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Surface-engineered bacterial cellulose as template for crystallization of calcium phosphate.

Bacterial cellulose (BC), produced by Acetobacter xylinum, and cotton linters as reference were surface modified by ozone-induced graft polymerization of acrylic acid and used as a template for crystallization of calcium phosphate. The grafting was verified using attenuated total reflection-infrared radiation (ATR-IR) and electron spectroscopy for chemical analysis (ESCA). ATR-IR revealed an additional absorption band at 1700 cm(-1), corresponding to the carbonyl group in polyacrylic acid. ESCA figures show, apart from the characteristic peaks for cellulose, additional peaks at 285 eV and 289 eV that correspond to groups in acrylic acid. The grafting yield is higher on cotton linters compared with BC, which most likely has to do with differences in crystallinity and reactivity of the different cellulose materials. No morphology difference directly caused by grafting could be seen with scanning electron microscopy (SEM), which might indicate that acrylic acid was grafted as a thin film on the surface of the cellulose micro fibrils. Calcium phosphate was formed on the surface-modified cellulose by first pre-soaking the materials in a saturated Ca(OH)2 and later in simulated body fluid (SBF). The atomic ratio of calcium phosphate was determined by ESCA to be about 1.5 for the different materials. Energy dispersive spectroscopy (EDS) was used to map and verify that the crystals were calcium phosphate. Secondary ion mass spectroscopy (SIMS) was also used to verify the presence of calcium phosphate complex onto BC. SEM images showed the difference in dimension, distribution and morphology of the crystals depending on the materials. Smaller and a greater number of crystals were obtained on the surface-modified BC and larger and fewer crystals on surface-modified cotton linters. Structural and grafting differences between the celluloses may lead to differences in nucleation sites and possibly differences in the morphology of the Ca-P crystals. The BC-calcium phosphate composite is expected to be useful as a scaffold for bone tissue regeneration.

Biocompatible Materials↗

Preparation and surface properties of PEO-sulfonate grafted polyurethanes for enhanced blood compatibility.

In order to improve the thromboresistance of the commercial polyurethane(PU), its surface modification was accomplished by three new different methods and their surface characteristics were investigated using ATR-FTIR, ESCA, SEM, and dynamic contact angle measurements. Sulfonations using propane sultone were performed directly onto PU or onto hydrophobic dodecanediol (DDO) grafted PU or onto hydrophilic poly(ethyleneoxide) (PEO) grafted PU. ESCA data coincided well with ATR-IR results, as more 0 at. % for PEO grafted PUs and the presence of S for the sulfonated PUs were revealed. At SEM observation the surfaces of PU-DDO and PU-PEO were relatively smooth, whereas all the sulfonated PU surfaces showed excellent smoothness and homogeneity. The hydrophilicity of the surfaces was considerably increased after PEO grafting or sulfonation. In addition, all the sulfonated PU surfaces, particularly PU-PEO-SO3, which has further hydrophilicity, exhibited complete wetting behavior due to the negatively charged SO3 groups.

Alkanesulfonates↗

Antithrombogenicity of hydrophilic polyurethane-hydrophobic polystyrene IPNs. I. Synthesis and characterization.

A series of interpenetrating polymer networks (IPNs) composed of hydrophilic polyurethane (PU) and hydrophobic polystyrene (PS) were prepared by the simultaneous polymerization method. The PU network was synthesized via the isocyanate-terminated PU prepolymer based on polyethylene glycol (PEG), a highly hydrophilic oligomer, and hexamethylene diisocyanate (HDI). The bulk and surface characteristics of these materials were analyzed by differential scanning calorimetry (DSC), tensile testing, scanning electron microscopy (SEM), attenuated total reflectance-Fourier transform infrared (ATR-FTIR), electron spectroscopy for chemical analysis (ESCA), and contact angle measurement. The PU/PS IPNs prepared in this study exhibited phase separated structures, which had dispersed PS domains in the continuous PU matrix, in both the bulk and surface showing two transition temperatures. The IPN containing 50 wt% of PS showed good mechanical properties. The enrichment of PU phase in the surface was revealed by SEM, ATR-FTIR, ESCA, and contact angle measurement.

Biocompatible Materials↗

Interactions of plasma proteins with a novel polysaccharide surfactant physisorbed to polyethylene.

A polysaccharide surfactant, dextran-[1,6 bis(2-hydroxypropyl-1-amine)hexane]-dextran, (D-H-D) was prepared by reacting dextran (Mw = 8200) with epichlorohydrin followed by reaction with 1,6-hexanediamine. The D-H-D polymer product was characterized by gel permeation chromatography (GPC), and 13C-nuclear magnetic resonance spectroscopy (13C-NMR). D-H-D was physisorbed on polyethylene (PE) from aqueous solution, and the adhesion stability and resistance to protein adsorption was examined under static and dynamic flow conditions, using a modified rotating disk system. Modified surfaces were characterized by attenuated total reflectance Fourier transformed infrared spectroscopy (ATR-FTIR), electron spectroscopy for chemical analysis (ESCA) and by water contact angles. Under applied shear stresses of up to 73 dyn cm-2, the adhesion of D-H-D on PE was sufficient to inhibit desorption by water (> 90% D-H-D on PE was retained) and 5% SDS surfactant solution (approximately 83% D-H-D retained), as determined by ATR-FTIR. Under similar shear stress conditions, albumin adsorption on D-H-D modified PE was reduced by over 90%, and protein adsorption from fresh human plasma was reduced by approximately 70% compared with unmodified PE. The results are discussed in terms of interfacial forces, and the suitability of this approach for studying protein-surface interactions and for developing a novel class of protein-resistant biomaterials.

Adsorption↗

The pattern of astigmatism in a Canadian preschool population.

PURPOSE: To measure magnitude, type, and central tendency of astigmatism found in a county-wide population of Canadian preschool children (mean age, 48.1 months). METHODS: Noncycloplegic autorefractive measures were taken in 1179 children attending a preschool health fair operated by their county board of health. Spherocylinder measures were transformed into three independent components. RESULTS: The equivalent sphere showed considerable variation between retinoscopy and autorefraction that was attributed to the variable overaccommodation induced by the autorefractor. Astigmatic components were not affected. Small discrepancies between the two techniques were similar to those in adults and were not of sufficient magnitude to affect validity. With-the-rule (WTR) astigmatism of at least 0.25 D was the most frequent form (45%) followed by against-the-rule (ATR; 40%) and oblique (15%). The 95th percentile for cylinder magnitude was found at 1.25 D. Astigmatisms beyond this value were predominately WTR. The mean (negative) cylinder magnitude was 0.08 Dx 015 degrees. CONCLUSIONS: When spherocylinder values are transformed into a mathematical continuum rather than WTR and ATR classifications, the true central tendency of the population is better defined and is close to zero. Astigmatisms of more than 1.25 D in the preschool child exceed the 95th percentile in this population and were more frequently WTR.

Adult↗

Prevalence of refractive errors in a rural South Indian population.

PURPOSE: To report the prevalence of refractive errors in a rural south Indian population. METHODS: Four thousand eight hundred subjects (age, >39 years) from rural south India were enumerated for a population-based study. All participants underwent complete ophthalmic evaluation. Subjects who were phakic in the right eye with best corrected visual acuity of 20/40 or better were included for analysis. Association of refractive errors with age, sex, cataract, and diabetes mellitus were analyzed. RESULTS: Of the 3924 responders, 2508 were eligible. The unadjusted prevalence of emmetropia (spherical equivalent [SE], -0.50 to +0.50 diopter sphere [DS]), myopia (SE < -0.50 DS), high myopia (SE < -5.00 DS), and hyperopia (SE > 0.50 DS) were 50.60%, 26.99%, 3.71%, and 18.70% and age and gender adjusted for the rural Tamil Nadu population were 46.77%, 30.97%, 4.32%, and 17.94%, respectively. The prevalence of emmetropia decreased significantly with age (P < 0.0001), and the prevalence of myopia and high myopia increased significantly with age (P < 0.001) and were significantly associated with nuclear sclerosis (P < 0.001). The prevalence of hyperopia increased until 60 years of age and then decreased. Hyperopia was more common among women than men (P < 0.001) and was negatively associated with nuclear sclerosis (P < 0.001) and positively with diabetes mellitus (P = 0.008). Of the participants with astigmatism (cylindrical error greater than 0.50 DC), 9.80% had with-the-rule (WTR) and 77.44% against-the-rule (ATR) astigmatism. The prevalence of WTR and ATR astigmatism significantly decreased (P < 0.001) and increased (P = 0.006) with age, respectively. CONCLUSIONS: The pattern of refractive errors in this rural south Indian population is similar to those reported in other tropical regions of the world.

Adult↗

Reduced platelet adhesion on the surface of polyurethane bearing structure of sulfobetaine.

Poly(etherurethane)s are widely used as blood-contacting biomaterials due to their good biocompatibility and mechanical properties. Nevertheless, their blood compatibility is still not adequate for the more demanding applications. Surface modification is an effective way to improve the blood compatibility and retain the bulk properties of biomaterials. The purpose of present study was to design and synthesize a novel nonthrombogenic biomaterial by modifying the surface of poly(etherurethane) with zwitterionic monomer. Films of polyurethane were grafted with sulfobetaine by a three-step procedure. In the first step, the film surfaces were treated with hexamethylene diisocyanate (HDI) in toluene at 50 degrees C in the presence of di-n-butyl tin dilaurate (DBTDL) as a catalyst. The extent of the reaction was measured by ATR-IR spectra; a maximum number of free NCO group was obtained after a reaction time of 90 min. In the second step, the hydroxyl group of 4-dimethylamino-1-butanol (DMAB) was allowed to react in toluene with isocyanate groups bound on the surface. In the third step, sulfobetaine was formed on the surface through the ring-opening reaction between tertiary amine of DMAB and 1,3- propane-sultone (PS). It was characterized by ATR-IR, XPS. The data showed that the grafted surfaces were composed of sulfobetaine. The results of the contact angle measurements showed that they were strongly hydrophilic. The state of platelet adhesion and shape variation for the attached platelets was described. The modified surface shows excellent blood compatibility feature by the low platelet adhesion.

Betaine↗

In vivo and in vitro stability of modified poly(urethaneurea) blood sacs.

In the present study, we investigate the in vivo and in vitro stability of modified poly(urethaneurea) (BioSpan MS/0.4) blood sacs. Blood sacs were utilized primarily in left ventricular assist devices that were implanted in calves for times ranging from 5 to 160 days. Cyclic testing in vitro was also conducted on similar sacs. Various analytical methods were employed to characterize the sacs after in vivo or in vitro service and corresponding retained "control" sacs. These methods included ATR-FTIR spectroscopy, scanning electron microscopy and gel permeation chromatography. In general, the characteristics of implanted and in vitro cycled sacs were similar to their control sacs. Thermal and microtensile properties were unchanged after testing. The same was true for the ATR-FTIR spectra, indicating relative chemical stability for the time frames explored here. The only significant changes occurred in molecular weight and gross surface morphology. A modest increase in weight average molecular weight was observed for most implanted blood sacs, indicating some type of chain extension or branching reaction in vivo. Although the surface morphologies of implanted blood sacs were often similar to their control sacs, we sometimes observed limited pitting on the nonblood contacting surfaces in regions of the sac that experience maximum bending during service.

Animals↗

Alpha thalassaemia-mental retardation, X linked.

X-linked alpha thalassaemia mental retardation (ATR-X) syndrome in males is associated with profound developmental delay, facial dysmorphism, genital abnormalities and alpha thalassaemia. Female carriers are usually physically and intellectually normal. So far, 168 patients have been reported. Language is usually very limited. Seizures occur in about one third of the cases. While many patients are affectionate with their caregivers, some exhibit autistic-like behaviour. Patients present with facial hypotonia and a characteristic mouth. Genital abnormalities are observed in 80% of children and range from undescended testes to ambiguous genitalia. Alpha-thalassaemia is not always present. This syndrome is X-linked recessive and results from mutations in the ATRX gene. This gene encodes the widely expressed ATRX protein. ATRX mutations cause diverse changes in the pattern of DNA methylation at heterochromatic loci but it is not yet known whether this is responsible for the clinical phenotype. The diagnosis can be established by detection of alpha thalassaemia, identification of ATRX gene mutations, ATRX protein studies and X-inactivation studies. Genetic counselling can be offered to families. Management is multidisciplinary: young children must be carefully monitored for gastro-oesophageal reflux as it may cause death. A number of individuals with ATR-X are fit and well in their 30s and 40s.

Abnormalities, Multiple↗

Neurokinin subtype receptors mediating substance P contraction in immature rabbit airways.

Two-week-old rabbit tracheal smooth muscle (TSM) and bronchial smooth muscle (BSM) segments were placed in organ baths, and isometric contractions to substance P (SP) were obtained. In the presence of phosphoramidon (PHOS), a neutral endopeptidase inhibitor, BSM segments were significantly more reactive and sensitive to SP than TSM segments. Neither neostigmine (NEO) nor atropine (ATR) eliminated these regional differences. Airway contractile responses to: 1) Senktide (NK-3 agonist); 2) neurokinin A (NKA, a NK-2 agonist); and 3) Septide (a highly selective NK-1 agonist) were separately obtained. In the presence of PHOS and NEO, Senktide was virtually inactive in both BSM and TSM. In the presence of PHOS, NEO, and ATR, NKA was equipotent in all airway segments; in contrast, the Septide response was significantly more reactive in BSM than in TSM segments. After inhibition of NK-1 activity with GR 82334, a competitive NK-1 receptor antagonist, the regional differences in SP reactivity were greatly diminished. This latter indication of a NK-1 contribution was confirmed using Septide-mediated inactivation of NK-1 receptors whereby the regional differences in airway sensitivity to SP were eliminated. These findings indicate that both endogenous neutral endopeptidase activity as well as NK-1 and NK-2 receptor influences may modulate the contractile responses to SP in immature rabbit airways.

Animals↗

Ontogeny of vascular angiotensin II receptor subtype expression in ovine development.

Angiotensin II (ANG II) increases arterial pressure in fetal sheep and may modulate cardiovascular adaptation before and after birth. The type 1 angiotensin II receptor (AT1R) predominates in adult vascular smooth muscle (VSM) and mediates vasoconstriction. In contrast, AT2R predominate in fetal tissues and are not known to mediate contraction. Although sheep are commonly used to study cardiovascular development, the ontogeny and distribution of VSM ATR subtypes is unknown. We examined ATR binding characteristics and subtype expression across the umbilicoplacental vasculature and in aorta, carotid, and mesenteric arteries from fetal (n = 44; 126-145 d gestation) and postnatal (n = 65; 1-120 d from birth) sheep using plasma membranes from tunica media and tissue autoradiography. Binding density (Bmax) was similar throughout the umbilicoplacental vasculature (p = 0.5), but only external umbilical arteries and veins and primary placental arteries expressed AT1R, whereas subsequent placental branches and fetal placentomes expressed only AT2R. Systemic VSM Bmax and binding affinity did not change significantly during development (p > 0.1). Fetal systemic VSM, however, expressed only AT2R, and binding was insensitive to GTPgammaS. Transition to AT1R in systemic VSM began 2 wk postnatal and was completed by 3 mo. Before birth, umbilical cord vessels are the primary site of AT1R expression in fetal sheep, and AT2R seem to predominate in systemic VSM until 2-4 wk postnatal.

Animals↗

Susceptibility of the trauma symptom inventory to malingering.

This study examined the sensitivity and specificity of the Trauma Symptom Inventory (TSI; Briere, 1995), a self-report measure of psychological sequelae of potentially traumatic events, to malingering. An optimal cutting score for a validity scale--Atypical Responding (ATR)--designed to identify exaggeration or other unusual response sets was developed in an analogue sample of 155 college students and subsequently applied to TSI profiles from several samples of patients with various psychiatric disorders. Use of a cross-validated T-score cutoff of 61 and below on the ATR scale produced good sensitivity (81%) and specificity (92%) rates in the analogue sample. Participants in the analogue sample who reported a history of traumatic experiences were no more able to successfully malinger trauma symptoms than were participants without such histories. Furthermore, false-positive rates in the clinical samples were generally low, suggesting that relatively few genuinely symptomatic individuals would be misclassified as malingering.

Adult↗

Risk of false positives when identifying malingered profiles using the trauma symptom inventory.

The Trauma Symptom Inventory (TSI; Briere, 1995) is a 100-item self-report measure of posttraumatic symptomatology that includes an Atypical Response (ATR) validity scale designed to differentiate honest from malingered profiles. In this study, using an analogue design to experimentally manipulate honest and malingered responses on the TSI, we found that proposed ATR cut scores produce a significant risk of false positives. Furthermore, the functioning of proposed cut scores worsened when we used posttraumatic stress disorder relevant samples and low estimates of malingering base rates. In light of these findings, the TSI should be used with caution when assessing claims of posttraumatic stress in forensic or disability settings.

Adult↗

Lipid synthesis inhibitors: effect on epidermal lipid conformational changes and percutaneous permeation of levodopa.

A combination of lipid synthesis inhibitors was used to enhance the in vitro and in vivo permeation of levodopa (LD) across rat epidermis, and their influence on epidermal lipids was investigated using attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy. Rat epidermis was treated with ethanol and a combination of atorvastatin (750 microg/7 cm2), cerulenin (20 microg/7 cm2), and beta-chloroalanine (600 microg/7 cm2) for sustaining the reduced content of epidermal cholesterol, fatty acids (as triglycerides), and ceramide (as sphingosine), respectively, in viable rat skin. This treatment resulted in significant (P < .05) synthesis inhibition of skin lipids up to 48 hours and 6-fold enhancement in the in vitro permeation of LD. The effective plasma concentration of LD was achieved within 1 hour and maintained over 48 hours after topical application to rat epidermis treated with a combination of these lipid synthesis inhibitors. ATR-FTIR studies of inhibitor(s)-treated rat epidermis revealed a significant decrease (P < .05) in peak height and area for both asymmetric and symmetric C-H stretching absorbances, suggesting extraction of lipids. However, an insignificant (P < .05) shift in the frequency of these peaks suggested no fluidization of epidermal lipids by lipid synthesis inhibitors. A direct correlation was observed between epidermal lipid synthesis inhibition, decrease in peak height or area, and percutaneous permeation of LD. Skin lipid synthesis inhibition by a combination of lipid synthesis inhibitors seems to offer a feasible approach for enhancing the transcutaneous delivery of LD.

Administration, Topical↗

Formulation of anastrozole microparticles as biodegradable anticancer drug carriers.

The purpose of this study was to develop poly(d,l-lactic-co-glycolic acid) (PLGA)-based anastrozole microparticles for treatment of breast cancer. An emulsion/extraction method was used to prepare anastrozole sustained-release PLGA-based biodegradable microspheres. Gas chromatography with mass spectroscopy detection was used for the quantitation of the drug throughout the studies. Microparticles were formulated and characterized in terms of encapsulation efficiency, particle size distribution, surface morphology, and drug release profile. Preparative variables such as concentrations of stabilizer, drug-polymer ratio, polymer viscosity, stirring rate, and ratio of internal to external phases were found to be important factors for the preparation of anastrozole-loaded PLGA microparticles. Fourier transform infrared with attenuated total reflectance (FTIR-ATR) analysis and differential scanning calorimetry (DSC) were employed to determine any interactions between drug and polymer. An attempt was made to fit the data to various dissolution kinetics models for multiparticulate systems, including the zero order, first order, square root of time kinetics, and biphasic models. The FTIR-ATR studies revealed no chemical interaction between the drug and the polymer. DSC results indicated that the anastrozole trapped in the microspheres existed in an amorphous or disordered-crystalline status in the polymer matrix. The highest correlation coefficients were obtained for the Higuchi model, suggesting a diffusion mechanism for the drug release. The results demonstrated that anastrozole microparticles with PLGA could be an alternative delivery method for the long-term treatment of breast cancer.

Anastrozole↗

Acetylcholine and norepinephrine: compared actions thyroid metabolism.

Acetylcholine (ACh; 5 X 10(-4) M), like norepinephrine (NE; 6 X 10(-6) M), as shown previously, stimulated iodide organification by mouse thyroids in vitro, while at the same time it inhibited TSH- or (Bu)2cAMP-induced T4 release. However, thyroid cAMP was not changed by ACh, suggesting that ACh, like NE, exerted its effects at a step beyond cAMP production. Also, while ACh increased cGMP concentrations, (Bu)2cGMP and 8-bromo-cGMP were not effective on thyroid function in this system. Neurotransmitters, then, presumably do not exert their action through cyclic nucleotide stimulation ACh-induced stimulation of organification and inhibition of release was reversed by 10(-5) M atropine (ATR) but not by 10(-5) M d-tubocurarine, indicating that muscarinic receptors were involved. ATR also reversed inhibition of T4 release induced by NE, suggesting that the presynaptic cholinergic pathway may be responsible for stimulation of postsynaptic cholinergic and adrenergic neurotransmitters in the thyroid gland.

Acetylcholine↗

Rapid determination of vitamin C by NIR, MIR and FT-Raman techniques.

Rapid spectroscopic determination of vitamin C in food and pharmaceutical products using infrared and Raman techniques was proposed. In this study, near-infrared (NIR), Fourier transform near-infrared (FT-NIR), Fourier transform infrared-attenuated total reflectance (FTIR-ATR), diffuse reflectance (DRIFTS), Fourier transform infrared-photoacoustic (FTIR-PAS) and FT-Raman spectroscopy were used in conjunction with partial least squares (PLS) regression to quantify vitamin C in powdered mixtures and solutions. Results indicate that the methods adopted have high prediction correlation. R2 values were 0.999 for FTIR-ATR, 0.976 for DRIFTS, 0.966 for FTIR-PAS, 0.988 for NIR, 0.992 for FT-NIR and 0.95 for FT-Raman, with an overall prediction error of 0.2-3.0%. The time required to complete an experiment ranged from 5 s (NIR) to 3 min (FT-Raman). The FTIR and FT-Raman techniques can be complementary tools for qualitative and quantitative characterization of the samples. Infrared and Raman techniques can be used to quantify vitamin C in foods and pharmaceutical products.

Ascorbic Acid↗

DNA damage checkpoint maintenance through sustained Chk1 activity.

The G2 DNA damage checkpoint prevents mitotic entry in the presence of DNA damage. This requires the activation of the phosphoinositide-3-kinase-related protein kinases ATR and ATM in human cells and the ATR homologue Rad3 in the fission yeast Schizosaccharomyces pombe. Rad3 activates the effector protein kinase Chk1 by phosphorylation. However, in fission yeast, inactivation of Rad3 following checkpoint activation has no impact on checkpoint duration. This demonstrates that Rad3 is not required for checkpoint maintenance and that the processes of checkpoint initiation and maintenance are distinct. Chk1 is required for checkpoint initiation but its role in checkpoint maintenance has not been investigated. We show here that Chk1 kinase activity is rapidly induced following irradiation and is maintained for the duration of a checkpoint arrest. On entry to mitosis, there is a transient decrease in Chk1 activity and phosphorylation, but Chk1 activity remains higher than that observed in unirradiated cells. We have generated temperature-sensitive alleles of chk1, which phenocopy chk1 deletion at the non-permissive temperature. Using these alleles, we have shown that inactivation of Chk1 during a checkpoint arrest leads to premature checkpoint termination, resulting in catastrophic mitoses that are a hallmark of checkpoint failure. Therefore, unlike Rad3, Chk1 is an important determinant of both checkpoint initiation and maintenance.

Amino Acid Sequence↗