Diabetes mellitus and its influence on the incidence and process of diabetic retinopathy.

OBJECTIVES: The main aim of the study was to show the effect of diabetes in relation to the gender of the patients, duration of the disease, and on the incidence of diabetic retinopathy. In this study, we investigated the prevalence of these two diseases, pathological ocular changes and progression of disease occurrence in relation to the duration of the disease with respect to their impact on the quality of vision of the patient. METHODS: The prospective observational cross-sectional study included 3,951 patients (1,838 males, 2,113 females) with diabetes mellitus from 7 districts of eastern Slovakia. Patients with diabetes mellitus and diabetic retinopathy were identified by special screening in the number of 2,093 (1,094 females and 999 males). Subjects were divided by sex and by duration of diabetes into 5 groups: patients with diabetes under 5, 10, 15, 20, and over 20 years. We differentiated between proliferative and non-proliferative forms of diabetic retinopathy and monitored changes in visual quality. Manifestations of pathological changes were recorded using special examination methods in the eye clinic. We observed a decrease in vision by two lines, pathological changes on the retina and the occurrence of practical blindness. RESULTS: Of the total number of diabetic patients examined, diabetic retinopathy was also present in more than half of the patients. The major form represented in the patients was the non-proliferative form of retinopathy. The obtained results confirmed that the representation of patients with diabetic retinopathy increases with increasing duration of diabetes. Similarly, pathological changes characteristic of this type of late complication of diabetes were also more frequent, such as deterioration of visual acuity, the appearance of aneurysms, hard exudates macular edema, and gradual loss of vision, which can result in practical blindness. CONCLUSION: The percentage of people with diabetic retinopathy increases with the duration of diabetes, as well as the increased frequency of pathological late complication of diabetes, including deterioration of visual acuity, the development of aneurysms, hard exudates, macular oedema, and gradual loss of vision, which can result in practical blindness. Early diagnosis of the disease and introduction of appropriate treatment would alleviate the symptoms of the disease in more than half of the patients, so more frequent preventive check-ups with an ophthalmologist should be performed in diabetic patients to avoid detection of the disease in its late stages.

Monitoring of selected anthropometric and biochemical parameters in men and women in Bardejov District.

OBJECTIVES: Metabolic and cardiovascular diseases represent a significant and ever-increasing problem, accounting for nearly one-third of all deaths and leading to significant morbidity. Lifestyle changes, which introduce new risk factors for cardiovascular disease, are leading to an increase in the risk of cardiovascular disease worldwide. The burden of cardiovascular disease can be reduced by careful risk reduction, and as such, primary prevention is an important priority for all. There is strong consensus among international guidelines regarding the necessity of smoking cessation, weight optimization, and the importance of exercise, while guidelines differ slightly in their approach to hypertension and vary greatly in their approach to the optimal lipid profile, which remains a controversial issue. METHODS: The work is focused on the analysis of biochemical (glucose, cholesterol and triacylglycerols) and anthropometric (body mass index) parameters in selected probands (n = 140), including men (n = 70) and women (n = 70) in the Bardejov district. Probands were divided into groups depending on age (18-38, 39-50, 51-60, and over 61 years). For statistical evaluation, the program Statistica ver. 12 and ANOVA were used. RESULTS: The results of the study showed statistically significantly higher values of BMI, total cholesterol (TC), triacylglycerides (TG), and glucose (GLU) (p < 0.001) between men and women in the age groups (18-38 years, 39-50 years). In probands from the age group 51-60 years and over 61 years, we found a high level of cholesterol above the reference limit, but a significant difference between men and women was not confirmed. Correlation analysis confirmed the positive correlation between glucose and BMI. CONCLUSION: Differences in risk-factor (e.g., biochemical and anthropometrics) burden translate into marked differences in the lifetime risk of cardiovascular disease. These differences are consistent across age groups.

Assessment of foot arch shape and health status of adult population from Eastern Slovakia.

BACKGROUND: The purpose of this study was to determine the foot arch shape and the associated health status in a selected sample of individuals belonging to the adult majority population of eastern Slovakia and to evaluate which of the observed factors are involved in the occurrence and development of foot arch abnormalities. METHODS: The weight and height of probands were measured according to standard anthropometric methods using a personal scale and an anthropometer. The BMI index was calculated for each proband from the measured data. A Pod4Foot Classic podoscope was used to obtain the plantograms. The Chippaux-Smirak index method was used to evaluate the plantograms. The study includes a questionnaire containing questions about factors affecting the foot arch shape. RESULTS: The arch of the right foot was supported by the effect of wearing shoes at home (p = 0.013). There was an association between wearing high-heeled shoes and foot arch disorder, both on the right (p = 0.011) and left (p = 0.045) foot. There was no significant relationship between the prevalence of overweight or obesity, between wearing orthopaedic insoles in shoes without a prescription, wearing orthopaedic footwear at home, between sport or static and active work and foot arch disorder in our study. CONCLUSIONS: Most of the probands had a bilaterally normal arched foot, which is a positive result. Nowadays, there are a large number of orthopaedic devices on the market that help to shape the arch of the foot properly, but it is not correct to use these devices arbitrarily, without a doctor's prescription and a diagnosis of foot arch deviation.

Use of bioimpedance in prevention of sarcopenia in the elderly.

OBJECTIVES: Physiological ageing is associated with major and progressive changes in body composition, particularly in the decline of fat-free mass, which puts older individuals at risk of geriatric syndromes such as sarcopenia and sarcopenic obesity. Bioimpedance analysis noninvasively allows the determination of body composition, thus being able to rapidly assess primary risk factors leading to sarcopenia prediction. METHODS: We conducted a study of 180 probands, 120 females (66.7%) with a mean age of 76.23 (SD = 9.29) years and 60 males (33.3%) with a mean age of 74.01 (SD = 8.99) years in cooperation with facilities for the elderly and with the inpatient department of the clinics of J.A. Reimann Hospital in Presov. Body height, body weight, hip circumference, and waist circumference were determined by the anthropometric method, from which the values of the body mass index and waist-to-hip ratio were calculated. Active body mass, total body water, extracellular body water, intracellular body water, cell mass, body mass fat, body fat index, fat-free mass index, impedance at frequencies of 50 kHz, resistance, reactance, and phase angle were determined using the Quadscan 4000 Touch bioimpedance instrument. Appendicular skeletal muscle mass (ASMM) and muscle mass (ASMM/Ht2) were calculated. The data obtained from anthropometric and bioimpedance measurements were processed in MS Excel 2000 and STATISTICA ver. 12. The difference of means in the studied groups was tested by the t-test. RESULTS: Presarcopenia was diagnosed in 12 (6.66%) probands out of 180 probands, of which were 3 (5%) men and 9 women (7.5%). Phase angle, ASMM and ASMM/Ht2 values were significantly lower (p < 0.001) in men and women with presarcopenia. CONCLUSION: By introducing bioimpedance measurements into practice, it is possible to obtain results in a non-invasive way revealing possible presarcopenia in the elderly.

Initial screening of the rs104893657 variant of the PAX8 gene in women with hypothyroidism from Northeastern Slovakia.

OBJECTIVE: Thyroid diseases are among the most common endocrinopathies and metabolic disorders. Hypothyroidism is caused by insufficient production of thyroid hormones with a higher prevalence in women. Causes for the development of endocrine diseases may be mutations in genes that encode peptide hormones. The aim of this scientific study was to determine the genotype and allele frequencies of the rs104893657 variant of the PAX8 gene and to determine the genotype versus phenotype association. METHODS: The study population consisted of 135 women from northeastern Slovakia who were divided on the basis of screening into two groups: a control group without diagnosed hypothyroidism (CG = 67) and a group of women with hypothyroidism (HY = 68). Biochemical markers - thyroid-stimulating hormone (TSH), prealbumin (PREA), calcium (Ca), phosphorus (P), and alkaline phosphatase (ALP) were determined using Cobas Integra 400 plus, Cobas e411 analysers (Roche). Genotyping was performed using TaqMan® SNP Genotyping Assay instrument 7500 Fast Real-Time PCR Systems (Applied Biosystem). RESULTS: Student's t-test revealed a statistically significant difference between CG and HY in biochemical parameters: TSH (p < 0.001), P (p = 0.008). By Chi-square test we found no statistically significant difference in the representation of genotypes (p = 0.788) in the rs104893657 polymorphism of PAX8 gene. The T allele was not associated with hypothyroidism in Slovak women (p = 0.548). In CC genotype we found statistically significant difference between CG and HY in parameters TSH (p < 0.001) and P (p = 0.006). CONCLUSION: The mutant T allele was detected at low frequency in both groups of women studied. The association of the T allele with the development of hypothyroidism in Slovak women was not confirmed. The results of this work provide initial information on the distribution of genotypes and alleles in the studied variant of PAX8 gene in the Slovak female population.

Level of biochemical parameters in patients with type 2 diabetes mellitus depending on the genotype of the FokI polymorphism in the vitamin D3 receptor (VDR gene).

OBJECTIVES: Diabetes mellitus type 2 (T2DM) is a multifactorial and polygenic disorder characterised by chronic hyperglycaemia accompanied by impaired lipid, carbohydrate, and protein metabolism. The disease is associated with several genetic polymorphisms, including the FokI polymorphism in the vitamin D receptor (VDR) gene. METHODS: We conducted a study of 327 probands (191 T2DM patients, 136 controls), with a mean age 65.06 (SD ± 10.88) years of patients with T2DM and 58.89 (SD ± 6.59) years in the healthy probands. We investigated the association between FokI polymorphism and biochemical parameters in T2DM patients in the Slovak population. Anthropometric measurements, biochemical, and genetic analysis were statistically evaluated by Statistica ver.13 software using t-tests. RESULTS: Biochemical analysis confirmed significantly higher mean values of total cholesterol (TC), triglyceride (TG), glucose (GLU), and uric acid (UA) (p < 0.001) in T2DM probands and statistically significantly lower values of high-density lipoprotein (HDL), cholesterol and vitamin D (p < 0.001). Allele frequencies and genotype distributions of the FokI (rs2228570) polymorphism were not significantly different between T2DM patients and controls (p = 0.909). Patients with T2DM and TT genotype had the highest glucose level of 11.39 (SD ± 2.32) uU/ml (p < 0.001). CONCLUSION: Our study did not provide evidence for an association of the investigated FokI polymorphism of the VDR gene with T2DM in the Slovak population. Further research is needed to evaluate the impact of single nucleotide polymorphisms (SNPs) in the VDR gene, focusing on related genetic analyses in a larger T2DM cohort.

Health literacy and patient rights awareness in ethnic Hungarian mothers in Eastern Europe.

OBJECTIVES: The paper proposes to identify the determinants of patients' rights awareness in mothers and to examine the relationship of health literacy with awareness of those rights. METHODS: Our results are based on data from a convenience sample of 894 non-health professional ethnic Hungarian mothers from Hungary, Romania and Slovakia. Health literacy is measured with the HLS-EU-16 questionnaire. RESULTS: Analysis of variance reveals a significant association of health literacy with patient rights awareness. Our results show that health literacy is the highest among patients who filed a complaint through formal channels and/or took legal measures to restore their rights upon violation. A logistic regression model is built to identify the likelihood of having high patient rights awareness, that is, acting formally for the restoration of rights upon infringement. The model controls for covariates. When controlled for covariates, the likelihood of having high patient rights awareness increases with age, and is higher for mothers with highest education, for inhabitants of larger towns, as well as for those with adequate health literacy. CONCLUSIONS: The findings of our study have implications for health policy, as they reveal significant inequalities in patient rights culture.

Self-rated health status and hygienic behaviour of pregnant women in Hungary during the pandemic.

OBJECTIVES: The aim of our research was to investigate the self-rated health status, health behaviour and hygiene habits of pregnant women, and to explore the influencing factors during the pandemic. METHODS: The sample included 1,200 pregnant women who gave birth in the most progressive institutions of the three examined counties in Hungary; 839 questionnaires were returned, based on which the willingness to answer can be said to be 69.9%. After data cleaning, 640 questionnaires were added to the database. Descriptive statistical analyses and correlation tests were performed, during which we used the Pearson's chi-square test and Fisher's exact test. RESULTS: Socio-demographic factors show a significant correlation with the perceived state of health and the degree of responsibility for health, however, the existence of a chronic disease does not affect either the self-assessed state of health or hygienic behaviour. Factors influencing hygienic behaviour include residence, education, assessed income, and gestational age. We found no correlation between health status and hygienic attitude. CONCLUSIONS: Our results draw attention to the fact that among pregnant women - especially pregnant women with low socioeconomic status - education about hygiene habits is of particular importance. Teamwork is needed in this area, which should start during the period of preparation for pregnancy, but at the latest in the early stages of pregnancy.

Examining the sleeping habits of preschool and elementary school children in Southern Slovakia.

OBJECTIVES: Our research aimed to examine children's sleeping habits from preschool to the end of elementary school age. Developing proper sleeping habits in childhood is essential, as it is decisive for the rest of our lives. METHODS: A total of 339 children (160 males and 179 females) took part in the research, of which 145 were preschool-age children (3-7 years old), 72 lower-grade elementary school children (6-11 years old), and 122 upper-grade elementary school children (12-16 years old). The questionnaire was completed in a paper form (elementary school students) and online (kindergarten children). RESULTS: The research results show that most of the children spend enough time sleeping following the recommendations. In case of the kindergarten children, the younger ones also sleep in the afternoon on weekends (average of 3.66 years, 28.3%), and the older ones do not sleep in the afternoon either in kindergarten during the week or at home at the weekend (average of 5.22 years, 46.2%). The use of blue light typically increases with age; 39% of the preschoolers, 61% of the 6-11-year-olds, and 67% of the 12-16-year-olds use it before falling asleep. Sleep aids and rituals are used by 87.6% of the preschoolers, 67.4% of the 6-11-year-olds, and 34.4% of the 12-16-year-olds, because significantly more preschoolers find it more difficult to fall asleep than older children. At night, 40% of the preschoolers wake up at least once (due to biological needs - 46.3%), 32% of the 6-11-year-olds wake up at night (due to nightmares - 42.3%), and 41% of the 12-16-year-olds also wake up all night (due to biological needs - 31.9%, and due to noise - 29.8%). CONCLUSION: Although the children get enough sleep, significantly more upper-grade school children feel tired in the morning. It is essential to help sleep and eliminate factors that prevent falling asleep to create a healthy circadian rhythm in the life of children.

Nutrition habits and physical activity of disabled adolescents in Hungary.

OBJECTIVES: The overall aim of the research is to gain insight into the lifestyle and health behaviour of young people with disabilities aged 12-18 years, and to learn about the factors shaping their health. In this study, our aim was to investigate the eating habits and physical activity of adolescents with disabilities and the influencing factors. METHODS: The study's source population was young people aged 12-18 with disabilities in the Szabolcs-Szatmár-Bereg County. A total of 239 completed questionnaires were processed and analysed. Boys were overrepresented in the study. RESULTS: The results show that inadequate eating habits also exist in the population we studied, the consumption of certain foods correlates with the education level of parents. A more secure social background is a protective factor in terms of physical activity. The survey was made with the aim of intervention much needed. With precise knowledge of the health behaviour of the target group, our goal is to develop a complex intervention programme adapted to the needs of children with disabilities. CONCLUSION: The eating habits of adolescents with disabilities in the Szabolcs-Szatmár-Bereg County are unsatisfactory. Experience and results show that differentiated health promotion of young people with disabilities is necessary.

In vitro patterning of pluripotent stem cell-derived intestine recapitulates in vivo human development.

The intestine plays a central role in digestion, nutrient absorption and metabolism, with individual regions of the intestine having distinct functional roles. Many examples of region-specific gene expression in the adult intestine are known, but how intestinal regional identity is established during development is a largely unresolved issue. Here, we have identified several genes that are expressed in a region-specific manner in the developing human intestine. Using human embryonic stem cell-derived intestinal organoids, we demonstrate that the duration of exposure to active FGF and WNT signaling controls regional identity. Short-term exposure to FGF4 and CHIR99021 (a GSK3ß inhibitor that stabilizes ß-catenin) resulted in organoids with gene expression patterns similar to developing human duodenum, whereas longer exposure resulted in organoids similar to ileum. When region-specific organoids were transplanted into immunocompromised mice, duodenum-like organoids and ileum-like organoids retained their regional identity, demonstrating that regional identity of organoids is stable after initial patterning occurs. This work provides insights into the mechanisms that control regional specification of the developing human intestine and provides new tools for basic and translational research.

Sox9 plays multiple roles in the lung epithelium during branching morphogenesis.

Lung branching morphogenesis is a highly orchestrated process that gives rise to the complex network of gas-exchanging units in the adult lung. Intricate regulation of signaling pathways, transcription factors, and epithelial-mesenchymal cross-talk are critical to ensuring branching morphogenesis occurs properly. Here, we describe a role for the transcription factor Sox9 during lung branching morphogenesis. Sox9 is expressed at the distal tips of the branching epithelium in a highly dynamic manner as branching occurs and is down-regulated starting at embryonic day 16.5, concurrent with the onset of terminal differentiation of type 1 and type 2 alveolar cells. Using epithelial-specific genetic loss- and gain-of-function approaches, our results demonstrate that Sox9 controls multiple aspects of lung branching. Fine regulation of Sox9 levels is required to balance proliferation and differentiation of epithelial tip progenitor cells, and loss of Sox9 leads to direct and indirect cellular defects including extracellular matrix defects, cytoskeletal disorganization, and aberrant epithelial movement. Our evidence shows that unlike other endoderm-derived epithelial tissues, such as the intestine, Wnt/ß-catenin signaling does not regulate Sox9 expression in the lung. We conclude that Sox9 collectively promotes proper branching morphogenesis by controlling the balance between proliferation and differentiation and regulating the extracellular matrix.

Rearrangements of Cycloalkenyl Aryl Ethers.

Rearrangement reactions of cycloalkenyl phenol and naphthyl ethers and the acid-catalyzed cyclization of the resulting product were investigated. Claisen rearrangement afforded 2-substituted phenol and naphthol derivatives. Combined Claisen and Cope rearrangement resulted in the formation of 4-substituted phenol and naphthol derivatives. In the case of cycloocthylphenyl ether the consecutive Claisen and Cope rearrangements were followed by an alkyl migration. The mechanism of this novel rearrangement reaction is also discussed.

Persistence and toxin production by Clostridium difficile within human intestinal organoids result in disruption of epithelial paracellular barrier function.

Clostridium difficile is the leading cause of infectious nosocomial diarrhea. The pathogenesis of C. difficile infection (CDI) results from the interactions between the pathogen, intestinal epithelium, host immune system, and gastrointestinal microbiota. Previous studies of the host-pathogen interaction in CDI have utilized either simple cell monolayers or in vivo models. While much has been learned by utilizing these approaches, little is known about the direct interaction of the bacterium with a complex host epithelium. Here, we asked if human intestinal organoids (HIOs), which are derived from pluripotent stem cells and demonstrate small intestinal morphology and physiology, could be used to study the pathogenesis of the obligate anaerobe C. difficile. Vegetative C. difficile, microinjected into the lumen of HIOs, persisted in a viable state for up to 12 h. Upon colonization with C. difficile VPI 10463, the HIO epithelium is markedly disrupted, resulting in the loss of paracellular barrier function. Since similar effects were not observed when HIOs were colonized with the nontoxigenic C. difficile strain F200, we directly tested the role of toxin using TcdA and TcdB purified from VPI 10463. We show that the injection of TcdA replicates the disruption of the epithelial barrier function and structure observed in HIOs colonized with viable C. difficile.

A large-scale assessment of hand hygiene quality and the effectiveness of the "WHO 6-steps".

BACKGROUND: Hand hygiene compliance is generally assessed by observation of adherence to the "WHO five moments" using numbers of opportunities as the denominator. The quality of the activity is usually not monitored since there is no established methodology for the routine assessment of hand hygiene technique. The aim of this study was to objectively assess hand rub coverage of staff using a novel imaging technology and to look for patterns and trends in missed areas after the use of WHO's 6 Step technique. METHODS: A hand hygiene education and assessment program targeted 5200 clinical staff over 7 days at the National University Hospital, Singapore. Participants in small groups were guided by professional trainers through 5 educational stations, which included technique-training and UV light assessment supported by digital photography of hands. Objective criteria for satisfactory hand hygiene quality were defined a priori. The database of images created during the assessment program was analyzed subsequently. Patterns of poor hand hygiene quality were identified and linked to staff demographic. RESULTS: Despite the assessment taking place immediately after the training, only 72% of staff achieved satisfactory coverage. Failure to adequately clean the dorsal and palmar aspects of the hand occurred in 24% and 18% of the instances, respectively. Fingertips were missed by 3.5% of subjects. The analysis based on 4642 records showed that nurses performed best (77% pass), and women performed better than men (75% vs. 62%, p<0.001). Further risk indicators have been identified regarding age and occupation. CONCLUSION: Ongoing education and training has a vital role in improving hand hygiene compliance and technique of clinical staff. Identification of typical sites of failure can help to develop improved training.

Complex Relationships between Milking-Induced Changes in Teat Structures and Their Pre-Milking Dimensions in Holstein Cows.

The study aimed to explore the relationship between teat structure dimensions and their short-term reaction to milking, to find the optimal dimensions of teat structures in relation to milking-induced teat tissue changes. Teat structures (teat length, canal length, thickness at barrel and apex, wall and cistern width) were measured by ultrasonography before and after milking for 38 Holstein cows at the beginning, middle, and end of lactation. We found that milking-induced changes in teat structures significantly depended on their pre-milking size. Furthermore, we observed that some teat structures and their changes were interconnected, and some did not affect each other. For example, changes in the barrel thickness and cistern width were affected by all structures, while the canal and apex did not influence each other. We deduced that more favorable changes were observed for teats of medium length, medium barrel and apex thickness, with teat canals of medium length, but with wider cisterns and thinner walls. The results of this study may help improve research in the area of milking-induced changes in teat morphology. Our findings could help understand potential health risks to animals in relation to teat morphology, milking equipment, and machine settings.

Comparing and Correcting Spectral Sensitivities between Multispectral Microscopes: A Prerequisite to Clinical Implementation.

Multispectral, multiplex immunofluorescence (mIF) microscopy has been used to great effect in research to identify cellular co-expression profiles and spatial relationships within tissue, providing a myriad of diagnostic advantages. As these technologies mature, it is essential that image data from mIF microscopes is reproducible and standardizable across devices. We sought to characterize and correct differences in illumination intensity and spectral sensitivity between three multispectral microscopes. We scanned eight melanoma tissue samples twice on each microscope and calculated their average tissue region flux intensities. We found a baseline average standard deviation of 29.9% across all microscopes, scans, and samples, which was reduced to 13.9% after applying sample-specific corrections accounting for differences in the tissue shown on each slide. We used a basic calibration model to correct sample- and microscope-specific effects on overall brightness and relative brightness as a function of the image layer. We tested the generalizability of the calibration procedure and found that applying corrections to independent validation subsets of the samples reduced the variation to 2.9 ± 0.03%. Variations in the unmixed marker expressions were reduced from 15.8% to 4.4% by correcting the raw images to a single reference microscope. Our findings show that mIF microscopes can be standardized for use in clinical pathology laboratories using a relatively simple correction model.

Genetic structure of the paternal lineage of the Roma people.

According to written sources, Roma (Romanies, Gypsies) arrived in the Balkans around 1,000 years ago from India and have subsequently spread through several parts of Europe. Genetic data, particularly from the Y chromosome, have supported this model, and can potentially refine it. We now provide an analysis of Y-chromosomal markers from five Roma and two non-Roma populations (N = 787) in order to investigate the genetic relatedness of the Roma population groups to one another, and to gain further understanding of their likely Indian origins, the genetic contribution of non-Roma males to the Roma populations, and the early history of their splits and migrations in Europe. The two main sources of the Roma paternal gene pool were identified as South Asian and European. The reduced diversity and expansion of H1a-M82 lineages in all Roma groups imply shared descent from a single paternal ancestor in the Indian subcontinent. The Roma paternal gene pool also contains a specific subset of E1b1b1a-M78 and J2a2-M67 lineages, implying admixture during early settlement in the Balkans and the subsequent influx into the Carpathian Basin. Additional admixture, evident in the low and moderate frequencies of typical European haplogroups I1-M253, I2a-P37.2, I2b-M223, R1b1-P25, and R1a1-M198, has occurred in a more population-specific manner.

Nonadhesive Alginate Hydrogels Support Growth of Pluripotent Stem Cell-Derived Intestinal Organoids.

Human intestinal organoids (HIOs) represent a powerful system to study human development and are promising candidates for clinical translation as drug-screening tools or engineered tissue. Experimental control and clinical use of HIOs is limited by growth in expensive and poorly defined tumor-cell-derived extracellular matrices, prompting investigation of synthetic ECM-mimetics for HIO culture. Since HIOs possess an inner epithelium and outer mesenchyme, we hypothesized that adhesive cues provided by the matrix may be dispensable for HIO culture. Here, we demonstrate that alginate, a minimally supportive hydrogel with no inherent cell instructive properties, supports HIO growth in vitro and leads to HIO epithelial differentiation that is virtually indistinguishable from Matrigel-grown HIOs. In addition, alginate-grown HIOs mature to a similar degree as Matrigel-grown HIOs when transplanted in vivo, both resembling human fetal intestine. This work demonstrates that purely mechanical support from a simple-to-use and inexpensive hydrogel is sufficient to promote HIO survival and development.

In Vitro Induction and In Vivo Engraftment of Lung Bud Tip Progenitor Cells Derived from Human Pluripotent Stem Cells.

The current study aimed to understand the developmental mechanisms regulating bud tip progenitor cells in the human fetal lung, which are present during branching morphogenesis, and to use this information to induce a bud tip progenitor-like population from human pluripotent stem cells (hPSCs) in vitro. We identified cues that maintained isolated human fetal lung epithelial bud tip progenitor cells in vitro and induced three-dimensional hPSC-derived organoids with bud tip-like domains. Bud tip-like domains could be isolated, expanded, and maintained as a nearly homogeneous population. Molecular and cellular comparisons revealed that hPSC-derived bud tip-like cells are highly similar to native lung bud tip progenitors. hPSC-derived epithelial bud tip-like structures survived in vitro for over 16 weeks, could be easily frozen and thawed, maintained multilineage potential, and successfully engrafted into the airways of immunocompromised mouse lungs, where they persisted for up to 6 weeks and gave rise to several lung epithelial lineages.