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Application of NIR spectroscopy in polymorphic analysis: study of pseudo-polymorphs stability.

The accelerated transformation of three azithromycin pseudo-polymorphs (viz. the anhydrous, monohydrate (MH), and dihydrate (DH) forms) at a high temperature and moisture level was examined by near infrared spectroscopy (NIRS). The most marked spectral differences between the pseudo-polymorphs occurred in the 1800-2200 nm region, which corresponds to the first overtone for water. The qualitative analysis of the NIR spectra for the pseudo-polymorphs following storage in a stove at 60 degrees C at 100% relative humidity for 60 days suggests that the crystalline forms (viz. the MH and DH) are stable, whereas the amorphous (anhydrous) form evolves to the DH. This was confirmed by determining the amounts of water and DH present in anhydrous azithromycin and the MH by use of partial least-squares regression (PLSR). The method used to quantify the DH in MH samples was developed and validated in accordance with the standards of the International Conference of Harmonization (ICH) and the European Medicines Agency (EMEA) with a view to its subsequent application by the pharmaceutical industry. The limits of detection (LD) and quantitation (LQ) for the DH in MH provided by the NIRS method were consistent with those obtained by X-ray diffraction (XRD) methodology. This testifies to the accuracy of the proposed method.

Azithromycin↗

A kinetic analysis of cell division, and induction and stability of recA protein in U.V. Irradiated ion+ and ion-strains of Escherichia coli K12.

Kinetic analysis of induction of recA protein synthesis after U.V. irradiation does not show correspondence with the kinetics of division inhibition in ion+ and ion- strains. When the induction of recA protein after U.V. is drastically reduced by rifampicin treatment, no effect on the kinetics of division inhibition is observed.

Bacterial Proteins↗

The CellSoft computerized semen analysis system. I. Consistency of measurements and stability of results in relation to sample size analyzed.

The CellSoft TM computer-assisted digital image analysis system was evaluated for reproducibility of measurements and for the sample size needed to obtain stable results for seven parameters characterizing human spermatozoa. The parameters were sperm density, velocity, linearity, percent motile cells, maximum and mean amplitude of sperm head's lateral displacement (ALH max and ALH mean) and beat cross frequency. Thirty men were assigned to three equally large groups according to sperm density, and the groups were studied separately. Consistency of measurements was highly acceptable for all parameters (coefficient of variation ranging from 0.9% to 3.7%). The average minimum number of sample size needed for stable results varied between the groups and within each group dependent upon the parameter studied. Velocity and linearity seemed to require the lowest average cell number, while percentage motile cells and ALH max needed the highest sample size. Our current recommendation of sample sizes is outlined in detail and limitations and future utility of CellSoft is discussed.

Evaluation Studies as Topic↗

Stabilization of pericardial tissue by dye-mediated photooxidation.

Bovine pericardial tissue was stabilized through a dye-mediated photooxidation reaction. Shrink temperature analysis of the stabilized tissue indicated a material with similar properties to untreated pericardial tissue and unlike identical tissue treated with glutaraldehyde. Photooxidized tissue was resistant to extraction when compared with untreated tissue or control tissues treated in the absence of light or dye. Photooxidized tissue was also resistant to enzymatic digestion by pepsin and to chemical digestion by cyanogen bromide (CNBr). In contrast, untreated or control treated tissues were readily digested by these reagents. Reduction of photooxidized tissue with beta-mercaptoethanol prior to CNBr digestion partially restored susceptibility of the tissue to CNBr digestion, indicating the photooxidation of methionine residues. Soluble collagen derived from bovine pericardium was used as a model compound for the photooxidation reaction. Polyacrylamide gel electrophoresis analysis indicated the photooxidative conversion of collagen into higher molecular weight aggregates consistent with intermolecular crosslink formation. Photooxidized tissue was stable to in vivo degradation when compared with control tissue. Results presented here indicate a crosslinked pericardial tissue produced by dye-mediated photooxidation possessing properties of chemical stability, enzymatic stability, in vivo stability, and biomechanical integrity suitable for use as a biomaterial.

Animals↗

Comparative fourier transform infrared and circular dichroism spectroscopic analysis of alpha1-proteinase inhibitor and ovalbumin in aqueous solution.

Alpha1-proteinase inhibitor (alpha1Pi) and ovalbumin are both members of the serpin superfamily. They share about a 30% sequence identity and exhibit great similarity in their three-dimensional structures. However, no apparent functional relationship has been found between the two proteins. Unlike alpha1Pi, ovalbumin shows no inhibitory effect to serine proteases. To see whether or not a conformational factor(s) may contribute to the functional difference, we carried out comparative analysis of the two proteins' secondary structure, thermal stability, and H-D exchange using FT-IR and CD spectroscopy. FT-IR analysis reveals significant differences in the amide I spectral patterns of the two proteins. Upon thermal denaturation, both proteins exhibit a strong low-wavenumber beta-sheet band at 1624 cm(-1) and a weak high-wavenumber beta-sheet band at 1694 cm(-1), indicative of intermolecular aggregate formation. However, the midpoint of the thermal-induced transition of alpha1Pi (approximately 55 degrees C) is 18 degrees C lower than that of ovalbumin (approximately 73 degrees C). The thermal stability analysis provides new insight into the structural changes associated with denaturation. The result of H-D exchange explains some puzzling spectral differences between the two proteins in D2O reported previously.

Circular Dichroism↗

Beta-turn propensities as paradigms for the analysis of structural motifs to engineer protein stability.

The thermodynamic stability of a protein provides an experimental metric for the relationship of protein sequence and native structure. We have investigated an approach based on an analysis of the structural database for stability engineering of an immunoglobulin variable domain. The most frequently occurring residues in specific positions of beta-turn motifs were predicted to increase the folding stability of mutants that were constructed by site-directed mutagenesis. Even in positions in which different residues are conserved in immunoglobulin sequences, the predictions were confirmed. Frequently, mutants with increased beta-turn propensities display increased folding cooperativities, suggesting pronounced effects on the unfolded state independent of the expected effect on conformational entropy. We conclude that structural motifs with predominantly local interactions can serve as templates with which patterns of sequence preferences can be extracted from the database of protein structures. Such preferences can predict the stability effects of mutations for protein engineering and design.

Amino Acid Sequence↗

Highly potent inhibitors of TNF-alpha production. Part II: metabolic stabilization of a newly found chemical lead and conformational analysis of an active diastereoisomer.

Design and synthesis of metabolically stabilized inhibitors of TNF-alpha production, which could be new drug candidates, are reported. Conformational analysis of an active diastereoisomer was performed based on biological evaluations of the conformationally fixed indane derivatives 17 and 18. Structure-activity relationships (SARs) based on biological evaluations of the optically active derivatives are also discussed. Full details including chemistry are reported.

Animals↗

Parameters affecting plasmid stability in Bacillus subtilis.

For the analysis of parameters affecting plasmid stability in Bacillus subtilis, we used a pUB 110-derived shuttle plasmid containing direct and inverted nucleotide repeats (DRs and IRs). Deletions of up to 6 kb were found to occur between DRs of 7 to 16 bp. IRs as small as 43 or 58 bp were shown to stimulate the formation of these deletions in their neighbourhood. However, these structural features (DRs and IRs) per se were not responsible for plasmid instability. The unstable recombinant plasmids, but not their deletion-carrying (delta) derivatives, were found to impair the growth of the host and to accumulate high amounts of linear plasmid multimers [high mol. wt. (hmw) DNA]. We propose that the accumulation of hmw DNA may be the major reason for the selective pressure against recombinant plasmids, and the enrichment of delta-plasmids. Host mutations and other parameters increasing the stability of recombinant plasmids in B. subtilis are described.

Bacillus subtilis↗

A demographic-economic explanation of political stability: Mauritius as a microcosm.

"This paper examines current models of economic and political development--social modernization theory, political and economic characteristics of stable regimes, and cross country analysis of political stability--and tests them on the Indian Ocean Island of Mauritius. The analysis continues with a causal explanation for political stability in Mauritius' recent history, derived from an examination of economic policies and demographic patterns. Political change in Mauritius over the past sixty years seems to be explained best by a model for political stability which integrates specific economic and demographic factors. The model, applicable to development in other third world nations, revises Malthus' conclusion that population and economic conditions move in an oscillatory relationship and replaces it with a more comprehensive theory, suggesting that political stability is a function of both economic development and a repeating cyclical relationship between economics and population."

Africa↗

Online Social Anxiety in the Digital Age: Transitions, Predictors, and Mental Health Associations in Emerging Adulthood.

BACKGROUND: Online social anxiety (OSA), a multidimensional form of social evaluative anxiety in online social contexts, disproportionately affects emerging adults who constitute the largest active group of media users and face heightened psychological sensitivity due to growing pressures and immature sociocognitive regulation during the transition to adulthood. However, its heterogeneity, transitions, and longitudinal associations with mental health outcomes remain underexplored. METHODS: This study utilized data from two waves of a three-wave longitudinal survey, with 849 Chinese participants (Meanage = 21.6 years; 50.4 percent female) assessed at 4-month intervals. Individuals were classified using latent profile analysis and the stability and changes of profiles were assessed via latent transition analysis (LTA). Multinomial logistic regressions were conducted separately at baseline and follow-up to identify correlates of profile membership. Predictors of profile transitions were examined using manual three-step LTA models, and associations between latent transition patterns and follow-up mental health outcomes were examined using BCH-LTA distal outcome analyses controlling for the corresponding baseline symptom level. RESULTS: Four profiles of OSA were identified: low, privacy-sensitive, moderate-high, and high OSA. Extreme profiles (low/high OSA) showed high stability (80.4 percent and 79.1 percent), while privacy-sensitive OSA exhibited the lowest stability (55.1 percent). Profile memberships were influenced by social-cognitive biases and digital interaction, particularly fear of negative evaluation and online interpersonal trust, whereas profile transitions were mainly associated with anxiety. Transitions toward less severe OSA profiles were generally associated with better subsequent mental health, whereas transitions toward more severe profiles corresponded to poorer outcomes, particularly for offline social anxiety. CONCLUSION: OSA was heterogeneous in its manifestation, severity and transitions. Personalized and early interventions targeting profile-specific vulnerabilities are critical to prevent the worsening of OSA and mitigate its psychological burden.

Humans↗

Lumbosacral stability of consolidated anteroposterior fusion after instrumentation removal determined by roentgen stereophotogrammetric analysis and direct surgical exploration.

STUDY DESIGN: The intervertebral stability of bony consolidated anteroposterior lumbosacral spondylodesis is evaluated by roentgen stereophotogrammetric analysis and direct surgical exploration before and after removal of the internal fixator. OBJECTIVES: To determine the remaining in vivo stability of spinal arthrodesis solely retained by a bony integrated carbon fiber cage. SUMMARY OF BACKGROUND DATA: Roentgen stereophotogrammetric analysis studies on posterolateral lumbar fusions demonstrate primary spinal stability after additional dorsal instrumentation, which is retained during bony fusion healing. Animal models show a persistent stabilizing effect of the fixator despite the presence of bony fusion. Although direct surgical inspection is the most reliable method to evaluate fused vertebrae, roentgen stereophotogrammetric analysis has also proven to be a highly accurate method to evaluate spinal stability. METHODS: In 10 patients lumbosacral fusion was performed using carbon interbody implants and an internal fixator. Ten months after initial surgery (range 7-15 months) the internal fixation was removed to reduce local soft tissue impingement as soon as bony fusion was achieved. Fusion site exploration in the course of instrumentation removal was performed by applying distraction, compression, and torque to the grafted area under fluoroscopic control. Any motion indicated a pseudarthrosis. Lumbosacral stability was evaluated by serial roentgen stereophotogrammetric analysis after fusion and after instrumentation removal. RESULTS: During instrumentation removal the mechanical stress test under fluoroscopic control did not indicate pseudarthrosis. After instrumentation removal, roentgen stereophotogrammetric analysis measurements revealed a nonsignificant increase in lumbosacral micromotions within the fused segment with 0.14, 0.31, and 0.44 mm in the transverse, vertical, and sagittal axes, respectively. CONCLUSIONS: The internal fixator could be removed without endangering the stability of the fusion. Direct surgical exploration confirmed the adequacy of roentgen stereophotogrammetric analysis as a reliable in vivo method to evaluate lumbosacral stability after anteroposterior fusion.

Adult↗

Glutamine stability in biological tissues evaluated by fluorometric analysis.

Although glutamine has been considered unstable during storage and therefore difficult to quantitate, recent results suggest this amino acid is stable at low pH ranges. We evaluated the stability of glutamine in plasma and tissue extracts, using fluorometric analysis. The measured concentration of glutamine detected varied linearly up to 0.8 mmol/L for the aqueous solution (r2 = 98.7, P = 0.0001) with a mean (+/- SD) coefficient of variation of 2.41% +/- 0.79%. When glutamine was dissolved in 50 g/L trichloroacetic acid (TCA), the values were essentially unaltered. Glutamine in an aqueous solution and stored at -70 degrees C was stable for at least 16 days; glutamine in TCA was stable for 6-8 days, then decreased to a concentration significantly lower than that of the aqueous solution. The expected and observed concentrations in plasma were equal (r2 = 0.99975) for increasing amounts of added glutamine. Glutamine concentrations in plasma were stable for > 1 year when stored at -70 degrees C. The glutamine of a transplantable rat sarcoma and a normal rat liver could be extracted with 50 g/L TCA with high efficiency (88.6% +/- 1.9% and 90.2% +/- 0.04%, respectively); the extracted glutamine is stable in TCA for at least 7 days without neutralization when stored at -70 degrees C. Fluorometric analysis of glutamine required only a small quantity of plasma (25 microL) or tissue (200 mg) and is a convenient method for quantifying this important amino acid.

Drug Stability↗

Dynamic flight stability in the desert locust Schistocerca gregaria.

Here we provide the first formal quantitative analysis of dynamic stability in a flying animal. By measuring the longitudinal static stability derivatives and mass distribution of desert locusts Schistocerca gregaria, we find that their static stability and static control responses are insufficient to provide asymptotic longitudinal dynamic stability unless they are sensitive to pitch attitude (measured with respect to an inertial or earth-fixed frame) as well as aerodynamic incidence (measured relative to the direction of flight). We find no evidence for a 'constant-lift reaction', previously supposed to keep lift production constant over a range of body angles, and show that such a reaction would be inconsequential because locusts can potentially correct for pitch disturbances within a single wingbeat. The static stability derivatives identify three natural longitudinal modes of motion: one stable subsidence mode, one unstable divergence mode, and one stable oscillatory mode (which is present with or without pitch attitude control). The latter is identified with the short period mode of aircraft, and shown to consist of rapid pitch oscillations with negligible changes in forward speed. The frequency of the short period mode (approx. 10 Hz) is only half the wingbeat frequency (approx. 22 Hz), so the mode would become coupled with the flapping cycle without adequate damping. Pitch rate damping is shown to be highly effective for this purpose - especially at the small scales associated with insect flight - and may be essential in stabilising locust flight. Although having a short period mode frequency close to the wingbeat frequency risks coupling, it is essential for control inputs made at the level of a single wingbeat to be effective. This is identified as a general constraint on flight control in flying animals.

Animals↗

Long-term stability of food patterns identified by use of factor analysis among Swedish women.

Limited data exist on the reproducibility of food patterns measured by factor analysis as well as on the use of stability of patterns over time. Our primary objective was to explore the long-term stability of food patterns derived by confirmatory factor analysis among 33,840 women participating in the Swedish Mammography Cohort. Our secondary objective was to compare factor solutions from confirmatory factor analysis with those derived by use of exploratory factor analysis. Diet was assessed by a food frequency questionnaire in 1987 and 1997, and food patterns were derived by the use of exploratory and confirmatory factor analysis. We observed four major food patterns (Healthy, Western/Swedish, Alcohol, and Sweets) at both time points. Correlations between confirmed food patterns in 1987 and 1997 were 0.37 for the Healthy pattern, 0.27 for the Western/Swedish pattern, 0.54 for the Alcohol pattern, and 0.44 for the Sweets pattern (P < 0.0001 for all associations). Patterns derived by the use of exploratory factor analysis were strongly associated with those derived by the use of confirmatory factor analysis (r >/= 0.90, P < 0.0001, for all associations). The patterns derived in this study were similar to those derived in other studies, indicating reproducibility of food patterns across populations. Our study suggests that food patterns measured by the use of confirmatory factor analysis are reproducible over time, and weaker correlations observed may reflect natural changes in eating behavior, the food supply, and/or perceptions of what is considered healthy. Testing whether patterns measured over a long time period can be used in analytic studies is the next step in assessing the validity of this method.

Alcohol Drinking↗

Sequences of B-chain/domain 1-10/1-9 of insulin and insulin-like growth factor 1 determine their different folding behavior.

Although insulin and insulin-like growth factor-1 (IGF-1) belong to one family, insulin folds into one thermodynamically stable structure, while IGF-1-folds into two thermodynamically stable structures (native and swap forms). We have demonstrated previously that the bifurcating folding behavior of IGF-1 is mainly controlled by its B-domain. To further elucidate which parts of the sequences determine their different folding behavior, by exchanging the N-terminal sequences of mini-IGF-1 and recombinant porcine insulin precursor (PIP), we prepared four peptide models: [1-9]PIP, [1-10]mini-IGF-1, [1-4]PIP, and [1-5]mini-IGF-1 by means of protein engineering, and their disulfide rearrangement, V8 digestion, circular dichroic spectra, disulfide stability, and in vitro refolding were investigated. Among them only [1-9]PIP, like mini-IGF-1/IGF-1, was expressed in yeast as two isomers: isomer 1 (corresponding to swap IGF-1) and isomer 2 (corresponding to native IGF-1), which are supported by the experimental results of disulfide rearrangements, peptide mapping of V8 endoprotenase digests, circular dichroic analysis, in vitro refolding, and disulfide stability analysis. The other peptide models, [1-10]mini-IGF-1, [1-4]PIP, and [1-5]mini-IGF-1, fold into one stable structure as PIP does, which indicates that sequence 1-4 of mini-IGF-1 is important for the folding behavior of mini-IGF-1/IGF-1 but not sufficient to lead to a bifurcating folding. The results demonstrated that the folding information, by which mini-IGF-1/IGF-1-folds into two thermodynamically structures, is encoded/written in its sequence 1-9, while sequences 1-10 of B chain in insulin/PIP play an important role in the guide of its unique disulfide pairing during the folding process.

Amino Acid Sequence↗

[Sex as a factor in drug consumption].

Based upon the hypothesis on the diverse dependency of drug consumption on the sex of population as its influencing factor, this relationship was closely studied. The dynamics of the dependency of large urban population has been analysed for a period of 4 years. Important differences in the dependency on sex and age have been found. The fine structure of this dependency including the analysis of its stability have been determined. The experiment was based on mathematical analysis methods.

Adolescent↗

[The latest development of the research on Chinese medicine by molecular vibrational spectroscopy].

The latest development of the molecular vibrational spectroscopy (Fourier Transform Raman Spectroscopy and Fourier Transform Infraed Spectroscopy) and the latest application of the non-destructive identification, quantitative analysis, thermal-stability monitoring and Chinese medicine optimizing were described in this paper. Some Chinese crude herbs can be directly and quickly identified by Fourier Transform Raman spectroscopy (FT - Raman) and Diffused Reflection Fourier Transform Infrared Spectroscopy (DR-FTIR). The content of Chinese herbs, medicine and medicament can be quantified without destruction by Diffused Reflection Fourier Transform and Near Infrared Spectroscopy (DR-NIR-FTIR). The mechanism of interaction between the Chinese medicine and some cells or bacteria can be in situ monitored by FT-Raman and HATR-FTIR. Best medicine could be selected and medicament could be optimized according to the injury degree of the cell or bacteria. The decomposing and degenerating process of medicines can also be timely monitored by FT-Raman and the temperature controlled FTIR. These are the latest use of molecular spectroscopy in Chinese medicine. The fast indentification, quantitative analysis monitoring, thermal stability predicting and the optimizing of Chinese herbs, medicine and medicament can be guided by the above mentioned methods. It is proved that these methods are nondestructive to samples, fast accurate, and easy to use.

Drug Stability↗

Solid-state stability studies of 13-cis-retinoic acid and all-trans-retinoic acid using microcalorimetry and HPLC analysis.

The solid-state stabilities of 13-cis-retinoic acid and all-trans-retinoic acid in the presence and absence of oxygen were investigated. The samples were first evaluated using microcalorimetry. The rate laws of different samples under different conditions were deduced from the shapes of the heat flow curves, and the activation energies of the reactions were determined from Arrhenius plots. Under an air atmosphere, the decomposition of 13-cis-retinoic acid is an autocatalytic reaction, while all-trans-retinoic acid undergoes a zero-order process. The degradation of the two compounds at a selected elevated temperature was also determined utilizing HPLC analysis. This technique confirmed the decomposition kinetics. Hence, their half-lives and shelf lives at room temperature could be calculated. Under a nitrogen atmosphere, the microcalorimetric experiment showed a first-order phenomenon for both samples, but HPLC analysis showed no degradation, suggesting that the two samples, in the absence of oxygen, undergo only a physical change.

Air↗