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The preparation of stable hypervalent metal complexes containing Ag(III) has historically been challenging due to their propensity for reduction under ambient conditions. This work explores the preparation of a tripotassium silver bisperiodate complex as a tetrahydrate via chemical oxidation of the central silver atom and orthoperiodate chelation. The isolation of the chelate complex in high yield and purity was achieved via acidimetric titration. The comprehensive physiochemical characterization of the tribasic silver bisperiodate included single crystal X-ray diffraction, thermogravimetric and differential scanning calorimetry, and infrared and ultraviolet–visible spectroscopy. Infrared and UV–visible absorption spectra (λmax 255 and 365 nm) were in good agreement with historically prepared pentabasic diperiodatoargentate chelate complexes. The C2/c monoclinic distorted square planar structure of the bis-chelate complex affords a mutually supportive framework to both Ag(III) and I(VII), conferring stability under both thermal and long-term ambient conditions. Thermal analysis of the tribasic silver bisperiodate complex identified an endothermic mass loss, ΔH = +278.35 kJ/mol, observed at 139.0 °C corresponding to a solid-state reduction of silver from Ag(III) to Ag(I). Under ambient conditions, no significant degradation was observed over a 12 month period (P = 0.30) for the silver bisperiodate complex in a solid state, with an observed half-life of τ1/2 = 147 days in a pH-neutral aqueous solution.

Postharvest quality, quantification of betalains, phenolic compounds, and antioxidant activity of peel, pulp, and juice of fruits of three prickly pears (Opuntia ficus-indica L. Mill.) cultivars of Colegio de Postgraduados in México, were measured. The red and orange cultivars showed outstanding features of postharvest quality (size, texture, TSS and pulp and juice content) highest content of betalains and phenolic compounds. Therefore, highest antioxidant activity. In general, the highest content of bioactive compounds was detected in the peel, besides the content in pulp and juice did not show statistically significant differences. Phenolic content is very high in comparison with other fruits. Antioxidant activity was measured by three assays:FRAP, ABTS, and DPPH. Three cultivars showed a high correlation between antioxidant activity and phenolic compounds. The methodologies used in this work are a very useful tool for the quantification of bioactive compounds in O. ficus-indica fruit tissues.

Background and objective Electrospun chitosan membranes (ESCM) modified with short-chain fatty acids have the ability to control the release of simvastatin (SMV), an anti-cholesterol drug with osteogenic potential, for guided bone regeneration (GBR) applications. This study evaluated in vivo osteogenic effects of rapid short release of SMV (4 weeks) vs long sustained release (8 weeks) from acetic anhydride (AA)-and hexanoic anhydride (HA)-modified ESCMs, respectively. Methods AA ESCMs loaded with 10 or 50 µg SMV and HA ESCMs loaded with 50 µg SMV were evaluated for biocompatibility and bone formation at 4 and 8 weeks, in 5 mm critical size rat calvarial defects, using histological evaluation and micro-CT analysis. Results No severe inflammatory response was noticed around the ESCMs. Less hydrophobic AA membranes showed signs of resorption by week 4 and were almost completely resorbed by week 8 whereas the more hydrophobic HA membranes resorbed slowly, remaining intact over 8 weeks. In micro-CT analysis, 10 µg SMV-loaded AA membranes did not show significant bone formation as compared to non-loaded AA membranes at either evaluation time points. 50 µg SMV-loaded AA membranes stimulated significantly more bone formation than non-loaded AA membranes by week 4 (%bone = 31.0 ± 5.9% (AA50) vs 18.5 ± 13.7% (AA0)) but showed no difference at week 8. HA membranes with 50 µg SMV showed significantly more bone formation as compared to corresponding non-loaded membranes by week 8 (%bone = 61.7 ± 8.9% (HA50) vs 33.9 ± 29.7% (HA0)), though such an effect was not significant at week 4. Conclusion These results indicate that modified ESCMs may be used to control the release of SMV and promote bone healing in GBR applications.

Inspired by the primary events that take place in Photosystem II (PSII), we designed and synthesized a heteroleptic Ir(III) complex featuring an attached naphthoquinone (NQ) as an electron transfer (ET) auxiliary reminiscent of the plastoquinone electron acceptor in PSII. In this design, NQ is covalently attached to the 2,2′-bipyridyl (bpy) ligand of [Ir(dF(CF3)ppy)2(bpy)][PF6], (dF(CF3)ppy = 2-(2,4-difluorophenyl)-5-(trifluoromethyl)pyridine). Following excitation of the photocatalyst ([Ir(dF(CF3)ppy)2(bpy-NQ)][PF6]), reduced NQ (NQ•‒) was observed in transient absorption spectroscopy. This novel catalyst has potential applications in oxidative and reductive photocatalytic processes.

Novel approaches for N- and O-desulfonylation under room temperature (rt) and transition-metal-free conditions have been developed. The first methodology involves the transformation of a variety of N-sulfonyl heterocycles and phenyl benzenesulfonates to the corresponding desulfonylated products in good to excellent yields using only KOtBu in dimethyl sulfoxide (DMSO) at rt. Alternately, a visible light method has been used for deprotection of N-methyl-N-arylsulfonamides with Hantzsch ester (HE) anion serving as the visible-light-absorbing reagent and electron and hydrogen atom donor to promote the desulfonylation reaction. The HE anion can be easily prepared in situ by reaction of the corresponding HE with KOtBu in DMSO at rt. Both protocols were further explored in terms of synthetic scope as well as mechanistic aspects to rationalize key features of desulfonylation processes. Furthermore, the HE anion induces reductive dehalogenation reaction of aryl halides under visible light irradiation. Fil: Heredia, Micaela Denise. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina Fil: Guerra, Walter Damián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina Fil: Barolo, Silvia Maricel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina Fil: Fornasier, Santiago J.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina Fil: Rossi, Roberto Arturo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina Fil: Budén, María E.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina

Cyclic nucleotides determine a spatiotemporally regulated system that modulates uterine myometrial contractility and parturition timing. Adenylate and guanylate cyclases synthesize cAMP and cGMP upon β-adrenergic, natriuretic peptide, and nitric oxide stimulation. Before parturition, cAMP suppresses contraction-associated protein synthesis via PKA, and cGMP activates hyperpolarizing K+ current via PKG. At term, cAMP stimulates contraction-associated protein expression via EPAC just as decreasing PKG attenuates cGMP tocolysis. The cAMP-specific phosphodiesterase 4 (PDE4) facilitates myometrial contraction by degrading cyclic nucleotides. The precise roles of cGMP on myometrial activation and the effect of other PDEs in parturition are not yet clear. Strategies to reduce preterm birth could benefit from a better understanding of the myometrial cyclic nucleotide system.

In order to provide additional information on the behaviour of biochemical parameters related to stress responses to a specific long-term competition, we aimed to compare the stressful effects of a long-lasting competition on physiological variables in men and women. This is a prospective observational analytical study. Twenty-five professional athletes, 15 men and 10 women, travelled 460 km for 4 days in an international edition of the Ecomotion/Pro AR World. After the competition, we detected an increase in α-amylase and cortisol levels and a decrease in salivary immunoglobulin A (lgA) levels. The relative percentage changes in α-amylase, IgA and cortisol levels were significantly higher in women than in men, whereas women had lower relative percentage changes in glucose and lactate levels compared with men. There was a decrease in lymphocyte, eosinophil and monocyte counts, with relative percentage decreases in lymphocytes and monocytes being significantly higher in female athletes than in males. There were increases in the serum activities of total creatine kinase (CK), the creatine kinase myocardial isoform (CKMB), alanine aminotransferase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) at the end of the test, with significantly higher elevations of total CK, CKMB and LDH in men and ALT in women. Long-lasting competition induced stress, muscle damage, anaemia and changes in the immune system. Women had more intense responses of cortisol and leukocytes. Um zusätzliche Informationen zum Verhalten biochemischer Parameter in Verbindung mit stressbedingten Reaktionen auf einen speziellen Langstreckenwettkampf zu gewinnen, verglichen wir die stressbedingten Auswirkungen eines Langstreckenwettkampfs auf physiologische Variablen bei Männern und Frauen. Bei dieser Studie handelt es sich um eine prospektive Beobachtungsanalyse. 25 Profisportler (15 Männer und 10 Frauen) legten in einer internationalen Ausgabe der Ecomotion/Pro AR World in 4 Tagen 460 km zurück. Nach dem Wettkampf fanden sich erhöhte α-Amylase- und Kortisolspiegel sowie eine Abnahme der Immunglobulin-A(IgA)-Spiegel im Speichel. Die relativen prozentualen Veränderungen der α-Amylase-, IgA- und Kortisolspiegel waren bei Frauen signifikant höher als bei Männern. Frauen wiesen geringere relative Veränderungen der Glukose- und Laktatwerte auf als Männer. Es zeigte sich eine Abnahme der Lymphozyten-, Eosinophilen- und Monozytenzahlen, wobei die relative prozentuale Abnahme der Lymphozyten und Monozyten bei weiblichen Athleten signifikant höher ausfiel als bei männlichen. Am Ende des Tests wurden signifikant höhere Serumaktivitäten der Gesamtkreatinkinase (CK), der Kreatinkinase-Myokard-Isoform (CKMB), der Alanin-Aminotransferase (ALT), der Aspartat-Aminotransferase (AST) und der Laktatdehydrogenase (LDH) gemessen. Hierbei wiesen die Männer eine signifikant höhere Zunahme der Gesamt-CK-, CKMB- und LDH-Werte auf, während bei den Frauen der ALT-Wert signifikant stärker erhöht war. Der Langzeitwettkampf führte zu Stress, Muskelschäden, Blutarmut und Veränderungen im Immunsystem. Frauen zeigten intensivere Reaktionen bei den Kortisol- und Leukozytenwerten.

Nitric oxide (NO) is a major inhibitory neurotransmitter that mediates nonadrenergic noncholinergic (NANC) signaling. Neuronal NO synthase (nNOS) is activated by Ca(2+)/calmodulin to produce NO, which causes smooth muscle relaxation to regulate physiologic tone. nNOS serine1412 (S1412) phosphorylation may reduce the activating Ca(2+) requirement and sustain NO production. We developed and characterized a nonphosphorylatable nNOS(S1412A) knock-in mouse and evaluated its enteric neurotransmission and gastrointestinal (GI) motility to understand the physiologic significance of nNOS S1412 phosphorylation. Electrical field stimulation (EFS) of wild-type (WT) mouse ileum induced nNOS S1412 phosphorylation that was blocked by tetrodotoxin and by inhibitors of the protein kinase Akt but not by PKA inhibitors. Low-frequency depolarization increased nNOS S1412 phosphorylation and relaxed WT ileum but only partially relaxed nNOS(S1412A) ileum. At higher frequencies, nNOS S1412 had no effect. nNOS(S1412A) ileum expressed less phosphodiesterase-5 and was more sensitive to relaxation by exogenous NO. Under non-NANC conditions, peristalsis and segmentation were faster in the nNOS(S1412A) ileum. Together these findings show that neuronal depolarization stimulates enteric nNOS phosphorylation by Akt to promote normal GI motility. Thus, phosphorylation of nNOS S1412 is a significant regulatory mechanism for nitrergic neurotransmission in the gut.

Advanced local delivery systems are needed as adjunctive treatments for severe injuries with high infection rates, such as open fractures. Chitosan systems have been investigated as antimicrobial local delivery systems for orthopaedic infection but possess mismatches between elution and degradation properties. Derivatives of chitosan were chosen that have enhanced swelling ratios or tailorable degradation properties. A combination of trimethyl chitosan and poly(ethylene glycol) diacrylate chitosan was developed as an injectable local delivery system. Research objectives were elution of antimicrobials for 7 days, degradation as open fractures heal, and cytocompatibility. The derivative combination eluted increased active concentrations of vancomycin and amikacin compared to the non-derivatized chitosan paste, 6 vs. 5 days and 5 vs. 4 days, respectively. The derivative combination degraded slower than non-derivatized paste in an enzymatic degradation study, 14 vs. 3 days, which increased antimicrobial delivery duration. Cytocompatibility of the combination with fibroblast and pre-osteoblast cells exceeds the cell viability standard set in ISO 10993-5. Combination paste requires an increased ejection force of 9.40 N (vs. 0.64 N), but this force was within an acceptable injection force threshold, 80 N. These preliminary results indicate combination paste should be further developed into a clinically useful adjunctive local delivery system for infection prevention.

Gasotransmitters are endogenous small gaseous messengers exemplified by nitric oxide (NO), carbon monoxide (CO), and hydrogen sulfide (H2S or sulfide). Gasotransmitters are implicated in myriad physiologic functions including many aspects of reproduction. Our objective was to comprehensively review basic mechanisms and functions of gasotransmitters during pregnancy from conception to uterine involution and highlight future research opportunities. We searched PubMed and Web of Science databases using combinations of keywords nitric oxide, carbon monoxide, sulfide, placenta, uterus, labor, and pregnancy. We included English language publications on human and animal studies from any date through August 2018 and retained basic and translational articles with relevant original findings. All gasotransmitters activate cGMP signaling. NO and sulfide also covalently modify target protein cysteines. Protein kinases and ion channels transduce gasotransmitter signals, and co-expressed gasotransmitters can be synergistic or antagonistic depending on cell type. Gasotransmitters influence tubal transit, placentation, cervical remodeling, and myometrial contractility. NO, CO, and sulfide dilate resistance vessels, suppress inflammation, and relax myometrium to promote uterine quiescence and normal placentation. Cervical remodeling and rupture of fetal membranes coincide with enhanced oxidation and altered gasotransmitter metabolism. Mechanisms mediating cellular and organismal changes in pregnancy due to gasotransmitters are largely unknown. Altered gasotransmitter signaling has been reported for preeclampsia, intrauterine growth restriction, premature rupture of membranes, and preterm labor. However, in most cases specific molecular changes are not yet characterized. Nonclassical signaling pathways and the crosstalk among gasotransmitters are emerging investigation topics.

During pregnancy, estrogen (E2) stimulates uterine artery blood flow (UBF) by enhancing nitric oxide (NO)-dependent vasodilation. Cystathionine γ-lyase (CSE) promotes vascular NO signaling by producing hydrogen sulfide (H2S) and by maintaining the ratio of reduced-to-oxidized intracellular glutathione (GSH/GSSG) through l-cysteine production. Because redox homeostasis can influence NO signaling, we hypothesized that CSE mediates E2 stimulation of UBF by modulating local intracellular cysteine metabolism and GSH/GSSG levels to promote redox homeostasis. Using non-pregnant ovariectomized WT and CSE-null (CSE KO) mice, we performed micro-ultrasound of mouse uterine and renal arteries to assess changes in blood flow upon exogenous E2 stimulation. We quantified serum and uterine artery NO metabolites (NOx), serum amino acids, and uterine and renal artery GSH/GSSG. WT and CSE KO mice exhibited similar baseline uterine and renal blood flow. Unlike WT, CSE KO mice did not exhibit expected E2 stimulation of UBF. Renal blood flow was E2-insensitive for both genotypes. While serum and uterine artery NOx were similar between genotypes at baseline, E2 decreased NOx in CSE KO serum. Cysteine was also lower in CSE KO serum, while citrulline and homocysteine levels were elevated. E2 and CSE deletion additively decreased GSH/GSSG in uterine arteries. In contrast, renal artery GSH/GSSG was insensitive to E2 or CSE deletion. Together, these findings suggest that CSE maintenance of uterine artery GSH/GSSG facilitates nitrergic signaling in uterine arteries and is required for normal E2 stimulation of UBF. These data have implications for pregnancy pathophysiology and the selective hormone responses of specific vascular beds., Graphical abstract Image 1, Highlights • CSE-null mice exhibit abnormal estrogen augmentation of uterine artery blood flow. • Estrogen lowers uterine artery nitric oxide metabolites in CSE null mice. • CSE loss and estrogen additively impair uterine artery glutathione homeostasis. • Neither CSE loss nor estrogen influences renal artery blood flow or glutathione.

14 p.-4 fig.-2 tab.- 8 schem., We prepared a series of free NH and N-substituted dibenzonthiazines with potential anti-tumor activity from N-aryl-benzenesulfonamides. A biological test of synthesized compounds (59 samples) was performed in vitro measuring their antiproliferative activity against a panel of six human solid tumor cell lines and its tubulin inhibitory activity. We identified 6-(phenylsulfonyl)-6H-dibenzo[c,e][1,2]thiazine 5,5-dioxide and 6-tosyl-6H-dibenzo[c,e][1,2]thiazine 5,5-dioxide as the best compounds with promising values of activity (overall range of 2-5.4 μM). Herein, we report the dibenzothiazine core as a novel building block with antiproliferative activity, targeting tubulin dynamics., This study was supported by Agencia Córdoba Ciencia,Consejo Nacional de Investigaciones Científicas y Técnicas(CONICET),Secretaría de Ciencia y Tecnología,Universidad Nacional de Córdoba (SECyT),and Agencia Nacional de Promoción Científica y Técnica(ANPCyT).W. D. G. gratefully acknowledge receipt of fellowship from CONICET.J. M. P. thanks the Spanish Government for financial support through project PGC2018-094503-B-C22(MCIU/AEI/FEDER,UE). The biochemical work was supported by grants Ministerio de Ciencia e Innovación PID2019-104545RB-I00/AEI/10.13039/501100011033,PIEs 201920E111 and 202020E301 from Consejo Superior de Investigaciones Científicas,grant “Bases moleculares de la regulación de microtúbulos y sus implicaciones en la neurotoxicidad producida por fármacos ”from Fundación TATIANA and European Union H2020-MSCA-ITN-2019 860070TUBINTRAIN grant to JFDP and a generous donation from Club Deportivo Escuela Hungaresa de Pontevedra.

In this study, the antifungal activity of AgNPs was tested against C. tropicalis (pathogen fungi) and S. boulardii (probiotic). The effectiveness of the AgNPs was assessed by comparing their antifungal activity with a triazole antifungal drug fluconazole and amphotericin B. The AgNPs have a polygonal-like shape (average size of 35 ± 15 nm) with 1.2% wt. of metallic silver stabilized with 18.8% wt. of polyvinylpyrrolidone (PVP) in 80% wt. of distilled water. The results revealed that 35 μg/mL of fluconazole inhibits 55–60% of both fungal cell growth. As for amphotericin B, 5 μg/mL is sufficient to inhibit more than 95% of both fungal cells. For AgNPs, 25 μg/mL was needed to inhibit 90% of the C. tropicalis cell growth, but remarkably, 50% of the S. boulardii cell population remains viable, which can potentiate cell reproduction. Our results could initiate the development of AgNPs possessing selective specificity against pathogenic fungal species.

BACKGROUND AND PURPOSE: The enteric neurotransmitter nitric oxide (NO) regulates gastrointestinal motility by relaxing smooth muscle. Pharmacological cAMP induction also relaxes gastrointestinal smooth muscle, but it is uncertain whether cAMP augments or suppresses enteric NO signalling. In other organ systems, cAMP can increase neuronal NO production by stimulating protein kinase A (PKA) to phosphorylate neuronal NOS (nNOS) Serine‐1412 (S1412). We hypothesized that cAMP also increases nNOS S1412 phosphorylation by PKA in enteric neurons to augment nitrergic relaxation of mouse ileum. EXPERIMENTAL APPROACH: We measured contractile force and nNOS S1412 phosphorylation in ileal rings suspended in an organ bath. We used forskolin to induce cAMP‐dependent relaxation of wild type, nNOS(S1412A) knock‐in and nNOSα‐null ileal rings in the presence or absence of PKA, protein kinase B (Akt) and NOS inhibitors. KEY RESULTS: Forskolin stimulated phosphorylation of nNOS S1412 in mouse ileum. Forskolin relaxed nNOSα‐null and nNOS(S1412A) ileal rings less than wild‐type ileal rings. PKA inhibition blocked forskolin‐induced nNOS phosphorylation and attenuated relaxation of wild type but not nNOS(S1412A) ileum. Akt inhibition did not alter nNOS phosphorylation with forskolin but did attenuate relaxation of wild type and nNOS(S1412A). NOS inhibition with l‐NAME eliminated the effects of PKA and Akt inhibitors on relaxation. CONCLUSION AND IMPLICATIONS: PKA phosphorylation of nNOS S1412 augments forskolin‐induced nitrergic ileal relaxation. The relationship between cAMP/PKA and NO is therefore synergistic in enteric nitrergic neurons. Because NO regulates gut motility, selective modulation of enteric neuronal cAMP synthesis may be useful for the treatment of gastrointestinal motility disorders.

Rendering is a global industry that recycles by-products resulting from butchering operations, which process billions of animals per year. About 50% of the weight of livestock is not consumed by humans and must be processed by rendering operations, which cook and separate the material into its protein and fat components. These products serve as a sustainable food source for livestock, feedstocks for oleochemicals, and raw material for biodiesel refineries. Due to the scale and nature of the raw materials and the cooking process, rendering operations emit a significant, but as yet poorly quantified, VOC load. Assessing this VOC load is important in order to calibrate the industry’s contribution to global VOC emissions, and to help address nuisance odor problems. We conducted VOC air sampling of two facilities in California, USA during the winter and summer seasons. VOC and reduced sulfur analyses were conducted using 8 h ambient air samples. Analyses for amines, ammonia, aldehydes/ketones, and volatile fatty acids were conducted using sampling pumps. These analyses detected 43 compounds at the facilities, and the number and concentration of detectable compounds were seasonally dependent. The compounds present at the highest concentrations included: ammonia (1600–2800 ppb, i.e., winter–summer levels), acetic acid (80–320 ppb, along with twelve other fatty acids ranging from ~0.5–140 ppb), acetone (55–241 ppb, along with nine other aldehyde/ketone products ranging from 0.4–60 ppb), and ethanol (15–81 ppb). These constituents have low odor thresholds and thus contribute to nuisance odor problems. Further, the overall VOC contribution arising from rendering facilities on a global scale is as yet very poorly characterized. This analysis will be useful to guide the development of new odor abatement strategies and strategies for the reduction of VOC emissions associated with this critical industry.

To address a need for highly X-ray opaque biodegradable materials, a series of poly(e-caprolactone) derivatives were prepared using oxime post-polymerization modification to conjugate two different iodinated hydroxylamines. These materials were synthesized with up to 32 wt. % iodine content and demonstrated improved X-ray contrast relative to both the ketone precursors and previously reported monoiodinated materials. © 2016 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2016

A novel approach for the synthesis of tetracyclic indoles and 7-azaindoles is reported. The strategy involves four steps, with a fast rt intramolecular α-arylation of ketones as key step. The reaction was inspected synthetically to achieve the synthesis of 11 novel tetracyclic structures with moderate to very good yields (39-85%). Theoretical combined with experimental studies led us to propose a probable polar mechanism (concerted S

The free radical nitric oxide (NO(•)) regulates diverse physiological processes from vasodilation in humans to gas exchange in plants. S-Nitrosoglutathione (GSNO) is considered a principal nitroso reservoir due to its chemical stability. GSNO accumulation is attenuated by GSNO reductase (GSNOR), a cysteine-rich cytosolic enzyme. Regulation of protein nitrosation is not well understood since NO(•)-dependent events proceed without discernible changes in GSNOR expression. Because GSNORs contain evolutionarily conserved cysteines that could serve as nitrosation sites, we examined the effects of treating plant (Arabidopsis thaliana), mammalian (human), and yeast (Saccharomyces cerevisiae) GSNORs with nitrosating agents in vitro. Enzyme activity was sensitive to nitroso donors, whereas the reducing agent dithiothreitol (DTT) restored activity, suggesting that catalytic impairment was due to S-nitrosation. Protein nitrosation was confirmed by mass spectrometry, by which mono-, di-, and trinitrosation were observed, and these signals were sensitive to DTT. GSNOR mutants in specific non-zinc-coordinating cysteines were less sensitive to catalytic inhibition by nitroso donors and exhibited reduced nitrosation signals by mass spectrometry. Nitrosation also coincided with decreased tryptophan fluorescence, increased thermal aggregation propensity, and increased polydispersity-properties reflected by differential solvent accessibility of amino acids important for dimerization and the shape of the substrate and coenzyme binding pockets as assessed by hydrogen-deuterium exchange mass spectrometry. Collectively, these data suggest a mechanism for NO(•) signal transduction in which GSNOR nitrosation and inhibition transiently permit GSNO accumulation.

Polyethylenimine (PEI) functionalized kaolinite clay was successfully prepared, characterized, and assessed for the remediation of volatile organic compounds (VOCs) comprising the aldehyde, carboxylic acid, and disulfide functional group classes. A gas chromatographic vapor capture assay evaluated the capability of unmodified and modified clay material to capture representative aldehyde, carboxylic acid, and disulfide VOCs in a laboratory setting. Unmodified kaolinite (Kao) clay was moderately or poorly effective at remediating these VOCs, while the poly(amine) functionalized Kao was capable of capturing VOCs in the vapor phase with reductions up to 100%. Sample cartridge tubes were packed with PEI-functionalized clay in order to assess their ability to reduce the detectable volatile fatty acid load at an open-air rendering plant in a relevant field test for applying these materials in a packed-bed scrubber application. The PEI-Kao packed cartridges were capable of significantly reducing the detectable concentration of volatile fatty acid effluent from the rendering operation. These volatile fatty acids are major contributors to nuisance odors associated with rendering.

Recent studies suggest cysteine S-nitrosation of S-nitrosoglutathione reductase (GSNOR) could regulate protein redox homeostasis. "Switch" assays enable discovery of putatively S-nitrosated proteins. However, with few exceptions, researchers have not examined the kinetics and biophysical consequences of S-nitrosation. Methods to quantify protein S-nitrosothiol (SNO) abundance and formation kinetics would bridge this mechanistic gap and allow interpretation of the consequences of specific modifications, as well as facilitate development of specific S-nitrosation inhibitors. Here, we describe a rapid assay to estimate protein SNO abundance with intact protein electrospray ionization mass spectrometry. Originally designed using recombinant GSNOR, these methods are applicable to any purified protein to test for or further study nitrosatable cysteines.

N-Arylation reaction is obtained when 2′-halo-[1,1′-biphenyl]-2-amines are irradiated in basic medium. On the basis of photochemical, photophysical experiments and computational studies we propose that carbazoles are formed by intermolecular electron transfer via SRN1 mechanism. In general, biphenylamines with an EDG like Me or OMe behave in the same way as H giving both, cyclized and reduced products. On the other hand, biphenylamines containing EWG like CN, COOEt or CF3gave only the corresponding carbazole. Herein, we report for the first time the chain length for the propagation cycle of intramolecular SRN1 reactions and explain that differences in the distribution of products suggest differences regarding the overall mechanism involved. Fil: Guerra, Walter Damián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina Fil: Buden, Maria Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina Fil: Barolo, Silvia Maricel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina Fil: Rossi, Roberto Arturo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina Fil: Pierini, Adriana Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina

Plant LOSS OF GDU 2 (LOG2) and Mammalian Mahogunin Ring Finger 1 (MGRN1) proteins are RING-type E3 ligases sharing similarity N-terminal to the RING domain. Deletion of this region disrupts the interaction of LOG2 with the plant membrane protein GLUTAMINE DUMPER1 (GDU1). Phylogenetic analysis identified two clades of LOG2/MGRN1-like proteins in vertebrates and plants. The ability of MGRN1 to functionally replace LOG2 was tested. MGRN1 ubiquitylates GDU1 in vitro and can partially substitute for LOG2 in the plant, partially restoring amino acid resistance to a GDU1-myc over-expression, log2-2 background. Altogether, these results suggest a conserved function for the N-terminal domain in evolution.

Intercellular amino acid transport is essential for the growth of all multicellular organisms, and its dysregulation is implicated in developmental disorders. By an unknown mechanism, amino acid efflux is stimulated in plants by overexpression of a membrane-localized protein (GLUTAMINE DUMPER 1 (GDU1)) that requires a ubiquitin ligase (LOSS OF GDU 2 (LOG2). Here we further explore the physiological consequences of the interaction between these two proteins. LOG2 ubiquitin ligase activity is necessary for GDU1-dependent tolerance to exogenous amino acids, and LOG2 self-ubiquitination was markedly stimulated by the GDU1 cytosolic domain, suggesting that GDU1 functions as an adaptor or coactivator of amino acid exporter(s). However, other consequences more typical of a ligase-substrate relationship are observed: disruption of the LOG2 gene increased the in vivo half-life of GDU1, mass spectrometry confirmed that LOG2 ubiquitinates GDU1 at cytosolic lysines, and GDU1 protein levels decreased upon co-expression with active, but not enzymatically inactive LOG2. Altogether these data indicate LOG2 negatively regulates GDU1 protein accumulation by a mechanism dependent upon cytosolic GDU1 lysines. Although GDU1-lysine substituted protein exhibited diminished in vivo ubiquitination, overexpression of GDU1 lysine mutants still conferred amino acid tolerance in a LOG2-dependent manner, consistent with GDU1 being both a substrate and facilitator of LOG2 function. From these data, we offer a model in which GDU1 activates LOG2 to stimulate amino acid export, a process that could be negatively regulated by GDU1 ubiquitination and LOG2 self-ubiquitination.

A new and general synthetic route to prepare dibenzosultams is here reported. This approach involves the synthesis of N-aryl-2-halobenzenesulfonamides (3), followed by intramolecular C–C photoinduced arylation under soft conditions without the use of “Transition Metal”. The photostimulated reactions exhibit very good tolerance to different substituent groups with good to excellent isolated yields (42–98%) of products. Moreover, it is shown that LED (λ = 395 nm) is an efficient light energy source to initiate efficiently the reactions. Theoretical inspection of the mechanism was made to probe the involvement of the radical-anion SRN1 process.

A one-pot synthesis of benzo[f]indole-4,9-diones from 1,4-naphthoquinone with α-aminoacetals has been developed. This method provides a straightforward synthesis of benzo[f]indole-4,9-diones via intramolecular nucleophilic attack of aminoquinones to aldehydes under mild reaction conditions. The detailed mechanism was also investigated.

A new and general synthetic route to prepare dibenzosultams is here reported. This approach involves the synthesis of N-aryl-2-halobenzenesulfonamides (3), followed by intramolecular C-C photoinduced arylation under soft conditions without the use of "Transition Metal". The photostimulated reactions exhibit very good tolerance to different substituent groups with good to excellent isolated yields (42-98%) of products. Moreover, it is shown that LED (= 395 nm) is an efficient light energy source to initiate efficiently the reactions. Theoretical inspection of the mechanism was made to probe the involvement of the radical-anion SRN1 process. Fil: Guerra, Walter Damián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina Fil: Rossi, Roberto Arturo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina Fil: Pierini, Adriana Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina Fil: Barolo, Silvia Maricel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina

A one-pot synthesis of benzo[f]indole-4,9-diones from 1,4-naphthoquinone with α-aminoacetals has been developed. This method provides a straightforward synthesis of benzo[f]indole-4,9-diones via intramolecular nucleophilic attack of aminoquinones to aldehydes under mild reaction conditions. The detailed mechanism was also investigated.

Amino acids serve as transport forms for organic nitrogen in the plant, and multiple transport steps are involved in cellular import and export. While the nature of the export mechanism is unknown, overexpression of GLUTAMINE DUMPER1 (GDU1) in Arabidopsis (Arabidopsis thaliana) led to increased amino acid export. To gain insight into GDU1’s role, we searched for ethyl-methanesulfonate suppressor mutants and performed yeast-two-hybrid screens. Both methods uncovered the same gene, LOSS OF GDU2 (LOG2), which encodes a RING-type E3 ubiquitin ligase. The interaction between LOG2 and GDU1 was confirmed by glutathione S-transferase pull-down, in vitro ubiquitination, and in planta coimmunoprecipitation experiments. Confocal microscopy and subcellular fractionation indicated that LOG2 and GDU1 both localized to membranes and were enriched at the plasma membrane. LOG2 expression overlapped with GDU1 in the xylem and phloem tissues of Arabidopsis. The GDU1 protein encoded by the previously characterized intragenic suppressor mutant log1-1, with an arginine in place of a conserved glycine, failed to interact in the multiple assays, suggesting that the Gdu1D phenotype requires the interaction of GDU1 with LOG2. This hypothesis was supported by suppression of the Gdu1D phenotype after reduction of LOG2 expression using either artificial microRNAs or a LOG2 T-DNA insertion. Altogether, in accordance with the emerging bulk of data showing membrane protein regulation via ubiquitination, these data suggest that the interaction of GDU1 and the ubiquitin ligase LOG2 plays a significant role in the regulation of amino acid export from plant cells.

The bioartificial chitosan–poly(vinyl alcohol) blends were toughened by means of a dehydrothermal treatment (DHT), to facilitate the formation of hydrogen bonds between the macromolecules. The materials were characterized by stress–strain test, contact angle measurement, spotlight Fourier transform infrared spectroscopy and chemical imaging, weight loss in water, swelling in water vapor saturated atmosphere, Alamar blue test to evaluate the indirect cytotoxicity, and the diffusive permeation, through membranes made with the blends, of D(+)glucose, vitamin B12, and bovine serum albumin. The results were compared with those of the blends crosslinked by glutaraldehyde (GTA). The Young's modulus ranges between 10.56 and 16.12 MPa, and it is higher for the blends subjected to DHT than for those crosslinked by GTA, a fact explainable by the elasticity of the latter, due to the flexible bridges connecting the different chains. The contact angles indicate a scarce wettability, and then a scarce hydrophilicity, which is confirmed by the chemical imaging of the surfaces, made in the total reflection (microATR) mode, of the films toughened by DHT: the ν(OH) band in the 4000–3000 cm-1 is nearly absent in all the regions of the maps. Moreover, the correlation maps indicate a homogeneous distribution of the two components within the blends. The weight loss in water is generally less than 15%, and increases with the content of the ionizable macromolecule chitosan in the blends, a trend shown also by the swelling after exposure to water vapors. Alamar blue test shows that none of the eluates, after being in contact with the materials up to seven days, appears cytotoxic toward murine fibroblasts. As concerning the diffusive permeation, it appears good for D(+)glucose, quite good for vitamin B12, and scarce for bovine serum albumin. In conclusion, the chitosan–poly(vinyl alcohol) blends studied appear to be suitable for their use as biomaterials.

Purpose One third of the world’s population suffers from gastrointestinal (GI) dysmotility, a common feature of which is disrupted nitric oxide (NO) signaling and spastic contractility. In the healthy gut, neuronal NO synthase (nNOS) induces cGMP-dependent GI smooth muscle relaxation, which facilitates digestion. However, regulation of nNOS activity is poorly understood. PKA or Akt phosphorylation of nNOS S1412 enhances hypothalamic and penile NO synthesis. We hypothesized that nNOS S1412 phosphorylation promotes GI motility. Methods We quantified the contractile force of ileal segments from C57Bl/6 wildtype and nNOSS1412A mutant mice in an organ bath under non-adrenergic, non-cholinergic (NANC) conditions. Relaxation stimuli—neuronal depolarization (EFS; 20V, 2msec pulsewidth) or adenylate cyclase activation (FSK)—were applied to ileal segments in the presence of kinase, nNOS, or cGMP pathway inhibitors. We measured nNOS S1412 phosphorylation by western blot. We determined non-NANC ileal contraction velocity with the gastrointestinal motility monitor. Results EFS (1Hz) and FSK (300nM) increased nNOS S1412 phosphorylation by 310% (p Conclusions nNOS S1412 phosphorylation promotes GI motility by decreasing spastic contractility in murine ileal segments. Neuronal depolarization elicits Akt-dependent phosphorylation, while adenylate cyclase activation elicits PKA-dependent phosphorylation of nNOS. Common GI motility disorders like irritable bowel syndrome may benefit from therapies that upregulate nNOS S1412 phosphorylation.

Bioartificial biodegradable materials were prepared mixing chitosan (CHI) and poly(vinyl alcohol) (PVA), then manufactured as films, and finally cross-linked with glutaraldehyde (GTA), both in the absence and in the presence of the edible hexa-alcohol sorbitol (SOR), as a plasticizer. The release of the components into water was tested by high performance liquid chromatography (HPLC); no release of CHI and scarce release of PVA were found. The water uptake was tested by measuring the swelling of the materials, after incubating them for 20 h in an atmosphere saturated with water vapor at 37°C. The swelling percentage increases with increasing CHI content in the blends, although it is the less hydrophilic polymer. This behavior was attributed to the difficulty of water to diffuse through the crystalline PVA structure, which is partially altered in the blends. The addition of SOR enhances the water sorption, as expected. The biodegradability of the materials was tested using the specific enzyme chitosanase, and was found to depend on the blend composition, as well as to be enhanced by the addition of SOR. The initial degradation rates were calculated; the maximum rates were found when the CHI to PVA ratio was 80:20 for all systems. The results of the enzymatic degradation generally agree with those of the swelling. The cross-linked blends were also tested as drug-delivery systems. The drugs chosen were the vitamin L-ascorbic acid (AsA) and the anti-cancer drug paclitaxel (PTX). The effective diffusion coefficients, D eff , were evaluated for the release of both the drugs from each material. Those of AsA are greater, of many powers of ten, than those of PTX, owing mainly to the hydrophilic nature of the first drug and to the hydrophobic of the second one. In conclusion, these materials seem available for biomedical use.

Oxidesulfurization (ODS) of benzothiophenic compounds prevailing in diesel was conducted with hydrogen peroxide in presence of various catalysts, using a model diesel and actual diesel fuel. ODS activities of dibenzothiophenes (DBTs) in hexadecane for a series of V2O5 catalysts supported on alumina, titania, ceria, niobia and silica, were evaluated. Results show that the oxidation activity of DBTs depends on the support used. It was observed that the sulfone yield is not proportional to textural properties or V content. For all catalysts, ODS of benzothiophene (BT), dibenzothiophene (DBT), 4-methyl dibenzothiophene (4-MDBT) and 4,6-dimethyl dibenzothiophene (4,6-DMDBT) decreased in the following order: DBT > 4-MDBT > 4,6-DMDBT > BT. This trend does not depend on the catalyst used or the textural properties of the catalysts and supports. In presence of indole ODS activities diminish, except with catalysts supported on alumina-titania mixed oxide, whereas with V2O5/TiO2 catalyst the performance is the highest. ODS of Mexican diesel fuel was carried out in presence of this catalyst and S level was diminished in about 99%.

Aldehyde and carboxylic acid volatile organic compounds (VOCs) present significant environmental concern due to their prevalence in the atmosphere. We developed biodegradable functional nanoparticles comprised of poly(d,l-lactic acid)-poly(ethylene glycol)-poly(ethyleneimine) (PDLLA-PEG-PEI) block co-polymers that capture these VOCs by chemical reaction. Polymeric nanoparticles (NPs) preparation involved nanoprecipitation and surface functionalization with branched PEI. The PDLLA-PEG-PEI NPs were characterized by using TGA, IR, (1) H NMR, elemental analysis, and TEM. The materials feature 1°, 2°, and 3° amines on their surface, capable of capturing aldehydes and carboxylic acids from gaseous mixtures. Aldehydes were captured by a condensation reaction forming imines, whereas carboxylic acids were captured by acid/base reaction. These materials reacted selectively with target contaminants obviating off-target binding when challenged by other VOCs with orthogonal reactivity. The NPs outperformed conventional activated carbon sorbents.

The starting materials are generally prepared following the approaches shown for (IIIa) and (XIII).

The aim of this work was the development of new synthetic polymeric systems, functionalized by surface chemical modification with bioactive peptides, for myocardial tissue engineering. Polycaprolactone and a poly(ester-ether-ester) block copolymer synthesized in our lab, polycaprolactone-poly(ethylene oxide)-polycaprolactone (PCL-PEO-PCL), were used as the substrates to be modified. Two pentapeptides, H-Gly-Arg-Gly-Asp-Ser-OH (GRGDS) from fibronectin and H-Tyr-Ile-Gly-Ser-Arg-OH (YIGSR) from laminin, were used for the functionalization. Polymeric membranes were obtained by casting from solutions and then functionalized by means of alkaline hydrolysis and subsequent coupling of the bioactive molecules through 1-(3-dimethylaminopropyl)-3-ethylcarbodimide hydrochloride/N-hydroxysuccinimide chemistry. The hydrolysis conditions, in terms of hydrolysis time, temperature, and sodium hydroxide concentration, were optimized for the two materials. The occurrence of the coupling reaction was demonstrated by infrared spectroscopy, as the presence on the functionalized materials of the absorption peaks typical of the two peptides. The peptide surface density was determined by chromatographic analysis and the distribution was studied by infrared chemical imaging. The results showed a nearly homogeneous peptide distribution, with a density above the minimum value necessary to promote cell adhesion. Preliminary in vitro cell culture studies demonstrated that the introduction of the bioactive molecules had a positive effect on improving C2C12 myoblasts growth on the synthetic materials.

Schizochytrium sp. is a marine microalga that has been developed as a commercial source for docosahexaenoic acid (DHA, C22:6 (omega-3), enriched biomass, and oil. Previous work suggested that the DHA, as well as docosapentaenoic acid (DPA, C22:5 omega-6), that accumulate in Schizochytrium are products of a multi-subunit polyunsaturated fatty acid (PUFA) synthase (1). Here we show data to support this view and also provide information on other aspects of fatty acid synthesis in this organism. Three genes encoding subunits of the PUFA synthase were isolated from genomic DNA and expressed in E. coli along with an essential accessory gene encoding a phosphopantetheinyl transferase (PPTase). The resulting transformants accumulated both DHA and DPA. The ratio of DHA to DPA was approximately the same as that observed in Schizochytrium. Treatment of Schizochytrium cells with certain levels of cerulenin resulted in inhibition of 14C acetate incorporation into short chain fatty acids without affecting labeling of PUFAs, indicating distinct biosynthetic pathways. A single large gene encoding the presumed short chain fatty acid synthase (FAS) was cloned and sequenced. Based on sequence homology and domain organization, the Schizochytrium FAS resembles a fusion of fungal FAS beta and alpha subunits.

An efficient and simple protocol for the preparation of a series of 9H-carbazoles by photostimulated SRN1 substitution reactions is presented. Substituted 9H-carbazoles were synthesized in low to excellent yields (up to 96%) through an intramolecular C-N bond formation of 2′-halo[1,1′-biphenyl]-2-amines by the photoinitiated SRN1 mechanism under mild and "transition-metal-free" conditions. The biphenylamines used as substrates were obtained with isolated yields ranging from 21% to 84% by two approaches: (A) the cross-coupling Suzuki-Miyaura reaction and (B) the radical arylation of anilines. Some key aspects of the proposed mechanism were evaluated at the B3LYP/6-311+G∗ level. Fil: Guerra, Walter Damián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina Fil: Rossi, Roberto Arturo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina Fil: Pierini, Adriana Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina Fil: Barolo, Silvia Maricel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina

Eight biomedical glasses and three commercial glasses, as finely divided powders, were tested as initiators for the ring-opening polymerization of ϵ-caprolactone in bulk and in vacuo at 185°C. All the glass powders were able to initiate the polymerization, along with Pyrex, which was totally inert toward the monomer as the inner surface of a phial. The obtained polymers were examined with Fourier infrared transform spectroscopy and atomic force microscopy. The molecular weights were measured by viscometry in CHCl3. The presence of a fraction of the polymer firmly linked to the glass was quantitatively checked by the determination of the weight loss from the residues of the extraction with CHCl3 after calcination in a kiln at 945°C. The molecular weights and weight losses per unit surface were elaborated mathematically so that a possible correlation between these properties and the atomic compositions of the glasses could be better investigated. Two possible initiation mechanisms, induced by the hydroxyls present on the glass surface, were proposed: one for free poly(ϵ-caprolactone) and one for poly(ϵ-caprolactone) linked to the glass. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 87: 1579–1586, 2003

Phosphorylation of the C-terminal Akt/PKA consensus site activates endothelial NOS (eNOS) and decreases its calcium/calmodulin requirement, but no definite physiological role for phosphorylation of the homologous site in neuronal NOS (nNOS) has been described. Therefore, we created a knock-in mouse in which nNOS Serine-1412 (S1412) is mutated to non-phosphorylatable alanine (nNOSS1412A). We tested whether nNOSS1412A ileal rings have altered nitrergic inhibitory non-adrenergic, non-cholinergic (NANC) signaling in a physiological organ bath. Under NANC conditions, electrical field stimulation (EFS; 20V, 2msec, 1Hz, 30sec train) relaxed WT to a greater degree than nNOSS1412A ileum (73 ± 20% inhibition vs 31 ± 17%; N=3). Similarly, pharmacological treatment with forskolin (FSK) also relaxed WT more effectively than nNOSS1412A ileum (IC50 95% CI 35-44nM; N=42 vs 78-107nM; N=29). Application of selective nNOS (NANT) or pan-NOS antagonists (L-NAME) reduced FSK dependent relaxation identically in WT and nNOSS1412A ilea (IC50 ~150nM for both genotypes), confirming that a portion of FSK relaxation was due to nNOS activity. Akt and PKA are putative nNOS S1412 kinases. While there was no difference between WT and nNOSS1412A relaxation with FSK using the Akt inhibitor MK-2206, PKA inhibitors (H-89, Myr-PKI, and Rp-cAMPs) only decreased FSK relaxation in WT ileum (~3-fold increase in IC50) but not in nNOSS1412A ileum, suggesting a critical link between PKA and nNOS. Together, our data identify a novel NANC neuroinhibitory mechanism in which PKA phosphorylation of nNOS S1412 leads to calcium independent neuronal NO production and smooth muscle relaxation. This previously unrecognized NO physiology might be important in other cell types and organ systems as well.

This work presents a relationship between the thermal properties in different polyethylene samples analyzed by differential scanning calorimetry (DSC). The morphology and structural changes were studied by transmission electron microscopy (TEM). A preparative method involving surface etching was used to obtain surface replicas. The main morphological features of the samples, characterized by lamellar structure, obtained in this work by TEM give values of mean lamellar thickness from 900 to 500 Ĺ in the highest branch content and molecular mass. Enthalpies of melting allowed to calculate crystallinity; given values in the range from 47 to 68%.

Composites between alumina and the biore-sorbable (poly(e-caprolactone)-block-poly(oxyethylene)-block-poly(e-caprolactone) copolymer were obtained by reacting e-caprolactone with preformed poly(ethylene glycol), in the presence of ceramic alumina powder, at 185°C under vacuum. The mechanical properties, tested by compression and flexural strengths and Young's modulus, show that the copolymer interacts poorly with the alumina grains. Both scanning electron and atomic force microscopy show a scare wettability between alumina and copolymer, as well as the aggregation of alumina micro-particles into clusters of big size. Both mechanical and morphological tests seem to indicate a stronger interaction between the alumina micro-particles than between the alumina surface and the reaction mixture during the polymerization, as well as a compacting effet by alumina on the forming copolymer. The FT-IR spectra of the composites show both copolymer and alumina absorption bands. The FT-IR analysis on the fractions of an extraction which CHCl 3 indicates the presence of traces of poly(e-caprolactone), stably linked to alumina. The polymerization of e-caprolactone with alumina alone in the same conditions gives poly(e-caprolactone), mainly free and in minor part linked to the alumina surface. Two polymerization mechanisms, simultaneously occuring, are proposed. The most relevant result of this work is the lack of chemical inertness of alumina towards e-caprolactone, which leads to reconsider also the use of alumina as a biochemically inert material.

The 5-fluorouracil release by biodegradable ɛ-caprolactone and L-lactide copoly(ester-ether-ester)s was tested. The drug-copolymer mixture was formed by fusion in thin sheets, which were dipped in Dulbecco's PBS for time intervals ranging from one hour to two months. Each experiment shows a fast initial release, which subsequently slows down and stops at a limiting value, depending on the copolymer composition. This behavior was attributed to an extraction of the drug present on the sheet surface, due only to its shape, and to hydrogen bonds between the drug and the copolymers. The results obtained lead to a possibility of using such copolymers as “time-delayed” drug-releasing systems, when formed in specimens with smaller surface-to-volume ratio, which could minimize the fast initial extraction. © 2001 Kluwer Academic Publishers

Fibers made by a bioresorbable poly(e-caprolactone)-block-poly(oxyethylene)-block-poly(e-caprolactone) copolymer, having a number average molecular mass of about 200 000 Da and an average molar composition of 66% oxycaproyl units and 34% oxyethylene units, were melt-spun, with the aim at using them as suture threads. Their properties were investigated by the stress-strain test and by differential scanning calorimetry (DSC). The results obtained show that the properties of this material depend very strongly on the alignment of its macromolecules. In particular, the only partial alignment, obtainable by a relatively moderate drawing just after the extrusion, leads to values of elongation at break too high for use of the fibers as suture threads. The DSC analysis reveals interesting properties of the material, but also confirms their strong dependence on the extrusion procedure and on the mechanical treatment. In conclusion, the results of this preliminary study show that the spinning technique must be improved, and that further investigations are necessary to ascertain the possibility of using these poly(ester-ether-ester)s for the fabrication of suture threads. © 1999 Kluwer Academic Publishers

Two chitosan-containing polyelectrolyte complexes, chitosan-poly(acrylic acid) and chitosan-poly(styrenesulphonate), were synthesized by polymerizing acrylic acid and sodium styrenesulphonate in the presence of chitosan and chitosan hydrochloride, respectively. The complexes were studied by optical microscopy and tested for cytotoxicity by the Neutral Red uptake, Kenacid Blue R-Binding and 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide assays. The optical microscopy confirmed the differences in crystallinity and structure already found for the two polycomplexes by other characterization techniques. The cytoxicity tests showed different influences on the cell activity by the extracts of the two polyelectrolyte complexes. Such results were discussed and correlated to the different structures of the two materials. © 1998 Chapman & Hall

The release of drugs, entrapped into the poly(allylammonium acrylate) (PAAC) polyelectrolyte complex structure, was tested for vitamin B 12 (VitB12) and tetracycline (TCY). The PAAC-drug “composites” were obtained by synthesizing PAAC in the presence of either VitB12 or TCY. Weighed samples of the “composites” were sintered as tablets, which were dipped into Dulbecco’s PBS (pH 7.3); the release was followed spectrophotometrically. The tests show very low limiting release values, with different kinetic behaviours for TCY and VitB12. The results are discussed and attributed to strong ionic interactions between the polyelectrolyte complex and the dipolar ions VitB12 and TCY.

In previous papers, we studied the hydrolytic degradation of six poly(ester-ether-ester) block copolymers, i.e. three poly(e-caprolactone)-block-poly(oxyethylene)-block-poly(e-caprolactone) copolymers and three poly(l-lactide)-block-poly(oxyethylene)-block-poly(l-lactide) copolymers. Their degradation products, 6-hydroxyhexanoic acid and l-lactic acid, have now been found to modulate endothelin release by human umbilical vein endothelial cells, with no significant alteration of the vasoconstrictor-vasodilator balance previously determined. The influence of the same degradation products on the cell proliferation has also been determined and discussed.