BACKGROUND: Mesenchymal stromal cells (MSCs) have potent immunomodulatory and neuroprotective properties, and have been tested in neurodegenerative diseases resulting in meaningful clinical improvements. Regulatory guidelines specify the need to perform preclinical studies prior any clinical trial, including biodistribution assays and tumourigenesis exclusion. We conducted a preclinical study of human bone marrow MSCs (hBM-MSCs) injected by intrathecal route in Non-Obese Diabetic Severe Combined Immunodeficiency mice, to explore cellular biodistribution and toxicity as a privileged administration method for cell therapy in Friedreich's Ataxia. METHODS: For this purpose, 3 × 10⁵ cells were injected by intrathecal route in 12 animals (experimental group) and the same volume of culture media in 6 animals (control group). Blood samples were collected at 24 h (n = 9) or 4 months (n = 9) to assess toxicity, and nine organs were harvested for histology and safety studies. Genomic DNA was isolated from all tissues, and mouse GAPDH and human β2M and β-actin genes were amplified by qPCR to analyze hBM-MSCs biodistribution. RESULTS: There were no deaths nor acute or chronic toxicity. Hematology, biochemistry and body weight were in the range of normal values in all groups. At 24 h hBM-MSCs were detected in 4/6 spinal cords and 1/6 hearts, and at 4 months in 3/6 hearts and 1/6 brains of transplanted mice. No tumours were found. CONCLUSION: This study demonstrated that intrathecal injection of hBM-MSCs is safe, non toxic and do not produce tumors. These results provide further evidence that hBM-MSCs might be used in a clinical trial in patients with FRDA.
Animals ; Biochemistry ; Body Weight ; Bone Marrow ; Brain ; Cell- and Tissue-Based Therapy ; Culture Media ; DNA ; Friedreich Ataxia ; Heart ; Hematology ; Humans ; Injections, Spinal ; Mesenchymal Stromal Cells ; Methods ; Mice ; Neurodegenerative Diseases ; Neuroprotection ; Reference Values ; Severe Combined Immunodeficiency ; Spinal Cord

A comparative in vivo pharmacokinetic (PK) study of tilmicosin (TIL) was conducted in 6 crossbred healthy pigs and 6 crossbred pigs infected with Haemophilus (H.) parasuis following oral administration of a single 40 mg/kg dose. The infected model was established by intranasal inoculation and confirmed by clinical signs, blood biochemistry, and microscopic examinations. Plasma TIL concentrations were determined by a validated high-performance liquid chromatography method with ultraviolet detection at 285 nm. PK parameters were calculated by using WinNonlin software. After TIL administration, the main PK parameters of TIL in healthy and H. parasuis-infected pigs were as follows: Area under the concentration-time curve, maximal drug concentration, half-life of the absorption phase, half-life of the distribution phase, and half-life of the elimination phase were 34.86 ± 9.69 vs. 28.73 ± 6.18 µg · h/mL, 1.77 ± 0.33 vs. 1.67 ± 0.28 µg/mL, 2.27 ± 0.45 vs. 2.24 ± 0.44 h, 5.35 ± 1.40 vs. 4.61 ± 0.35 h, and 43.53 ± 8.17 vs. 42.05 ± 9.36 h, respectively. These results of this exploratory study suggest that there were no significant differences between the PK profiles of TIL in the healthy and H. parasuis-infected pigs.
Absorption ; Administration, Oral ; Biochemistry ; Chromatography, Liquid ; Haemophilus parasuis* ; Haemophilus* ; Half-Life ; Methods ; Pharmacokinetics* ; Plasma ; Swine*

Dendrobium moniliforme (L.) Sw., an herb of the Orchidaceae family, has long been used in traditional medicine to strengthen bones, nourish the stomach, and promote the production of bodily fluid. Recently, polysaccharides isolated from Dendrobium have been used in functional foods and nutraceutical products. A traditional method to process Dendrobium is to soak fresh stems in an ethanol solution, which is the most important factor to ensure high yields of aqueous-extractable polysaccharides. The present study was carried out to investigate the potential acute toxicity of D. moniliforme aqueous extract (DMAE), by a single oral dose in Sprague-Dawley rats. The test article was orally administered once by gavage to male and female rats at doses of 0, 2,500, and 5,000 mg/kg body weight (n=5 male and female rats for each dose). Throughout the study period, no treatment-related deaths were observed and no adverse effects were noted in clinical signs, body weight, food consumption, serum biochemistry, organ weight, or gross findings at any dose tested. The results show that a single oral administration of DMAE did not induce any toxic effects at a dose below 5,000 mg/kg in rats, and the minimal lethal dose was considered to be over 5,000 mg/kg body weight for both sexes. With respect to cytotoxicity, the cell viability of human embryonic kidney (HEK293) cells was less than 50% when the cells were treated with 10 mg/mL aqueous extract for 24 h.
Administration, Oral ; Animals ; Biochemistry ; Body Weight ; Cell Survival ; Dendrobium* ; Dietary Supplements ; Ethanol ; Female ; Functional Food ; Humans ; In Vitro Techniques* ; Kidney ; Male ; Medicine, Traditional ; Methods ; Orchidaceae ; Organ Size ; Polysaccharides ; Rats ; Rats, Sprague-Dawley ; Stomach

Base Sequence ; Binding Sites, Antibody ; Biochemistry ; methods ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; Genetic Vectors ; Immunoglobulin Fc Fragments ; genetics ; metabolism ; Immunoglobulin G ; genetics ; metabolism ; Ligands ; Molecular Sequence Data ; Plasmids ; Protein Binding ; Recombinant Fusion Proteins ; genetics ; isolation & purification ; Staphylococcal Protein A ; genetics ; metabolism

BACKGROUND: The Samsung LABGEO PT10 (Samsung Electronics, Korea) has been developed as a point-of-care testing (POCT) chemistry analyzer. We evaluated the performance of the Samsung LABGEO PT Biochemistry Test 15 (Samsung Electronics) and the HbA1c Test (Samsung Electronics), which are dedicated cartridges for the LABGEO PT10. METHODS: Based on the Clinical and Laboratory Standards Institute guidelines, the precision, linearity, and methodology were evaluated for seven chemistry analytes (cholesterol, triglycerides, HDL cholesterol, blood urea nitrogen, creatinine, amylase, and hemoglobin A1c). All the analytes, except for hemoglobin A1c, were obtained from three different types of samples: whole blood, plasma, and serum, to evaluate matrix effects. RESULTS: In the precision analysis, both within-run and total-run coefficients of variation were less than 10% for the seven analytes. Dose curves for the seven analytes were linear in the clinically relevant concentration ranges. The methodology study yielded correlation coefficients > or =0.98 for the seven comparisons of the LABGEO PT10 cartridge tests with other methods. Except for HDL cholesterol, the percentage differences between test results obtained from whole blood, plasma, and serum, were within +/-10%. The concentrations of HDL cholesterol measured in whole blood samples were 0.9 mg/dL and 5.6 mg/dL higher than those measured in plasma and serum specimens, respectively. CONCLUSIONS: The LABGEO PT10 showed suitable analytical performance with respect to precision and linearity and demonstrated a good correlation with automated chemistry analyzer. With the additional benefits of a short turnaround time and ease of use, the LABGEO PT10 is an acceptable POCT chemistry analyzer.
Amylases ; Biochemistry ; Blood Urea Nitrogen ; Chemistry* ; Cholesterol, HDL ; Creatinine ; Methods ; Plasma ; Point-of-Care Systems ; Triglycerides

A fully-automated biochemical analysis system is developed, intending for primary medical units. It features high reliability, high usability, strong adaptability, low operation cost, low maintenance cost, and low requirements for operators.
Automation ; instrumentation ; methods ; Biochemistry ; instrumentation ; methods ; Equipment Design ; Equipment and Supplies, Hospital ; Hospitals, Rural

In this paper the system structure The automatic biochemistry analyzer is a necessary instrument for clinical diagnostics. First of is analyzed. The system problems description and the fundamental principles for dispatch are brought forward. Then this text puts emphasis on the modeling for the automatic biochemistry analyzer control system. The objects model and the communications model are put forward. Finally, the implementation method is designed. It indicates that the system based on the model has good performance.
Autoanalysis ; instrumentation ; methods ; Biochemistry ; instrumentation ; methods ; Equipment Design ; Models, Theoretical

OBJECTIVETo establish a methodology of protein misfolding cyclic amplification (PMCA) and utilize in the detection of PrP(Sc) in brain tissues from prion diseases.

METHODSDifferent amounts of Scrapie 263K agent bulk were mixed with brain homogenates of health hamsters and treated with repeated incubation/sonication for 10 to 15 cycles. The proteinase K-resistant PrP(Sc) was evaluated with Western Blot.

RESULTSIn this experimental situation, 263K agent replicated rapidly in vitro, utilizing hamsters' brains as the medium. With the established PrP(Sc)-PMCA technique, PrP(Sc) signals in the preparations containing less than 10(-5) diluted 263K bulk could be detected. Compared with conveniently used immuno-blot assay, the sensitivity of PrP(Sc)-PMCA for PrP(Sc) was 10(5) to 10(6)-fold increased. It has been also shown that homogenates of cerebellar and brain stem could be used as the medium for PrP(Sc) replication.

CONCLUSIONA rapidly replicating method for PrP(Sc), PrP(Sc)-PMCA, was successfully established, providing a new approach for early diagnosis of prion diseases and research on the biological features of prion.

Animals ; Biochemistry ; methods ; Blotting, Western ; Brain ; metabolism ; pathology ; Cricetinae ; PrPSc Proteins ; analysis ; chemistry ; Prion Diseases ; diagnosis ; metabolism ; Protein Folding ; Reproducibility of Results ; Sensitivity and Specificity

OBJECTIVETo establish a method of determining alpha-glucosidase activity in seminal plasma by semi-automatic biochemistry analyzer.

METHODSAlpha-glucosidase activity in seminal plasma from 51 men with normal semen parameters in routine semen analysis were detected by semi-automatic biochemistry analyzer and manual glucose oxidase method, respectively. Then, the intra-assay and inter-assay coefficient variation (CV) and normal reference value were calculated. In the meanwhile, the correlation between the two methods was analyzed.

RESULTSThe intra-assay CVs of 2 seminal plasma samples with different alpha-glucosidase activity detected by semi-automatic biochemistry analyzer were 12.63% and 9.13%, and the inter-assay CVs were 10.67% and 13.49%, respectively. The normal reference value for seminal alpha-glucosidase activity detected with semi-automatic biochemistry analyzer ranged from 102.28 to 555.08 U/L. There was a significantly positive correlation between the semi-automatic biochemistry analyzer and the manual glucose oxidase method (r = 0.792, P < 0.01).

CONCLUSIONThe method of determining alpha-glucosidase activity in seminal plasma by semi-automatic biochemistry analyzer, with its simplicity, less cost of time and reagents, and more reliable result, could be applied to clinical laboratory medicine.

Adult ; Biochemistry ; instrumentation ; methods ; Humans ; Male ; Reference Values ; Reproducibility of Results ; Semen ; enzymology ; alpha-Glucosidases ; analysis ; metabolism

Octenyl succinic anhydride (OSA) modified early Indica rice starch was prepared in aqueous slurry systems using response surface methodology. The paste properties of the OSA starch were also investigated. Results indicated that the suitable parameters for the preparation of OSA starch from early Indica rice starch were as follows: reaction period 4 h, reaction temperature 33.4 degrees C, pH of reaction system 8.4, concentration of starch slurry 36.8% (in proportion to water, w/w), amount of OSA 3% (in proportion to starch, w/w). The degree of substitution was 0.0188 and the reaction efficiency was 81.0%. The results of paste properties showed that with increased OSA modification, the starch derivatives had higher paste clarity, decreased retrogradation and better freeze-thaw stability.
Biochemistry ; methods ; Chemical Phenomena ; Chemistry, Physical ; Freezing ; Hydrogen-Ion Concentration ; Light ; Models, Statistical ; Oryza ; metabolism ; Starch ; chemistry ; Succinic Anhydrides ; analysis ; chemistry ; Temperature