Patients affected by inflammatory bowel disease (IBD) exhibit compromised intestinal barrier function and decreased mucus accumulation, as well as increased inflammation, fibrosis and cancer risk, with symptoms often being exacerbated in women during pregnancy. Here we replicate these IBD hallmarks in human-derived organ-on-a-chip devices lined by colon epithelial cells from individuals with IBD when interfaced with matched fibroblasts, cultured under flow, exposed to peristalsis-like motions and perfused with circulating immune cells. Use of heterotypic tissue recombinants revealed that IBD fibroblasts are the primary drivers of multiple IBD symptoms. In the IBD chip, inflammation and fibrosis are accentuated by peristalsis-like motions and, in female-derived chips, also by exposure to pregnancy-associated hormones. When exposed to carcinogens, the IBD chip shows increased inflammation, gene mutations and chromosome duplication, in contrast to healthy chips. These data suggest that the intestinal stroma, sex hormones and peristalsis-associated mechanical deformations have a key role in driving inflammation, fibrosis and disease progression in male and female individuals with IBD.

Flavonoids, a diverse class of polyphenols found in many plant-based foods, are increasingly recognized for their health benefits, including antioxidant and anti-inflammatory activities. Recent evidence links higher dietary flavonoid diversity with reduced all-cause mortality and improved outcomes in chronic diseases. Yet, due to concerns about non-specific binding, their exclusion from drug development pipelines has limited deeper mechanistic understanding. Here, we show how flavonoids may promote cellular resilience by forming supramolecular assemblies that interact with proteins. Using molecular dynamics simulations (MDS) and in vitro assays, we found that different flavonoids self-assemble into ordered structures that influence protein structural dynamics and fiber formation. These structures have differential effects on enzyme activity and cell viability under stress. The ability to undergo supramolecular assembly may be important as flavonoids protect human cells against ultraviolet radiation-induced damage through a non-antioxidant mechanism. These findings suggest that supramolecular assembly and structural heterogeneity of flavonoids may underlie their diverse bioactivities and help to explain how the diversity of dietary flavonoids can support adaptive changes in cellular biochemistry, enhance resilience to environmental stressors, and improve human health.

Inflammatory bowel disease (IBD) patients exhibit compromised intestinal barrier function and decreased mucus accumulation, as well as increased inflammation, fibrosis, and cancer risk, with symptoms often being exacerbated in women during pregnancy. Here, we show that these IBD hallmarks can be replicated using human Organ Chips lined by IBD patient-derived colon epithelial cells interfaced with matched fibroblasts cultured under flow. Use of heterotypic tissue recombinants revealed that IBD fibroblasts are the primary drivers of multiple IBD symptoms. Inflammation and fibrosis are accentuated by peristalsis-like motions in IBD Chips and when exposed to pregnancy-associated hormones in female IBD Chips. Carcinogen exposure also increases inflammation, gene mutations, and chromosome duplication in IBD Chips, but not in Healthy Chips. These data enabled by human Organ Chip technology suggest that the intestinal stroma, sex hormones, and peristalsis-associated mechanical deformations play a key role in driving inflammation, fibrosis, and disease progression in male and female IBD patients.

Abstract In mice and one human case study, Borrelia burgdorferi (Bb), the causative agent of Lyme disease (LD), disrupts germinal center architecture and function. Bb proteins also mimic self-antigens. Thus, the antibody response may be of poor quality and target self antigens. To understand this pathology and its relevance to human infection-induced autoimmune diseases, we infected human lymphoid organ chips (Goyal et al., Adv Sci., 2022) created from healthy volunteers with Bb. We discovered unexpected and severe suppression of the adaptive immune system by Bb. T cell and B cell activation were absent in all of the donors (n>10) activated by Bb. Further, T cell proliferation was suppressed by Bb. Yet, some donors displayed hypergammaglobulinemia (hyperIg), seen in many antibody-mediated autoimmune diseases. We collected the exosomes secreted from the lymphoid organ chips and performed mass spectrometry to identify proteins associated with Bb infection and hyperIg. A heat-killed, fixed suspension of Staph. aureus Cowan I (SAC), commonly used to benchmark patient immune responses, was used as a control. We identified complement and coagulation factors, chaperone proteins, and immunological synapse proteins as key players in discriminating between successful activation, immunosuppression, and hyperIg (Bb). Our studies establish an innovative way to study human infectious diseases and their role in autoimmunity and reveal novel therapeutic targets to treat post-treatment Lyme disease.

IntroductionAutonomic and sensory neuropathy have been observed in both prediabetes and manifest diabetes mellitus. However, there is a lack of available data regarding whether patients at a moderate or high risk of developing diabetes, yet without a current diagnosis of prediabetes or diabetes, exhibit an increased prevalence of neuropathy.MethodsFINDRISC (Finnish Diabetes Risk Score) was used to classify individuals at risk (≥12 points, n = 44; control <12 points, n = 28). HbA1c levels >5.6% served as exclusion criteria, and patients with known medical conditions predisposing to neuropathy were also excluded. Cardiac autonomic function (Ewing tests) and peripheral sensory neuropathy (Neurometer and Q-sense) were assessed by standardized protocols, and their potential association with increased FINDRISC points was analyzed using a regression model.ResultsMean age was 46.7 ± 14.3 years in the control and 55.7 ± 14.1 years in the increased risk group. Male/female ratio did not differ. Individuals with increased risk of diabetes were more obese (BMI: 29.9 ± 12.5 kg/m2 vs. 25.9 ± 8.9 kg/m2). Additionally, hypertension was more frequent among them (68.2% vs. 17.9%), and their lipid parameters were also less favorable. Parasympathetic neuropathy was present in both groups (56.8% vs. 32.1%, respectively). Sympathetic neuropathy was not found. Sensory nerve dysfunction was of low prevalence in the high-risk group and did not occur in healthy controls. In multiple logistic regression analysis, HbA1c exhibited an independent association with parasympathetic neuropathy (OR: 5.9; 95% CI: 1.08–32.68; p < 0.041).DiscussionAn increased risk of developing prediabetes/diabetes does not appear to have a strong correlation with an increased likelihood of developing autonomic or sensory neuropathy. However, the etiology behind the occurrence of parasympathetic autonomic neuropathy in healthy individuals remains unknown.

Modulation of the cervix by steroid hormones and commensal microbiome play a central role in the health of the female reproductive tract. Here we describe organ-on-a-chip (Organ Chip) models that recreate the human cervical epithelial-stromal interface with a functional epithelial barrier and production of mucus with biochemical and hormone-responsive properties similar to living cervix. When Cervix Chips are populated with optimal healthy versus dysbiotic microbial communities (dominated by Lactobacillus crispatus and Gardnerella vaginalis, respectively), significant differences in tissue innate immune responses, barrier function, cell viability, proteome, and mucus composition are observed that are similar to those seen in vivo. Thus, human Cervix Organ Chips represent physiologically relevant in vitro models to study cervix physiology and host-microbiome interactions, and hence may be used as a preclinical testbed for development of therapeutic interventions to enhance women's health.

Abstract The rates of tertiary lymphoid organ (TLO) formation in pancreatic cancer are very low but patients with TLO have dramatically better survival rates. Our goal is to understand how pancreatic cancer suppresses TLO so that we can improve patient survival and reduce recurrence. We integrated human cell lines from pancreatic and lung cancer with either high or low PDL1/L2 expression into our previously described lymphoid follicle (LF) chip (Goyal et al., Adv. Sci, 2022), where peripheral blood mononuclear cells are induced to form 3D TLO-like structures. The hot lung cancer cell line, which expressed high levels of PDL1/L2 and a “hot” phenotype in published murine studies, produced a high levels of cytokines and stimulated increased TLO formation in the LF chip. In contrast, the cold lung and pancreatic cell lines did not induce the cytokine signature or TLOs. Importantly, TLO formation correlated with increased B cell activation, anti-tumor CD8 activity and tumor cell death. We have coupled this in vitro study with analysis of the pancreatic cancer transcriptome from the Cancer Genome Atlas to identify therapeutic targets to promote TLO formation in pancreatic cancer. In ongoing studies, we are assessing the functionality of the lung cancer TLO in vitro and in vivo and identifying therapeutics that may improve TLO formation in pancreatic cancer. Citation Format: Yunhao Zhai, Pranav Prabhala, Lucas Barck, Min Wen Ku, Aditya Patil, Viktor Horvath, Russell A. Gould, Donald E. Ingber, Girija Goyal. Tertiary lymphoid organogenesis and lymphocyte activation in human organ chips [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Pancreatic Cancer; 2023 Sep 27-30; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(2 Suppl):Abstract nr B063.

Aims: To estimate prevalence of diagnosed (dDM) and undiagnosed diabetes (uDM) in Hungary and investigate determinants of uDM. Methods: Data was obtained from the nationally representative H-UNCOVER study. As laboratory measurements were available for 11/19 Hungarian counties, n = 5,974/17,787 people were eligible. After exclusions, 5,673 (representing 4,976,097 people) were included. dDM was defined by self-reporting, while uDM as negative selfreporting and elevated fasting glucose (>= 7 mmol/l) and/or HbA1c (>= 48 mmol/mol). Logistic regression for complex samples was used to calculate comparisons between dDM and uDM adjusted for age and BMI. Results: Diabetes prevalence was 12.0 %/11.9 % (women/men, 95 %CI:10.7-13.4 %/10.7-13.2 %), while 2.2 %/2.8 % (1.7-2.8 %/2.2-3.6 %) of women/men were uDM. While the proportion of uDM vs. dDM was similar for women >= 40, men in their forties had the highest odds for uDM. Neither unemployment (women/men OR:0.58 [0.14-2.45]/0.50 [0.13-1.92]), nor education level (tertiary vs. primary; women/men OR: 1.16 [0.53-2.56]/ 0.53 [0.24-1.18]) were associated with uDM. The risk of uDM was lower in both sexes with chronic morbidities. Conclusions: We report higher prevalence of diabetes and undiagnosed diabetes than previous Hungarian estimates. The finding that socioeconomic factors are not associated to uDM suggests that universal health care could provide equitable access to diabetes diagnosis.

Flavonoids are phytonutrients commonly found in plant-based foods and are generally known for their health benefits. However, their utility as potential therapeutics has not been explored because their presence in drug development tests can lead to false positives due to non-specific binding. Here, we employed molecular dynamic simulations (MDS) to examine flavonoid behavior and discovered that they form highly organized supramolecular assemblies that physically interact with disordered regions of enzymatic proteins and can physically interlink multiple protein molecules. These flavonoid assemblies adopt secondary structural patterns like those found in proteins and nucleic acids, and they physically influence molecular movement and tertiary protein structure, thereby modulating the biochemical activities of a diverse range of enzymes. Moreover, in the presence of flavonoids, human cells are protected against injury caused by ultraviolet radiation. These findings unveil a novel form of biochemical regulation wherein small molecules can modulate the function of larger proteins by forming supramolecular assemblies which results in enhanced molecular and cellular resilience. Single Sentence Summary Molecular dynamic simulations led to the discovery that flavonoid phytonutrients can self-assemble into highly ordered supramolecular structures that interact with enzymatic proteins, slow biochemical activities, and protect cells against injury. ### Competing Interest Statement The authors have declared no competing interest.

A hiperglikémia a cukorbetegség szövődményeihez vezet. A UKPDS hosszú távú adatai látványosan igazolták valamennyi vizsgált makro- és mikrovaszkuláris végpont csökkenését. Részben az elmúlt évek gyógyszerfejlesztési eredményeinek köszönhetően a vércukorszint kontrollja mellett a 2-es típusú diabetes mellitusban szenvedő betegek kezelésében a kardiovaszkuláris kockázat mérséklésére helyeződött a hangsúly. Ugyanakkor az ún. kardiovaszkuláris biztonságossági vizsgálatok tervezése során az volt a célkitűzés, hogy az antidiabetikus hatás a vizsgálatok két ágán közel azonos mértékű legyen. A glikémiás kontroll javítása valamennyi forgalomban lévő antidiabetikus szer, így a régóta piacon lévő dipeptidil-peptidáz-4-inhibitorok esetében érvényesül, tehát a vércukorcsökkentő hatás arányában ezek is mérséklik a kardiovaszkuláris kockázatot. A DPP-4-gátlók biztonságossági profillal rendelkeznek, nem okoznak hipoglikémiát, testtömegsemlegesek, ugyanakkor biztonságosan alkalmazhatóak idős, veseelégtelen betegekben. A posztprandiális vércukor hipoglikémiamentes csökkentése révén mérséklik a vércukor-variabilitást, ami ugyancsak kedvező prognosztikus tényező. Továbbá, figyelembe véve a hipoglikémia és az autonóm neuropathia összefüggéseit, kiválóan alkalmasak autonóm neuropathiában szenvedő cukorbetegek kezelésére.

Summary: Background: Sulfadoxine-pyrimethamine (SP) antimalarial therapy has been suggested to potentially increase the birth weight of infants in pregnant women in sub-Saharan Africa, independently of malarial infection. Here, we utilized female intestinal organoid-derived cells cultured within microfluidic Organ Chips to investigate whether SP could directly impact intestinal function and thereby improve the absorption of essential fats and nutrients crucial for fetal growth. Methods: Using a human organ-on-a-chip model, we replicated the adult female intestine with patient organoid-derived duodenal epithelial cells interfaced with human intestinal endothelial cells. Nutrient-deficient (ND) medium was perfused to simulate malnutrition, resulting in the appearance of enteric dysfunction indicators such as villus blunting, reduced mucus production, impaired nutrient absorption, and increased inflammatory cytokine secretion. SP was administered to these chips in the presence or absence of human peripheral blood mononuclear cells (PBMCs). Findings: Our findings revealed that SP treatment effectively reversed multiple intestinal absorptive abnormalities observed in malnourished female Intestine Chips, as validated by transcriptomic and proteomic analyses. SP also reduced the production of inflammatory cytokines and suppressed the recruitment of PBMCs in ND chips. Interpretation: Our results indicate that SP could potentially increase birth weights by preventing enteric dysfunction and suppressing intestinal inflammation. This underscores the potential of SP as a targeted intervention to improve maternal absorption, subsequently contributing to healthier fetal growth. While SP treatment shows promise in addressing malabsorption issues that can influence infant birth weight, we did not model pregnancy in our chips, and thus its usefulness for treatment of malnourished pregnant women requires further investigation through clinical trials. Funding: The Bill and Melinda Gates Foundation, and the Wyss Institute for Biologically Inspired Engineering at Harvard University, and the HDDC Organoid Core of the P30 DK034854.

Drug repurposing requires distinguishing established drug class targets from novel molecule-specific mechanisms and rapidly derisking their therapeutic potential in a time-critical manner, particularly in a pandemic scenario. In response to the challenge to rapidly identify treatment options for COVID-19, several studies reported that statins, as a drug class, reduce mortality in these patients. However, it is unknown if different statins exhibit consistent function or may have varying therapeutic benefit. A Bayesian network tool was used to predict drugs that shift the host transcriptomic response to SARS-CoV-2 infection towards a healthy state. Drugs were predicted using 14 RNA-sequencing datasets from 72 autopsy tissues and 465 COVID-19 patient samples or from cultured human cells and organoids infected with SARS-CoV-2. Top drug predictions included statins, which were then assessed using electronic medical records containing over 4,000 COVID-19 patients on statins to determine mortality risk in patients prescribed specific statins versus untreated matched controls. The same drugs were tested in Vero E6 cells infected with SARS-CoV-2 and human endothelial cells infected with a related OC43 coronavirus. Simvastatin was among the most highly predicted compounds (14/14 datasets) and five other statins, including atorvastatin, were predicted to be active in > 50% of analyses. Analysis of the clinical database revealed that reduced mortality risk was only observed in COVID-19 patients prescribed a subset of statins, including simvastatin and atorvastatin. In vitro testing of SARS-CoV-2 infected cells revealed simvastatin to be a potent direct inhibitor whereas most other statins were less effective. Simvastatin also inhibited OC43 infection and reduced cytokine production in endothelial cells. Statins may differ in their ability to sustain the lives of COVID-19 patients despite having a shared drug target and lipid-modifying mechanism of action. These findings highlight the value of target-agnostic drug prediction coupled with patient databases to identify and clinically evaluate non-obvious mechanisms and derisk and accelerate drug repurposing opportunities.

Objectives: Our aim in this study was to compare the efficacy and safety of commercially available fixed-ratio combinations (FRCs) of glucagon-like peptide-1 receptor agonists (GLP-1RAs) and basal insulins by a network meta-analysis of randomized controlled trials (RCTs) of people with type 2 diabetes.Methods: We present a systematic review and network meta-analyses of RCTs of individuals with type 2 diabetes randomized to FRCs or to their components for >24 weeks. All reports were obtained from PubMed or ClinicalTrials.gov up to February 28, 2022. The primary outcome was glycated hemoglobin (A1C) level attained. Secondary outcomes included fasting plasma glucose, change in body weight, and incident hypoglycemia. Treatment effects were estimated as mean difference (MD) and standard error (SE), or as odds ratio (OR) with 95% confidence interval (CI) using the fixed combination of insulin glargine 100 IU/mL and lixisenatide (iGlarLixi) as reference.Results: We included 29 RCTs from among the 1,404 articles identified. No direct comparisons between FRCs were found. After excluding some insulin-capped trials to reach model consistency, both FRCs were more efficacious regarding A1C than their components, but no difference between FRCs was found (MD,-0.10%; SE, 0.10%). The effect of the fixed combination of insulin degludec and liraglutide (IDegLira) (MD,-0.47 mmol/L; SE, 0.24 mmol/L) and basal insulins was similar to that of iGlarLixi (reference) on fasting glucose, whereas GLP-1RAs had lower efficacy than iGlarLixi. Weight gain was lower with GLP-1RAs and IDegLira (MD,-0.72 kg; SE, 0.32 kg) than with iGlarLixi (reference) and higher with basal insulins. Incident hypo-glycemia (based on different definitions) was least frequent with GLP-1RAs, followed by IDegLira (OR, 0.78; 95% CI, 0.39 to 1.57), iGlarLixi (reference), and basal insulins.Conclusions: For A1C, both FRCs were more efficacious over their individual components, with similar efficacies of the 2 FRCs.& COPY; 2023 The Author(s). Published by Elsevier Inc. on behalf of Canadian Diabetes Association. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

Cardiovascular autonomic neuropathy (CAN) is a frequent complication of diabetes mellitus and is associated with increased morbidity and mortality in patients with diabetes. Hence, early and correct diagnosis of CAN is crucial. Standard cardiovascular reflex rests (CARTs) have been the gold standard of CAN assessment. Originally, CARTs consisted of five reflex tests, but measuring diastolic blood pressure response to sustained handgrip exercise has no longer been suggested as an established clinical test. Increasing body of evidence suggests that isometric handgrip test should no longer be used for the evaluation of sympathetic dysfunction during cardiovascular autonomic neuropathy assessment in diabetic patients. The associations of isometric handgrip test results with parameters of hypertension and markers of hypertension-related target-organ damage in diabetic and non-diabetic individuals point toward its potential role as a screening tool to identify patients with high cardiovascular risk. The current review summarizes historical view of standard cardiovascular reflex tests and latest data on isometric handgrip test.

Modulation of mucus production by the human ectoand endo-cervical epithelium by steroid hormones and associated interactions with commensal microbiome play a central role in the physiology and pathophysiology of the female reproductive tract. However, most of our knowledge about these interactions is based on results from animal studies or in vitro models that fail to faithfully mimic the mucosal environment of the human cervix. Here we describe microfluidic organ-on-a-chip (Organ Chip) models of the human cervical mucosa that recreate the cervical epithelial-stromal interface with a functional epithelial barrier and produce abundant mucus that has compositional, biophysical, and

Hypoglycaemia increases the risk of falls, unconsciousness, seizures and dementia in the elderly population. Besides, deepening hypoglycaemia precipitates QT -prolongation, premature cardiac beats and conduction abnormalities. The old diabetic patient (female, age: 84 ys; HbA1c 7.0%; BMI: 26.8 kg/m2) was admitted to hospital due to recurring episodes of collapse. During the last event of losing consciousness, severe hypoglycaemia was detected (glucose: 1.7 mmol/l). On clinical admission, the electrocardiogram (ECG) recording showed QT -interval prolongation and Mobitz type II 2nd degree AV block being in accordance with findings of a Holter test performed in the preceding weeks. The decade -long used sulfonylurea (SU) was omitted and the DPP4-inhibitor sitagliptin was added on to metformin as glucose -lowering medication. Neither cardiac conduction abnormalities nor arrhythmias returned during the patient's hypoglycaemia -free hospital observation and Holter monitoring. Long-lasting severe hypoglycaemia as a side -effect of SU treatment could account for the cardiac arrhythmia. As for differential diagnosis, carotid ultrasound revealed significant stenosis of the left carotid artery (80-85%) and echocardiographic imaging was negative. Diabetic sensorimotor polyneuropathy and mild -to -moderate cardiovascular autonomic neuropathy was established. However, the role of orthostatic hypotension could be excluded.