Effect of dietary glycine supplementation on productive performance, egg quality, stress response, and fatty liver incidence in laying hens raised under heat stress conditions.

The current experiment aimed to investigate the effect of dietary glycine (Gly) supplementation on productive performance, egg quality, stress response, and fatty liver incidence in laying hens raised under heat stress (HS) conditions. A total of two hundred eighty 24-wk-old Lohmann Brown-Lite laying hens were randomly allotted to 1 of 4 dietary treatments with 7 replicates. The negative control (NC) diet was prepared to meet or exceed the nutrient and energy requirement for Lohmann Brown laying hens, whereas the positive control (PC) diet was formulated to increase AMEn by 100 kcal/kg compared with the NC diet. Two additional diets were prepared by supplementing 0.341% and 0.683% Gly to the NC diet. All hens were exposed to cyclic HS at 31.4 ± 1.17°C for 8 h/d and 26.7 ± 1.10°C for the remaining time for a 12-wk trial. Results indicated that increasing supplementation of Gly in diets tended (linear, P = 0.088) to decrease the FCR of laying hens. Increasing supplementation of Gly in diets increased (linear, P < 0.05) eggshell lightness and decreased (linear, P < 0.05) egg yolk color. Moreover, a tendency for a quadratic association (P < 0.10) of serum aspartate aminotransferase and alanine aminotransferase concentrations with increasing supplementation of Gly was observed. Increasing supplementation of Gly in diets decreased (linear, P < 0.05) blood heterophil:lymphocyte ratio of laying hens. Hens fed the NC diet showed higher fatty liver incidence (P < 0.05) than those fed the PC diet, but increasing supplementation of Gly decreased (linear, P < 0.05) fatty liver incidence of laying hens. In conclusion, increasing supplementation of Gly up to 0.683% in diets decreases FCR, stress response, and fatty liver incidence in laying hens raised under HS conditions.

Sinus augmentation with poly(ε)caprolactone-ß tricalcium phosphate scaffolds, mesenchymal stem cells and platelet rich plasma for one-stage dental implantation in minipigs.

PURPOSE: This study evaluated the efficacy of a tube-shaped poly(ε) caprolactone - ß tricalcium phosphate (PCL-TCP) scaffold with the incorporation of human umbilical cord-derived mesenchymal stem cells (hUCMSCs) and platelet-rich plasma (PRP) for bone regeneration in the procedure of single-stage sinus augmentation and dental implantation in minipigs. METHODS: Implants were placed in the bilateral sides of the maxillary sinuses of 5 minipigs and allocated to a PCL-TCP+hUCMSCs+PRP group (n=5), a PCL-TCP+PRP group (n=5), and a PCL-TCP-only group (n=6). After 12 weeks, bone regeneration was evaluated with soft X-rays, micro-computed tomography, fluorescence microscopy, and histomorphometric analysis. RESULTS: Four implants failed (2 each in the PCL-TCP+hUCMSCs+PRP and PCL-TCP+hUCMSC groups). An analysis of the grayscale levels and bone-implant contact ratio showed significantly higher mean values in the PCL-TCP+hUCMSCs+PRP than in the PCL-TCP group (P=0.045 and P=0.016, respectively). In fluoromicroscopic images, new bone formation around the outer surfaces of the scaffolds was observed in the PCL-TCP+hUCMSCs+PRP group, suggesting a tenting effect of the specially designed scaffolds. Bone regeneration at the scaffold-implant interfaces was observed in all 3 groups. CONCLUSIONS: Using a tube-shaped, honeycombed PCL-TCP scaffold with hUCMSCs and PRP may serve to enhance bone formation and dental implants' osseointegration in the procedure of simultaneous sinus lifting and dental implantation.

Mesenchymal stem cells and platelet-rich plasma-impregnated polycaprolactone-ß tricalcium phosphate bio-scaffold enhanced bone regeneration around dental implants.

BACKGROUND: Finding a material that supports bone regeneration is the concern for many investigators. We supposed that a composite scaffold of poly(ε) caprolactone and ß-tricalcium phosphate (PCL-TCP) would entail desirable characteristics of biocompatibility, bioresorbability, rigidity, and osteoconductivity for a proper guided bone regeneration. Furthermore, the incorporation of mesenchymal stem cells (MSCs) and platelet-rich plasma (PRP) would boost the bone regeneration. We conducted this study to evaluate the bone regeneration capacity of PCL-TCP scaffold that is loaded with MSCs and PRP. MATERIALS AND METHODS: Five miniature pigs received 6 implants in 6 created-mandibular bony defects in the right and left lower premolar areas. The bony defects were managed according to the following three groups: the PCL-TCP scaffold loaded with MSCs and PRP (MSCs+PRP+PCL-TCP) group (n = 10), PCL-TCP scaffold loaded with PRP (PRP+PCL-TCP) group (n = 10), and PCL-TCP scaffold group (n = 10). After 12 weeks, the bone regeneration was assessed using fluorochrome bone labeling, µCT bone morphogenic analysis, and histomorphometric analysis. RESULTS: All of the three groups supported the bone regeneration around the dental implants. However, the PCL-TCP scaffold loaded with MSCs and PRP (MSCs+PRP+PCL-TCP) group showed non-significant higher bone surface, bone specific surface, and bone surface density than the other two groups as revealed by the µCT bone morphogenic analysis. Histologically, the same group revealed higher bone-implant contact ratio (BIC) (p = 0.017) and new bone height formation (NBH, mm) (p = 0.0097) with statistically significant difference compared to the PCL-TCP scaffold group. CONCLUSIONS: PCL-TCP scaffold is compatible for bone regeneration in bone defects surrounding dental implants. Moreover, the incorporation of MSCs and PRP optimized the bone regeneration process with respect to the rate of scaffold replacement, the height of the regenerated bone, and implant stability.

Postulated release profile of recombinant human bone morphogenetic protein-2 (rhBMP-2) from demineralized dentin matrix.

Demineralized dentin matrix (DDM) has been used as a recombinant human bone morphogenetic protein-2 (rhBMP-2) carrier in many clinical trials. To optimize the clinical safety and efficacy of rhBMP-2 with DDM, efforts have been made to improve the delivery of rhBMP-2 by 1) lowering the administered dose, 2) localizing the protein, and 3) prolonging its retention time at the action site as well as the bone forming capacity of the carrier itself. The release profile of rhBMP-2 that is associated with endogenous BMP in dentin has been postulated according to the type of incorporation, which is attributed to the loosened interfibrillar space and nanoporous dentinal tubule pores. Physically adsorbed and modified, physically entrapped rhBMP-2 is sequentially released from the DDM surface during the early stage of implantation. As DDM degradation progresses, the loosened interfibrillar space and enlarged dentinal tubules release the entrapped rhBMP-2. Finally, the endogenous BMP in dentin is released with osteoclastic dentin resorption. According to the postulated release profile, DDM can therefore be used in a controlled manner as a sequential delivery scaffold for rhBMP-2, thus sustaining the rhBMP-2 concentration for a prolonged period due to localization. In addition, we attempted to determine how to lower the rhBMP-2 concentration to 0.2 mg/mL, which is lower than the approved 1.5 mg/mL.

Evaluation of the LH780 hematology analyzer for detection and therapeutic monitoring of malaria: Cross-reactivity with nucleated RBCs.

We evaluated the diagnostic usefulness of the LH780 Coulter blood cell counter for detection and therapeutic monitoring of malaria including cross-reactivity with nucleated RBC (NRBC) samples. A total of 405 patients (43 patients with Plasmodium vivax malaria and the control group of 120 healthy subjects, 111 patients with fever, and 131 patients with NRBCs) were analyzed with routine CBC using the LH780. We analyzed the CBC results according to three selected parameters: an abnormal peak in the WBC histogram before 35fL, the presence of red dots in the nonwhite cell zone of 2D WBC Diff Dataplot, and platelet-related flags suggesting platelet clumps or giant platelets. Of the 43 malaria samples collected at diagnosis, an abnormal peak (≥2.2mm) was present in 93.0% (95% confidential interval (CI), 80.9-98.5%). Of all samples, 97.7% (95% CI, 87.7-99.9%) exhibited red dots, and platelet-related flags were observed in 81.4% (95% CI, 66.6-91.6%). The specificity of these three selected parameters was 83.1% (95% CI, 78.9-86.9%), 77.3% (95% CI, 72.7-81.6%), and 90.1% (95% CI, 86.5-92.9%), respectively. The abnormal peak (≥2.2mm) showed moderate correlation with parasite level (r=0.79). The three selected LH780 parameters were useful for identifying malaria in healthy subjects and febrile patients, but unsatisfactory for discriminating malaria in NRBC samples. The parameters showed a substantial proportion of false positives in the NRBC group, ranging from 26.7% to 49.6%. Therefore, microscopic confirmation will be necessary for application of these parameters for malaria screening and treatment monitoring.

Study of erythrocyte aggregation at pulsatile flow conditions with backscattering analysis.

In vivo red blood cell aggregation will vary under pulsatile flow but few studies have been reported due to various difficulties in generating physiological flow conditions and detecting RBC aggregation. The present study developed a microfluidic system that generates cyclic pulsatile flow through a microchannel. Backscattered light signals from human blood were recorded over time and analyzed for RBC aggregation in pulsatile flow. Four different blood samples (control, normal RBCs in PBS, hardened RBCs in autologous plasma, and hardened RBCs in PBS) were examined. In a cyclic pulsatile flow condition, light intensity-time curve for the control and hardened RBCs in plasma exhibited apparent critical shear stresses that were similar to the respective values measured at a single pulse flow condition. During entire cycles of pulsatile flow, the measured critical shear stress remained nearly constant. We conclude that the critical shear stress can be observed in cyclic pulsatile flow and would be an important index to represent in-vivo pulsatile blood flow rheology.

Investigation of critical shear stress with simultaneous measurement of electrical impedance, capacitance and light backscattering.

Recent electrical investigation of hemorheology provided useful information on the kinetics of red blood cell (RBC) aggregation. However, because of the inconsistent results in the electrical measurements, we need to understand the electrical characteristics of RBC aggregation at various flow conditions. In the present study, AC electrical-capacitance (EC) and -impedance (EI) and light backscattering (LB) were simultaneously measured for transient shear-decreasing blood flow in a microchannel. EI, EC and LB signals of RBCs in plasma show similar time-varying curves, both yielding either a peak or a minimal point in the optimal frequency range (10~500 kHz). Critical shear stress (CSS) determined from EC showed the nearly same results as that determined from LB, with yielding hematocrits-independence and dextran-concentration dependence. However, the high concentration of fibrinogen caused electrical saturation, which resulted in different results of CSS determined from between LB and EC. These results suggest that electrical properties of RBC suspensions should be further examined to replace the optical method of measurement of RBC aggregation.

Comparison of light-transmission and -backscattering methods in the measurement of red blood cell aggregation.

Light-transmission and light-backscattering methods are commonly used to determine red-blood-cell (RBC) aggregation. Even though the results reveal good correlations between the parameters that are measured by these two methods, the methods themselves yield quite different values. The objective of this research is to investigate and delineate the characteristics of the two optical methods. We measured RBC aggregation by using a newly developed microchip-based aggregometer. An orthogonal polarization technique, wherein multiple scattering causes polarized light to be depolarized and passed through an orthogonal polarizer, was applied to the backscattering method. Our results were also compared to those of conventional aggregometers [laser-assisted optical rotational cell analyzer (LORCA)], and revealed that the backscattering method yielded higher aggregation indices than the transmission method and LORCA. However, the backscattering method with orthogonal polarization yielded the same values of aggregation indices as the transmission method. These agreements between the two methods were also found in measurements of RBC aggregability in various concentrations of dextran solutions.

Temperature-dependent threshold shear stress of red blood cell aggregation.

Red blood cell (RBC) aggregation is becoming an important hemorheological parameter, which exhibits a unique temperature dependence. However, further investigation is still required for understanding the temperature-dependent characteristics of hemorheology that includes RBC aggregation. In the present study, blood samples were examined at 3, 10, 20, 30, and 37 degrees C. When the temperature decreases, the whole-blood and plasma viscosities increase, whereas the aggregation indices (AI, M, and b) yield contrary results. Since these contradictory results are known to arise from an increase in the plasma viscosity as the temperature decreases, aggregation indices that were corrected for plasma viscosity were examined. The corrected indices showed mixed results with the variation of the temperature. However, the threshold shear rate and the threshold shear stress increased as the temperature decreased, which is a trend that agrees with that of the blood viscosity. As the temperature decreases, RBC aggregates become more resistant to hydrodynamic dispersion and the corresponding threshold shear stress increases as does the blood viscosity. Therefore, the threshold shear stress may help to better clarify the mechanics of RBC aggregation under both physiological and pathological conditions.

Measurement of the temperature-dependent threshold shear-stress of red blood cell aggregation.

Red blood cell (RBC) aggregation is becoming an important hemorheological parameter, which typically exhibits temperature dependence. Quite recently, a critical shear-stress was proposed as a new dimensional index to represent the aggregative and disaggregative behaviors of RBCs. The present study investigated the effect of the temperature on the critical shear-stress that is required to keep RBC aggregates dispersed. The critical shear-stress was measured at various temperatures (4, 10, 20, 30, and 37 degrees C) through the use of a transient microfluidic aggregometry. The critical shear-stress significantly increased as the blood temperature lowered, which accorded with the increase in the low-shear blood viscosity with the lowering of the temperature. Furthermore, the critical shear-stress also showed good agreement with the threshold shear-stress, as measured in a rotational Couette flow. These findings assist in rheologically validating the critical shear-stress, as defined in the microfluidic aggregometry.