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Solid Organ Transplant (SOT) recipients are at significant higher risk for COVID-19 and due to immunosuppressive medication, the immunogenicity after vaccination is suboptimal. In the previous studies, booster method showed significant benefit in this population. In the current study, we compared using a mix-and-match method vs. same vaccine as a third dose in SOT recipients. This was a patient-blinded, single center, randomized controlled trial comparing BNT162b2 vs. JNJ-78436735 vaccine as the third dose after two doses of BNT162b2 vaccine. We included adult SOT recipients with functional graft who had received two doses of BNT162b2 vaccine. Participants were randomly assigned to receive either BNT162b2 or JNJ-78436735 in one-to-one ratio. Primary outcome was SARS-CoV-2 IgG positivity at 1 month after the third dose. Sixty SOT recipients, including 36 kidney, 12 liver, 2 lung, 3 heart, and 5 combined transplants, were enrolled, and 57 recipients were analyzed per protocol. There were no statistically significant differences between the two vaccine protocols for IgG positivity (83.3% vs. 85.2% for BNT162b2 and JNJ-78436735, respectively, p = 0.85, Odds Ratio 0.95, 95% Confidence Interval 0.23–4.00). Comparison of the geometric mean titer demonstrated a higher trend with BNT162b2 (p = 0.09). In this pilot randomized controlled trial comparing mix and match method vs. uniform vaccination in SOT recipients, both vaccines were safely used. Since this was a small sample sized study, there was no statistically significant difference in immunogenicity; though, the mix and match method showed relatively lower geometric mean titer, as compared to uniform vaccine. Further studies need to be conducted to determine duration of this immunogenicity.Clinical Trial Registration: https://clinicaltrials.gov/ct2/show/NCT05047640?term=20210641&draw=2&rank=1, identifier 20210641.

Cognitive dysfunction, sickness-like behavior, for instance, anxiety, and depression are common aspects of neuropsychiatry often associated with neurodegenerative disorders. Growing evidence suggests that high mobility group box 1 (HMGB1) may act as a proinflammatory cytokine that aggravates neurobehavioral dysfunction. However, the detailed underlying mechanism is still elusive. Here we focus on determining the relationship between lipopolysaccharide (LPS)-induced neuroinflammation (in both in vitro and in vivo models), cognitive dysfunction, sickness-like behavior and thus decode the impact of HMGB1 inhibition (using Glycyrrhizin; Gcy as an antagonist). Using a mice model of repeated LPS (1 mg/kg, i.p. for 4 days) injections, we found that LPS induced neurobehavioral deficit and a strong proinflammatory response with increased proinflammatory markers, including tumor necrosis factor-α (TNF-α), interleukin-1 beta (IL-1β), interleukin-6 (IL-6) and iNOS (inducible nitric oxide synthase) at 7 days after the final dose of LPS compared to control animals. Our findings suggest that neurobehavioral dysfunction strongly correlates with the proinflammatory immune response following LPS stimulation. In vitro Gcy pretreatment to LPS-activated BV2 microglia cells significantly reduced nitrite and reactive oxygen species production, along with diminished expression of classical proinflammatory cytokines (TNF-α, IL-1β, IL-6, iNOS). These key proinflammatory changes with LPS and Gcy treatment are also found in vivo mice model and correlate with improved cognitive function and reduced anxiety/depression. Together, these results show that blocking HMGB1 using Gcy abrogated the cognitive dysfunction, sickness-like behavior of anxiety and depression induced by LPS which can be a promising avenue for crucial neurobehavioral dysfunction.

Background: Sepsis and septic shock are still associated with a high mortality rate. Early-stage prediction of septic shock outcomes would be helpful to clinicians for designing their treatment protocol. In addition, it would aid the clinicians in patient management by understanding gender disparity in terms of clinical outcomes of septic shock by identifying whether there are sex-based differences in sepsis-associated mortality. Objective: This study aimed to test the hypothesis that gender-based metabolic heterogeneity is associated with sepsis survival and identify the biomarkers of mortality for septic shock in the Indian cohort. Method: The study is performed in an Indian population cohort diagnosed with sepsis/septic shock within 24 hours of admission. The study group includes 50 patients admitted in intensive care, of which 23 are females, and 27 are males. Univariate and multivariate analysis is performed to identify biomarkers for septic shock mortality and gender-specific metabolic fingerprint in septic shock-associated mortality. Results: Energy-related metabolites, ketone bodies, choline, and NAG were primarily responsible for differentiating survivors and non-survivors. The gender-based mortality stratification identified female-specific association of anti-inflammatory response, innate immune response and β oxidation and male-specific association of pro-inflammatory response to septic shock. Conclusion: These identified mortality biomarkers may help estimate the severity of the case, predicting outcome and treatment efficacy. The study underlines that gender is one of the most significant biological factors influencing septic shock metabolomic profiles. This understanding can be utilized to identify novel gender-specific biomarkers and innovative targets relevant for gender medicine.

Panchvalkal kwath (PK) is a bark formulation of five pharmacologically important plants, i.e., Ficus benghalensis, Ficus racemosa, Ficus religiosa, Thespesia populnea, and Ficus lacor. The Ayurvedic formulation is being used since ancient times to cure diabetes, bacterial infections and heal wounds. The present study aims to identify the metabolite profiles of PK which could explain its properties and its mode of action against specific diseases and disorders. The aqueous extract of Panchvalkal is prepared through a hot maceration process. The extract is subjected to preliminary identification of phytoconstituents and FTIR spectroscopy to recognize functional groups. GC-MS analysis reveals that the extract is enriched with 24-Norursa-3,12-diene (25.16%); Lup-20(29)-en-3-one (16.76%); 2-methyl-3-(4-propan-2-ylphenyl) propanal (7.04%); 2-(hydroxymethyl)-2-nitropropane-1,3-diol (11.21%) and 3,5-dihydroxy-6-methyl-2,3-dihydropyran-4-one (4.15%). The presence of three new phytocompounds that are 4-(hydroxymethyl)-7-methyl-1,3-dioxepane-5,6-diol; 1-(4-isopropylphenyl)-2-methylpropylacetate and 4,4,6 A,6B,8A,11,11,14B-octamethyl-1,4,4A,5,6,6A,8,8a,910,11,12,12a,12b,13,14,14a,14b-ctadecahydro-3(2H)-picenone are detected in the extract. Metabolite profiles of the extract also constitute isoeugenol, stigmasterol, ergosterol, ocimene, myrcene, squalene, sphingosine, betulin, methyl ferulate and cis-jasmone, which are unraveled by 1 D 1H and 2 D 1H-13C HSQC NMR spectroscopy. This article focuses on the presence of different phytocompounds in PK in order to demonstrate its efficacy as a therapeutic formulation for a variety of diseases.

Background During the ongoing Coronavirus disease of 2019 (COVID-19) pandemic, there have been increasing reports of viral, bacterial and fungal co-infections. Two COVID-19-associated fungal infections (CFIs) have been identified – COVID-19 associated pulmonary aspergillosis (CAPA) and COVID-19 associated mucormycosis (CAM), but incidence and occurrence in solid organ transplant recipients (SOTRs) is limited. We describe our experience with CFIs in SOTRs with COVID-19. Methods In a single center retrospective study at a large volume transplant center in South Florida, USA, we included adult SOTRs (≥18 years) diagnosed with COVID-19 between March 1st 2020 and January 31st 2022, with subsequent diagnosis of CFI. We collected information related to demographics, comorbidities, COVID-19 diagnosis and therapeutics, and CFI diagnostics and management. Data obtained was analyzed descriptively. Results We identified 612 SOTRs with COVID-19, of which 23 (3.8%) were diagnosed with CFIs. The patients were predominantly male (17/23, 73.9%), with median age of 59 years (range 43-79) [Table 1]. Twenty (86.9%) were kidney transplant recipients. Majority of SOTRs had lymphopenia (18/23, 78.3%) with elevated inflammatory markers at time of COVID-19 diagnosis. They received most commonly remdesivir and corticosteroids for COVID-19, with 22 (95.6%) needing intensive care unit admission and 19 (82.6%) needing continuous renal replacement therapy. CFIs were diagnosed at median 21 days (range, 3-161) after initial COVID-19 diagnosis. Probable CAPA was diagnosed in most patients (16/23, 69.6%), with CAM noted in 1 patient [Table 2]. 34.8% (8/23) had specific fungal species identified, with elevated fungal markers noted in 95.6% (22/23). Concurrent or prior cytomegalovirus DNAemia was noted in 26.1% (6/23). Patients were followed for median 70 days (range, 19-572), with median hospitalization duration 56 days (range, 7-204). Mortality was noted in 73.9% (17/23). Table 1:COVID-19 related clinical characteristics in study patients (N=23)Table 2:CFI-related clinical characteristics in study patients (N=23) Conclusion Fungal co-infections were noted in a small proportion of our SOTRs, with poor outcomes. Transplant physicians should have a high suspicion for early diagnosis and treatment of CFI. Further studies are needed to determine predictors for CFI and role for anti-fungal prophylaxis. Disclosures All Authors: No reported disclosures.

An early metabolic signature associated with the responsiveness to treatment can be useful in the better management of septic shock patients. This would help clinicians in designing personalized treatment protocols for patients showing non-responsiveness to treatment.We analyzed the serum on Day 1 (n = 60), Day 3 (n = 47), and Day 5 (n = 26) of patients with septic shock under treatment using NMR-based metabolomics. Partial least square discriminant analysis (PLS-DA) was performed to generate the list of metabolites that can be identified as potential disease biomarkers having statistical significance (that is, metabolites that had a VIP score 1, and p value 0.05, False discovery rate (FDR) 0.05).Common significant metabolites amongst the three time points were obtained that distinguished the patients being responsive (R) and non-responsive (NR) to treatments, namely 3 hydroxybutyrate, lactate, and phenylalanine which were lower, whereas glutamate and choline higher in patients showing responsiveness.The study gave these metabolic signatures identifying patients' responsiveness to treatment. The results of the study will aid in the development of targeted therapy for ICU patients.

Background: Growth retardation, malnutrition, and failure to thrive are some of the consequences associated with congenital heart diseases. Several metabolic factors such as hypoxia, anoxia, and several genetic factors are believed to alter the energetics of the heart. Timely diagnosis and patient management is one of the major challenges faced by the clinicians in understanding the disease and provide better treatment options. Metabolic profiling has shown to be potential diagnostic tool to understand the disease. Objective: The present experiment was designed as a single center observational pilot study to classify and create diagnostic metabolic signatures associated with the energetics of congenital heart disease in cyanotic and acyanotic groups. Methods: Metabolic sera profiles were obtained from 35 patients with cyanotic congenital heart disease (TOF) and 23 patients with acyanotic congenital heart disease (ASD and VSD) using high resolution 1D 1H NMR spectra. Univariate and multivariate statistical analysis were performed to classify particular metabolic disorders associated with cyanotic and acyanotic heart disease. Results: The results show dysregulations in several metabolites in cyanotic CHD patients versus acyanotic CHD patients. The discriminatory metabolites were further analyzed with area under receiver operating characteristic (AUROC) curve and identified four metabolic entities (i.e. mannose, hydroxyacetone, myoinositol, and creatinine) which could differentiate cyanotic CHDs from acyanotic CHDs with higher specificity. Conclusion: An untargeted metabolic approach proved to be helpful for the detection and distinction of disease-causing metabolites in cyanotic patients from acyanotic ones and can be useful for designing better and personalized treatment protocol.

Lung transplantation may be appropriate, and may offer benefit only to a carefully selected subset of morbidly ill patients afflicted by coronavirus disease-2019. Identifying the appropriate recipient for the allocation of scarce resources, by safely navigating through the challenges that are unique to lung transplantation for coronavirus disease-2019-associated acute respiratory distress syndrome, demands a conscientious and meticulous approach. Categorizing the respiratory failure in coronavirus disease-2019 may facilitate the process of evaluation for the purpose of transplant. The progress in rescue transplants over the past two decades has greatly improved our ability to successfully perform high-risk lung transplantation.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has been raging since the end of 2019 and has shown worse outcomes in solid organ transplant (SOT) recipients. The clinical differences as well as outcomes between respiratory viruses have not been well defined in this population.This is a retrospective cohort study of adult SOT recipients with nasopharyngeal swab or bronchoalveolar lavage PCR positive for either SARS-CoV-2, seasonal coronavirus, respiratory syncytial virus (RSV) or influenza virus from January 2017 to October 2020. The follow up period was 3 months. Clinical characteristics and outcomes were evaluated.A total of 377 recipients including 157 SARS-CoV-2, 70 seasonal coronavirus, 50 RSV and 100 influenza infections were identified. The most common transplanted organ was kidney 224/377 (59.4%). Lower respiratory tract infection (LRTI) was found in 210/377 (55.7%) and the risk factors identified with multivariable analysis were SARS-CoV-2 infection, steroid use, and older age. Co- and secondary infections were seen in 77/377 (20.4%) recipients with bacterial pathogens as dominant. Hospital admission was seen in 266/377 (67.7%) recipients without significant statistical difference among viruses, however, ICU admission, mechanical ventilation and mortality were higher with SARS-CoV-2 infection. In the multivariable model, the risk factors for mortality were SARS-CoV-2 infection and older age.We found higher incidence of ICU admission, mechanical ventilation, and mortality among SARS-CoV-2 infected recipients. Older age was found to be the risk factor for lower respiratory tract infection and mortality for SARS-CoV-2, coronaviruses, RSV and influenza virus groups.

Solid-state nuclear magnetic resonance (ssNMR) spectroscopy has grown to be one of the most precise and increasingly used spectroscopic tools to provide high-resolution structural insight into biological materials. This chapter focuses on recent developments in multidimensional ssNMR methods for biological materials such as bone, cartilage, tooth enamel, dentin, keratins, and other species such as tropical and deep-water corals and silk fibers at the atomic scale. The chapter discusses the current development in ssNMR methodologies used for the structure determination and dynamics study of such biological materials. The chapter also emphasizes the molecular-level structural information as well as studies related to the mechanisms of action of these biological materials.

Solid organ transplant (SOT) recipients are at high risk for severe disease with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Emerging variants of concern have disproportionately affected this population. Data on severity and outcomes with the Omicron variant in SOT recipients are limited. Thus we conducted this single-center, retrospective cohort study of SOT recipients diagnosed with SARS-CoV-2 infection from December 18, 2021 to January 18, 2022, when prevalence of the Omicron variant was more than 80%-95% in the community. Univariate and multivariate logistic regression analysis was performed to identify risk factors for hospital admission. We identified 166 SOT patients: 112 (67.5%) kidney, 22 (13.3%) liver, 10 (6.0%) lung, seven (4.2%) heart, and 15 (9.0%) combined transplants. SARS-CoV-2 vaccine series was completed in 59 (35.5%) recipients. Ninety-nine (59.6%) and 13 (7.8%) recipients received casirivimab/imdevimab and sotrovimab, respectively. Fifty-three (32%) recipients required hospital admission, of which 19 (35.8%) required intensive care unit level of care. Median follow-up was 50 (interquartile range, 25-59) days, with mortality reported in six (3.6%) patients. Risk factors identified for hospital admission were African American race (p.001, odds ratio [OR] 4.00, 95% confidence interval [CI] 1.84-8.70), history of coronary artery disease (p = .031, OR 3.50, 95% CI 1.12-10.87), and maintenance immunosuppression with corticosteroids (p = .048, OR 2.00, 95% CI 1.01-4.00). In conclusion, contrary to that in the general population, we found a higher hospital admission rate in SOT recipients with omicron variant infection. Further studies to investigate the efficacy of newer treatments are necessary, even as outcomes continue to improve.

Compared segments of the East and West Indian passive margins have different evolutions and crustal architecture. The East Indian margin is less magmatic. It results from a crust first–mantle second break-up scenario of a continent experiencing two rift events. The West Indian margin is more magmatic. It results from a mantle first–crust second break-up scenario of a continent experiencing four rift events. The architecture across both margins can be divided into stretching, thinning and hyperextension zones. The East Indian margin is characterized by oceanward-dipping listric normal faults that accommodate thinning in the thinning and hyperextension zones, and a zone of exhumed mantle separating continental and oceanic crusts. The West Indian margin in contrast is characterized by landward-dipping listric faults that accommodate magma-assisted thinning in the thinning and hyperextension zones, and no exhumed mantle. The final break-up affects the lithospheric mantle layer in the East Indian case and the crustal layer in the West Indian case. Although the temperature-dependent rheologies of these two last unbroken layers are somewhat different, seismic interpretation suggests that they are both broken by upward-convex normal faults, which succeeded the development of listric faults. They appear to be the first spontaneously formed faults in the break-up-delivering process, although their nucleation may be magma-assisted. The main difference between the controlling factors of the aforementioned break-up scenarios affecting similar lithospheres at similar extension rates is the cumulative length of time of the pre-break-up rift events, which is 62 and 115 myr for the East and West Indian margins, respectively.

Congenital heart disease (CHD) is one of the most important causes of the death of children and young adults. Most of the patients do not survive past their teen years. This occurs either due to delay in diagnosis or no diagnosis at all. In recent times, several studies have shown the importance of biomarkers in the prediction of such defects. These biomarkers give the real time snapshot of the on going processes inside the cells and can significantly support the diagnosis of CHD. The present experiment was designed as an observational single centre pilot study to identify and establish the diagnostic metabolic signatures associated with the congenital heart diseases. Metabolic profiles of sera collected from 35 cyanotic congenital heart disease patients and 15 controls were obtained using high-resolution 1D 1H CPMG and NMR spectra. The metabolic profiles were compared using multivariate statistical analysis to identify the disease specific metabolic disturbances associated with cyanotic heart disease. The results show perturbation in several metabolites in cyanotic CHD patients versus controls. The discriminatory metabolites were further analysedwith area under receiver operating characteristic (AUROC) curve and identified five metabolic entities (i.e.valine, glucose, glutamine, creatinineand PUFA) which could differentiate cyanotic CHDs from controls with higher specificity.In conclusion, untargeted metabolic approach proved to be helpful in identifying and differentiating disease causing metabolites in cyanotic cases from controls.

Diagnosis and management of patients with septic shock is still a significant challenge for clinicians with its high mortality amongst hospitalized patients. Septic shock is a heterogeneous condition and is usually accompanied by various underlying disease conditions. Dissecting the specific metabolic changes induced by these underlying disease conditions through metabolomics has shown the potential to improve our understanding of the disease's relevant pathophysiological mechanisms, leading to improved treatment. This study has shown the metabolic alterations caused due to co-morbid conditions like diabetes, hypertension, CAD, and CKD in septic shock. It has also shown the distinct metabolic profiles of septic shock patients with underlying respiratory illnesses and encephalopathy. Metabolic profiling of sera obtained from 50 septic shock patients and 20 healthy controls was performed using high-resolution 1D 1H CPMG and diffusion-edited NMR spectra. Univariate and multivariate statistical analyses were performed to identify the potential molecular biomarkers. Noted dysregulations in amino acids, carbohydrates, and lipid metabolism were observed in septic shock patients. Further stratification within the septic shock patients based on co-morbid conditions and primary diagnosis has shown their role in causing metabolic alterations. Evaluation of these compounds during treatment will help design a personalized treatment protocol for the patients, improving therapeutics.

Bone is a dynamic tissue composed of organic proteins (mainly type I collagen), inorganic components (hydroxyapatite), lipids, and water that undergoes a continuous rebuilding process over the lifespan of human beings. Bone mineral is mainly composed of a crystalline apatitic core surrounded by an amorphous surface layer. The supramolecular arrangement of different constituents gives rise to its unique mechanical properties, which become altered in various bone-related disease conditions. Many of the interactions among the different components are poorly understood. Recently, solid-state nuclear magnetic resonance (ssNMR) has become a popular spectroscopic tool for studying bone. In this article, we present a study probing the interaction of water molecules with amorphous and crystalline parts of the bone mineral through

Body fluids, cells, and tissues contain a wide variety of metabolites that consist of a mixture of various low-molecular-weight compounds, including amino acids, peptides, lipids, nucleic acids, and organic acids, which makes comprehensive analysis more difficult. Quantitative nuclear magnetic resonance (NMR) spectroscopy is a well-established analytical technique for analyzing the metabolic profiles of body fluids, cells, and tissues. It enables fast and comprehensive detection, characterization, a high level of experimental reproducibility, minimal sample preparation, and quantification of various endogenous metabolites. In recent times, NMR-based metabolomics has been appreciably utilized in diverse branches of medicine, including microbiology, toxicology, pathophysiology, pharmacology, nutritional intervention, and disease diagnosis/prognosis. In this review, the utility of NMR-based metabolomics in clinical studies is discussed. The significance of in vitro NMR-based metabolomics as an effective tool for detecting metabolites and their variations in different diseases are discussed, together with the possibility of identifying specific biomarkers that can contribute to early detection and diagnosis of disease.

Metabolic reprogramming, a key hallmark of cancer, plays a pivotal role in fulfilling the accelerated biological demands of tumor cells. Such metabolic changes trigger the production of several proinflammatory factors, thereby inciting cancer development and its progression. Serine protease inhibitor Kazal Type 1 (SPINK1), well known for its oncogenic role and its upregulation via acute-phase reactions, is highly expressed in multiple cancers including colorectal cancer (CRC). Here, we show accumulation of lipid droplets in CRC cells stained with Oil Red O upon SPINK1 silencing. Furthermore, NMR spectroscopy analysis revealed an accretion of monounsaturated fatty acids (MUFAs) and phosphatidylcholine in these CRC cells, while the levels of polyunsaturated fatty acids remained unaltered. This alteration indicates the presence of MUFAs with the triglycerides in the lipid droplets as observed in SPINK1-silenced CRC cells. Considering the role of MUFAs in the anti-inflammatory response, our data hint that suppression of SPINK1 in CRC leads to activation of an anti-inflammatory signaling milieu. Conclusively, our study uncovers a connection between lipid metabolism and SPINK1-mediated CRC progression, hence paving the way for further exploration and better prognosis of SPINK1-positive CRC patients.

Absorption spectra from 950-1170 cm-1 are measured at 500 Hz with a swept-wavelength external cavity quantum cascade to determine H2O temperatures and column densities in propagating H2/O2 flames and compared to computational fluid dynamic models.

Visualizing and quantifying cellular heterogeneity is of central importance to study tissue complexity, development, and physiology and has a vital role in understanding pathologies. Mass spectrometry-based methods including imaging mass cytometry (IMC) have in recent years emerged as powerful approaches for assessing cellular heterogeneity in tissues. IMC is an innovative multiplex imaging method that combines imaging using up to 40 metal conjugated antibodies and provides distributions of protein markers in tissues with a resolution of 1 μmMATISSE results in improved quality and quantity of segmented cells when compared to IMC-only segmentation in sections of heterogeneous tissues. Additionally, MATISSE enables more complete and accurate identification of epithelial cells, fibroblasts, and infiltrating immune cells in densely packed cellular areas in tissue sections. MATISSE has been designed based on commonly used open-access tools and regular fluorescence microscopy, allowing easy implementation by labs using multiplex IMC into their analysis methods.MATISSE allows segmentation of densely packed cellular areas and provides a qualitative and quantitative improvement when compared to IMC-based segmentation. We expect that implementing MATISSE into tissue section analysis pipelines will yield improved cell segmentation and enable more accurate analysis of the tissue microenvironment in epithelial tissue pathologies, such as autoimmunity and cancer.

Hydroxychloroquine and chloroquine are medications that have been used for a long time. Their most common use is for the treatment and prophylaxis of malaria. However, these antimalarial drugs are known to also have anti-inflammatory and antiviral effects and are used for several chronic diseases such as systemic lupus erythematosus with low adverse effects. The antiviral action of hydroxychloroquine and chloroquine has been a point of interest to different researchers due to its mechanism of action. Several in vitro studies have proven their effectiveness on severe acute respiratory syndrome virus and currently both in vitro and in vivo studies have been conducted on 2019 novel coronavirus (covid-19). The purpose of this article is to review the history and mechanism of actions of these drugs and the potential use they can have on the current covid-19 pandemic.

Drilling and reaching to deeper target zones through an overpressured overburden formation in a structurally complex geologic setting requires robust geologic and geomechanical analysis to mitigate risk and control operational costs. These types of geologic conditions are present in the Krishna-Godavari Basin, where a series of horst and grabens defined by deep-seated faults and persistent high sedimentation rates through geologic time, result in the development of challenging conditions for exploratory drilling. We have developed possible overpressure mechanisms across the central part of the Krishna-Godavari Basin and its interplay through fault-related lateral pressure transfer. The basin sits over a horst, which is one of the many northeast–southwest-trending en echelon horst and graben structures comprising sediments from the lower Cretaceous to Holocene. In the study area, Paleocene formations in the horst are overpressured (12–12.2 ppg). Three wells were drilled through this formation and reached the target without any drilling issues in the central and eastern part. However, the same formation in the western part of the horst (adjacent to the graben) has higher overpressure of approximately 14 ppg, which complicates the drilling operations because it requires an additional intermediate casing to reach the target reservoir safely. A detailed analysis of the overpressure mechanisms across the horst area to the adjacent deep graben revealed that the disequilibrium compaction signatures are related to the burial history and overburden thickness. The major difference between horst and graben area is the magnitude of overpressure, with an average of 16 ppg across the graben area. The larger overpressures experienced toward the western part of the horst indicate a secondary source of pressure from the adjacent deep graben. The fault stress analysis in this region presents a feasible lateral pressure transfer through critically stressed faults/fractures from the deep graben to the western part of the horst structure. The current model accounts the common pore pressure estimation method along with other critical geologic information to predict such overpressure related challenges in the upcoming future wells in a similar geologic setup to plan safe and cost-effective wells.

Summary Exploring tissue heterogeneity on a single-cell level by imaging mass cytometry (IMC) remains challenging because of its limiting resolution. We previously demonstrated that combining higher resolution fluorescence with IMC data in the analysis pipeline resulted in high-quality single-cell segmentation. Here, we provide a step-by-step workflow of this MATISSE pipeline, including instructions regarding the staining procedure, and the analysis route to generate single-cell data. For complete details on the use and execution of this protocol, please refer to Baars et al., 2021., Graphical abstract, Highlights • High-plex tissue staining combining isotope-labeled antibodies with DNA intercalator • Imaging mass cytometry (IMC) and fluorescent microscopy in a single workflow • Combined data processing pipeline of IMC and fluorescent images • The MATISSE pipeline generates high-quality single-cell segmentation of IMC data, Exploring tissue heterogeneity on a single-cell level by imaging mass cytometry (IMC) remains challenging because of its limiting resolution. We previously demonstrated that combining higher resolution fluorescence with IMC data in the analysis pipeline resulted in high-quality single-cell segmentation. Here, we provide a step-by-step workflow of this MATISSE pipeline, including instructions regarding the staining procedure, and the analysis route to generate single-cell data.

Drusen deposition on sub-retinal pigment epithelium is the causal factor for age-related macular degeneration for the old-aged individuals. These deposits contain hydroxyapatite-cholesterol spherules on which several proteins and lipids accumulate to cover the retina and choroid, causing blurred vision and blindness. Amyloid-β, the known culprit in Alzheimer's disease, is one among the few major proteins known to occur in these deposits. In the present article, we report preliminary analyses of interactions between amyloid-β and hydroxyapatite-cholesterol composites using Thioflavin-T binding kinetics, solid-state NMR and transmission electron microscopy (TEM). Thioflavin-T fluorescence kinetics shows that amyloid-β (1-42) aggregates only under certain conditions of concentration of cholesterol in the hydroxyapatite-cholesterol composites prepared by two different methods. These results were confirmed by 1D 13C CPMAS solid-state NMR. TEM imaging revealed that there is an exposure of the cholesterol surface in the composites prepared by sonication method. These imaging experiments explain the dependence of aggregation kinetics on the exposure and availability of cholesterol surface in the composites to a certain extent.