Abstract: Muscle regeneration after injury (as irradiation) is of great importance. However, the molecular and cellular mechanisms are still unclear. Cytokines are believed to play fundamental role in the different stages of muscle regeneration. They are secreted by many cell populations, but the predominant producers are macrophages and helper T cells. On the other hand, it has been shown that adipose tissue derived stromal/stem cell (ASC) injection could improve muscle regeneration. Stem cells probably induce the coordinated modulations of gene expression in different macrophage cells. Therefore, we investigated the patterns and timing of changes in gene expression of different cytokines occurring upon stem cells loading. Muscle regeneration was studied in an irradiated muscle of minipig animal model in presence or absence of ASC treatment (irradiated and treated with ASCs, IRR+ASC; irradiated not-treated with ASCs, IRR; and non-irradiated no-IRR). We characterized macrophage populations by immunolabeling in the different conditions. In our study, we found mostly M2 and a few M1 macrophages in the IRR+ASC samples. However, only few M2b macrophages were noticed in the IRR muscles. In addition, we found intensive fibrosis in the IRR samples. With in situ hybridization and immunolabeling, we analyzed the cytokine expression of the different macrophages and we showed that M2d macrophage are the most abundant in the IRR+ASC samples. By in situ hybridization, strong expression of the transforming growth factor β (TGF-β) was observed in the IRR+ASC but very week in the IRR samples. But when we analyzed TGF-β level with immunolabeling the expression was very different: many M2 macrophages showed week expression in IRR+ASC and few cells expressing stronger level in IRR muscles. Therefore, we investigated the MMP expressions in the different muscles. Our data showed that the M2 macrophages of the IRR+ASC muscle expressed MMP2 proteins. Our working hypothesis is that MMP2 expression of the M2 macrophages can decrease fibrosis in the IRR+ASC muscle by capturing TGF-β.
Abstract: This study was aimed to measure effective transverse
relaxation rates (R2*) in the liver and muscle of normal New Zealand
White (NZW) rabbits. R2* relaxation rate has been widely used in
various hepatic diseases for iron overload by quantifying iron contents
in liver. R2* relaxation rate is defined as the reciprocal of T2*
relaxation time and mainly depends on the constituents of tissue.
Different tissues would have different R2* relaxation rates. The signal
intensity decay in Magnetic resonance imaging (MRI) may be
characterized by R2* relaxation rates. In this study, a 1.5T GE Signa
HDxt whole body MR scanner equipped with an 8-channel high
resolution knee coil was used to observe R2* values in NZW rabbit’s
liver and muscle. Eight healthy NZW rabbits weighted 2 ~ 2.5 kg were
recruited. After anesthesia using Zoletil 50 and Rompun 2% mixture,
the abdomen of rabbit was landmarked at the center of knee coil to
perform 3-plane localizer scan using fast spoiled gradient echo
(FSPGR) pulse sequence. Afterwards, multi-planar fast gradient echo
(MFGR) scans were performed with 8 various echo times (TEs) to
acquire images for R2* measurements. Regions of interest (ROIs) at
liver and muscle were measured using Advantage workstation.
Finally, the R2* was obtained by a linear regression of ln(sı) on TE.
The results showed that the longer the echo time, the smaller the signal
intensity. The R2* values of liver and muscle were 44.8 ± 10.9 s-1 and
37.4 ± 9.5 s-1, respectively. It implies that the iron concentration of
liver is higher than that of muscle. In conclusion, the more the iron
contents in tissue, the higher the R2*. The correlations between R2*
and iron content in NZW rabbits might be valuable for further
exploration.
Abstract: Haemodialysis (HD) is a procedure saving patient lives around the world, unfortunately it brings numerous complications. Oxidative stress is one of the major factors which lead to erythrocytes destruction during extracorporeal circulation. Repeated HD procedures destroy blood elements and the organism is not able to keep up with their production. 30 HD procedures on healthy sheep were performed to evaluate effects of such treatment. Oxidative stress study was performed together with an analysis of basic blood parameters and empirical assessment of dialyzer condition after the procedure. A reversible decline in absolute leukocyte count, during first 30 min of HD, was observed. Blood clots were formed in the area of the blood inlet and outlet of the dialyzer. Our results are consistent with outcomes presented throughout the literature specifically with respect to the effects observed in humans and will provide a basis to evaluate methods for blood protection during haemodialysis.
Abstract: Transdermal delivery of ondansetron hydrochloride (OdHCl) can prevent the problems encountered with oral ondansetron. In previously conducted studies, effect of amount of polyvinyl pyrrolidone, permeation enhancer and casting solvent on the physicochemical properties on OdHCl were investigated. It is feasible to develop ondansetron transdermal patch by using ethyl cellulose and polyvinyl pyrrolidone with dibutyl pthalate as plasticizer, however, the desired flux is not achieved. The primary aim of this study is to use dimethyl succinate (DMS) and propylene glycol that are not incorporated in previous studies to determine their effect on the physicochemical properties of an OdHCl transdermal patch using ethyl cellulose and polyvinyl pyrrolidone. This study also investigates the effect of permeation enhancer (eugenol and phosphatidylcholine) on the release of OdHCl. The results showed that propylene glycol is a more suitable plasticizer compared to DMS in the fabrication of OdHCl transdermal patch using ethyl cellulose and polyvinyl pyrrolidone as polymers. Propylene glycol containing patch has optimum drug content, thickness, moisture content and water absorption, tensile strength, and a better release profile than DMS. Eugenol and phosphatidylcholine can increase release of OdHCl from the patches. From the physicochemical result and permeation profile, a combination of 350mg of ethyl cellulose, 150mg polyvinyl pyrrolidone, 3% of total polymer weight of eugenol, and 40% of total polymer weight of propylene glycol is the most suitable formulation to develop an OdHCl patch. OdHCl release did not increase with increasing the percentage of plasticiser. DMS 4, PG 4, DMS 9, PG 9, DMS 14, and PG 14 gave better release profiles where using 300mg: 0mg, 300mg: 100mg, and 350mg: 150mg of EC: PVP. Thus, 40% of PG or DMS appeared to be the optimum amount of plasticiser when the above combination where EC: PVP was used. It was concluded from the study that a patch formulation containing 350mg EC, 150mg PVP, 40% PG and 3% eugenol is the best transdermal matrix patch compositions for the uniform and continuous release/permeation of OdHCl over an extended period. This patch design can be used for further pharmacokinetic and pharmacodynamic studies in suitable animal models.
Abstract: Total liquid ventilation can support gas exchange in animal models of lung injury. Clinical application awaits further technical improvements and performance verification. Our aim was to develop a liquid ventilator, able to deliver accurate tidal volumes, and a computerized system for measuring lung mechanics. The computer-assisted, piston-driven respirator controlled ventilatory parameters that were displayed and modified on a real-time basis. Pressure and temperature transducers along with a lineal displacement controller provided the necessary signals to calculate lung mechanics. Ten newborn lambs (
Abstract: Total liquid ventilation can support gas exchange in animal models of lung injury. Clinical application awaits further technical improvements and performance verification. Our aim was to develop a liquid ventilator, able to deliver accurate tidal volumes, and a computerized system for measuring lung mechanics. The computer-assisted, piston-driven respirator controlled ventilatory parameters that were displayed and modified on a real-time basis. Pressure and temperature transducers along with a lineal displacement controller provided the necessary signals to calculate lung mechanics. Ten newborn lambs (
Abstract: In this paper we evaluated the efficacy of
photodynamic treatment of infected wounds on pig animal model by
diffuse reflectance spectrometry. The study was conducted on fifteen
wounds contaminated with Staphylococcus aureus bacteria that were
incubated for 30 min with methylene blue solution (c = 3.3 x 10-3 M)
and exposed to laser radiations (λ = 670 nm, P = 15 mW) for 15 min.
The efficiency of photodynamic inactivation of bacteria was
evaluated by microbiological exams and diffuse reflectance
spectrometry. The results of the microbiological exams showed that
the bacterial concentration has decreased from 6.93±0.138
logCFU/ml to 3.12±0.108 logCFU/ml. The spectral examination
showed that the diffuse reflectance of wounds contaminated with
Staphylococcus aureus has decreased from 5.06±0.036 % to
3.36±0.025 %. In conclusion, photodynamic therapy is an effective
method for the treatment of infected wounds and there is a correlation
between the CFU count and diffuse reflectance.
Abstract: Snake bite cases in Malaysia most often involve the
species Naja-naja and Calloselasma rhodostoma. In keeping with the
need for a rapid snake venom detection kit in a clinical setting, plate
and dot-ELISA test for the venoms of Naja-naja sumatrana,
Calloselasma rhodostoma and the cobra venom fraction V antigen
was developed. Polyclonal antibodies were raised and further used to
prepare the reagents for the dot-ELISA test kit which was tested in
mice, rabbit and virtual human models. The newly developed dot-
ELISA kit was able to detect a minimum venom concentration of
244ng/ml with cross reactivity of one antibody type. The dot-ELISA
system was sensitive and specific for all three snake venom types in
all tested animal models. The lowest minimum venom concentration
detectable was in the rabbit model, 244ng/ml of the cobra venom
fraction V antigen. The highest minimum venom concentration was
in mice, 1953ng/ml against a multitude of venoms. The developed
dot-ELISA system for the detection of three snake venom types was
successful with a sensitivity of 95.8% and specificity of 97.9%.