Tissue Surface Area Research Articles

Isolation of Adipose Tissue-Derived Stem Cells: Enzymatic Digestion in Combination with Mechanical Distortion to Increase Adipose Tissue-Derived Stem Cell Yield from Human Aspirated Fat.

Mesenchymal stem cells (MSCs) are of great interest due to their properties of immune modulation, tissue regeneration, and multipotent differentiation. Future developments of clinical applications, however, require a higher yield of MSCs, lower number of passages of cells in culture, and shorter time from harvest to use. Optimization and standardization of techniques for mesenchymal adipose tissue-derived stem cell isolation offers solutions to current bottlenecks as a larger amount of MSCs can be isolated. These improvements result in shorter expansion time, fewer passages, less donor material needed, and higher MSC yield. This paper describes an MSC isolation method combining enzymatic digestion with mechanic disruption. This protocol is a standardized and easy-to-implement method for reaching significantly higher MSC yields compared to conventional enzymatic isolation protocols. Based on the results presented, we hypothesize that the combined enzymatic and mechanical method increases the surface area of the adipose tissue, facilitating digestion by enzymes. This approach reduces the amount of adipose tissue and in vitro expansion time needed to reach sufficient amounts of MSCs for clinical purposes. Importantly, the method does not require increased amounts of collagenase, nor does it impair the viability or differentiability of the MSCs. Using this protocol increases MSC yield by a factor of three. As a consequence, these results indicate that the physiological concentration of MSCs in adipose tissue is higher than previously assumed. © 2018 by John Wiley & Sons, Inc.

Lower Subcutaneous or Visceral Adiposity Assessed by Abdominal Computed Tomography Could Predict Adverse Outcome in Patients With Crohn's Disease.

Changes in body composition have been described in patients with Crohn's disease, but their predictive performances on disease evolution remain undefined. The aims of our study were to assess, in patients with Crohn's disease requiring abdominal computed tomography, body composition by computed tomography, and to study the outcome according to various body composition parameters at the time of the computed tomography. Patients with Crohn's disease who underwent abdominal computed tomography for suspected complications were retrospectively included. The definition of adverse outcome included death or need for surgery within 6 months of the computed tomography. Skeletal muscle index and visceral and subcutaneous adiposity indexes were calculated from tissue surface areas measured at the third lumbar vertebra, divided by the height squared. The prevalence of underweight was 26.8% and the prevalence of sarcopenia was 33.6%. After gender adjustment, skeletal muscle index tended to be reduced in patients with adverse outcome, compared with patients without surgery or death [p = 0.07]. Moreover, subcutaneous adiposity index and visceral adiposity index were significantly lower in patients with surgery or death [p = 0.009 and p < 0.001, respectively]. These differences were almost equivalent in both genders for the subcutaneous adiposity index but were clearly stronger in men for the visceral adiposity index. Subcutaneous and visceral adiposity indexes correlate inversely with adverse outcome in patients with Crohn's disease. Alteration of body composition assessed by computed tomography in these patients appears to be a marker of disease severity.

A Randomized Clinical Trial of Flex 19G Needles versus 22G Needles for Endobronchial Ultrasonography in Suspected Lung Cancer

Background: A flexible 19-gauge (Flex 19G) needle has been developed for endobronchial ultrasonography. Objectives: We aimed to evaluate quantitative and qualitative specimen characteristics of Flex 19G in a randomized controlled setting for patients with suspected lung cancer. Methods: We undertook a single-center, randomized, controlled trial. A computer-generated randomization assigned all enrolled patients 1: 1 to undergo endobronchial ultrasonography using a Flex 19G or a 22-gauge (22G) needle for lymph node tissue sampling. Pathologists were blinded to the group assignment. The primary end point was histological tissue core procurement. The secondary end points were diagnostic yield, specimen bloodiness and overall quality, tissue surface area and performance for next-generation sequencing (NGS), and procedure-related complications. Results: Between June 2016 and February 2017, we randomly allocated a total of 78 patients: 39 patients to Flex 19G and 39 patients to 22G. No superiority in tissue core procurement was observed for Flex 19G compared to 22G (67 vs. 72%, p = 0.81). No significant difference was observed in diagnostic yield and overall specimen quality, but transbronchial needle aspiration specimens by Flex 19G were bloodier and had a larger tissue surface area. NGS was successful for clinically relevant genes in 96% and for all 26 genes tested in 81% of the samples. There was no difference in clinically relevant complications. Conclusions: No superiority is observed for Flex 19G in histological tissue core procurement rate. The Flex 19G needle could be considered when a larger tissue surface is of special interest.

Effect of long-term nitrite administration on browning of white adipose tissue in type 2 diabetic rats: A stereological study

IntroductionNitric oxide (NO) deficiency is associated with obesity and type 2 diabetes. Nitrite, a NO donor, is considered as a new therapeutic agent in diabetes. This study aims at determining effects of long-term nitrite administration on browning of white adipose tissue (WAT) in type 2 diabetic rats. MethodsMale rats were divided into 4 groups: Control, control + nitrite, diabetes, and diabetes + nitrite. Sodium nitrite (50 mg/L in drinking water) was administered for 3 months. Body weight was measured weekly. Fasting serum levels of glucose and nitric oxide metabolites (NOx) were measured monthly. Histological evaluations and measurement of cyclic guanosine monophosphate (cGMP) and NOx levels in adipose tissue were done at the end of the study. ResultsNitrite decreased serum glucose concentration and body weight gain in diabetic rats by 27.6% and 37.9%, respectively. In diabetic rats, nitrite increased NOx and cGMP levels in inguinal WAT by 95.7% and 33.1%, respectively. Numerical density in WAT of nitrite-treated diabetic rats was higher than in diabetic ones (995 ± 83 vs. 2513 ± 256 cell/mm3, P < 0.001); in addition, total surface area (4.84 ± 0.32 vs. 44.26 ± 9.7, mm2, P < 0.001) and volume of inguinal beige adipose tissue (7.2 ± 0.49 vs. 66.4 ± 14.51 mm3, <0.001) were higher in nitrite-treated diabetic rats compared to diabetic ones. ConclusionsFavorable effects of long-term nitrite administration in obese type 2 diabetic rats is, at least in part, due to browning of WAT and also associated with increased NOx and cGMP level in adipose tissue. These findings may have potential applications for management of diabesity.

Regeneration of Activated Carbon Fiber by the Electro-Fenton Process.

An electro-Fenton (EF) based technology using activated carbon (AC) fiber as cathode and BDD as anode has been investigated for both regeneration of AC and mineralization of organic pollutants. The large specific surface area and low intraparticle diffusion resistance of AC tissue resulted in high maximum adsorption capacity of phenol (PH) (3.7 mmol g-1) and fast adsorption kinetics. Spent AC tissue was subsequently used as the cathode during the EF process. After 6 h of treatment at 300 mA, 70% of PH was removed from the AC surface. The effectiveness of the process is ascribed to (i) direct oxidation of adsorbed PH by generated hydroxyl radicals, (ii) continuous shift of adsorption equilibrium due to oxidation of organic compounds in the bulk, and (iii) local pH change leading to electrostatic repulsive interactions. Moreover, 91% of PH removed from AC was completely mineralized, thus avoiding adsorption of degradation byproducts and accumulation of toxic compounds such as benzoquinone. Morphological and chemical characteristics of AC were not affected due to the effect of cathodic polarization protection. AC tissue was successfully reused during 10 cycles of adsorption/regeneration with regeneration efficiency ranging from 65 to 78%, in accordance with the amount of PH removed from the AC surface.

3D fiber deposited polymeric scaffolds for external auditory canal wall.

The external auditory canal (EAC) is an osseocartilaginous structure extending from the auricle to the eardrum, which can be affected by congenital, inflammatory, and neoplastic diseases, thus reconstructive materials are needed. Current biomaterial-based approaches for the surgical reconstruction of EAC posterior wall still suffer from resorption (biological) and extrusion (synthetic). In this study, 3D fiber deposited scaffolds based on poly(ethylene oxide terephthalate)/poly(butylene terephthalate) were designed and fabricated to replace the EAC wall. Fiber diameter and scaffold porosity were optimized, leading to 200 ± 33 µm and 55% ± 5%, respectively. The mechanical properties were evaluated, resulting in a Young's modulus of 25.1 ± 7.0 MPa. Finally, the EAC scaffolds were tested in vitro with osteo-differentiated human mesenchymal stromal cells (hMSCs) with different seeding methods to produce homogeneously colonized replacements of interest for otologic surgery. This study demonstrated the fabrication feasibility of EAC wall scaffolds aimed to match several important requirements for biomaterial application to the ear under the Tissue Engineering paradigm, including shape, porosity, surface area, mechanical properties and favorable in vitro interaction with osteoinduced hMSCs. This study demonstrated the fabrication feasibility of outer ear canal wall scaffolds via additive manufacturing. Aimed to match several important requirements for biomaterial application to ear replacements under the Tissue Engineering paradigm, including shape, porosity and pore size, surface area, mechanical properties and favorable in vitro interaction with osteo-differentiated mesenchymal stromal cells.

Examining the association between adiposity and biochemical recurrence after radical prostatectomy.

Herein, we examined the association between adiposity, as measured by computed tomography (CT), and biochemical recurrence (BCR) after radical prostatectomy (RP). Using axial CT images, preoperative fat mass index (FMI) was calculated for 698 men who underwent RP from 2007-2010 by using measurements of total surface area of adipose tissue at the L3 level. Obesity was classified according to National Health and Nutrition Examination Survey (NHANES) standards for obesity (FMI >9 kg/m2). The associations between obesity and the distribution of adiposity (visceral vs. subcutaneous) with BCR were examined using the Kaplan-Meier method and Cox proportional hazards regression analyses. Obese men were older than non-obese men (63.0 vs. 60.7 years; p<0.001), but were similar with regards to all other clinical and pathological characteristics. With a median followup of six years, 152 patients were diagnosed with BCR. Five-year BCR-free survival was similar between obese and non-obese patients (80.6% vs. 82.1%; p=0.27). Furthermore, in multivariable analyses, obesity was not independently associated with the risk of BCR (hazard ratio [HR] 1.02; 95% confidence interval [CI] 0.73-1.43). Similar results were obtained when analyzing FMI as a continuous variable (HR 1.02; 95% CI 0.94-1.09 for each 1 kg/m2 increase in FMI). Additionally, neither visceral adiposity, subcutaneous adiposity, or visceral-to-subcutaneous adiposity ratio were associated with BCR (all p>0.05) in multivariable analyses. Neither total abdominal adiposity nor the distribution of adiposity were independently associated with BCR after RP in this study. As such, the presence of obesity may not be a marker of increased oncological risk after RP.

Contribution of dental tissues to sex determination in modern human populations.

Accurate sex estimation is an essential step for the reconstruction of the biological profile of human remains. Earlier studies have shown that elements of the human permanent dentition are sexually dimorphic. The aims of this study are to determine the degree of sexual dimorphism in the dental tissue volumes and surface areas of mandibular canines and to explore its potential for reliable sex determination. The teeth included in this study (n = 69) were selected from anthropological collections from Spain, South Africa and Sudan. In all cases, the sex of the individuals was known. The teeth were scanned and three-dimensional (3D) measurements (volumes and surfaces areas) were obtained. Finally, a dsicriminant function analysis was applied. Our results showed that sexual dimorphism in canine size is due to males having greater amounts of dentine, whereas enamel volume does not contribute significantly to overall tooth size dimorphism. Classification accuracy of the multivariable equations tested on slightly worn teeth ranged from 78 to 90.2% for the crossvalidation, and from 71.43 to 84.62% for the hold-out sample validation. When all functions were applied together, the sex was correctly assigned 92.30% of the time. Our results suggest that the 3D variables from mandibular canine dental tissues are useful for sex determination as they present a high degree of dimorphism. The results obtained show the importance of 3D dental tissue measurements as a methodology in sex determination, which application should be considered as a supplemental method to others.

Colony size and turbulent flow speed modulate the calcification response of the coral Pocillopora verrucosa to temperature

For poikilothermic organisms in aquatic environment, temperature strongly affects metabolism and modulates the extent to which it is mass transfer limited. The effects of temperature on metabolism are translated into organism performance depending on the organism size, morphology, and the flow regime to which they are exposed. Environmental forcing of metabolism is particularly germane to tropical reef corals, because they increasingly are exposed to upward temperature perturbations to which they display variable responses that remain incompletely understood. In this study, we tested the effects of colony size and seawater turbulence on the response of the common Indo-Pacific branching coral, Pocillopora verrucosa, to different seawater temperatures. Using whole-colony calcification as a response variable, 12 tanks (each 150 l) were used in two trials lasting 14 days to contrast the effects of seawater turbulence (two levels) and temperature (25.5 vs 29.5 °C) on colonies varying in size from ~ 4 to 13-cm diameter. Turbulence in the tanks was measured as the root mean square (rms) turbulent flow speed (q rms), which quantified the variation in speed of complex and non-linear flow, regardless of direction. Treatments contrasted low (q rms = 0.63 cm s−1) and high (q rms = 2.07 cm s−1) turbulence that were ecologically relevant for shallow coral reefs. The effects of turbulent flow speed and temperature on calcification (mg day−1) differed among colony sizes: at 25.5 °C, calcification increased with tissue surface area at a slower rate under high, compared to low turbulent flow speeds; at low turbulent flow speeds, calcification increased with tissue surface area more slowly at 29.5 versus 25.5 °C. These results show that the response of branching corals to temperatures depends on colony size and turbulent flow speed. This outcome suggests that spatial heterogeneity in turbulent flow regimes across a coral reef may provide opportunities for branching corals to exploit colony size to modulate susceptibility to rising temperature.

Visceral Adiposity Is an Independent Determinant of Hypercoagulability as Measured by Thrombin Generation in Morbid Obesity.

Introduction Increased visceral adipose tissue (VAT) has been shown to be associated with the development of insulin resistance, type 2 diabetes, stroke, and ischemic heart disease. It remains unknown whether fat distribution impacts on coagulation markers and/or the risk of venous thrombosis. This study evaluates markers of hypercoagulability in class III obesity (body mass index [BMI] >40 kg/m 2 ) compared with nonobese controls. We further investigated whether hypercoagulability was influenced by VAT, metabolic syndrome, and metabolic markers, including adiponectin. Patients and Methods Ninety patients were recruited from the obesity clinic at King's College Hospital from November 2009 to December 2011. The inclusion criteria were class III obesity (BMI ≥40 kg/m 2 ) and age 18 to 65 years. A control group (healthy ambulatory participants, with a BMI < 30 kg/m 2 ) was recruited from volunteers responding to advertisement. Abdominal VAT and subcutaneous adipose tissue surface areas were determined by evaluation of a single-slice CT at spinal vertebra L4. Results Thrombin generation revealed a significantly increased peak and endogenous thrombin potential in patients compared with controls. Lag time and time to peak (ttP) were also significantly prolonged in patients. VAT was found to have the strongest association with thrombin generation parameters: lag time (β = 0.378; p < 0.001), peak thrombin (0.378; p = 0.04), and ttP (β = 0.373; p = 0.001). BMI was found to be a predictor for lag time only (β = 0.313; p = 0.003). SAT was not associated with any of the thrombin generation parameters (data not shown). VAT was found to be an independent determinant of peak thrombin, lag time, and ttP. The study suggests not only fat mass but also fat distribution, particularly visceral adiposity, mediates hypercoagulability in obesity.

Comparative leaf growth strategies in response to low-water and low-light availability: variation in leaf physiology underlies variation in leaf mass per area in Populus tremuloides.

Developmental phenotypic plasticity can allow plants to buffer the effects of abiotic and biotic environmental stressors. Therefore, it is vital to improve our understanding of how phenotypic plasticity in ecological functional traits is coordinated with variation in physiological performance in plants. To identify coordinated leaf responses to low-water (LW) versus low-light (LL) availability, we measured leaf mass per area (LMA), leaf anatomical characteristics and leaf gas exchange of juvenile Populus tremuloidesMichx. trees. Spongy mesophyll tissue surface area (Asmes/A) was correlated with intrinsic water-use efficiency (WUEi: photosynthesis, (Aarea)/stomatal conductance (gs)). Under LW availability, these changes occurred at the cost of greater leaf tissue density and reduced expansive growth, as leaves were denser but were only 20% the final area of control leaves, resulting in elevated LMA and elevated WUEi. Low light resulted in reduced palisade mesophyll surface area (Apmes/A) while spongy mesophyll surface area was maintained (Asmes/A), with no changes to WUEi. These leaf morphological changes may be a plastic strategy to increase laminar light capture while maintaining WUEi. With reduced density and thickness, however, leaves were 50% the area of control leaves, ultimately resulting in reduced LMA. Our results illustrate that P. tremuloides saplings partially maintain physiological function in response to water and light limitation by inducing developmental plasticity in LMA with underlying anatomical changes. We discuss additional implications of these results in the context of developmental plasticity, growth trade-offs and the ecological impacts of climate change.

MP33-14 THE ROLE OF CHRONIC PROSTATIC INFLAMMATION IN PROSTATE CARCINOGENESIS: A RETROSPECTIVE FOLLOW-UP STUDY

You have accessJournal of UrologyProstate Cancer: Detection & Screening III1 Apr 2017MP33-14 THE ROLE OF CHRONIC PROSTATIC INFLAMMATION IN PROSTATE CARCINOGENESIS: A RETROSPECTIVE FOLLOW-UP STUDY Liwei Jia, Shawn Silver, and Gregory MacLennan Liwei JiaLiwei Jia More articles by this author , Shawn SilverShawn Silver More articles by this author , and Gregory MacLennanGregory MacLennan More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2017.02.1010AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Chronic inflammation is a well-known contributor to several human cancers. The aim of this study was to investigate the role of chronic prostatic inflammation in prostatic carcinogenesis, with a specific focus on high grade cancers. METHODS A retrospective search was performed to identify prostate biopsy cases accessioned in our institution between 01/01/2007 and 12/31/2009. Pathology reports of each case were reviewed. We reviewed the slides of all patients with benign initial prostate biopsies, followed by at least one set of prostate biopsies longer than one year after the initial biopsies. The presence and extent of chronic inflammation and other pathological findings were assessed and recorded. We defined chronic inflammation as involvement of 1% or more of the surface area of the prostatic tissue by a chronic inflammatory cell infiltrate. We also recorded the presence or absence of postatrophic hyperplasia (PAH) and inflamed atrophy (IA), a lesion defined by the presence of clusters of atrophic glands surrounded by a prominent and readily identifiable lymphocytic infiltrate. Cases in which prostatic adenocarcinoma was subsequently diagnosed were evaluated for Gleason pattern and score. RESULTS Among 1006 patients who underwent prostate biopsies, a total of 244 cases with available slides were diagnosed initially as benign prostatic tissue, of which two patient subsets were identified, including 206 patients with and 38 patients without chronic inflammation in the initial biopsy. Post-atrophic hyperplasia (PAH) and inflamed atrophy (IA) were more frequently identified in patients with chronic inflammation. In follow up biopsies performed at least 1 year following the initial benign biopsies, 75 patients (36%) with chronic inflammation were found to have adenocarcinoma, whereas cancer was found in only 6 patients (16%) without chronic inflammation in the initial biopsies. Of those who were found to have cancer after the initial biopsies showing only chronic inflammation, 46% had a Gleason 3 + 3 pattern (grade group 1), 20% had a Gleason 3 + 4 pattern (grade group 2), and 13% had a Gleason 4 + 3 pattern (grade group 3). Significantly, high grade cancers with Gleason score of 8 or higher (grade groups 4 or 5) were identified in 21% (16 of 75) of patients with chronic inflammation in the initial benign biopsies, whereas no patient without chronic inflammation in the initial benign biopsies were found to have cancer with a Gleason score of 8 or higher (grade groups 4 or 5). CONCLUSIONS Our retrospective follow up study suggests a strong association between chronic prostatic inflammation and the development of prostatic adenocarcinoma. © 2017FiguresReferencesRelatedDetails Volume 197Issue 4SApril 2017Page: e422 Advertisement Copyright & Permissions© 2017MetricsAuthor Information Liwei Jia More articles by this author Shawn Silver More articles by this author Gregory MacLennan More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

A Single US Center Experience of Transbronchial Lung Cryobiopsy for Diagnosing Interstitial Lung Disease With a 2-Scope Technique.

Background:Transbronchial lung cryobiopsy (TBLC) has been used to establish the diagnosis of interstitial lung disease (ILD) in recent years. The technique and diagnostic yield vary among institutions. We report a new 2-scope technique and the results of TBLC in our institution.Methods:This is a retrospective chart review of patients who underwent TBLC for evaluation of ILD. Bronchoscopy with TBLC was performed by a board-certified interventional pulmonologist with a 2-scope technique under general anesthesia.Results:A total of 74 patients underwent TBLC with a 2-scope technique. Their mean age was 54±14 years. The mean tissue surface area was 63.54±6.76 mm2. The average anesthesia time was 80.66 minutes. The diagnostic yield was 87.84%. The most common diagnosis was sarcoidosis pneumothorax, which occurred in 5 cases (7%). There was 1 case with bronchoscopic-related respiratory failure associated with significant bleeding. Death occurred in 3 cases (4%), which is comparable to recent mortality data for “elective” surgical lung biopsy for ILD (1.7% to 4.2%).Conclusion:TBLC with a 2-scope technique could be an alternative method for diagnosing various types of ILD in patients unfit for surgical lung biopsy. Further prospective studies should clarify its role in the diagnostic armamentarium for undiagnosed ILDs.

In vivo imaging of coral tissue and skeleton with optical coherence tomography.

Application of optical coherence tomography (OCT) for in vivo imaging of tissue and skeleton structure of intact living corals enabled the non-invasive visualization of coral tissue layers (endoderm versus ectoderm), skeletal cavities and special structures such as mesenterial filaments and mucus release from intact living corals. Coral host chromatophores containing green fluorescent protein-like pigment granules appeared hyper-reflective to near-infrared radiation allowing for excellent optical contrast in OCT and a rapid characterization of chromatophore size, distribution and abundance. In vivo tissue plasticity could be quantified by the linear contraction velocity of coral tissues upon illumination resulting in dynamic changes in the live coral tissue surface area, which varied by a factor of 2 between the contracted and expanded state of a coral. Our study provides a novel view on the in vivo organization of coral tissue and skeleton and highlights the importance of microstructural dynamics for coral ecophysiology.

A Modified Approach to Extensive Oromandibular Reconstruction Using Free Fibula Flaps.

In select patients with advanced disease resulting in large composite tissue defects, consideration is often given to multiple flap reconstruction. The authors propose an alternative option. Using virtual surgical planning the authors demonstrate how modest sacrifice in projection translates into a substantial decrease in the volume and surface area of soft tissue needed, in turn maximizing soft tissue coverage with a single fibula free flap. The authors used 3-dimensional virtual surgery to simulate angle-to-angle reconstructions using free fibula flaps. The reference 3-segment reconstruction was done using symphyseal projection to the plane perpendicular to the anterior nasal spine, a customary landmark. Additional simulations were then performed using recessed projections 0.5 mm, 1 cm, 1.5 cm, and 2 cm posterior to anterior nasal spine plane. Program analytics were used to calculate the surface area and volume of the floor of mouth. With projection recessed by 1 cm, surface area decreased 22% to 14 cm. With projection recessed by 2 cm, surface area decreased 44% to 10 cm. With a 3-segment construct converted to a 2-segment construct, surface area decreased 22% to 14 cm. This demonstrates for the first time an official analysis of an intraoperative modification that sacrifices little and gains a lot. Ultimately, 1 compound flap can be used in extensive reconstructions with increased confidence that it will not be overly stressed.

Modeling the Effect of Curvature on the Collective Behavior of Cells Growing New Tissue

The growth of several biological tissues is known to be controlled in part by local geometrical features, such as the curvature of the tissue interface. This control leads to changes in tissue shape that in turn can affect the tissue’s evolution. Understanding the cellular basis of this control is highly significant for bioscaffold tissue engineering, the evolution of bone microarchitecture, wound healing, and tumor growth. Although previous models have proposed geometrical relationships between tissue growth and curvature, the role of cell density and cell vigor remains poorly understood. We propose a cell-based mathematical model of tissue growth to investigate the systematic influence of curvature on the collective crowding or spreading of tissue-synthesizing cells induced by changes in local tissue surface area during the motion of the interface. Depending on the strength of diffusive damping, the model exhibits complex growth patterns such as undulating motion, efficient smoothing of irregularities, and the generation of cusps. We compare this model with in vitro experiments of tissue deposition in bioscaffolds of different geometries. By including the depletion of active cells, the model is able to capture both smoothing of initial substrate geometry and tissue deposition slowdown as observed experimentally.

Aberrant lung remodeling in a mouse model of surfactant dysregulation induced by modulation of the Abca3 gene

The lipid transporter, ATP binding cassette class A3 (ABCA3), plays a critical role in the biogenesis of alveolar type 2 (AT2) cell lamellar bodies (LBs). A relatively large number of mutations in the ABCA3 gene have been identified in association with diffuse parenchymal lung disease (DPLD), the most common of which is a missense mutation (valine substitution for lysine at residue 292 (ABCA3E292V)) that leads to functional impairment of the transporter in vitro. The consequences of ABCA3E292V gene expression in vivo are unknown. To address this question, we developed mouse models expressing ABCA3E292V knocked-in to the endogenous mouse locus. The parental (F1) mouse line (mAbca3E292V) that retained an intronic pgk-Neo selection cassette (inserted in reverse orientation) (mAbca3E292V–rNeo) demonstrated an allele dependent extracellular surfactant phospholipid (PL) deficiency. We hypothesize that this PL deficiency leads to aberrant parenchymal remodeling contributing to the pathophysiology of the DPLD phenotype. Compared to wild type littermates, baseline studies of mice homozygous for the pgk-Neo insert (mAbca3E292V–rNeo+/+) revealed nearly 50% reduction in bronchoalveolar lavage (BAL) PL content that was accompanied by quantitative reduction in AT2 LB size with a compensatory increase in LB number. The phenotypic alteration in surfactant lipid homeostasis resulted in an early macrophage predominant alveolitis which peaked at 8 weeks of age. This was followed by age-dependent development of histological DPLD characterized initially by peribronchial inflammatory cell infiltration and culminating in both an emphysema-like phenotype (which included stereologically quantifiable reductions in both alveolar septal surface area and volume of septal wall tissue) plus foci of trichrome-positive collagen deposition together with substantial proliferation of hyperplastic AT2 cells. In addition to spontaneous lung remodeling, mABCA3E292V–rNeo mice were rendered more vulnerable to exogenous injury. Three weeks following intratracheal bleomycin challenge, mAbca3–rNeo mice demonstrated allele-dependent susceptibility to bleomycin including enhanced weight loss, augmented airspace destruction, and increased fibrosis. Removal of the rNeo cassette from mAbca3 alleles resulted in restoration of BAL PL content to wild-type levels and an absence of changes in lung histology up to 32 weeks of age. These results support the importance of surfactant PL homeostasis as a susceptibility factor for both intrinsic and exogenously induced lung injury/remodeling.

Changes in the surface area of facial soft tissue before and after orthognathic surgery

Changes in the surface area of facial soft tissue before and after orthognathic surgery

Oral-Facial Tissue Reconstruction in the Regenerative Axolotl.

Absence of large amounts of orofacial tissues caused by cancerous resections, congenital defects, or trauma results in sequelae such as dysphagia and noticeable scars. Oral-neck tissue regeneration was studied in the axolotl (regenerative amphibian) following a 2.5-mm punch biopsy that simultaneously removed skin, connective tissue, muscle, and cartilage in the tongue and intermandibular region. The untreated wound was studied macroscopically and histologically at 17 different time points ranging from 0 to180 days (N = 120 axolotls). At 12 hr, the wound's surface was smoothened and within 1mm, internal lingual muscular modifications occurred; at the same distance, between days 4-7 lingual muscle degradation was complete. Immunofluorescence indicates complete keratinocytes migration by 48 hr. These cells with epidermal Leydig cells, appearing yellow, lead the chin's deep tissue outgrowth until its closure on the 14th day. Regeneration speeds varied and peaked in time for each tissue, (1) deep chin 84.3 μm/hr from 24 to 96 hr, (2) superficial chin 71.1 μm/hr from 7-14 days, and (3) tongue 86.0 μm/hr between 48 hr and 7 days. Immunofluorescence to Col IV showed basement membrane reconnected between days 30-45 coinciding with the chin's dermal tissue's surface area recovery. New muscle appeared at 21 days and was always preceded by the formation of a collagen bed. Both chin tissues regain all surface area and practically all components while the lingual structure lacks some content but is generally similar to the original. The methodology and high-resolution observations described here are the first of its kind for this animal model and could serve as a basis for future studies in oral and facial regenerative research.

25 - Visceral adipose tissue and perirenal fat surface area as a predictor of renal cortical neoplasm histopathology

25 - Visceral adipose tissue and perirenal fat surface area as a predictor of renal cortical neoplasm histopathology