On uniform stability and numerical simulations of complex valued neural networks involving generalized Caputo fractional order.

The dynamics and existence results of generalized Caputo fractional derivatives have been studied by several authors. Uniform stability and equilibrium in fractional-order neural networks with generalized Caputo derivatives in real-valued settings, however, have not been extensively studied. In contrast to earlier studies, we first investigate the uniform stability and equilibrium results for complex-valued neural networks within the framework of a generalized Caputo fractional derivative. We investigate the intermittent behavior of complex-valued neural networks in generalized Caputo fractional-order contexts. Numerical results are supplied to demonstrate the viability and accuracy of the presented results. At the end of the article, a few open questions are posed.

Nonlinear optical studies of heme protein dynamics: implications for proteins as hybrid states of matter.

Protein structure is fundamentally related to function. However, static structures alone are insufficient to understand how a protein works. Dynamics play an equally important role. Given that proteins are highly associated aperiodic systems, it may be expected that protein dynamics would follow glass-like dynamics. However, protein functions occur on time scales orders of magnitude faster than the time scales typically associated with glassy systems. It is becoming clear that the reaction forces driving functions do not sample entirely the large number of configurations available to a protein but are highly directed along an optimized pathway. Could there be any correlation between specific topological features in protein structures and dynamics that leads to strongly correlated atomic displacements in the dynamical response to a perturbation? This review will try to provide an answer by focusing upon recent nonlinear optical studies with the aim of directly observing functionally important protein motions over the entire dynamic range of the protein response function. The specific system chosen is photoinduced dynamics of ligand dissociation at the active site in heme proteins, with myoglobin serving as the simplest model system. The energetics and nuclear motions from the very earliest events involved in bond breaking on the femtosecond time scale all the way out to ligand escape and bimolecular rebinding on the microsecond and millisecond time scale have been mapped out. The picture that is emerging is that the system consists of strongly coupled motions from the very instant the bond breaks at the active site that cascade into low frequency collective modes specific to the protein structure. It is this coupling that imparts the ability of a protein to function on time scales more commensurate with liquids while simultaneously conserving structural integrity akin to solids.

Observation of the cascaded atomic-to-global length scales driving protein motion.

Model studies of the ligand photodissociation process of carboxymyoglobin have been conducted by using amplified few-cycle laser pulses short enough in duration (<10 fs) to capture the phase of the induced nuclear motions. The reaction-driven modes are observed directly in real time and depict the pathway by which energy liberated in the localized reaction site is efficiently channeled to functionally relevant mesoscale motions of the protein.

Effects of non-steroidal anti-inflammatory drugs on the luminol and lucigenin amplified chemiluminescence of human neutrophils.

A panel of non-steroidal anti-inflammatory drugs commonly used for therapeutic purposes was assessed for their effects on the respiratory burst of isolated human polymorphonuclear neutrophils. Cells were stimulated with opsonised yeast and the production of reactive oxygen species was measured by amplified chemiluminescence with luminol and lucigenin which are two luminogenic agents measuring different cellular events. A special attention was devoted to the establishment of dose-effect curves and calculation of ED50. Some of the drugs tested (acemetacine, diclofenac, flufenamic acid and niflumic acid) were able to decrease both luminol and lucigenin chemiluminescence in a dose-dependent manner reflecting an inhibitory effect on the respiratory burst. The most potent derivative was flufenamic acid (ED50 8 and 78 microM, respectively, with luminol and lucigenin), followed by diclofenac (21 and 98 microM), niflumic acid (97 and 227 microM) and acemetacine (585 and 427 microM). In contrast, several other drugs (flurbiprofen, ibuprofen, ketoprofen, piroxicam) stimulated both luminol and lucigenin chemiluminescence, suggesting a pro-oxidant activity. Acetylsalicylic acid (up to 1250 microM) was a modest inhibitor (maximum 25% inhibition) showing no dose-dependent effect and tolmetin (up to 125 microM) had no significant effect in both systems. The results were in agreement using both luminogenic agents, except for indomethacin, naproxen and tenoxicam which showed different kinds of effects. The unspecific and complex nature of the measurement systems used did not allow to give a complete mechanistic interpretation of the results, but the comparison with literature data gave some pertinent explanations for both anti- and pro-oxidant effects.

Scientific and ethical issues of preimplantation diagnosis.

Preimplantation diagnosis (PID) offers couples at high risk of having offspring affected with a genetic disorder the possibility of an early prenatal diagnosis. For many couples this approach will give the opportunity to avoid a selective termination of affected pregnancies. Substantial advances were made in PID since the report, in 1990, of the first birth obtained after PID. Yet, many technical hazards have to be solved for PID to become a standard clinical tool. The very close correlation existing between the forthcoming developments in the fields of PID and human genome mapping will improve the reliability and efficiency of genetic diagnosis. In the near future, the procedure may also become easier and safer. As a consequence, the indications for PID could be extended to other genetic defects, such as multifactorial diseases. They could also be extended to cases with no medical background, such as social gender selection or behavioural traits. In this perspective, it is now time for both the medical and scientific communities to identify the ethical issues related to these potential new indications.

Dose-response relationship to inhaled endotoxin in normal subjects.

Exposure to endotoxin and to its purified derivative lipopolysaccharide (LPS) is related to several occupational pulmonary diseases and to severe domestic asthma. An inhalation of a given dose of pure LPS produces both a systemic and a bronchial inflammatory response. Information on the dose-response relationship to inhaled LPS in normal subjects is a prerequisite to define the safety threshold of exposure. In the present study, the clinical and inflammatory responses to rising doses of inhaled LPS was evaluated. Nine normal volunteers were challenged weekly by inhalation with saline, 0.5, 5, and 50 microg LPS (Escherichia coli). The response determinators are the clinical symptoms, fever, FEV1, blood polymorphonuclear neutrophils (PMNs) with their level of activation (measured by luminol enhanced-chemiluminescence), and both the blood and the urine concentrations of the C-reactive protein (CRP). To assess the bronchial inflammatory response, an induced sputum was obtained 6 h after each dose of LPS, and the total and differential cell counts as well as the MPO, ECP, and TNF-alpha concentrations were measured. Compared with the saline, an inhalation of 0.5 microg LPS induces a significant decrease in the PMN luminol-enhanced chemiluminescence (p < 0.01), which could reflect a process of margination and/or extravascular sequestration of activated PMN. Inhalation of 5 microg LPS is associated with a significant rise in blood CRP (p < 0.01) and PMNs (p < 0.001) and in sputum PMNs (p < 0.05), monocytes (p < 0.05), and MPO (p < 0.05). Inhalation of 50 microg LPS was characterized by a significant increase in temperature (p < 0.01), blood PMNs (p < 0.001), blood and urine CRP (p < 0.01 and < 0.01), and sputum PMNs (p < 0.001), monocytes (p < 0.05), lymphocytes (p < 0.05), MPO (p < 0.01), TNF-alpha (p < 0.01), and ECP (p < 0.01) while five subjects develop symptoms. In normal subjects, the response to inhaled LPS is dose-related, the most sensitive markers of LPS-induced inflammation being the blood PMNs count with their level of activation, the blood CRP concentration, and the sputum PMNs count. The no-response threshold to an acute inhalation of LPS is less than 0.5 microg.

Effect of the mucoactive drug nacystelyn on the respiratory burst of human blood polymorphonuclear neutrophils.

In lung diseases such as chronic obstructive pulmonary disease (COPD) or cystic fibrosis, the activation of phagocytic cells produces high amounts of cytotoxic reactive oxygen species (ROS) that are partly implicated in the pathogenic process. In this study, the ex vivo antioxidant activity of nacystelyn (NAL), a recently developed mucoactive thiol-containing agent, was investigated using the respiratory burst of human blood polymorphonuclear neutrophils (PMNs). The ROS generation was induced by serum-opsonized zymosan and assessed with luminol- and lucigenin-enhanced chemiluminescence (ECL). The activity of NAL was compared with N-acetylcysteine (ACC) and captopril, other thiol-containing pharmacological agents having documented antioxidant properties. The three drugs significantly inhibited the ECL response of activated PMNs in the presence of luminol, a luminogenic agent which mostly reflects the production of hydroxyl and hypohalite radicals. NAL was more efficient than the other two drugs: the concentrations producing a 50% inhibition (IC50) of total luminol-ECL were 290 microM, 1580 microM and 760 microM for NAL, ACC and captopril, respectively. The inhibition of the lucigenin-ECL response of activated PMNs was less marked for all compounds suggesting a poorer reactivity with superoxide radicals. These findings demonstrate that NAL, at concentrations obtainable in vivo by inhalation, impairs the PMNs chemiluminescence response related to hydroxyl and hypohalite radicals production. As those radicals are highly cytotoxic, NAL appears as a promising agent in the prevention of oxidative lung damage caused by an active inflammatory response.

Epitope mapping on intact, heated and reduced molecular variants of human chorionic gonadotrophin.

Monoclonal antibodies (MAbs) raised against purified human chorionic gonadotrophin (hCG) (n = 30) and synthetic peptides derived from hCG (n = 3) were able to recognize by Western blotting several hCG dimers (57-47 and 42 kDa), free beta-subunits (35-32, 26 and 16 kDa) and free alpha-subunit (21 kDa) which coexist in commercially available hCG preparations. According to differences in the immunoreactivity of hCG-related molecular forms observed under native or denaturing conditions such as boiling or reducing hCG samples before or after gel electrophoresis, nine classes of MAbs able to recognize different immunoreactive domains were determined. Three domains corresponded to continuous epitopes recognized by MAbs raised against hCG-related peptides. The six remaining domains, recognized by the other MAbs, contained discontinuous epitopes from which three were surface-oriented and three disguised in the holo-hormone. This solid-phase approach, combining native and denaturing conditions, represents a simple and powerful tool to screen the specificity of MAbs from varying sources and to investigate molecular variants of proteic hormone.

Total amount of circulating human chorionic gonadotrophin alpha and beta subunits in first trimester trisomies 21 and 18.

The aim of this study was to evaluate the variations in the balance between total (free and combined) circulating alpha and beta subunits of human chorionic gonadotrophin (hCG) in trisomy 21 and 18. Maternal serum samples were collected at 10 and 11 weeks of gestation from 22 singleton pregnancies with trisomy 21 (n = 17) and trisomy 18 (n = 5) and 66 chromosomally normal controls, matched for gestational age. The hCG and free alpha and beta subunits circulating levels were measured using specific immunoradiometric assays and converted in a common unit system obtained using calibration of the assays with intact and thermally dissociated purified hCG preparation. In trisomy 21, the only significant difference from controls was in the free beta hCG level which was increased. In trisomy 18, intact hCG, free beta hCG as well as total alpha hCG and total beta hCG levels were significantly lower whereas the free alpha hCG level was significantly higher than in controls. The decrease in total beta hCG was more pronounced than the decrease in total alpha hCG resulting in a significant increase in the total alpha- to beta hCG subunit ratio in trisomy 18. These findings suggest some modifications in the biosynthesis and/or release rates of the hCG subunits in these trisomies.

Linear and non linear competition plots in the deoxyribose assay for determination of rate constants for reaction of nonsteroidal antiinflammatory drugs with hydroxyl radicals.

Performing the deoxyribose (DR) assay for determination of the rate constants for reaction of non steroidal antiinflammatory drugs with hydroxyl radicals led to some unusual competition plots. The molecules from the arylpropionic family of drugs: ibuprofen, flurbiprofen, ketoprofen and naproxen produced the linear relationship. However, acemetacin, diclofenac Na, flufenamic acid, indometacin, indometacin, niflumic acid, tolmetin Na and sulindac presented non linear competition plots manifesting at relatively low drug concentrations. This effect was corrected by increasing DR concentrations from 2.8 mM to 15 mM. The modification did not affect rate constants values for those derivatives which already presented a linear plot at 2.8 mM, but allowed to calculate rate constants for other compounds. It is suggested that the experimental conditions have to be adapted particularly for those derivatives with a relatively high rate constant for reaction with the radical species. The oxicam derivatives (tenoxicam and piroxicam) presented another kind of deviation that revealed a prooxidant effect in this system: non linear plots were also observed at relatively low drug concentrations, but in the opposite direction than for the other molecules. This last effect was independent of DR concentration but could be corrected by increasing ascorbate concentration in the system.

Temporal relationship between the human chorionic gonadotrophin peak and the establishment of intervillous blood flow in early pregnancy.

The purpose of this study was to investigate the temporal relationship between the early pregnancy peak of circulating human chorionic gonadotrophin (HCG) concentration and the establishment of maternal blood flow in the placental intervillous space. The presence of blood flow echoes within intervillous space was determined by colour Doppler imaging from 44 women with clinically uncomplicated pregnancy between 6 and 18 weeks gestation. Circulating HCG, free alpha- and beta HCG subunits, oestradiol and progesterone concentrations were immunoassayed in blood samples collected at the time of Doppler examination. A continuous intervillous blood flow was detected in all cases with a gestational age > or = 11.7 weeks (n = 18) but never before this time. Circulating concentrations of free alpha HCG, oestradiol and progesterone were linearly or exponentially correlated with gestational age (r = 0.860, 0.903 and 0.538 respectively, all with P < 0.001), indicating steady increase of these hormones with advancing gestation. However, the best fitted lines were found to be parabolic for HCG (r = 0.771, P < 0.001) and beta HCG (r = 0.695, P < 0.001), their highest points corresponding to 11.24 and 10.74 weeks gestational age respectively. The close temporal relationship between the Doppler advent of intervillous maternal blood flow and the HCG peak suggests that the establishment of the intervillous blood flow is associated with the decline in circulating HCG concentrations.

Coelomic fluid chorionic gonadotrophin and protein concentrations in normal and complicated first trimester human pregnancies.

Coelomic fluid and maternal serum samples were collected from 43 normal pregnancies and 18 missed abortions between 7 and 12 weeks of gestation. The samples were analysed for the concentrations of intact human chorionic gonadotrophin (HCG), free alpha HCG, free beta HCG and total protein. The relationships between the biological findings and the ultrasound and pathological features were assessed by regression analysis. In normal pregnancies, intact HCG, free alpha HCG and free beta HCG concentrations were respectively 1.3, 185 and 33 times higher in coelomic fluid than in maternal serum. The coelomic concentrations of intact HCG and free alpha HCG decreased significantly with advancing gestation. No relationship was found between coelomic fluid and maternal serum concentrations of the different variables. These findings suggest that in normal pregnancies, the concentration of HCG in the coelomic fluid, as in maternal serum, is mainly influenced by cytotrophoblastic differentiation and that the metabolic clearance of HCG molecules is slower in the coelomic cavity than in maternal serum. In missed abortions, the serum concentrations of intact HCG, alpha HCG and free beta HCG and the coelomic concentration of total protein were significantly lower than in normal pregnancies. In three out of nine anembryonic pregnancies diagnosed by ultrasound, embryonic remnants were present at histological examination. The coelomic concentration of total protein was extremely low in all missed abortions with advanced trophoblastic necrosis whereas the HCG concentration was low when embryonic remnants were absent. These findings support the concept that embryonic and placental development are closely related in the first trimester of human pregnancy, placental biological functions persisting only for a limited period of time after embryonic demise.

Placental production of human chorionic gonadotrophin alpha and beta subunits in early pregnancy as evidenced in fluid from the exocoelomic cavity.

Levels of human chorionic gonadotrophin (hCG) and of its free alpha and beta subunits were measured using specific immunoradiometric assays in exocoelomic fluid (ECF) and maternal serum (MS) collected from five pregnant women at 6.6-8 weeks of gestation. Mean levels of hCG and its free subunits were significantly (P < 0.001) higher in ECF than in MS: 3.5-fold for hCG, 600-fold for free alpha hCG and 38-fold for beta hCG. There was no correlation between either hCG levels or levels of its free subunits in ECF and MS. On a molar basis, the quantity of free alpha hCG subunit expressed as a percentage of the total (free+combined) amount was 83% in ECF and 2.7% in MS (P < 0.001). The amount of free beta hCG subunit as a percentage of the total was 22% in ECF and 3.5% in MS (P < 0.001). The ratio of the total amounts of alpha- and beta hCG subunits amounted to 4.6 in ECF and 0.99 in MS (P < 0.001). The heterogeneity of hCG was further investigated by polyacrylamide gel electrophoresis followed by immunoblotting. Several bands with molecular mass ranging from 42 to 57 kDa, corresponding to hCG dimers, were immunodetected in ECF and MS with anti-alpha hCG and anti-beta hCG monoclonal antibodies. A free 35 kDa beta hCG immunoreactive band was found in ECF and MS. A free alpha hCG immunoreactive band was observed at 23 kDa in ECF and at 21 kDa in MS. These findings suggest that the exocoelomic cavity is a reservoir where hCG and its subunits produced by trophoblast accumulate directly.(ABSTRACT TRUNCATED AT 250 WORDS)

Origin and significance of the heterogeneity of protein hormones.

The heterogeneity of circulating protein hormone is the result of multiple steps including gene expression, mRNA maturation, post-translational processing and peripheral catabolism. As a consequence of these cumulative events, it seems difficult to evaluate the endocrine function by using specific radioimmunoassays for each circulating variant of a protein hormone. Moreover, the discovery of new molecular variants of protein hormones with unknown biological significance complicates the standardization and the clinical interpretation of immunoassays.

Total amounts of circulating human chorionic gonadotrophin alpha and beta subunits can be assessed throughout human pregnancy using immunoradiometric assays calibrated with the unaltered and thermally dissociated heterodimer.

The aim of the present study was to determine the variations in the balance between total (free plus combined) circulating alpha and beta subunits of human chorionic gonadotrophin (hCG) throughout human pregnancy. The equivalence between the International Units (IU) of hCG (IRP 75/537) and those assigned to the alpha (IRP 75/569) and beta (IRP 75/551) free subunits was experimentally determined by using intact and thermally dissociated hCG. Heat exposure (2 min at 100 degrees C) of hCG preparations resulted in a complete dissociation of hCG into free, soluble and intact alpha and beta subunits. The hCG and alpha and beta subunit contents of unaltered and heated hCG preparations were assessed by specific immunoradiometric assays. The amount of immunoreactive subunits dissociated by heat from hCG could then be evaluated on a molar basis. Circulating hCG and its free alpha and beta subunits were immunoassayed in 836 blood samples collected from healthy pregnant women at different gestational ages. After conversion of hCG and its subunits into a common IU system, the gestational profiles of the total amounts (free plus combined) of alpha- and beta hCG subunits increased together and peaked at 9-10 weeks of gestation. Thereafter, total alpha and beta subunits decreased and subsequently remained stable until term. The decline in total alpha hCG subunit was less marked than that of total beta hCG subunit. The alpha- to beta hCG ratio was equimolar until 10 weeks of gestation when it increased almost fourfold until term (P < 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS)

Purification, primary structure and molecular cloning of a rat ribosomal protein showing homology with yeast ribosomal protein YL34.

1. A new 17-kDa mammalian ribosomal protein (PR17) was purified to homogeneity from the rat exocrine pancreas. The purification procedure was based on acidic extraction of a heat-denatured homogenate, ammonium-sulfate precipitation, hydrophobic chromatography on phenyl-Sepharose and analytical reverse-phase HPLC on mu Bondapak C18. Fractions of interest were collected using an antiserum directed against the first (1-14) moiety of somatostatin (1-28). 30 micrograms pure RP17 were obtained from 1 g fresh pancreas. 2. A short 111-b cDNA encoding RP17 was amplified from rat pancreatic first-strand cDNA template by using two 64-fold degenerate heptadecamer primers in the DNA-polymerase-chain reaction. From the sequence of amplified cDNA, an unambiguous oligonucleotide probe was designed to screen a rat pancreatic cDNA library. A cDNA clone coding for RP17 was isolated, whose nucleotide sequence, with an open reading frame coding for 155 amino acids (molecular mass of 17,199 Da), confirmed the partial amino acid sequences directly obtained from the purified protein. 3. Northern-blot analysis showed that a similar 0.75-kb transcript was present in rat pancreas, in the rat pancreatic acinar cell line AR 4-2J and in the human neuroblastoma cell-line NB-OK-1, the highest level being in the latter two preparations, despite similar levels of RP17 in all three preparations, as tested with a rabbit antiserum directed against purified RP17. 4. The N-terminal sequence of both RP17 and the ribosomal protein YL43 from Saccharomyces cerevisiae (39 amino acid residues) showed a high degree of identity (77%), indicating that RP17 is a mammalian homolog of yeast ribosomal protein YL43.