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Immunochemical evaluation of radioiodinated protein A.

The [125I]iodoprotein A iodinated by the chloramine-T method using carrier-free 125I- to various specific activities (0.02-2.0 125I/mole protein A) displayed 85% binding activity to excess solid-phase rabbit IgG. On storage, however, the solid-phase IgG binding activity of the high specific activity [125I]iodoprotein A (approximately 2 125I/protein A) decreased rapidly while that of the low specific activity [125I]iodoprotein A (approximately 0.2 125I/protein A) remained essentially unchanged. Analysis of aged high specific activity [125I]iodoprotein A by thin-layer chromatography revealed that free 125I- was released from the [125I]iodoprotein A during storage. Removal of the released 125I- was accomplished by using anion exchange resin. High specific activity [125I]iodoprotein A was found to be more sensitive than the low specific activity [125I]iodoprotein A in quantitating aggregated IgG while the latter was as useful as the former in hybridoma screening. When protein A was labeled with the Bolton-Hunter reagent, the [125I]iodoprotein A had an IgG binding property and stability similar to the [125I]iodoprotein A labeled with the chloramine-T method. Since the chloramine-T method can yield high specific activity [125I]iodoprotein A and repurification of the aged preparation can easily be achieved by using anion exchange resin, we concluded that chloramine-T is more efficient than Bolton-Hunter reagent and that both methods yield materials with comparable immunochemical properties.

Animals↗

Metal ion coordination of macromolecular bioligands: formation of zinc(II) complex of hyaluronic acid.

The coordination of zinc(II) ion to hyaluronate (Hya), a natural copolymer, in aqueous solution at pH 6 was investigated by potentiometric and circular dichroism (CD) spectroscopic methods, and by monitoring the changes in macroscopic properties by high-precision measurements. The zinc(II)-selective electrode, and CD measurements proved the binding of zinc(II) by Hya. A number of Hya fragments (Mr approximately 3.3 x 10(3)-1.4 x 10(6)) were studied to estimate the contributions of the polyelectrolyte effect, the solvation and host-guest interactions to the extra stabilization of the macromolecular zinc(II) complexes as compared with the monomeric unit. The zinc(II) ion activity increase reflected a stability decrease for the fragments with Mr < 4 x 10(4). This molecular weight differs from that where cleavage of the Hya skeleton starts (approximately 5 x 10(5), according to the size-exclusion gel, and anion-exchange chromatographic behavior of the Hya fragments) and from that where the polyelectrolyte effect stops (approximately 6 x 10(3)). The excess volumes and Bingham shear yield values of the solutions revealed the transformation of the coherent random coil structure stabilized by intermolecular association in the NaHya to an intramolecular association producing the globular structure of the ZnHya molecule, with a smaller but more strongly bound solvate water sheet.

Chromatography, Gel↗

Use of a high molecular weight fluorescein (fluorexon) ophthalmic strip in assessments of tear film break-up time in contact lens wearers and non-contact lens wearers.

PURPOSE: To assess whether fluorexon strips could be used in tear stability [tear film break-up time (TBUT)] measures, as compared with fluorescein strips. METHODS: The pre-ocular tear film stability was assessed in 40 adults (19-29 years of age), half of whom were adapted soft contact lens wearers, using the dye-assisted tear film break-up time (TBUT) method. The measures were carried out between 10.00 and 17.00 hours, with contact lens wearers removing their lenses just before the assessments. For all subjects, fluorescein or fluorexon ophthalmic strips (pre-wetted with preservative-free saline) were used to add dye to the tear film, the TBUT was assessed three times and then the eye lightly irrigated with saline. The measures were repeated 45 min later with the other dye, with half the subjects receiving fluorescein first and the other half receiving fluorexon first. RESULTS: Overall, the mean TBUT times were 12.2 +/- 5.9 s for fluorescein and 12.6 +/- 5.6 s for fluorexon (p = n.s.), but the non-lens wearers had longer TBUT times (14.6 and 14.5 s for the two dyes) compared with contact lens wearers (9.8 and 10.7 s, p < 0.01). While the inter-subject variability (as S.D.) was larger for non-contact lens wearers, the intra-subject variability [as coefficient of variation (COV)] was greater in the contact lens wearers when fluorescein was used (p < 0.001). When analysed as a function of the time of day that the measures were made, there were clear trends for both the fluorescein and fluorexon. TBUT values to increase in non-contact lens wearers and decrease in contact lens wearers. These trends failed to reach statistical significance, but the results with the contact lens wearers showed less variation than the non-contact lens wearers. CONCLUSION: The use of a fluorexon dye-impregnated strip (pre-wetted with preservative-free saline) yielded comparable results with a fluorescein strip for assessments of tear film stability based on TBUT. The studies also indicate that further research is needed on changes in TBUT values during the day.

Adult↗

Energetics of intersubunit and intrasubunit interactions of Escherichia coli adenosine cyclic 3',5'-phosphate receptor protein.

Escherichia coli cAMP receptor protein (CRP) regulates the expression of a large number of catabolite-sensitive genes. The mechanism of CRP regulation most likely involves communication between subunits and domains. A specific message, such as the activation of CRP, may be manifested as a change in the interactions between these structural entities. Hence, the elucidation of the regulatory mechanism would require a quantitative evaluation of the energetics involved in these interactions. Thus, a study was initiated to define the conditions for reversible denaturation of CRP and to quantitatively assess the energetics involved in the intra- and intersubunit interactions in CRP. The denaturation of CRP was induced by guanidine hydrochloride. The equilibrium unfolding reaction of CRP was monitored by three spectroscopic techniques, namely, fluorescence intensity, fluorescence anisotropy, and circular dichroism. The spectroscopic data implied that CRP unfolds in a single cooperative transition. Sedimentation equilibrium data showed that CRP is dissociated into its monomeric state in high concentrations of denaturant. Unfolding of CRP is completely reversible, as indicated by fluorescence and circular dichroism measurements, and sedimentation data indicated that a dimeric structure of CRP was recovered. The functional and other structural properties of renatured and native CRP have also been examined. Quantitatively identical results were obtained. Results from additional studies as a function of protein concentration and from computer simulation demonstrated that the denaturation of CRP induced by guanidine hydrochloride proceeds according to the following pathway: (CRP2)Native<-->2(CRP)Native<-->2(CRP)Denatured. The delta G values for dissociation (delta Gd) and unfolding (delta G(u)) in the absence of guanidine hydrochloride were determined by linear extrapolation, yielding values of 12.0 +/- 0.6 and 7.2 +/- 0.1 kcal/mol, respectively. To examine the effect of the DNA binding domain on the stability of the cAMP binding domain, two proteolytically resistant cAMP binding cores were prepared from CRP in the presence of cAMP by subtilisin and chymotrypsin digestion, yielding S-CRP and CH-CRP, respectively. Results from an equilibrium denaturation study indicated that the denaturation of both CH-CRP and S-CRP is also completely reversible. Both S-CRP and CH-CRP exist as stable dimers with similar delta Gd values of 10.1 +/- 0.4 and 9.5 +/- 0.4 kcal/mol, respectively. Results from this study in conjunction with crystallographic data [McKay, D. B., Weber, I. T., & Stietz, T. A. (1982) J. Biol. Chem. 257, 9518-9524] indicate that the DNA binding domain and the C-helix are not the only structural elements that are responsible for subunit dimerization.(ABSTRACT TRUNCATED AT 400 WORDS)

Binding Sites↗

Trehalose glass-facilitated thermal reduction of metmyoglobin and methemoglobin.

The reduction of ferric derivatives of hemeproteins in solution typically requires moderate to strong reducing agents. Reducing sugars are not adequate to reduce ferric myoglobins or hemoglobins under solution conditions favorable to protein stability. We find that embedding aquo-met derivatives of horse myoglobin and human adult hemoglobin in a glucose-doped glassy matrix derived from trehalose facilitates an efficient thermally initiated reduction that yields a five-coordinate high-spin ferrous heme. The trehalose glass plays a central role by stabilizing the reduction-prone bis-histidine heme (hemichrome) intermediate under the high-temperature conditions that favor the open reducing form of glucose. Due to glass-imposed limitations on conformational reorganization, this process has clear applications in biophysics where it can be used to generate nonequilibrium ferrous derivatives having the initial conformation of the aquo-met derivative. Since the glassy matrix can be redissolved to release the embedded protein, this technique is not only a basis for a relatively benign method of reducing hemoglobin-based blood substitutes that have undergone autoxidation during storage but may also be a way to reactivate stored proteins that have undergone oxidation.

Animals↗

Retention versus sacrifice of the posterior cruciate ligament in total knee replacement for treatment of osteoarthritis and rheumatoid arthritis.

BACKGROUND: The functional and clinical results to support the choice whether or not to retain the posterior cruciate ligament (PCL) during total knee arthroplasty have not been gathered and analysed so far. There are at least some trials showing no difference. OBJECTIVES: To identify the difference in functional, clinical, and radiological outcome between retention and sacrifice of the PCL in total knee arthroplasty in patients with osteoarthrosis and other non-traumatic diseases. SEARCH STRATEGY: A search was conducted in MEDLINE(Through PubMed; 1966 - March 2004), EMBASE (1980 - March 2004), Cochrane Central Register of Controlled Trials (CENTRAL Issue 2004 - 1), and Current Contents (1996 - March 2004). Also, references of selected articles were checked and citation tracking on the articles selected was performed. SELECTION CRITERIA: Randomised controlled trials comparing retention to sacrifice of the PCL during total knee arthroplasty with regard to functional, radiological and clinical outcome in patients with osteoarthritis and other non-traumatic diseases were selected by two independent reviewers. DATA COLLECTION AND ANALYSIS: Methodological quality was assessed with the checklist by van Tulder and the Jadad list. Data was collected with a predeveloped form. Meta-analysis was performed with subgroup analyses on age, gender, disease severity, and follow-up time, if allowed by adequate power. MAIN RESULTS: Eight randomised controlled trials were found. Two treatment options were compared against PCL retention: PCL sacrifice without additional stabilisation (post and cam mechanism) (2 studies), and PCL sacrifice with posterior stabilized design (5 studies). One study included all three options. Range of motion was found to be 8.1 degrees higher in the posterior stabilized group compared to the PCL retention group (p=0.01, 95% confidence interval [1.7, 14.5]), although the heterogeneity was high (I(2 )= 66.3%). PCL resection without substituting the PCL with a posterior stabilised prosthesis showed no difference compared to PCL retention (p=0.31, I(2) = 83.2%). On clinical scores, only Hospital for Special Surgery score revealed a significant difference of 1.6 points (p=0.03, 95% confidence interval [-3.1, -0.1]) between PCL retention versus PCL sacrifice and substitution combined favouring the latter group. The necessary subgroup analyses could not be performed for the clinical scores. AUTHORS' CONCLUSIONS: These results should be interpreted with caution as the methodological quality of the studies was highly variable. We conclude that there is, so far, no solid base for the decision to either retain or sacrifice the PCL with or without use of a posterior stabilized design during total knee arthroplasty. The technique of PCL retention is difficult because the normal configuration and tension need to be reproduced with ligament tensioners. Knowledge of the technique needs to be improved before it can yield superior results compared to the more straightforward techniques of PCL sacrifice or use of a posterior stabilized design. Also, studies evaluating the effect of both techniques should address the right outcome parameters such as range of motion, contact position, and anterior-posterior stability. Suggestions are given to improve future research on this specific topic of knee arthroplasty.

Arthritis, Rheumatoid↗

Insertion of natural intron 6a-6b into a human cDNA-derived gene therapy vector for cystic fibrosis improves plasmid stability and permits facile RNA/DNA discrimination.

BACKGROUND: The gene therapy vector pCMV-CFTR containing human CFTR cDNA shows high segregational instability during growth in Escherichia coli. METHODS: By host strain screening and optimization of fermentation, satisfactory levels of pCMV-CFTR production were achieved. However, the vector was also vulnerable to structural instability manifested by the appearance during fermentation of a more stable mutant form in which the bacterial insertion sequence IS1 had transposed into exon 7 of plasmidborne CFTR. The instability of pCMV-CFTR is attributable to transcription from an upstream cryptic promoter leading to the production of CFTR peptide fragments known to be toxic when expressed in E. coli. To address this, we inserted the 1.1 kb natural human 6a-6b intron into pCMV-CFTR. RESULTS: The new vector pCMV-CFTR-int6ab is more stable in E. coli than either pCMV-CFTR or the IS1 mutant, grows to high cell density giving higher DNA yields and expresses CFTR appropriately in transfected cells. Thus, the intron has a stabilizing effect comparable to the IS1 insertion yet retains full functionality for gene therapy. We describe a PCR assay using primers directed to sequences flanking the intron that allows differentiation between DNA and mature mRNA. The T936C mutation present only in vector DNA has also been exploited to allow transgene CFTR to be distinguished and its dose-dependent expression to be detected in human cellular backgrounds. CONCLUSIONS: Instability of a plasmid vector for gene therapy has been minimized by rational modification. The introduction of an intron for this purpose offers the additional advantage of providing a discriminatory RT-PCR assay.

Amino Acid Sequence↗

Luminescent CdSe/CdS core/shell nanocrystals in dendron boxes: superior chemical, photochemical and thermal stability.

The surface ligands, generation-3 (G3) dendrons, on each semiconductor nanocrystal were globally cross-linked through ring-closing metathesis (RCM). The global cross-linking of the dendron ligands sealed each nanocrystal in a dendron box, which yielded box-nanocrystals. Although the dendron ligands coated CdSe nanocrystals (CdSe dendron-nanocrystals) were already quite stable, the stability of CdSe box-nanocrystals against chemical, photochemical, and thermal treatments were dramatically improved in comparison to that of the original dendron-nanocrystals. Furthermore, the box structure of the ligands monolayer coupled with the stable inorganic CdSe/CdS core/shell nanocrystals resulted in a class of extremely stable nanocrystal/ligands complexes. The band edge photoluminescence of the core/shell dendron-nanocrystals and box-nanocrystals were partially remained, and could be further brightened through controlled chemical oxidation or photooxidation. Practically, the stability of the box-nanocrystals is sufficient for most fundamental studies and technical applications. The box-nanocrystals may represent a general solution for the commonly encountered instability for many types of colloidal nanocrystals. The size distribution of the empty dendron boxes formed by the dissolution of the inorganic nanocrystals in concentrated HCl was very narrow. The empty boxes as new types of polymer capsules are soluble in solution, mesoporous, and with a very thin but stable peripheral. Those nanometer-sized cavities should be of interest for many purposes in the field of solution host-guest chemistry.

Journal Article↗

[Treatment of patients with fractures of the odontoid of the C2 vertebra].

The study was undertaken to improve an approach to treating patients with fractures of the odontoid of the C2 vertebra. Forty-nine patients were admitted to hospital for fractures of the odontoid process on January 14, 1989 to January 3, 2004. Physical, target X-ray study of the cervical spine, computed tomography of the C1-2 vertebrae, magnetic resonance imaging of the neck were performed on admission. Type II odontoid fractures were found in 17 patients, Type III in 32, of them 5 patients were observed to have an axial fracture concurrent with Jefferson's fracture. Forty-one of the 49 patients were operated on. Eight patients underwent rigid external fixation with reposition using a Halo apparatus. In all cases, a Halo apparatus was used before surgery to correct dislocation and to fix the upper cervical part of the vertebral column. According to the type of a fracture, its duration, reducibility of atlas-axial dislocation, the patient's status, the following procedures were performed: 1) posterior combined spondylodesis of the C1-C2 vertebrae (Halifax braces) alone and in combination with transpharyngeal resection of the odontoid process; 2) transdental spiral fixation; 3) occipital spondylodesis; 4) isolated Halo reposition and fixation. In all the cases, the results of treatment were assessed as good. The effect of reposition and stabilization was achieved. The prehospital pain syndrome and neurological deficit virtually entirely regressed. It was concluded that active surgical policy in injuries of the upper cervical spine may yield the optimum clinical and orthopedic results that a differential approach is required to choose a method for decompression and stabilization depending on the type of an upper cervical vertebral injury.

Adolescent↗

[Isolation, purification and stability of a glutamin(asparagin)ase preparation from Pseudomonas boreopolis 526].

A technique for purification of glutamine asparaginase from Pseudomonas boreopolis 526 is described which provides a 37% yield of the enzyme homogeneous according to electrophoresis in polyacrylamide gel in the presence of sodium dodecyl sulfate. The effect of pH, freezing, thawing and lyophilic drying on the stability of glutamine asparaginase was studied. The enzyme is rather stable at pH 4.8 and 4 degrees C. Lyophilic drying of the homogeneous enzyme without addition of stabilizers resulted in a decrease of its activity an is accompanied by formation of protein conglomerates with molecular weights of 280,000 and 660,000 D. Freezing and thawing decreased the activity of the nature enzyme by 40-50%.

Amidohydrolases↗

Constitutive overexpression of cytosolic glutamine synthetase (GS1) gene in transgenic alfalfa demonstrates that GS1 may be regulated at the level of RNA stability and protein turnover.

Glutamine synthetase (GS) catalyzes the ATP-dependent condensation of NH4+ with glutanate to yield glutamine. Gene constructs consisting of the cauliflower mosaic virus (CaMV) 35S promoter driving a cytosolic isoform of GS (GS1) gene have been introduced into alfalfa (Medicago sativa). Although transcripts for the transgene were shown to accumulate to high levels in the leaves, they were undetectable in the nodules. However, significant amounts of beta-glucuronidase activity could be detected in nodules of plants containing the CaMV 35S promoter-beta-glucuronidase gene construct, suggesting that the transcript for the GS1 transgene is not stable in the root nodules. Leaves of alfalfa plants with the CaMV 35S promoter-GS1 gene showed high levels of accumulation of the transcript for the transgene when grown under low-nitrogen conditions and showed a significant drop in the level of GS1 transcripts when fed with high levels of NO3-. However, no increase in GS activity or polypeptide level was detected in the leaves of transgenic plants. The results suggest that GS1 is regulated at the level of RNA stability and protein turnover.

Cytosol↗

Ter(9,9-diarylfluorene)s: highly efficient blue emitter with promising electrochemical and thermal stability.

Novel ter(9,9-diarylfluorene)s were synthesized by a Suzuki-coupling reaction of 2-bromofluorene (1) and 2,7-fluorenediboronic ester derivatives (3) with high isolated yields (63-86%). The X-ray structure analysis of ter(9,9'-spirobifluorene) (4aa) revealed that the conjugated chromophore adopts a helical conformation. This conformation effectively releases the steric interaction between the fluorene moieties and prevents inter-chromophore interactions. The introduction of aryl groups at the C9 position of fluorene was highly beneficial to the thermal and morphological stability of these oligomers. These terfluorenes exhibit intense blue fluorescence with excellent quantum yields both in solution ( approximately 100%) and in solid state (66-90%), and possess interesting reversible redox properties. Highly efficient blue light-emitting OLED devices were fabricated using 4aa and 4cc as emitters as well as hole transporters. The devices exhibit low turn-on voltage ( approximately 3 V) and high EL external quantum efficiency (2.5-3%).

Journal Article↗

Microemulsion and diafiltration approaches: an attempt to maximize the global yield of DNA-loaded nanospheres.

The yield of DNA-loaded nanospheres in its widest definition includes encapsulation efficiency and the integrity of the loaded molecules plus the production yield of fabricated nanospheres. The former aspect could be considerably improved by adopting the microemulsion concept to enhance the stability of the primary emulsion during the preparation of nanospheres by the double emulsion solvent-removal method. The droplet size of the mentioned emulsion was monitored by means of photon electron correlation spectroscopy and could serve as an index for emulsion fineness and stability. DNA stability as a function of applied mechanical stress was monitored by horizontal agarose gel electrophoresis. The impact of the primary emulsion on nanosphere porosity was assessed as well. Regarding the second aspect of the global yield of nanospheres, i.e. production yield, a modified diafiltration technique was adopted for the washing and recovery processes in comparison with the traditional and for the conservation of particle size characteristics of the recovered nanospheres.

1-Butanol↗

Labeling and quality control of 188Re-lanreotide.

Lanreotide was labelled with 188Re obtained from 188W/188Re generator, using stannous ion as reducing agent, ascorbic acid as stabilizers and hydroxy ethylidene bisphosphonate (HEDP) as intermediary ligand at different molar ratios, pH and incubation times. Best yields (>95%) were obtained using molar ratios SnF2/lanreotide, ascorbic/lanreotide and HEDP/lanreotide of 40, 12 and 260, respectively, pH 1-2 with an incubation at 100 degrees C for 30 min. Quality control evaluation and stability of the radiolabel compound was done by the following selected methods: chromatography in Whatman 3 MM with MEK and NaCl 0.15 M as solvents, ITLC-SG with ethanol-HCl 0.01N (90:10); reverse phase extraction cartridge (Sep-pak C18, Waters Associated) and RP-HPLC with radiometric and UV detection (220 nm) using MCH-5 n-capp column with linear gradient from 90% H2O (TFA 0.1%): 10% ACN (TFA 0.1%) up to 10% H2O (TFA 0.1%):90% ACN (TFA 0.1%) in 30 min, at flow 1 ml/min. Biodistribution in normal mice showed that 188Re-lanreotide is excreted mainly through the hepatobiliary system: more than 70% I.D. is present in gallbladder and intestines at 2 hr post injection. The stability of the 188Re-peptide bond by cysteine challenge test at 37 degrees C, during 2 and 24 hr of incubation time, reveals that approximately 300 and 100 molar ratio cys/peptide is required to displace 50% of the 188Re from the complex. In vitro stability of 188Re-lanreotide at room temperature (Rt) was demonstrated during 24 hr Future works must be done in order to investigate its binding capacity to somatostatin receptors.

Animals↗

Thermally-induced delayed fluorescence of photosystem I and II chlorophyll in thermophilic cyanobacterium Synechococcus elongatus.

Stationary delayed fluorescence (DF) of chlorophyll in isolated membrane preparations from thermophilic cyanobacterium Synechococcus elongatus was investigated as a function of temperature. Two peaks at different temperatures were observed. The low-temperature peak (54-60 degrees C) coincided with the main maximum of the thermally-induced delayed fluorescence of chlorophyll in intact cells and PSII-particles with active oxygen-evolving system. The high-temperature peak (78 degrees C) coincided with the minor band of delayed light emitted by intact cells. It was also observed in the delayed fluorescence emission from a PSI-enriched fraction preparation. The intensities of the DF peaks were dependent on the presence of inhibitors, donors and acceptors that cause specific effects on electron transport of the two photosystems. The low-temperature and high-temperature peaks were related to PSII and PSI, respectively. The manifestation of delayed fluorescence from PSI and PSII at different temperatures seems to be a specific property of thermophilic cyanobacteria. The reason for this may be a high thermal stability of the photosystems and the lack of the PSII antenna complex in isolated membranes. Consequently, the relative yield of delayed fluorescence from PSI markedly increases. Thermally-induced fluorescence seen in membranes of cyanobacteria showed a high sensitivity to structural and functional membrane alterations induced by pH changes, different electron transport stabilizing agents or different concentrations of MgCl2.

Chlorophyll↗

Quality control phantom for digital chest radiography.

PURPOSE: To develop and test a chest phantom for routine quality control testing of digital radiography systems. MATERIALS AND METHODS: The phantom was constructed from sheets of copper, aluminum, and acrylic, which were cut and arranged to yield a radiographic projection resembling that of a human thorax. Regional test objects allowed quantitative assessment of optical density, contrast detail, and spatial resolution. Validation tests were performed to assess image stability in a stable imaging environment and sensitivity to changes in image quality when they occur. RESULTS: The phantom yielded consistent pseudoclinical images when used in a routine quality control program and facilitated detection of simulated problems that were induced in imaging system performance. CONCLUSION: The chest phantom enables quantitative, full-system testing of digital radiography system as they are used clinically for chest radiography.

Humans↗

Comparison of scanning electron microscopy, dynamic light scattering and analytical ultracentrifugation for the sizing of poly(butyl cyanoacrylate) nanoparticles.

Nanoparticles represent promising carriers for controlled drug delivery. This work focuses on the size and molecular mass characterization of polyalkylcyanoacrylate nanoparticles formed by anionic emulsion polymerization of butylcyanoacrylate in the presence of poloxamer 188 as a stabilizer. Three different methods were used to determine the size and size distribution of the particle populations: scanning electron microscopy (SEM), dynamic light scattering (DLS), and analytical ultracentrifugation (ANUC). SEM on freeze-dried and Au-shadowed samples showed a relatively narrow distribution of virtually spherical particles with a mean diameter of 167 nm. DLS yielded a monomodal distribution with hydrodynamic diameters around 199 nm (in the absence of additional stabilizer) or 184 nm (in the presence of 1% poloxamer 188). The size distribution determined by ANUC using sedimentation velocity analysis was somewhat more complex, the size of the most abundant particles being around 184 nm. Molar particle mass distributions centered around 2.3x10(9) g/mol. The advantages and disadvantages of the three sizing techniques are discussed.

Enbucrilate↗

[Optimizing cultural conditions of measles vaccine working seed lot with orthogonal experiments].

OBJECTIVE: To improve the quality of measles vaccine and find out good combination of cultural factors of measles vaccine virus seeds. METHODS: Orthogonal experiments were made to confirm optimized cultural factors. 5 lots of optimized measles vaccine working seed lot were prepared. RESULTS: The titers of the optimized vaccine seed improved significantly (> or = 0.69 +/- 0.35 LgCCID50/1.0 ml, P < 0.05), compared with 5 lots of traditional working seed lot. With these seeds we trial-prepared 5 lots of lyophilized vaccines respectively, the titer and stability met the Chinese Requirements of Biological Products. CONCLUSION: The orthogonal experiment is a kind of good method for optimizing cultural conditions of measles vaccine working seed lot. The yield and quality of measles vaccine can be improved in the optimized working seed lot with high titer and good stability.

Culture Media↗