Influence of dextran solution and corneal collagen crosslinking on Corneal Biomechanical Parameters Evaluated by Corvis ST

Purpose: To analyze the influence of dextran solution and corneal collagen crosslinking (CXL) on corneal biomechanical parameters evaluated by Corneal Visualization Scheimpflug Technology (Corvis ST). Materials and Methods: Forty porcine eyes were included in this study. Twenty porcine eyes were soaked in dextran solution for 30 minutes (10 eyes in 2% dextran solution and 10 eyes in 20% dextran solution). CXL treatment was performed in 10 porcine eyes, the other 10 porcine eyes were regarded as a control group. Each eye was fixed on an experimental inflation platform to carry out Corvis measurements at different IOPs. Corneal biomechanical parameters were calculated based on Corvis measurement. Statistical analysis was used to analyze the influence of dextran solution and CXL on corneal biomechanical parameters based on Corvis parameters. Results: Corneal energy absorbed area (Aabsorbed) decreased after being soaked in dextran solution under IOP of 15 mmHg; Corneal elastic modulus (E) decreased after being soaked in 2% dextran solution and increased after being soaked in 20% dextran solution; SP-A1 increased after CXL. Conclusion: Both dextran solution and CXL can change corneal biomechanical properties; SP-A1 may be used as an effective parameter for the evaluation of CXL.

Immobilization of Chaetomium erraticum dextranase (CED) by adsorption on carboxylated multi walled carbon nanotubes (c-MWCNT)

In this study, CED was immobilized onto c-MWCNT by adsorption. Optimization of immobilization conditions (immobilization buffer's pH and molarity, c-MWCNT amount, and immobilization time) was resulted in 100% immobilization yield and 114.13% activity yield. Further, characterization of FCED and ICED was also studied. After immobilization, the optimum pH shifted from 5.0 to 6.0, while the optimum temperature (55 °C) did not change. Furthermore, kinetic constants for FCED and ICED were also determined using the Lineweaver-Burk plot. The Km value for both FCED and ICED were 54.35 g / L, while Vmax values for FCED and ICED were 2.77 μmol reducing sugar / L.mg.min and 3.19 μmol reducing sugar / L.mg.min, respectively. Moreover, there was no reduction in the initial activity of ICED after 20 consecutive uses and 30 days of storage at optimal storage conditions. Finally, 17.15% and 17.53% of the dextran in 10% dextran solution (pH 6.0) were converted to reduced sugars (IMOs and Glucose) in 12 hours using FCED and ICED, respectively. Consequently, it can be concluded that ICED obtained in this study can be effectively used for industrial production of IMOs and for hydrolysis of dextran.

Ultrasound Standing Wave-Based Cell-to-liquid Separation for Measuring Viscosity and Aggregation of Blood Sample

When quantifying mechanical properties of blood samples flowing in closed fluidic circuits, blood samples are collected at specific intervals. Centrifugal separation is considered as a required procedure for preparing blood samples. However, the use of centrifuge is associated with several issues, including the potential for red blood cell (RBC) lysis, clotting activation, and RBC adhesions in the tube. In this study, an ultrasonic transducer is employed to separate RBCs or diluent from blood sample. The ultrasonic radiation force is much smaller than the centrifugal force acting in centrifuge, it can avoid critical issues occurring under centrifuge. Then, the RBC aggregation and blood viscosity of the blood sample are obtained using the microfluidic technique. According to the numerical results, ultrasonic transducers exhibited a maximum quality factor at an excitation frequency of 2.1 MHz. Periodic pattern of acoustic pressure fields were visualized experimentally as a column mode. The half wavelength obtained was as 0.5 λ = 0.378 ± 0.07 mm. The experimental results agreed with the analytical estimation sufficiently. An acoustic power of 2 W was selected carefully for separating RBCs or diluent from various blood samples (i.e., Hct = 20% ~ 50%; diluent: plasma, 1x phosphate-buffered saline (PBS), and dextran solution). The present method was employed to separate fixed blood samples which tended to stack inside the tube while using the centrifuge. Fixed RBCs were collected easily with an ultrasonic transducer. After various fixed blood samples with different base solutions (i.e., glutaraldehyde solution, 1x PBS, and dextran solution) were prepared using the present method, RBC aggregation and the viscosity of the blood sample are successfully obtained. In the near future, the present method will be integrated into ex vivo or in vitro fluidic circuit for measuring multiple mechanical properties of blood samples for a certain longer period.

Urinary TCP1-eta: A Cortical Damage Marker for the Pathophysiological Diagnosis and Prognosis of Acute Kidney Injury

Acute kidney injury (AKI) is a serious syndrome with increasing incidence and health consequences, and high mortality rate among critically ill patients. Acute kidney injury lacks a unified definition, has ambiguous semantic boundaries, and relies on defective diagnosis. This, in part, is due to the absence of biomarkers substratifying AKI patients into pathophysiological categories based on which prognosis can be assigned and clinical treatment differentiated. For instance, AKI involving acute tubular necrosis (ATN) is expected to have a worse prognosis than prerenal, purely hemodynamic AKI. However, no biomarker has been unambiguously associated with tubular cell death or is able to provide etiological distinction. We used a cell-based system to identify TCP1-eta in the culture medium as a noninvasive marker of damaged renal tubular cells. In rat models of AKI, TCP1-eta was increased in the urine co-relating with renal cortical tubule damage. When kidneys from ATN rats were perfused in situ with Krebs-dextran solution, a portion of the urinary TCP1-eta protein content excreted into urine disappeared, and another portion remained within the urine. These results indicated that TCP1-eta was secreted by tubule cells and was not fully reabsorbed by the damaged tubules, both effects contributing to the increased urinary excretion. Urinary TCP1-eta is found in many etiologically heterogeneous AKI patients, and is statistically higher in patients partially recovered from severe AKI. In conclusion, urinary TCP1-eta poses a potential, substratifying biomarker of renal cortical damage associated with bad prognosis.

Intermolecular Interaction of Aqueous Dextran with Urea

In this paper, the authors have given information regarding intermolecular interactions of aqueous dextran solution in urea. The behavior of dextran in urea has been examined by the help of ultrasonic interferometer working at frequency 5MHz at different temperatures ranging from 303 K to 323 K in 5K interval. Ultrasonic speed, density, viscosity measurements have been used for the evaluation of thermodynamic parameters like Gibb’s free energy (ΔG) as well as acoustical parameters are acoustic impedance (Z), isentropic compressibility (β), intermolecular free length (Lf ) and relaxation time (τ), etc. The results have been used to throw light on the nature of the interaction among solute and solvent, interpreted in the light of structural rearrangement occurs in the aqueous dextran and urea solution.

Effects of immediate versus delayed ex-vivo lung perfusion in a porcine cardiac arrest donation model

Objective:Ex-vivo lung perfusion is a promising tool to evaluate and recondition marginal donor lungs usually after a cold static preservation. The concept of continuous organ perfusion is supposed to reduce ischemic damage; however, the optimal perfusion protocol has not been established yet. The aim of this study was to compare immediate ex-vivo lung perfusion (I-EVLP) to delayed ex-vivo lung perfusion (D-EVLP) after a certain cold static preservation period on lung function in a large animal model.Methods:In a porcine model, lungs were procured after circulatory death and 60 min of no-touch warm ischemia. Lungs were preserved with single-flush cold low potassium dextran solution and prepared either for I-EVLP (n = 8) or stored cold for 9 h with subsequent D-EVLP (n = 8). Functional outcomes and morphology were compared during 4 h of ex-vivo lung perfusion, using STEEN SolutionTMas perfusion solution.Results:Pulmonary functional data, perfusate activities of lactate dehydrogenase, alkaline phosphatase, and products of lipid peroxidation did not differ significantly. There was a trend toward lower wet–dry ratio (I-EVLP: 13.4 ± 2.9; D-EVLP: 9.1 ± 2.5) and higher ΔpO2in D-EVLP group (I-EVLP: 209 ± 51.6 mmHg; D-EVLP: 236.3 ± 47.3 mmHg).Conclusion:In this donation-after-circulatory-death model, 9 h of cold static preservation followed by ex-vivo lung perfusion results in comparable pulmonary function to I-EVLP as indicated by oxygenation capacities and wet–dry ratio. Our findings indicate that prolonged cold static preservation prior to ex-vivo lung perfusion is as safe and effective as I-EVLP in the procurement of donor lungs.

Biomechanical efficacy of corneal cross-linking using hypoosmolar riboflavin solution

Purpose: The use of hypoosmolar riboflavin solution has been suggested for cross-linking thin corneas. The aim of this study was to compare the biomechanical efficacy of corneal cross-linking using hypoosmolar dextran-free riboflavin solution (HCXL) versus isoosmolar standard corneal cross-linking treatment (CXL). Methods: A total of 24 postmortem porcine eyes with debrided corneas were subdivided into three treatment groups: Controls, the isoosmolar group with isoosmolar 0.1% riboflavin-20% dextran solution and the hypoosmolar group with dextran-free, 0.1% riboflavin solution. The samples were irradiated with UVA light of 365 nm wavelength and an irradiance of 3 mW/cm² for 30 min (dose 5.4 J/cm²). For the biomechanical measurements, 400-µm-deep anterior corneal flaps were created using a lamellar rotating microkeratome. Uniaxial stress–strain measurements were performed. Results: In the isoosmolar treatment group, stress and Young’s modulus at 8% strain were significantly increased by 67.97%, respectively, 62.62% versus the controls. In the hypoosmolar treatment group, stress and Young’s modulus at 8% strain were significantly increased by 81.21%, respectively, 51.40% versus the controls. There was no significant difference between the iso- and hypoosmolar groups in biomechanical efficacy. On histology, there was no edema in the anterior 200 µm of the corneas after stromal swelling by the hypoosmolar riboflavin solution. Conclusion: Corneal cross-linking using isoosmolar or hypoosmolar riboflavin solution induces a comparable biomechanical effect. This is explained by the localization of the maximum cross-linking effect in the anterior 200 µm of the cornea which are not affected by the swelling effect of hypoosmolar riboflavin solution.

3D Characterization of corneal deformation using ultrasound speckle tracking

The three-dimensional (3D) mechanical response of the cornea to intraocular pressure (IOP) elevation has not been previously reported. In this study, we use an ultrasound speckle tracking technique to measure the 3D displacements and strains within the central 5.5[Formula: see text]mm of porcine corneas during the whole globe inflation. Inflation tests were performed on dextran-treated corneas (treated with a 10% dextran solution) and untreated corneas. The dextran-treated corneas showed an inflation response expected of a thin spherical shell, with through-thickness thinning and in-plane stretch, although the strain magnitudes exhibited a heterogeneous spatial distribution from the central to more peripheral cornea. The untreated eyes demonstrated a response consistent with swelling during experimentation, with through-thickness expansion overriding the inflation response. The average volume ratios obtained in both groups was near 1 confirming general incompressibility, but local regions of volume loss or expansion were observed. These results suggest that biomechanical measurements in 3D provide important new insight to understand the mechanical response of ocular tissues such as the cornea.