Humidified Chamber Research Articles

First Report of Fusarium incarnatum-equiseti Species Complex Causing Fruit Rot on Muskmelon (Cucumis melo) in China

Muskmelon (Cucumis melo L.) is an economically important fruit crop in China. In September 2018, fruit rot was observed on approximately 20% of muskmelon fruits in Harbin, Heilongjiang Province, China. Brown water-soaked lesions were observed on the fruit side in contact with soil initially, which gradually extended to most of or the entire fruit. Internal decay was observed with white to dark brown mycelium on the fruit surface. Diseased muskmelon tissues were surface disinfested with 1% NaOCl for 3 min, 70% ethanol for 10 s, and then washed three times with sterile water. The disinfested tissues were cut into 1-cm pieces and placed on potato dextrose agar (PDA) amended with streptomycin sulfate (50 mg/liter) and incubated at 25°C for 1 week. Isolations were done on 10 fruit fragments, and the same fungus was obtained. The cultures were purified using the hyphal-tip technique and used for morphological and molecular analyses. Morphological characteristics were observed on 1-week-old PDA cultures grown at 28°C. The aerial mycelium changed from white to light yellow, and the back of the plate turned pale brown with time. Microconidia were single celled, hyaline, nonseptate, ovoid, and 8.5 to 10.6 × 3.2 to 4.4 μm. Hyaline macroconidia (mostly three-septate) were slightly curved at the apex and ranged from 19.4 to 35.2 × 3.8 to 7.7 μm. Chlamydospores with thick, roughened walls were abundant in clumps or chains, ellipsoidal or subglobose. Primers ITS1/ITS4 (White et al. 1990) and EF-1/EF-2 (O’Donnell et al. 2000) were used to amplify and sequence the internal transcribed spacer (ITS1-5.8S-ITS2) and translation elongation factor 1-α (TEF1) region. ITS and TEF1 gene sequences were deposited in NCBI GenBank nucleotide database with accession numbers MH910492 and MH920853, respectively. BLASTn analysis showed 99% nucleotide sequence identity with Fusarium incarnatum-equiseti species complex (FIESC). The TEF1 gene sequence of isolate NEAU-TG1 was compared with sequences in the FUSARIUM-ID database (Geiser et al. 2004), which indicated that the pathogen was most closely (99% identity) related to phylogenetic species within the FIESC. Pathogenicity of these isolates was confirmed by following Koch’s postulates. Ten muskmelon fruits of cultivar Xiangfei were surface disinfested with 2% NaOCl for 2 min and rinsed with sterile distilled water three times. Spores were produced on PDA for 7 days at 28°C and washed with sterile distilled water; the concentrations were adjusted to 1 × 10⁶ spores/ml using a hemocytometer. Ten muskmelon fruits were inoculated by injecting the suspension (2 ml, 1 × 10⁶ spores/ml) and covered with plastic bags for 24 h, and another 10 surface-disinfested fruits treated with sterile distilled water were used as a control. The fruits were placed in a humidified chamber (>95% relative humidity) at 25°C for 48 h after inoculation and kept in a growth chamber at 25°C with 12-h day/night cycle for 8 days. All inoculated fruits showed symptoms identical to those observed in the field. No disease occurred on the controls. The pathogen was reisolated from diseased fruits, and species identification was confirmed by the morphological and molecular method described above. Members of the FIESC can cause disease on plants, such as Allium cepa, Festuca arundinacea, Morchella importuna, and Cucumis trigonus. Furthermore, the members of the FIESC have been reported to produce type A and B trichothecene mycotoxins that cause toxicosis in humans and animals (Leslie and Summerell 2006; O’Donnell et al. 2009). To our knowledge, this is the first report of fruit rot caused by a member of the FIESC on muskmelon in China. This pathogen presents a threat to muskmelon production, and its accurate identification is necessary to develop effective management strategies.

Experimental and theoretical evaluation of bubbler humidifier for humidification-dehumidification water desalination system

The requirement of fresh drinking water is one of the most severe problems all over the globe. The humidification dehumidification systems have shown promising outcomes while being used for water desalination. The performance of humidification-dehumidification water desalination systems is generally presented in the form of productivity of the humidifier or dehumidifier. Humidification being the predecessor process plays a crucial role in the performance of the Humidification - Dehumidification system. This work presents the theoretical and experimental investigation of bubbler humidifier. The performance of the bubbler humidifier is studied under various operation conditions such as: hole diameter, bubble column height, air mass flow rate, temperature of air and temperature of water inside the humidification chamber. A mechanistic model for bubbler humidifier is presented and validated with the experimental results. The Relative Humidity (RH) of air was achieved in the range of 68% to 97% for various operation conditions while the highest RH of air was recorded 97% for 1 mm of hole diameter, 7 g/s of mass flow rate of air, 60 °C of temperature of water, 48 °C of temperature of air, 7 cm of Height of water Column.

Nasal High-Flow Nebulization for Lung Drug Delivery: Theoretical, Experimental, and Clinical Application

The use of nasal high-flow (NHF) therapy is rapidly spreading across acute care facilities. This raises the question of optimal delivery of inhaled medication to patients undergoing this noninvasive ventilatory support consisting in delivering heated and humidified high gas flow rates through nasal cannulas. In this article, we review experimental and clinical work evaluating the delivery of inhaled medication within the NHF circuit to target the lung without interrupting the ventilatory support. Using vibrating mesh nebulizers placed immediately upstream or downstream of the humidification chamber, with flow rates of 30-45 L/min in adults and 2-6 L/min in children and infants, about 1%-10% of the drug charged in the nebulizer may be delivered to the lungs. Compared with conventional facemask aerosol interfaces, this amount is significantly lower than amounts delivered to adults (i.e., up to 25% of the nominal dose), but similar to amounts delivered to children and infants, the latter having a predominantly nasal breathing. However, significant clinical effects have been shown in both populations when delivering bronchodilators through NHF. This interface is particularly well tolerated and may be useful to improve aerosol therapy tolerance in the pediatric setting. Thus, among patients undergoing NHF therapy, bronchodilators may be delivered through this route. Whereas other drugs may be delivered this way or if there is a patient-centered benefit to specifically use NHF for aerosol therapy among patients without ongoing ventilatory support, requires further evaluation and technological development.

Comparative evaluation of fracture resistance using two rehydration protocols for fragment reattachment in uncomplicated crown fractures.

Uncomplicated crown fracture is the most common traumatic dental injury. The International Association of Dental Traumatology has recommended fragment reattachment as the best method for restoring uncomplicated crown fractures of permanent teeth. Dehydration can affect fracture resistance after reattachment. However, a standard protocol for rehydration is still lacking. Hence, the aim of this study was to design a humidification chamber and assess its efficacy for improving the rehydration of tooth fragments and increasing fracture resistance after reattachment. Sixty mandibular bovine incisors with similar dimensions and free of any structural deformities were fractured and randomized into five groups: Group I, Control Group (sound teeth); Group II (dehydrated for 24hours); Group III (rehydrated in distilled water for 15minutes); Group IV (rehydrated in a humidification chamber for 15minutes); and Group V (restored with composite). A humidification chamber was designed and used for rehydration for 15minutes in Group IV. Fragments in Group III were immersed in distilled water for 15minutes. Reattachment procedures and materials remained the same in all groups. Fracture resistance was tested in a universal testing machine, and statistical analysis was done by Stata-14. The Control Group with sound teeth (Group I) exhibited a maximum value of 282±10.32N, while Group II (fragment reattached without rehydration) had the least fracture resistance, 49.75±5.2N. Rehydration by means of the humidification chamber protocol (Group IV) resulted in significantly higher fracture resistance (150.54±6.49N) than in Group III (rehydration by means of immersion). Fracture resistance after fragment reattachment was significantly affected by the rehydration of fragments for 15minutes in the humidification chamber. Fragment reattachment after rehydration showed better fracture resistance than the composite restorations.

Mathematical Analysis of the Biophysical Response of EGCG‐treated yperglycemic HMEC‐1 and UMR Cells in vitro using Electric Cell‐Substrate Impedance Sensing Technology

Hyperglycemia, the hallmark of diabetes mellitus, is a major risk factor for endothelial dysfunction and compromised bone cell metabolism. Not many studies have measured the biophysical response of hyperglycemic human microvascular endothelial cells (HMEC‐1) and bone‐like rat osteosarcoma cells, UMR 106‐01 BSP (UMR cells), treated with (−)‐Epigallocatechin‐3‐gallate (EGCG), a natural antioxidant and the active ingredient in green tea, using electric cell‐substrate impedance sensing (ECIS) technology. The aim of this study was to determine the biophysical response of HMEC‐1 and UMR cells under hyperglycemic conditions when treated with EGCG. Moreover, the derivation of predictive mathematical formulas was sought to analyze the ECIS‐generated biophysical data. We hypothesized that ECIS technology can be used as a tool to evaluate the effects of glucose and EGCG on cell membrane resistance, spreading and attachment in HMEC‐1 and UMR cell cultures by mathematical analysis. HMEC‐1 and UMR cells were separately cultured and plated at 4.0×104 and 2.5×105 cells/mL, respectively, in eight‐well (8W) ECIS array plates. Designated plate wells were treated with glucose with or without EGCG. The cell cultures were incubated in a humidified chamber at 37° C with infusion of 5% CO2. Membrane resistance, time of complete cell attachment, and rate of spreading of the cells were monitored and measured in vitro, using the ECIS machine. The one‐tailed correlation analysis was employed to statistically study the relationships between glucose concentration and the following parameters: maximum resistance, spreading rate, and complete attachment time in both HMEC‐1 and UMR cells. To verify statistical significance of the results, p‐values were obtained for each set of experiments. The experimental data showed that the higher concentrations of glucose lowered the maximum cell membrane resistance, cell spreading rate, and time of cell attachment. The following mathematical formulas were derived based on the ECIS data: 1) dR/dt=kR(1−R/L), 2) R(t)=L/(1+be−kt), and 3) ln(π/1−π) = kt−ln(b), (where dR/dt=rate of spreading, R(t)=cell membrane resistance at any time point, L=maximum resistance, k=rate constant, b=(L−R0)/R0, t=time in hours, and π=r(R(t)−R0/L). The formulas were found to be of predictive value. The p‐values obtained from the correlation test of glucose concentration and two of the parameters tested indicated statistical significance (L for HMEC‐1 cells: 0.061; dR/dt for HMEC‐1 cells: 0.027; L for UMR cells: p=0.041; dR/dt for UMR cells: p=0.047). The results suggest that the ECIS technology is a useful tool in the mathematical analysis of biophysical phenomena in HMEC‐1 and UMR 106 cells in vitro.Support or Funding InformationFunding for this study was provided by the Ryckman Student Research Endowment Fund of the Biology Department of La Sierra University in Riverside, CA.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

ROS Play a Role in Long‐term Gamma Radiation‐induced Heart Damage

Cardiovascular problems are common side effects of cancer therapy, and the heart is one of the major organs that are of concern for potential damage following radiation exposure. Radiation directly affects biological molecules. However, the most impact of radiation‐mediated cell death derives from the generated free radicals, which increase oxidative stress. Reactive oxygen species (ROS) are a hallmark in cardiovascular diseases (CVD), mainly because of its interplay with nitric oxide (NO). NO offers cardioprotection against ROS‐induced damage and ROS, in turn, limits the beneficial effects of NO. Furthermore, radiation induces adaptive alterations in the vasculature leading to vascular dysfunction. Although pathways triggering cardiovascular damage following radiation exposure involve vascular damage in humans, the long‐term mechanisms by which radiation‐induced oxidative stress affects the heart have not yet been clearly defined. We hypothesized that increased ROS contribute to long‐term heart damage following radiation exposure. Flash frozen heart (apex) samples from male C5BL/6J mice (5 months) sacrificed nine months' post radiation was provided by the Department of Surgery at Duke University. Briefly, live animals were subjected to whole body radiation (dose: 200 cGy) at the Brookhaven National Lab. Control animals received Sham treatment. Transverse sections (10 μm) of heart from irradiated and non‐irradiated mice were obtained in a cryostat. Representative images of cardiac fibers were revealed with H&E staining. Additionally, ROS and NO production were evaluated by confocal microscopy. Briefly, the fluorescent probes, Dihydroethidium (DHE) and DAF‐FM Diacetate, were diluted in PBS (5 μM) and the samples were incubated in a humidified chamber at 37° C for 30 minutes. Slides were imaged using a 20x objective and analyzed using ImageJ software (NIH, Bethesda, MD, USA). Protein expression for NOS isoforms was also assessed using a standard Western blotting technique. Our histological analysis indicates loss of cardiac tissue (heart ‐ apex) in irradiated animals compared to Sham animals, suggestive of anatomical remodeling. Confocal microscopy showed an increase in intracellular ROS in irradiated tissue (n=5; p≤0.05), but no significant difference was observed in NO production (n=5; p>0.05). Different NOS isoforms evaluation showed elevated iNOS protein expression levels in irradiated animals. In conclusion, gamma radiation stimulates sustained ROS production in heart tissue and activation of iNOS might be a mechanism for counterbalancing the disruption in the heart antioxidant response system.Support or Funding InformationNASA Florida space Grant Consortium 2018This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

EXPERIMENTAL RESEARCH ON SPRAY COOLING FEATURES OF PRESSURE NOZZLE IN HUMIDIFICATION CHAMBER

The normal operation of spray humidification air cooler hinges on the good contact between droplets and air in the humidification chamber. The spiral pressure nozzle was chosen for the experiment on inlet air spray cooling. During the experiment, the spray cooling performance and atomization features were examined, and the cooling effects of different nozzle layouts were compared.Through the analysis of heat and mass transfer, the critical pressure of the spiral pressure nozzle was determined, and the relevant phenomenon was explained based on the energy balance relationship in droplet atomization. Meanwhile, the cooling effect fitting correlation was acquired with evaporative cooling being the major cooling mechanism. The comparison between different nozzle layouts revealed that the cooling effect of row spacing: 500mm is superior to that of row spacing: 1,000mm under the condition of the same .T.L. In this way, the critical pressure and optimal layout of the spiral pressure nozzle in the humidification chamber were discovered.

Water content of delivered gases during Helmet Continuous Positive Airway Pressure in healthy subjects.

Introduction: During Continuous Positive Pressure Ventilation delivered through helmet, the patient inhales high flows of gas without adequate conditioning. However, the need to humidify the inspired gas during Helmet-CPAP, has not been sufficiently explored. Methods: Experimental design study. Six healthy individuals underwent High Flow Helmet CPAP with different gas flows (60 and 80 L/min) and FiO2 (0.35, 0.5, 0.7 and 1) generated by a Venturi system, with and without active humidification. The active humidifier setting was 26 ° at the humidification chamber and 28 ° at the helmet gas inlet. At each setting, measurements about temperature and relative humidity inside helmet were taken. Comfort level at each setting was evaluated using a visual analog scale rated from 0 to 10. Results: Without heated humidification, the mean value of absolute humidity in the eight combinations investigated was 5.9±2.1 mg H2O/L, with a mean temperature of 25.8±0.9°. With heated humidification mean absolute humidity was 15.0±3.5 mg H2O/L with mean temperature of 29.0±0.1°. The median comfort scale value was 6 (IQR: 5.25-6.75) during the phase without humidification vs 8 (7.25-8.0 - P<0.01) when active humidification was applied. Conclusions: In healthy subjects undergoing High Flow Helmet CPAP, heated humidifiers with heated wires tubes are necessary to avoid the under-humidification inside the helmet. To obtain patient’s comfort and airways mucosal humidification during continuous Helmet CPAP, the most desirable conditions are reached by heated humidifiers with a humidifying chamber temperature settled between 26-28°C. (www.actabiomedica.it)

Nasal high-flow bronchodilator nebulization: a randomized cross-over study

BackgroundThere is an absence of controlled clinical data showing bronchodilation effectiveness after nebulization via nasal high-flow therapy circuits.ResultsTwenty-five patients with reversible airflow obstruction received, in a randomized order: (1) 2.5 mg albuterol delivered via a jet nebulizer with a facial mask; (2) 2.5 mg albuterol delivered via a vibrating mesh nebulizer placed downstream of a nasal high-flow humidification chamber (30 L/min and 37 °C); and (3) nasal high-flow therapy without nebulization. All three conditions induced significant individual increases in forced expiratory volume in one second (FEV1) compared to baseline. The median change was similar after facial mask nebulization [+ 350 mL (+ 180; + 550); + 18% (+ 8; + 30)] and nasal high flow with nebulization [+ 330 mL (+ 140; + 390); + 16% (+ 5; + 24)], p = 0.11. However, it was significantly lower after nasal high-flow therapy without nebulization [+ 50 mL (− 10; + 220); + 3% (− 1; + 8)], p = 0.0009. FEV1 increases after facial mask and nasal high-flow nebulization as well as residual volume decreases were well correlated (p < 0.0001 and p = 0.01). Both techniques showed good agreement in terms of airflow obstruction reversibility (kappa 0.60).ConclusionAlbuterol vibrating mesh nebulization within a nasal high-flow circuit induces similar bronchodilation to standard facial mask jet nebulization. Beyond pharmacological bronchodilation, nasal high flow by itself may induce small but significant bronchodilation.

ガス供給と電源容量の問題が解決されhigh flow nasal cannula呼吸補助下に施設間陸路搬送した1例(A case of interhospital ground transport with a high flow nasal cannula for respiratory support enabled by the resolution of gas and power supply capacity problems)

要旨High flow nasal cannula（HFNC）療法は様々な医療場面での使用が増加しているが，HFNCを用いた施設間搬送の報告は少ない。これまで当院では，HFNCシステムへの酸素・混合ガス供給とドクターカーの電源容量の問題によりHFNC呼吸補助下の施設間陸路搬送が実施できなかった。しかし，新たなHFNCシステムの導入と，ドクターカーの電源容量増加と補助電源の確保により，HFNCを用いた施設間搬送が可能となった。今回，我々はHFNC呼吸補助下に施設間陸路搬送した症例を経験したので報告する。症例は11か月の男児。Respiratory syncytial virus（RSV）細気管支炎による急性呼吸不全のため当院へ転院し，pediatric intensive care unit（PICU）でHFNC療法が導入された。その後症状が軽快しHFNC呼吸補助下に医療機関へ転院搬送となった。搬送中，呼吸状態の悪化や重篤な副作用を認めず，安全に搬送が行われた。搬送にHFNCを用いる場合，デバイスによっては呼吸器設定に限界があること，屋外使用時はデバイスへの外気温の影響を考慮すること，および複数の医療機器を同時に使用する場合の総電源容量と搬送車両の電源容量を把握するなど留意すべき点がある。今後，症例を蓄積し施設間搬送におけるHFNC療法の安全性と有効性を評価する必要がある。

Measuring the detector-observed impact of optical blurring due to aerosols in a laboratory cloud chamber

Deleterious effects of the atmosphere on remotely acquired images includes absorption and scattering of light by aerosol particulates, which not only attenuates the signal but can potentially cause blurring due to forward-scattered light accepted by the imaging system. Proposed aerosol scattering models (e.g., Ishimaru) provide a method for simulating the contrast and spatial detail expected when imaging through atmospheres with significant aerosol optical depth. This work explores closure between modulation transfer functions (MTFs) obtained from directly measured images and MTFs calculated from theory using measured cloud properties. The closure experiments are performed in a laboratory cloud chamber in which cloud droplet number density and size distribution are directly measured. Images of a binary knife-edge target were taken with an optical detector on the other side of a water cloud generated through reduction of pressure in the humidified chamber. The key results of this closure experiment are: the theoretical expression for the aerosol MTF is likely overly simplistic and does not account for broad particle size distributions. The significance of optical blurring from light scattering by aerosol particles depends sensitively on the properties of both the particles and the imaging system.

In Vitro Determination of the Main Effects in the Design of High-Flow Nasal Therapy Systems with Respect to Aerosol Performance.

IntroductionThe use of concurrent aerosol delivery during high-flow nasal therapy (HFNT) may be exploited to facilitate the delivery of a variety of prescribed medications for inhalation. Until now, a systematic approach to determine the conditions required to yield an optimal emitted dose has not been reported. The aim of this study was to establish the effects of gas flow rate, input droplet size, and nebulizer position on the amount of aerosol exiting the nasal cannula during HFNT and thus becoming available for inhalation.MethodsTesting was completed according to a factorial statistical design of experiments (DOE) approach. Emitted dose was characterized with a vibrating mesh nebulizer (Aerogen Solo, Aerogen Ltd) for an adult model of HFNT at three clinically relevant gas flow rates, using three nebulizers producing varying input droplet sizes and placed at two different nebulizer positions.ResultsIncreasing the gas flow rate significantly lowered the emitted dose, with a dose of 7.10% obtained at 10 LPM, 2.67% at 25 LPM, and 1.30% at 40 LPM (p < 0.0001). There was a significant difference in emitted dose between nebulizers with different input droplet sizes, with increasing input droplet size associated with a reduced emitted dose (6.11% with an input droplet size of 3.22 µm, 2.76% with 4.05 µm, and 2.38% with 4.88 µm, p = 0.0002, Pearson’s r = − 0.2871). In addition, the droplet size exiting the nasal cannula interface was lower than that produced by the aerosol generator for all devices under test. Positioning the nebulizer immediately after the humidification chamber yielded a marginally greater emitted dose (3.79%) than when the nebulizer was placed immediately upstream of the nasal cannula (3.39%). Flow rate, input droplet size, and nebulizer position were at the 0.10 level of significance, indicating that all three factors had significant effects on emitted dose. According to the DOE model, flow rate had the greatest influence on emitted dose, followed by input droplet size and then nebulizer position.ConclusionOur findings indicate that in order to optimize the amount of aerosol exiting the nasal cannula interface during HFNT, it is necessary for gas flow rate to be low and the input droplet size to be small, while the nebulizer should be positioned immediately after the humidification chamber.FundingAerogen Limited.

A humidifier in the invasive mode during noninvasive respiratory support could increase condensation and thereby impair airway patency.

Humidifying noninvasively ventilated preterm infants is critical to prevent dehydration of respiratory mucosa, but over-humidification can result in impaired airway patency and lung mechanics. This neonatal bench study investigated the humidity delivered using invasive and noninvasive humidification modes during nasal continuous positive airway pressure. The study was conducted at the neonatal intensive care unit of Emma Children's Hospital, the Netherlands, in March 2014. A mannequin was connected to a CareFusion Infant Flow SiPAP system, combined with a Fisher & Paykel MR850 humidifier and a Carefusion Infant Flow LP Generator. We measured the temperature, relative humidity and absolute humidity at the humidification chamber's expiratory port and at the patient's mask. The absolute humidity at the mask was 35-40 mg H2 O/L at 38-39°C (relative humidity 74-80%) for the invasive mode of the humidifier and 23-27 mg H2 O/L at 34-35°C (relative humidity 63-70%) for the noninvasive mode. The absolute humidities exceeded the recommended values for the invasive mode of the humidifier, but not the noninvasive mode, and could be associated with increased condensation. The absolute humidity delivered by the humidifier in invasive mode could be associated with increased condensation, which has been associated with airway impairment.

A new approach to assess the effect of photodamage on corneocyte envelope maturity using combined hydrophobicity and mechanical fragility assays.

The maturity of the corneocyte envelope (CE) provides information about the barrier functionality of the stratum corneum (SC). Corneocytes are enclosed by the CE, a protein-lipid matrix, contributing to mechanical resistance and hydrophobicity of the SC. The aim of the work was to develop a novel and robust approach to characterize CE maturity based on rigidity, hydrophobicity and surface area. This offers an alternative approach to the Nile red staining and antigenicity of involucrin to characterize the CE. The photoexposed (PE) cheek and photoprotected (PP) post-auricular sites were selected for investigation. Nine tape strips were obtained from the cheek and post-auricular sites of healthy Caucasians. CEs on the first and last tape strip were subjected to sonication to assess rigidity, and Nile red staining to determine hydrophobicity per unit surface area. In addition, the presence of involucrin and lipids was assessed to determine CE maturity by examination of the red/green pixel ratio, percentage of involucrin expressing CEs and alternatively the ratio of fluorescence density. The CE rigidity was lower in the deeper SC layers of the cheek, whereas post-auricular CEs were mechanically more resistant. Post-auricular CEs from the superficial SC had a larger surface area with a stronger fluorescence signal than those from the cheek. Interestingly, those CEs from the deeper SC layers had similar surface areas in both anatomical sites but were significantly different in hydrophobicity. These three parameters can be summarized as a relative CE maturity index that expresses CE maturity more precisely with a higher sensitivity than the conventional involucrin and Nile red staining approach. CEs of the cheek surface are more mature than CEs in the deeper SC layer, whereas CEs obtained from the post-auricular surface are more mature than those from the cheek surface. The combined method developed allows characterization of CE maturity based on hydrophobicity per unit surface area and rigidity rather than a simple ratio of lipid to involucrin. A more robust and sensitive measurement has therefore been developed addressing the limitations of earlier protocols.

Energy recovery from a vapour compression refrigeration system using humidification dehumidification desalination

Energy consumption of a process can be reduced by properly utilizing the large amount of waste heat liberated into atmosphere. The aim of this research is to improve the coefficient of performance (COP) of a vapour compression refrigeration system, by recovering waste heat from the condenser and the evaporator using humidification dehumidification desalination process. To recover the heat, air from the blower is passed through a humidifier chamber in which water is sprayed over the gunny bag placed in the air passage. The humid air circulated around the condenser coil which sub-cools the refrigerant and it also absorbs the heat from the condenser, producing hot humid air. The hot humid air is then passed over another heat exchanger (dehumidifier coil) which heats the refrigerant coming out from the evaporator so the quality of the vapour improves. At the same time, condensation occurs in the hot humid air along the surface of evaporator coil and the distilled water is collected. Refrigerator system is tested with insert like twisted tape and cone type turbulators in the dehumidifier passage to augment the performance. These modifications improved the COP of the refrigeration system up to 7.6 with a distillate water output of 0.4 l/m2 h.

Using Electric Cell‐substrate Impedance Sensing (ECIS) Technology as a Tool for Mathematical Analysis of the Effects of Glucose on Bone‐like Osteosarcoma Cells (UMR 106‐01 BSP)

Mathematical modeling has been useful in studies of the progression of type II diabetes. However, not many studies have been done using electric cell‐substrate impedance sensing (ECIS) technology to validate mathematical modeling in bone cells exposed to high concentrations of glucose. In this study we hypothesized that ECIS technology can be used as a predictive tool to evaluate the effects of glucose on cell membrane impedance in UMR cell cultures by mathematical analysis through predictive mathematical modeling. Briefly, UMR cells were plated at 2.5 × 105 cells/mL in eight‐well (8W) ECIS array plates. Designated wells were treated with different concentrations of glucose (10, 5, and 3 mM). Cell cultures were incubated in a humidified chamber at 37° C with infusion of 5% CO2. Membrane resistance, time of attachment and rate of spreading of the cells were monitored and measured in vitro, using the ECIS machine. Based on the data generated by the ECIS machine, a mathematical formula was derived to predict the resistance of the UMR cell membrane for any concentration of glucose used. The formula was tested based on the generated data. F test ANOVA was employed to compare variances between all experimental trials; all values in the results are expressed as means ±SD. The results show that the highest concentration of glucose caused a decreased time of attachment, rate of spreading and an increased cell membrane resistance in the UMR cells. The following formulas were derived based on the ECIS data: dR/dt=kR0ekt and R[G]=R0e+sG (where dR/dt=rate of spreading, k=rate constant, R0=initial resistance, t=time, [G]=glucose concentration, and s=rate constant (1/[glucose]). The formulas were found to be of predictive value. The results suggest that the ECIS technology is a potentially useful tool in the mathematical analysis of physiological phenomena in bone‐like cells in vitro.Support or Funding InformationThis work was supported by the Ryckman Endowment Student Research Fund of the Biology Department at La Sierra University in Riverside, CA.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

The Development and Validation of an In Vitro Airway Model to Assess Realistic Airway Deposition and Drug Permeation Behavior of Orally Inhaled Products Across Synthetic Membranes.

Current in vitro approaches to assess lung deposition, dissolution, and cellular transport behavior of orally inhaled products (OIPs) have relied on compendial impactors to collect drug particles that are likely to deposit in the airway; however, the main drawback with this approach is that these impactors do not reflect the airway and may not necessarily represent drug deposition behavior in vivo. The aim of this article is to describe the development and method validation of a novel hybrid in vitro approach to assess drug deposition and permeation behavior in a more representative airway model. The medium-sized Virginia Commonwealth University (VCU) mouth-throat (MT) and tracheal-bronchial (TB) realistic upper airway models were used in this study as representative models of the upper airway. The TB model was modified to accommodate two Snapwell® inserts above the first TB airway bifurcation region to collect deposited nebulized ciprofloxacin-hydrochloride (CIP-HCL) droplets as a model drug aerosol system. Permeation characteristics of deposited nebulized CIP-HCL droplets were assessed across different synthetic membranes using the Snapwell test system. The Snapwell test system demonstrated reproducible and discriminatory drug permeation profiles for already dissolved and nebulized CIP-HCL droplets through a range of synthetic permeable membranes under different test conditions. The rate and extent of drug permeation depended on the permeable membrane material used, presence of a stirrer in the receptor compartment, and, most importantly, the drug collection method. This novel hybrid in vitro approach, which incorporates a modified version of a realistic upper airway model, coupled with the Snapwell test system holds great potential to evaluate postairway deposition characteristics, such as drug permeation and particle dissolution behavior of OIPs. Future studies will expand this approach using a cell culture-based setup instead of synthetic membranes, within a humidified chamber, to assess airway epithelia transport behavior in a more representative manner.

Global histone modifications predict the outcome of glaucoma surgery

PurposeConjunctival and subconjunctival fibrosis at the surgical site influences the outcome of glaucoma surgery. Since glaucoma surgery initiates a cascade of events that leads to up‐ or downregulation of various genes and the subsequent release of cytokines and growth factors, we analyzed global histone modifications at the site of glaucoma surgery and their relationship with the outcome of glaucoma surgery.Methods44 patients who were scheduled to undergo glaucoma surgery were enrolled in this study. A 4 × 4 mm section of Tenon's tissue with overlying conjunctiva was cut from the eye at the margin of the incision. Primary rabbit anti‐histone antibodies were applied in humidified chamber at the following dilutions: H3 K9 K14 K18 K23 K27Ac at 1:200, H4 K5 K8 K12 K16Ac at 1:200, H3 K4diMe at 1:800 (Abcam), and H4 R3diMe at 1:25 (Millipore). The sections were counterstained with Harris's haematoxylin, dehydrated, and mounted.ResultsUsing histochemical staining, we determined the number of cells that were positive for histone acetylation and demethylation of histones H3 and H4. The patients were divided into high‐ and low‐acetylated or ‐methylated groups accordingly. The low‐acetylation group for H3 K9 K14 K18 K23 K27 and H4 K5 K8 K12 K16 had poorer surgical outcomes based on Kaplan‐Meier plots. The preoperative intraocular pressure was significantly related to acetylation of both H3 K9 K14 K18 K23 K27 and H4 K5 K8 K12 K16 in univariate and multivariate regression analyses.ConclusionsOur findings suggest global histone modifications in the conjunctival and subconjunctival tissues predict glaucoma surgery outcomes. A low acetylation status indicated a poor prognosis for glaucoma surgery. Prognostication using histone modifications might have implications for epigenetic therapy to improve glaucoma surgery outcomes.

Cell Migration and Palatal Fusion

Cleft palate, one of the most common birth defects, occurs as a result of the developing secondary palatal shelves' failure to grow to midline, degrade the midline epithelial seam (MES), and fuse to leave a confluent shelf. The mechanism of MES degradation is largely unknown, but the role of ephrin signaling in palatal fusion suggests an induction of epithelial to mesenchymal transition (EMT). Our goal is to determine the role of ephrin signaling in MES degradation. We dissected palatal shelves from embryonic day 14.5 Ephrin‐B2/GFP mice, in which a nuclear‐localized green fluorescent protein (GFP) under control of the ephrin‐B2 (EB2) gene labels 100% of medial edge epithelia (MEE). The adhered shelves were placed oral‐side down in a glass‐bottomed dish, encased in low‐melt agarose, and submerged in culture medium. Palates were imaged same day for a total of 48 hours, using a Zeiss 880 laser scanning confocal with a humidified chamber. Displacement of Ephrin‐B2/GFP labeled cells was tracked using Imaris Software. After imaging, palates were sectioned and stained with hematoxylin and eosin. Fluorescently labeled MEE cells moved as an aggregate/sheet and not individually. Histology confirmed that the imaged palates fused. Cell expression of EB2/GFP was visible throughout the 48‐hour experiment. The data indicate that MES degradation involves MEE cell migration and suggest that elements of the EMT program are a part of palatal fusion. Continued expression of EB2/GFP throughout fusion suggests that MEE cells do not undergo apoptosis prior to seam degradation.Support or Funding InformationNIDCR R01DE022804

Assessment of skin barrier function and biochemical changes of ex vivo human skin in response to physical and chemical barrier disruption

Topical dermatotherapy is intended to be used on diseased skin. Novel drug delivery systems even address differences between intact and diseased skin underlining the need for pre-clinical assessment of different states of barrier disruption.Herein, we studied how short-term incubation in culture media compared to incubation in humidified chambers affects human skin barrier function and viability. On both models we assessed different types and intensities of physical and chemical barrier disruption methods with regard to structural integrity, biophysical parameters and cytokine levels.Tissue degeneration and proliferative activity limited the use of tissue cultures to 48h. Viability is better preserved in cultured tissue. Tape-stripping (50×TS) and 4h sodium lauryl sulfate (SLS) pre-treatment were identified as highly reproducible and effective procedures for barrier disruption. Transepidermal water loss (TEWL) values reproducibly increased with the intensity of disruption while sebum content and skin surface pH were of limited value. Interleukin (IL)-6/8 and various chemokines and proteases were increased in tape-stripped skin which was more pronounced in SLS-treated skin tissue extracts.Thus, albeit limited to 48h, cultured full-thickness skin maintained several barrier characteristics and responded to different intensities of barrier disruption. Potentially, these models can be used to assess pre-clinically the efficacy and penetration of anti-inflammatory compounds.