Evolving topological order in the postnatal visceral pleura.

BACKGROUND: Changes in epithelial cell shape reflects optimal cell packing and the minimization of surface free energy, but also cell-cell interactions, cell proliferation, and cytoskeletal rearrangements. RESULTS: Here, we studied the structure of the rat pleura in the first 15 days after birth. After pleural isolation and image segmentation, the analysis demonstrated a progression of epithelial order from postnatal day 1 (P1) to P15. The cells with the largest surface area and greatest shape variability were observed at P1. In contrast, the cells with the smallest surface area and most shape consistency were observed at P15. A comparison of polygonal cell geometries demonstrated progressive optimization with an increase in the number of hexagons (six-sided) as well as five-sided and seven-sided polygons. Analysis of the epithelial organization with Voronoi tessellations and graphlet motif frequencies demonstrated a developmental path strikingly distinct from mathematical and natural reference paths. Graph Theory analysis of cell connectivity demonstrated a progressive decrease in network heterogeneity and clustering coefficient from P1 to P15. CONCLUSIONS: We conclude that the rat pleura undergoes a striking change in pleural structure from P1 to P15. Further, a geometric and network-based approach can provide a quantitative characterization of these developmental changes.

The role of vasculature and angiogenesis in respiratory diseases.

In European countries, nearly 10% of all hospital admissions are related to respiratory diseases, mainly chronic life-threatening diseases such as COPD, pulmonary hypertension, IPF or lung cancer. The contribution of blood vessels and angiogenesis to lung regeneration, remodeling and disease progression has been increasingly appreciated. The vascular supply of the lung shows the peculiarity of dual perfusion of the pulmonary circulation (vasa publica), which maintains a functional blood-gas barrier, and the bronchial circulation (vasa privata), which reveals a profiled capacity for angiogenesis (namely intussusceptive and sprouting angiogenesis) and alveolar-vascular remodeling by the recruitment of endothelial precursor cells. The aim of this review is to outline the importance of vascular remodeling and angiogenesis in a variety of non-neoplastic and neoplastic acute and chronic respiratory diseases such as lung infection, COPD, lung fibrosis, pulmonary hypertension and lung cancer.

Vascularization of the adult mouse lung grafted onto the chick chorioallantoic membrane.

In the later stages of angiogenesis, the vascular sprout transitions into a functional vessel by fusing with a target vessel. Although this process appears to routinely occur in embryonic tissue, the biologic rules for sprout fusion and lumenization in adult regenerating tissue are unknown. To investigate this process, we grafted portions of the regenerating post-pneumonectomy lung onto the chick chorioallantoic membrane (CAM). Grafts from all 4 lobes of the post-pneumonectomy right lung demonstrated peri-graft angiogenesis as reflected by fluorescent plasma markers; however, fluorescent microsphere perfusion primarily occurred in the lobe of the lung that is the dominant site of post-pneumonectomy angiogenesis-namely, the cardiac lobe. Vascularization of the cardiac lobe grafts was confirmed by active tissue growth (p < .05). Functional vascular connections between the cardiac lobe and the CAM vascular network were demonstrated by confocal fluorescence microscopy as well as corrosion casting and scanning electron microscopy (SEM). Bulk transcriptional profiling of the cardiac lobe demonstrated the enhanced expression of many genes relative to alveolar epithelial cell (CD11b-/CD31-) control cells, but only the upregulation of Ereg and Fgf6 compared to the less well-vascularized right upper lobe. The growth of actively regenerating non-neoplastic adult tissue on the CAM demonstrates that functional lumenization can occur between species (mouse and chick) and across the developmental spectrum (adult and embryo).

Age-related changes in testicular histological structure, testosterone level, libido and semen traits in male Coturnix quail.

To investigate the effect of age in male quail on testicular weight and histology, sexual libido and semen characteristics, a study was performed on 100 quails at 10, 16, 22, 28 and 34 weeks of age. The body and testicular weights were significantly (p < .05) higher at 16 and 22 than at 28 weeks of age. The circumference and diameter of the seminiferous tubules were significantly (p < .05) higher at 28 and 34 than at 10 and 16 weeks of age. Histological evaluation of testicular slices revealed advanced and effective seminiferous tubes as early as 10 weeks, while spermatogenic activity peaked at 16 weeks of age. The highest semen volume, sperm motility and sperm concentration were observed at weeks of age and then decreased gradually with age. At the same time, the testosterone level and libido were significantly (p < .05) higher at 22 than at 10 weeks of age. Furthermore, the age was positive (p < .05), correlated to seminiferous tubule circumference and diameter, and negative (p < .05), correlated to sperm concentration. In conclusion, the age of quail markedly affected the testicular histological structure, libido, testosterone level and semen characteristics.

Topography of pleural epithelial structure enabled by en face isolation and machine learning.

Pleural epithelial adaptations to mechanical stress are relevant to both normal lung function and parenchymal lung diseases. Assessing regional differences in mechanical stress, however, has been complicated by the nonlinear stress-strain properties of the lung and the large displacements with ventilation. Moreover, there is no reliable method of isolating pleural epithelium for structural studies. To define the topographic variation in pleural structure, we developed a method of en face harvest of murine pleural epithelium. Silver-stain was used to highlight cell borders and facilitate imaging with light microscopy. Machine learning and watershed segmentation were used to define the cell area and cell perimeter of the isolated pleural epithelial cells. In the deflated lung at residual volume, the pleural epithelial cells were significantly larger in the apex (624 ± 247 µm2 ) than in basilar regions of the lung (471 ± 119 µm2 ) (p < 0.001). The distortion of apical epithelial cells was consistent with a vertical gradient of pleural pressures. To assess epithelial changes with inflation, the pleura was studied at total lung capacity. The average epithelial cell area increased 57% and the average perimeter increased 27% between residual volume and total lung capacity. The increase in lung volume was less than half the percent change predicted by uniform or isotropic expansion of the lung. We conclude that the structured analysis of pleural epithelial cells complements studies of pulmonary microstructure and provides useful insights into the regional distribution of mechanical stresses in the lung.

Inpatient Understanding of Their Care Team and Receipt of Mixed Messages: a Two-Site Cross-Sectional Study.

BACKGROUND: Patient understanding of their care, supported by physician involvement and consistent communication, is key to positive health outcomes. However, patient and care team characteristics can hinder this understanding. OBJECTIVE: We aimed to assess inpatients' understanding of their care and their perceived receipt of mixed messages, as well as the associated patient, care team, and hospitalization characteristics. DESIGN: We administered a 30-item survey to inpatients between February 2020 and November 2021 and incorporated other hospitalization data from patients' health records. PARTICIPANTS: Randomly selected inpatients at two urban academic hospitals in the USA who were (1) admitted to general medicine services and (2) on or past the third day of their hospitalization. MAIN MEASURES: Outcome measures include (1) knowledge of main doctor and (2) frequency of mixed messages. Potential predictors included mean notes per day, number of consultants involved in the patient's care, number of unit transfers, number of attending physicians, length of stay, age, sex, insurance type, and primary race. KEY RESULTS: A total of 172 patients participated in our survey. Most patients were unaware of their main doctor, an issue related to more daily interactions with care team members. Twenty-three percent of patients reported receiving mixed messages at least sometimes, most often between doctors on the primary team and consulting doctors. However, the likelihood of receiving mixed messages decreased with more daily interactions with care team members. CONCLUSIONS: Patients were often unaware of their main doctor, and almost a quarter perceived receiving mixed messages about their care. Future research should examine patients' understanding of different aspects of their care, and the nature of interactions that might improve clarity around who's in charge while simultaneously reducing the receipt of mixed messages.

Sexual behaviour response, testicular development and semen quality of New Zealand white rabbit bucks as influenced by dietary chitosan.

The present study was conducted to determine the effects of dietary chitosan supplementation on sexual behaviour responses, testicular development, and semen quality traits of New Zealand White (NZW) rabbit bucks. Twenty-four 5-week-old rabbit bucks were used in this experiment. Animals were grouped into four equal experimental groups: the control group was fed only on a basal diet, whereas the other groups were fed the basal diet supplemented with three levels of chitosan at 0.2, 0.4, or 0.6 g/kg, respectively. Also, bucks that received chitosan at 0.2 and 0.4 g/kg had a significantly earlier time of sexual libido (p ≤ .05) and had significantly higher ejaculate volume and sperm concentration than other groups (p ≤ .001). Furthermore, basic and sexual behaviours were significantly improved in bucks fed chitosan at 0.2 and 0.4 g/kg compared with other groups. Therefore, it could be concluded that using chitosan at 0.2 and 0.4 g/kg enhanced sexual behaviour, improved semen quality, and reproductive efficiency in the NZW rabbit bucks.

Morphological growth pattern of Phanerochaete chrysosporium cultivated on different Miscanthus x giganteus biomass fractions.

BACKGROUND: Solid-state fermentation is a fungal culture technique used to produce compounds and products of industrial interest. The growth behaviour of filamentous fungi on solid media is challenging to study due to the intermixity of the substrate and the growing organism. Several strategies are available to measure indirectly the fungal biomass during the fermentation such as following the biochemical production of mycelium-specific components or microscopic observation. The microscopic observation of the development of the mycelium, on lignocellulosic substrate, has not been reported. In this study, we set up an experimental protocol based on microscopy and image processing through which we investigated the growth pattern of Phanerochaete chrysosporium on different Miscanthus x giganteus biomass fractions. RESULTS: Object coalescence, the occupied surface area, and radial expansion of the colony were measured in time. The substrate was sterilized by autoclaving, which could be considered a type of pre-treatment. The fastest growth rate was measured on the unfractionated biomass, followed by the soluble fraction of the biomass, then the residual solid fractions. The growth rate on the different fractions of the substrate was additive, suggesting that both the solid and soluble fractions were used by the fungus. Based on the FTIR analysis, there were differences in composition between the solid and soluble fractions of the substrate, but the main components for growth were always present. We propose using this novel method for measuring the very initial fungal growth by following the variation of the number of objects over time. Once growth is established, the growth can be followed by measurement of the occupied surface by the mycelium. CONCLUSION: Our data showed that the growth was affected from the very beginning by the nature of the substrate. The most extensive colonization of the surface was observed with the unfractionated substrate containing both soluble and solid components. The methodology was practical and may be applied to investigate the growth of other fungi, including the influence of environmental parameters on the fungal growth.

A novel culture system for adult porcine intestinal crypts.

Porcine models are useful for investigating therapeutic approaches to short bowel syndrome and potentially to intestinal stem cell (ISC) transplantation. Whereas techniques for the culture and genetic manipulation of ISCs from mice and humans are well established, similar methods for porcine stem cells have not been reported. Jejunal crypts were isolated from murine, human, and juvenile and adult porcine small intestine, suspended in Matrigel, and co-cultured with syngeneic intestinal subepithelial myofibroblasts (ISEMFs) or cultured without feeder cells in various culture media. Media containing epidermal growth factor, noggin, and R-spondin 1 (ENR medium) were supplemented with various combinations of Wnt3a- or ISEMF-conditioned medium (CM) and with glycogen synthase kinase 3 inhibitor (GSK3i), and their effects were studied on cultured crypts. Cell lineage differentiation was assessed by immunohistochemistry and quantitative polymerase chain reaction. Cultured porcine cells were serially passaged and transduced with a lentiviral vector. Whereas ENR medium supported murine enteroid growth, it did not sustain porcine crypts beyond 5 days. Supplementation of Wnt3a-CM and GSK3i resulted in the formation of complex porcine enteroids with budding extensions. These enteroids contained a mixture of stem and differentiated cells and were successfully passaged in the presence of GSK3i. Crypts grown in media supplemented with porcine ISEMF-CM formed spheroids that were less well differentiated than enteroids. Enteroids and spheroids were transfected with a lentivirus with high efficiency. Thus, our method maintains juvenile and adult porcine crypt cells long-term in culture. Porcine enteroids and spheroids can be successfully passaged and transduced by using lentiviral vectors.

Mouse model of endoscopically ablated enteric nervous system.

BACKGROUND: Current transgenic animal models of Hirschsprung disease are restricted by limited survival and need for special dietary care. We used small animal colonoscopy to produce chemically ablated enteric nervous system in the distal colon and rectum of normal mice. MATERIALS AND METHODS: Adult C57BL/6 mice underwent colonoscopy with submucosal injection of 75-100 µL of saline (n = 2) or 0.002% (n = 2), 0.02% (n = 15), or 0.2% (n = 2) benzalkonium chloride (BAC). Each mouse received 1-3 injections in the distal colon and rectum. Mice were sacrificed on postprocedure day 7 or 28. Injection sites were analyzed histologically and with immunostaining for ß-tubulin III. RESULTS: Submucosal injection of 0.02% BAC resulted in megacolon and obliteration of 82 ± 8.8% of myenteric ganglia at the injection site on postprocedure day 7 compared with normal colon. This effect was sustained until day 28. Injection of 0.002% BAC had little effect on the myenteric neuronal network at these time points. Multiple injections of 0.002% or 0.02% BAC (up to three injections per mouse) were well tolerated. Injection of 0.2% BAC caused acute toxicity or death. CONCLUSIONS: A novel model of chemically ablated enteric nervous system in the mouse colon and rectum is introduced. This model can be valuable in evaluating targeted cell delivery therapies for Hirschsprung disease.

Anesthetic Management of Advanced Stage Ludwig's Angina: A Case Report and Review With Emphasis on Compromised Airway Management.

Ludwig's angina, although uncommon, remains a potentially life-threatening condition because of the risk of impending airway obstruction. Effective treatment is based on early recognition of the clinical process, with the appropriate use of parenteral antibiotics, securing the airway, and formal surgical drainage of the infection. Awake fiberoptic intubation under topical anesthesia may be the preferred method to secure the airway. Flexible nasotracheal intubation requires skill and experience. When fiberoptic bronchoscopy is not feasible, not available, or has failed, an elective awake cricothyrotomy and tracheostomy are the options. Furthermore, the introduction of newer advanced airway techniques, such as video-assissted laryngoscopy, may allow the clinician additional flexibility in nonsurgical airway management. We present a recent case of a patient with Ludwig's angina, successfully managed at our hospital, with a brief review of airway management options.

Diverticulitis occurs early after lung transplantation.

BACKGROUND: Lung transplantation recipients are at an increased risk for developing diverticulitis. However, the incidence and natural history of diverticulitis have not been well characterized. Our objective was to identify patient and transplant-related factors that may be associated with an increased risk of developing diverticulitis in this patient population. MATERIALS AND METHODS: This is a retrospective single institution study. All patients who received a lung transplant between May 2008 and July 2013 were evaluated using an existing lung transplantation database. Patient-related factors, the incidence and timing of diverticulitis, and outcomes of medical and surgical management were measured. RESULTS: Of the 314 patients who received a lung transplant, 14 patients (4.5%) developed diverticulitis. All episodes (100%) of diverticulitis occurred within the first 2 y after transplantation. Eight patients (57%) required surgery with a mortality rate of 12.5%. Six patients (43%) were managed medically and did not require surgery with a mean follow-up period of 442 d. CONCLUSIONS: Diverticulitis is common after lung transplantation and occurs with a higher incidence compared with the general population. Diverticulitis occurs early in the posttransplant period, and the majority of patients require surgery. Patients who respond promptly to medical treatment may not require elective resection. A greater awareness of the risk of diverticulitis in the early posttransplant period may allow for earlier diagnosis and treatment.

Loss and Return of Ticklishness in Functional Neurological Disorder.

BACKGROUND: People with functional neurological disorder (FND) have abnormalities in sensory processing. Loss of ticklishness has been rarely reported. OBJECTIVES: To describe associated clinical features in people with FND and loss of ticklishness and explore correlations with sensory changes. METHODS: Retrospective audit of clinical letters of people diagnosed with FND in a tertiary clinic and further cases identified in a general neurology clinic. RESULTS: Thirty-eight patients with loss of ticklishness are described, of which most had other functional sensory symptoms and signs. It was more often localized to one limb, rather than generalized, in those with pain or weakness. Dissociation for the affected body part was often described. CONCLUSIONS: Loss of ticklishness in FND is frequently described and offers insights into mechanisms of agency, sensory processing and interoception, which are known to be altered in FND.

Boosting Sinh Cosh Optimizer and arithmetic optimization algorithm for improved prediction of biological activities for indoloquinoline derivatives.

Quantitative Structure Activity Relation (QSAR) models are mathematical techniques used to link structural characteristics with biological activities, thus considered a useful tool in drug discovery, hazard evaluation, and identifying potentially lethal molecules. The QSAR regulations are determined by the Organization for Economic Cooperation and Development (OECD). QSAR models are helpful in discovering new drugs and chemicals to treat severe diseases. In order to improve the QSAR model's predictive power for biological activities of naturally occurring indoloquinoline derivatives against different cancer cell lines, a modified machine learning (ML) technique is presented in this paper. The Arithmetic Optimization Algorithm (AOA) operators are used in the suggested model to enhance the performance of the Sinh Cosh Optimizer (SCHO). Moreover, this improvement functions as a feature selection method that eliminates superfluous descriptors. An actual dataset gathered from previously published research is utilized to evaluate the performance of the suggested model. Moreover, a comparison is made between the outcomes of the suggested model and other established methodologies. In terms of pIC50 values for different indoloquinoline derivatives against human MV4-11 (leukemia), human HCT116 (colon cancer), and human A549 (lung cancer) cell lines, the suggested model achieves root mean square error (RMSE) of 0.6822, 0.6787, 0.4411, and 0.4477, respectively. The biological application of indoloquinoline derivatives as possible anticancer medicines is predicted with a high degree of accuracy by the suggested model, as evidenced by these findings.

Inpatients' understanding of the hospitalist role and common medical terminology.

Many patients are unable to identify members of their hospital care team and experience confusion regarding some medical terminology used during hospitalization, including descriptions of the structure of their inpatient care team. This cross-sectional study sought to (1) examine inpatients' understanding of the role of a hospitalist and (2) assess inpatients' familiarity with other medical terminology commonly used in the hospital. We surveyed 172 patients admitted to the hospital medicine service at two academic medical centers. We found that almost half (47%) of respondents were unfamiliar with the term and/or role of a hospitalist, while the remaining patients had varied understanding of the role. Several other medical terms were frequently misunderstood (such as "NPO," "PA," and "Attending"). Ongoing efforts are needed to improve communication to ensure that hospitalized patients understand the hospitalist's role and the medical terms shared with them.

Buckling forces and the wavy folds between pleural epithelial cells.

Cell shapes in tissues are affected by the biophysical interaction between cells. Tissue forces can influence specific cell features such as cell geometry and cell surface area. Here, we examined the 2-dimensional shape, size, and perimeter of pleural epithelial cells at various lung volumes. We demonstrated a 1.53-fold increase in 2-dimensional cell surface area and a 1.43-fold increase in cell perimeter at total lung capacity compared to residual lung volume. Consistent with previous results, close inspection of the pleura demonstrated wavy folds between pleural epithelial cells at all lung volumes. To investigate a potential explanation for the wavy folds, we developed a physical simulacrum suggested by D'Arcy Thompson in On Growth and Form. The simulacrum suggested that the wavy folds were the result of redundant cell membranes unable to contract. To test this hypothesis, we developed a numerical simulation to evaluate the impact of an increase in 2-dimensional cell surface area and cell perimeter on the shape of the cell-cell interface. Our simulation demonstrated that an increase in cell perimeter, rather than an increase in 2-dimensional cell surface area, had the most direct impact on the presence of wavy folds. We conclude that wavy folds between pleural epithelial cells reflects buckling forces arising from the excess cell perimeter necessary to accommodate visceral organ expansion.

Mannan oligosaccharides as a prebiotic for laying hens: effects on fertility, hatchability, productive performance, and immunity.

This experiment examined how adding mannan-oligosaccharides (MOS) to the diet affected fertility, hatching rates, egg production, carcass characteristics, cost-effectiveness, and immune function in laying hens. One hundred and twenty Mandarah chickens (30 hens and 3 roosters per group) were randomly chosen between 34 and 50 wk old and divided into four groups. The first group was the control group, which was given just the basal diet. The basal diet was given to the second, third, and fourth experimental groups along with three different levels of MOS (0.1, 0.2, and 0.5 g/kg diet, respectively). Results found that hens fed MOS at various levels laid eggs at a significantly higher rate, enhanced egg number, egg mass and feed conversion ratio than the control group (P < 0.05). MOS seemed to improve carcass quality. The best results for egg quality (Haugh unit) and testosterone levels were seen with a dose of 0.5 g/kg of MOS compared to the control birds (P < 0.05). All MOS levels led to higher estradiol-17ß (E2) levels and better economic efficiency (EE). MOS also improved the hens' immune systems as compared to the control group. Hens-fed MOS had significantly greater levels of antibodies against Influenza viruses (H9N2) and Infectious Bronchitis Virus (P < 0.05). Also, the spleen and thymus gland, both crucial immune system components, were slightly larger (P < 0.05). It's important to note that fertility rates, hatchability, and embryo mortality rates remained similar across all groups. So, our findings suggest that incorporating MOS into the birds' diet enhances their productivity, strengthens their immune system, improves EE, and contributes to the overall health of the hens.