR activity and impacts the response to ibrutinib. A meta-analysis revealed that lack of SHP1 protein expression as well as SHP1 promoter hypermethylation is strongly associated with NHL including DLBCL. On a tissue microarray of 95 DLBCL samples, no substantial difference in SHP1 expression was found between the GCB and non-GCB subtypes of DLBCL. However, we identified a strong reverse correlation between SHP1 expression and promoter methylation suggesting that promoter hypermethylation is responsible for SHP1 loss. SHP1 knockout in BCR-dependent GCB and ABC cell lines increased BCR signaling activities and sensitize lymphoma cells to the action of ibrutinib. Rescue of SHP1 in the knockout clones, on the other hand, restored BCR signaling and ibrutinib resistance. Further, pharmacological inhibition of SHP1 in both cell lines and patient-derived primary cells demonstrate that SHP1 inhibition synergized with ibrutinib in suppressing tumor cell growth. Thus, SHP1 loss may serve as an alternative biomarker to cell-of-origin to identify patients who potentially benefit from ibrutinib treatment. Our results further suggest that reducing SHP1 pharmacologically may represent a new strategy to augment tumor response to BCR-directed therapies. Schematic diagram summarizing the major findings. Left panel. When SHP1 is present and functional, it negatively regulates the activity of the BCR pathway. Right pane. When SHP1 is diminished or lost, cells depend more on the increased BCR signaling and making them vulnerable to BTK inhibitor, ibrutinib. Diagram was generated using BioRender.

Oncology phase I trial design and conduct: time for a change - MDICT Guidelines 2022.

In 2021, the Food and Drug Administration Oncology Center of Excellence announced Project Optimus focusing on dose optimization for oncology drugs. The Methodology for the Development of Innovative Cancer Therapies (MDICT) Taskforce met to review and discuss the optimization of dosage for oncology trials and to develop a practical guide for oncology phase I trials. Defining a single recommended phase II dose based on toxicity may define doses that are neither the most effective nor the best tolerated. MDICT recommendations address the need for robust non-clinical data which are needed to inform trial design, as well as an expert team including statisticians and pharmacologists. The protocol must be flexible and adaptive, with clear definition of all endpoints. Health authorities should be consulted early and regularly. Strategies such as randomization, intrapatient dose escalation, and real-world eligibility criteria are encouraged whereas serial tumor sampling is discouraged in the absence of a strong rationale and appropriately validated assay. Endpoints should include consideration of all longitudinal toxicity. The phase I dose escalation trial should define the recommended dose range for later testing in randomized phase II trials, rather than a single recommended phase II dose, and consider scenarios where different populations may require different dosages. The adoption of these recommendations will improve dosage selection in early clinical trials of new anticancer treatments and ultimately, outcomes for patients.

Prevalence of alcohol dependence syndrome in the patients of acute pancreatitis: A cross-sectional study.

Background: Acute pancreatitis (AP) is the main reason for mortality and morbidity. Numerous studies have shown a link between chronic alcohol usage and AP. However, there are few studies on the percentage of patients developing AP as a result of dependent patterns of drinking and associated risk factors. Aim: This study aimed to study the prevalence and impact of risk factors of alcohol dependence syndrome with AP patients hospitalized in tertiary care facilities. Materials and Methods: This is a cross-sectional observational study. Sociodemographic and clinical data were taken from patients with AP after consent. Eighty-five patients with AP who met the inclusion criteria were involved after each participant had clinical evaluation using the modified Marshall score, the Alcohol Use Disorder Identification Test, and the Severity of Alcohol Dependence Questionnaire (SAD-Q). The outcomes were tabulated and analyzed using Statistical Package for the Social Sciences (SPSS) software. Results: 38.8% of patients with AP were found to have features of alcohol dependence syndrome. Higher values of mean corpuscular volume (MCV), serum glutamic-oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), Gamma-glutamyl transferase (GGT), and uric acid were associated with a greater propensity to have AP. The severity of alcohol dependence syndrome and AP was not associated. Conclusion: AP is potentially a fatal disease. In this study, 38.8% of AP patients had alcohol dependence syndrome. There was no statistical association between the severity of AP and alcohol dependence syndrome, though high values of MCV, SGOT, SGPT, and GGT were at greater risk of developing AP. As a result, alcohol dependence syndrome should be examined in all individuals with AP.

Childhood trauma and bipolar affective disorder: Is there a linkage?

Bipolar affective disorder (BPAD) is a major psychiatric illness impairing the quality of life. The etiology of BPAD is influenced by different factors possibly related to gene-environment interactions. Approximately 30% to 50% of individuals with BPAD have experienced some traumatic event in childhood. Serious adverse experiences that children may suffer early in life are often described as childhood trauma (CT). It includes physical, emotional, and sexual abuse and physical and emotional neglect. CT is linked with an elevated risk of developing BPAD. Childhood adversities play a role in modulating the early onset of illness, increased number of depressive episodes, increased suicide attempts, and other clinical severity of BPAD. Hospital-based studies comprising heterogeneous populations had researched the specific role of each trauma subtype as a predisposing factor for BPAD. Identifying and addressing CT through early intervention methods may prevent the future development of chronic disorders like BPAD. This review article is an attempt to explore and highlight the existing literature regarding the association of different subtypes of CT with BPAD.