Retrospective study assessing the role of the androgen receptor in clear cell renal cell cancer patients treated with VEGFR inhibitors in monotherapy.

BACKGROUND AND PURPOSE: Despite that incorporating antiangiogenic in combination with immune-checkpoint inhibitors as the standard first-line treatment for advanced clear cell renal cell cancer (ccRCC) yields promising outcomes, these regimens often lead to significant toxicity. However, a subgroup of patients has shown responsiveness to VEGFR tyrosine-kinase inhibitors (TKIs) in monotherapy, leading to the question of whether employing combination therapies can significantly enhance overall survival in all patients over monotherapy. Thus, we aim to identify gene expression signatures that can predict TKI response within subpopulations that might benefit from single-agent therapies, to minimize unnecessary exposure to combination therapies and their associated toxicities, as well as to discover new potential therapeutic targets to improve ccRCC treatment. Based on prior data, the androgen receptor (AR) might meet both conditions. PATIENTS AND METHODS: We evaluated the association between AR expression, assessed through NanoString® technology-derived mRNA counts, and the clinical outcomes of 98 ccRCC patients treated with first-line antiangiogenics and determined its association with other genes implicated in ccRCC tumorigenesis. RESULTS: Higher AR-expression correlates significantly with better prognosis and survival based on the MSKCC risk score, and longer PFS. Furthermore, we have identified a gene set signature associated with AR-overexpression and several genes involved in angiogenesis and transcriptional targets of the hypoxia-inducible factor, a cornerstone of ccRCC. CONCLUSIONS: AR-overexpression and its association with other genes could favor a transcriptomic signature set to aid in identifying patients suitable for TKI in monotherapy, rather than aggressive combinations, enhancing thus, precision and personalized therapeutic decisions.

Turbine system vs engine generator in the production of energy by methane in the pacific of Mexico: a technical, economic, and environmental analysis.

This paper aims to analyze the potential of energy production using methane from organic waste as a sustainable alternative to mitigate the environmental impact of energy generation from fossil fuels and the generation of Municipal Solid Waste (MSW). To carry out the analysis, two technologies were evaluated from technical, economic, and environmental perspectives. The LandGEM model was used to estimate the methane generation potential. The amount of energy produced, along with the respective financial indicators, was also calculated. The results showed that for an average feed of 671,892 tons per year of waste, 6160 ft3/min of CH4 would be generated at a maximum peak in the tenth year, with an annual average of 4735 ft3/min. Additionally, 0.831 million metric tons of CO2 equivalent would be avoided. In terms of power generation, a combined cycle micro-turbine system with an installed capacity of 11.35 MW would be feasible and would yield an Internal Rate of Return (IRR) of 35% and a Net Present Value (NPV) of $11,608,006 USD. For an engine-generator system, it was not possible to verify profitability due to a significant increase in capital and O&M costs. These results are intended to provide reference information to assist in the decision-making process related to the implementation of projects aligned with the Sustainable Development Goals within the framework of the 2030 Agenda.

Copper nano metal-organic framework paper-based sensor for dual optical detection of biogenic amines to evaluate the food freshness.

The improvement of food safety and the reduction of food loss and waste require the development of new bioanalytical tools that provide chemical information about the composition of food that is of great value for improving traceability and extending the shelf life of food. Herein, a Cu-based metal-organic framework has been synthesized and immobilized onto cellulose paper disks for colorimetric and fluorescent detection and quantification of biogenic amines in food. The color of the nano metal-organic framework changes from green to brown in the presence of low amounts of biogenic amine vapors. Also, the fluorescence emission of the nano metal-organic framework greatly decreases after exposing the cellulose disks to amine vapors. The developed sensing paper disk exhibits a quick response to the presence of volatile biogenic amines, very low detection limits, and great selectivity. Also, the paper sensor was used for real-time monitoring of biogenic amines in bass samples at different temperature conditions, being a highly valuable method for evaluating food freshness and safeguarding food safety.

3D-printed tooth reduction guide for minimally invasive veneer preparations. A case report.

AIM: Current CAD/CAM techniques allow clinicians to design and 3D print tooth reduction guides that facilitate controlled preparations in the esthetic zone. The present case report describes the workflow for designing and printing a tooth reduction guide intended to assist ceramic veneer preparations. This 3D-printed guide divides the facial surface of the teeth into four areas and allows the measuring of all areas, including the incisal edge, with a periodontal probe. Using this 3D-printed appliance can simplify and expedite tooth preparation for ceramic veneers and reduce the risk of excessive tooth reduction. CLINICAL CONSIDERATIONS: A 45-year-old female patient presented at the clinic with the primary concern of the esthetics of her smile, which she wanted to improve. The initial evaluation revealed clinical signs of wear, spaces between the teeth, and the lack of anterior tooth symmetry. Therefore, to improve the appearance of her anterior teeth, a treatment plan that consisted of veneer restorations from the maxillary right second premolar to the left second premolar was formulated and presented. The patient accepted the option of undergoing minimally invasive anterior tooth preparations with a digital tooth reduction guide. MATERIALS AND METHODS: A diagnostic intraoral scan was performed and a tooth reduction guide was digitally designed and 3D printed. Subsequently, the tooth preparations were performed with the aid of the guide. An intraoral scan of the preparations was performed and used to design and manufacture lithium disilicate ceramic veneers using subtractive techniques. Subsequently, the restorations were finished, treated, and cemented under rubber dam isolation. CONCLUSIONS: The treatment fulfilled the patient's esthetic demands. This novel 3D-printed tooth reduction guide permits a more comprehensive assessment of the available tooth structure and provides ample access to more tooth surfaces than traditional silicone tooth reduction guides. Overall, this type of guide can significantly assist clinicians in performing tooth preparations in the esthetic zone.

Long-term nitrogen deposition reduces the diversity of nitrogen-fixing plants.

Biological nitrogen fixation is a fundamental part of ecosystem functioning. Anthropogenic nitrogen deposition and climate change may, however, limit the competitive advantage of nitrogen-fixing plants, leading to reduced relative diversity of nitrogen-fixing plants. Yet, assessments of changes of nitrogen-fixing plant long-term community diversity are rare. Here, we examine temporal trends in the diversity of nitrogen-fixing plants and their relationships with anthropogenic nitrogen deposition while accounting for changes in temperature and aridity. We used forest-floor vegetation resurveys of temperate forests in Europe and the United States spanning multiple decades. Nitrogen-fixer richness declined as nitrogen deposition increased over time but did not respond to changes in climate. Phylogenetic diversity also declined, as distinct lineages of N-fixers were lost between surveys, but the "winners" and "losers" among nitrogen-fixing lineages varied among study sites, suggesting that losses are context dependent. Anthropogenic nitrogen deposition reduces nitrogen-fixing plant diversity in ways that may strongly affect natural nitrogen fixation.

Beta cell-specific PAK1 enrichment ameliorates diet-induced glucose intolerance in mice by promoting insulin biogenesis and minimising beta cell apoptosis.

AIMS/HYPOTHESIS: p21 (CDC42/RAC1) activated kinase 1 (PAK1) is depleted in type 2 diabetic human islets compared with non-diabetic human islets, and acute PAK1 restoration in the islets can restore insulin secretory function ex vivo. We hypothesised that beta cell-specific PAK1 enrichment in vivo can mitigate high-fat-diet (HFD)-induced glucose intolerance by increasing the functional beta cell mass. METHODS: Human islets expressing exogenous PAK1 specifically in beta cells were used for bulk RNA-seq. Human EndoC-ßH1 cells overexpressing myc-tagged PAK1 were used for chromatin immunoprecipitation (ChIP) and ChIP-sequencing (ChIP-seq). Novel doxycycline-inducible beta cell-specific PAK1-expressing (ißPAK1-Tg) mice were fed a 45% HFD pre-induction for 3 weeks and for a further 3 weeks with or without doxycycline induction. These HFD-fed mice were evaluated for GTT, ITT, 6 h fasting plasma insulin and blood glucose, body composition, islet insulin content and apoptosis. RESULTS: Beta cell-specific PAK1 enrichment in type 2 diabetes human islets resulted in decreased beta cell apoptosis and increased insulin content. RNA-seq showed an upregulation of INS gene transcription by PAK1. Using clonal human beta cells, we found that PAK1 protein was localised in the cytoplasm and the nucleus. ChIP studies revealed that nuclear PAK1 enhanced pancreatic and duodenal homeobox1 (PDX1) and neuronal differentiation 1 (NEUROD1) binding to the INS promoter in a glucose-responsive manner. Importantly, the ißPAK1-Tg mice, when challenged with HFD and doxycycline induction displayed enhanced glucose tolerance, increased islet insulin content and reduced beta cell apoptosis when compared with ißPAK1-Tg mice without doxycycline induction. CONCLUSIONS/INTERPRETATION: PAK1 plays an unforeseen and beneficial role in beta cells by promoting insulin biogenesis via enhancing the expression of PDX1, NEUROD1 and INS, along with anti-apoptotic effects, that culminate in increased insulin content and beta cell mass in vivo and ameliorate diet-induced glucose intolerance. DATA AVAILABILITY: The raw and processed RNA-seq data and ChIP-seq data, which has been made publicly available at Gene Expression Omnibus (GEO) at https://www.ncbi.nlm.nih.gov/geo/ , can be accessed in GSE239382.

ATP-induced reconfiguration of the micro-viscoelasticity of cardiac and skeletal myosin solutions.

We study the high-frequency, micro-mechanical response of suspensions composed of cardiac and skeletal muscle myosin by optical trapping interferometry. We observe that in low ionic strength solutions, upon the addition of magnesium adenosine triphosphate (MgATP2-), myosin suspensions radically change their micro-mechanics properties, generating a viscoelastic fluid characterized by a complex modulus similar to a suspension of worm-like micelles. This transduction of energy, from chemical to mechanical, may be related to the relaxed states of myosin, which regulate muscle contractility and can be involved in the etiology of many myopathies. Within an analogous generic mechanical response, cardiac and skeletal myosin suspensions provide different stress relaxation times, elastic modulus values, and characteristic lengths. These discrepancies probably rely on the dissimilar physiological functions of cardiac and skeletal muscle, on the different MgATPase hydrolysis rates of cardiac and skeletal myosins, and on the observed distinct cooperative behavior of their myosin heads in the super-relaxed state. In vitro studies like these allow us to understand the foundations of muscle cell mechanics on the micro-scale, and may contribute to the engineering of biological materials whose micro-mechanics can be activated by energy regulators.

Shock Waves for Hair Growth and Regeneration in Patients With Female Pattern Hair Loss.

BACKGROUND: Female pattern hair loss (FPHL) is women's most common form of alopecia. The study aimed to evaluate the efficacy of extracorporeal shock waves for hair growth and regeneration in women with FPHL. METHODS: A prospective single-center noncontrolled study was conducted in patients with FPHL who underwent eight treatment sessions with C-Actor, a shock wave applicator. Outcome measures included total hair count, number of follicular units (FUs)-single FUs, double FUs, triple and bigger FUs-the average thickness, cumulative hair thickness, and changes in the Derived Sinclair scale, assessed with Tricholab (Fotofinder System). Patients completed a survey to assess their subjective perceptions of hair improvement regarding softness, hair loss, quality, and volume. RESULTS: Twenty women were included with a mean age of 43 (SD = 5.65). The mean increase in total hair count was 8.9% (p < 0.00001), in number of FUs 4.6% (p = 0.00031), in number of triple and bigger FUs 18.4% (p = 0.0004), in cumulative hair thickness 8.0% (p < 0.00001), with a decrease of 5.7% in mean Derived Sinclair scale value by 2.2-2.1 (p < 0.0001). After 3 months, 85% of patients perceived improvement in hair volume. CONCLUSION: Extracorporeal shock waves for hair growth and capillary regeneration in women with FPHL showed statistically significant outcomes, and all patients reported an improvement in hair characteristics.

Wearable Devices Based on Bioimpedance Test in Heart-Failure: Design Issues.

Heart-failure (HF) is a severe medical condition. Physicians need new tools to monitor the health status of their HF patients outside the hospital or medical supervision areas, to better know the evolution of their patients' main biomarker values, necessary to evaluate their health status. Bioimpedance (BI) represents a good technology for sensing physiological variables and processes on the human body. BI is a non-expensive and non-invasive technique for sensing a wide variety of physiological parameters, easy to be implemented on biomedical portable systems, also called "wearable devices". In this systematic review, we address the most important specifications of wearable devices based on BI used in HF real-time monitoring and how they must be designed and implemented from a practical and medical point of view. The following areas will be analyzed: the main applications of BI in heart failure, the sensing technique and impedance specifications to be met, the electrode selection, portability of wearable devices: size and weight (and comfort), the communication requests and the power consumption (autonomy). The different approaches followed by biomedical engineering and clinical teams at bibliography will be described and summarized in the paper, together with results derived from the projects and the main challenges found today.

Wearable Devices Based on Bioimpedance Test in Heart Failure: Clinical Relevance: Systematic Review.

Background: Heart failure (HF) represents a frequent cause of hospital admission, with fluid overload directly contributing to decompensations. Bioimpedance (BI), a physical parameter linked to tissue hydration status, holds promise in monitoring congestion and improving prognosis. This systematic review aimed to assess the clinical relevance of BI-based wearable devices for HF fluid monitoring. Methods: A systematic review of the published literature was conducted in five medical databases (PubMed, Scopus, Cochrane, Web of Science, and Embase) for studies assessing wearable BI-measuring devices on HF patients following PRISMA recommendations on February 4th, 2024. The risk of bias was evaluated using the ROBINS tool. Results: The review included 10 articles with 535 participants (mean age 66.7 ± 8.9 years, males 70.4%). Three articles identified significant BI value differences between HF patients and controls or congestive vs non-congestive HF patients. Four articles focused on the devices' ability to predict HF worsening-related events, revealing an overall sensitivity of 70.0 (95% CI 68.8-71.1) and specificity of 89.1 (95% CI 88.3-89.9). One article assessed prognosis, showing that R80kHz decrease was related to all-cause-mortality with a hazard ratio (HR) of 5.51 (95% CI 1.55-23.32; p = 0.02) and the composite all-cause-mortality and HF admission with a HR of 4.96 (95% CI 1.82-14.37; p = 0.01). Conclusions: BI-measuring wearable devices exhibit efficacy in detecting fluid overload and hold promise for HF monitoring. However, further studies and technological improvements are required to optimize their impact on prognosis compared to standard care before they can be routinely implemented in clinical practice. PROSPERO Registration: The search protocol was registered at PROSPERO (CRD42024509914).

Preliminary Metabolomics Study Suggests Favorable Metabolic Changes in the Plasma of Breast Cancer Patients after Surgery and Adjuvant Treatment.

Background/Objectives: The management of early breast cancer (BC) includes surgery, followed by adjuvant radiotherapy, chemotherapy, hormone therapy, or immunotherapy. However, the influence of these interventions in metabolic reprogramming remains unknown. This study explored alterations in the plasma metabolome of BC patients following distinct treatments to deepen our understanding of BC pathophysiology, outcomes, and the identification of potential biomarkers. Methods: We included 52 women diagnosed with BC and candidates for surgery as primary oncological treatment. Blood samples were collected at diagnosis, two weeks post-surgery, and one month post-radiotherapy. Plasma samples from 49 healthy women served as controls. Targeted metabolomics assessed 74 metabolites spanning carbohydrates, amino acids, lipids, nucleotide pathways, energy metabolism, and xenobiotic biodegradation. Results: Before treatment, the BC patients exhibited notable changes in carbohydrate, nucleotide, lipid, and amino acid metabolism. We noticed a gradual restoration of specific metabolite levels (hypoxanthine, 3-phosphoglyceric acid, xylonic acid, and maltose) throughout different treatments, suggesting a normalization of the nucleotide and carbohydrate metabolic pathways. Moreover, we observed increased dodecanoic acid concentrations, a metabolite associated with cancer protection. These variations distinguished patients from controls with high specificity and sensitivity. Conclusions: Our preliminary study suggests that oncological treatments modify the metabolism of patients towards a favorable profile with a decrease in the pathways that favor cell proliferation and an increase in the levels of anticancer molecules. These findings emphasize the pivotal role of metabolomics in recognizing the biological pathways influenced by each cancer treatment and the resulting metabolic consequences. Furthermore, it aids in identifying potential biomarkers for disease onset and progression.

Outcomes of controlled DCDD lung transplantation after thoraco-abdominal vs abdominal normothermic regional perfusion: The Spanish experience.

BACKGROUND: Thoraco-abdominal normothermic regional perfusion (TA-NRP) has emerged as a strategy for evaluating and recovering the heart in controlled donation after the circulatory determination of death (cDCDD). However, its impact on lung grafts remains largely unknown. We aimed to assess the impact of TA-NRP on the outcomes of recipients of cDCDD lungs. METHODS: This is a retrospective, multicenter, nationwide study describing the outcomes of cDCDD lung transplants (LTs) performed in Spain from January 2021 to November 2023. Patients were divided in 2 groups based on the recovery technique: TA-NRP with the simultaneous recovery of the heart vs abdominal NRP (A-NRP) without simultaneous heart recovery. The primary endpoint was the incidence of Primary Graft Dysfunction (PGD) grade 3 at 72 hours. Secondary endpoints included the overall incidence of PGD, days on mechanical ventilation, intensive care unit (ICU) and hospital length of stay, early survival rates, and mid-term outcomes. RESULTS: Two hundred and eighty three cDCDD LTs were performed during the study period, 28 (10%) using TA-NRP and 255 (90%) using A-NRP. No differences were observed in the incidence of PGD grade 3 at 72 hours between the TA-NRP and the A-NRP group (0% vs 7.6%; p = 0.231), though the overall incidence of PGD was significantly lower with TA-NRP (14.3% vs 41.5%; p = 0.005). We found no significant differences between the groups regarding other post-transplant outcome variables. CONCLUSIONS: TA-NRP allows the simultaneous recovery of both the heart and the lungs in the cDCDD scenario with appropriate LT outcomes comparable to those observed with the A-NRP approach.

Three-Month Mortality in Nonhaematological Patients with Chronic Pulmonary Aspergillosis: Differences between Subtypes.

Objectives: Chronic pulmonary aspergillosis (CPA) is a fungal lung infection characterised by the slowly progressing destruction of the lung parenchyma and has four main subtypes. The objective of this work was to evaluate the epidemiology of CPA in our area and evaluate the involvement of the different subtypes in mortality. Methods: This was a descriptive longitudinal retrospective study developed in three tertiary hospitals in Spain. Among all patients admitted with a pulmonary aspergillosis diagnosis, we selected those who fulfilled the criteria for chronic aspergillosis according to the criteria of Denning, excluding those with a haematological disorder. Results: Among 409 inpatients recorded as having a pulmonary aspergillosis infection, only 76 (18.5%) fulfilled the criteria for CPA, with an estimated incidence of 0.67 cases/100,000 inhabitants/year. The subtypes detected were subacute invasive aspergillosis (SAIA) in 33 (43.4%) patients, simple aspergilloma (SA) in 25 (32.9%) patients, cavitary chronic aspergillosis (CCPA) in 13 (17.1%) patients, and chronic fibrosis (CFPA) in five (6.5%) patients. The overall three-month mortality rate was 23%, which was higher in SAIA patients. The predictors of early mortality were age > 65 years (OR 3.0 CI 95 1.0-9.5 p = 0.043) and the SAIA subtype vs. other subtypes (OR 3.1 CI 95 1.0-9.5 p = 0.042). Conclusions: The incidence rate estimated was inferior to that previously reported. The three-month mortality in patients with CPA was high, with older age and the SAIA subtype being the variable independent predictors of a worse prognosis.

Malignant transformation of oral leukoplakia: Systematic review and comprehensive meta-analysis.

OBJECTIVES: To update the current evidence on the malignant transformation of oral leukoplakia (OL), including all studies published worldwide on the subject, selected with the maximum rigor regarding eligibility. MATERIALS AND METHODS: MEDLINE, Embase, Web of Science and Scopus were searched for studies published before June-2024, with no lower date limit. The risk of bias was analyzed using the Joanna Briggs Institute tool for meta-analyses of proportions. We carried out meta-analyses, explored heterogeneity across subgroups and identified risk factors with potential prognostic value. RESULTS: Fifty-five studies (41,231 with OL) were included. The pooled malignant transformation proportion for OL was 6.64% (95% CI = 5.21-8.21). The malignant transformation did not significantly vary by time periods (p = 0.75), 5.35% prior to 1978, 7.06% from 1979 to 2007 and 6.97% during more recent times. The risk factors that significantly had a higher impact on malignant transformation were the non-homogeneous leukoplakias (RR = 4.23, 95% CI = 3.31-5.39, p < 0.001), the larger size (RR = 2.08, 1.45-2.96, p < 0.001), leukoplakia located on the lateral border of tongue (malignant transformation = 12.71%; RR = 2.09, 95% CI = 1.48-2.95, p < 0.001), smoking (RR = 1.64, 95% CI = 1.25-2.15, p < 0.001), and the presence of epithelial dysplasia (RR = 2.75, 95% CI = 2.26-3.35, p < 0.001). CONCLUSIONS: OL presents a considerable malignant transformation probability that is especially increased in large non-homogeneous lesions in smokers, located on the lateral border of the tongue, with epithelial dysplasia.

Modelling the Distribution and Habitat Suitability of the European Wildcat (Felis silvestris) in North-Western Spain and Its Conservation Implications.

Human activities have resulted in severe habitat degradation and fragmentation at a global scale. Despite this scenario, some carnivore species that adapted to the new conditions are expanding, leading to close coexistence with humans and the emergence of potential conflicts. In this work, we used a European wildcat (Felis silvestris) observations database of more than 350 sightings over 17 years in NW Spain to build suitability models based on environmental, topographic, climatic, and human impact variables. MaxEnt was used to analyse the availability of suitable habitats for the species at a regional scale. Our results showed that less than one third of the suitable area for the species had confirmed wildcat presence. Elevation, the percentage of forested area, and footpath density were the three main variables conditioning wildcat presence, with the first two variables having positive effects and footpath density negatively affecting wildcat presence. The selection of high areas and forest areas by the species seems to be related to food availability, while the avoidance of footpaths seems to be related to the fact that main mortality causes are linked to human disturbances. The results enhance the understanding of the European wildcat ecology and provide insight into potential management plans to ensure the conservation of one of the main populations of the species throughout its range.

Adaptive and innate immune response of Leishmania infantum infection in Cirneco dell'Etna dog breed.

Leishmania spp. are an intracellular protozoa present in many countries around the world. In Europe, both the parasite and the disease it causes, leishmaniasis, are endemic in the Mediterranean basin. Clinical signs and severity of disease are highly variable depending on the host in both humans and dogs, traditionally considered the main reservoir of the parasite. The reason for these differences is not known, but it has been speculated that some hosts present immune response, related to activation of Th1 and Th17, capable of controlling the spread of the parasite, and that these immune responses are related to the genetic background of the host. The Ibizan hound, an autochthonous canine breed of the Mediterranean basin, has been postulated as a breed resistant to infection, but other canine breeds evolutionarily close to it and native to this region have not been studied. One of them is the Cirneco dell'Etna, native to the island of Sicily in southern Italy. In this study, the immune response against L. infantum infection in this canine breed was analysed. The results showed that infected dogs of this breed present high levels of several cytokines related to Th1 and Th17 immune response, and significant correlation between serum levels of cytokines related to disease resistance. Further studies are necessary in this canine breed to determine the mechanisms of immune response and genetic background related to L. infantum infection control.

Serological and molecular detection of Toxoplasma gondii infection in apparently healthy horses in eastern of Spain.

Toxoplasmosis is one of the most common parasitic zoonoses and represents a significant health risk for humans, especially for immunodeficient patients. The main transmission route is by oral uptake of oocysts and consumption of undercooked meat of infected animals. Different species have been evaluated as possible reservoirs of the parasite, but few studies have been carried out to examine the role of horses in transmission of the disease. Given the proximity of these animals to humans and the widespread consumption of their meat in many countries, including the Mediterranean basin, it is important to determine the prevalence of T. gondii infection in this species. In this study, blood samples from 105 horses were collected and the presence of T. gondii was evaluated by serological and molecular methods. Antibodies against T. gondii of 12 horses (11.43%) were detected by enzyme-linked immunosorbent assay (ELISA), whereas 29 horses (27.62%) showed positive for PCR. Seroprevalence was related to use of the animals, being higher in horses used for dressage than in others. Purebreds had higher seroprevalence than crossbred animals. No differences between breed, sex or age were found. The results of this study confirm the presence of T. gondii infection in horses, highlighting the need to analyse the meat of this species before human consumption and to control of this infection in horses, as they could be an important reservoir of this zoonotic parasite.

Characterization of Human Pancreatic Islet Cells using a Single-Cell Western Blot Platform.

OBJECTIVE: Islet transplantation is an effective treatment for type 1 diabetes. However, transplant success depends on quick islet assessment because islets deteriorate 2-3 days after isolation. A new tool, single-cell Western blot (scWestern), offers results within one day. In this study, we aimed to test the suitability of scWestern to detect protein markers for beta (insulin), alpha (glucagon), and delta (somatostatin) cells, the three major endocrine cell types in islets. METHODS: We characterized the antibody specificity, signal intensity, and cell identification on the scWestern platform, and then compared the islet cell composition analysis between scWestern and immunohistochemistry performed by the Integrated Islet Distribution Program (IIDP). RESULTS: Islet cell composition is comparable for alpha and beta cells, but not delta cells. Protein expression levels of insulin, glucagon, and somatostatin in individual islet cells varied greatly, highlighting cell type heterogeneity. Surprisingly, scWestern revealed double-hormonal cells (~1%), co-expressing insulin and somatostatin or insulin and glucagon, in non-diabetic and non-obese adult human islets, which was confirmed by confocal immunofluorescence microscopy. CONCLUSIONS: These results demonstrate that each alpha, beta, and delta cells express varying levels of peptide hormones, and a small subpopulation co-expresses double hormones in normal human islets. The scWestern platform will enable timely assessment of beta cell mass in isolated islets before clinical transplantation.