Ag-carried CMC/functional copolymer/ODA-Mt wLED-treated NC and their responses to brain cancer cells.

The subject of this work is synthesis and characterization of novel multifunctional nanocomposite (8/2A-NC) consisting (1) carboxymethyl cellulose (CMC) as a matrix biopolymer and poly (maleic acid-alt-acrylic acid) as a reactive synthetic partner matrix polymer; (2) octadecyl amine montmorillonite (ODA-MMT) reactive organoclay provide intercalated silicate layers structures and aqueous colloidal dispersing medium, and MMT as carriers and targeting agents for anticancer agents in drug delivery systems, respectively. ODA as a intercalated surfactant finely dispersed 8/2A NC and its compatibility with matrix polymers via the interfacial polarization (complexing) and functionalization of matrix polymers by amine (ODA) and carboxylic acids from both the CMC and copolymer; (3) silver nanoparticles (AgNPs) as in-situ generated onto matrix polymers with unique nano-size and morphology parameters was synthesized. Important material science and bioengineering aspects of these investigations included (a) novel approach in synthetic pathways; (b) effects of physical and chemical structural rearrangements; (c) effects of Light Emitting Dioda (LED)-treatment on the FT-IR spectra, XRD reflection parameters, SEM-TEM morphology and nano-size and diameter distribution of AgNPs onto matrix polymers; (d) positive effect of LED-treatment of 8/2A nanocomposite and its response to the MIAPaCa-2 and U87 human brain cancer cell lines were evaluated. Novel 8/2A-NC multifunctional drug consisting unique positive, intercalating and encapsulated core-shell morphology structures, nano-size (5.6 nm) and narrow diameter distribution (94%) of AgNPs onto matrix polymers [silver NPs (0.25%) in 8/2A NC (25%)] with highest volume of contact area compared with used cancer micro-cells show lowest cell viability as an excellent anticancer platform. 8/2A-NC is a novel multifunctional drug with intercalating and encapsulated core-shell morphology structures consisting of positively charged, non-randomly distributed AgNPs with a large contact area and low diameters (5-6 nm). The anticancer properties of (This factor is not conformed experimentally in work) this drug can be explained by the following structural factors: 8/2A-NC contains a combination of active sites from protonated hydroxyl, carboxyl and amine groups; Ag+-cations and ODA-MMT with high physical and chemical surface areas. We suggest this material be further explored for anti-cancer testing.

Multifunctional colloidal nanofiber composites including dextran and folic acid as electro-active platforms.

This work presents the fabrication and characterization of novel colloidal multifunctional polymer nanofiber composites (NFCs) from water dispersion blends of intercalated silicate layered nanocomposites of poly (2-vinyl-N-pyrrolidone)/octadecyl amine-montmorillonite (ODA-MMT) and dextran/ODA-MMT as matrix and partner polymer intercalated nanocomposites in the presence of NaOH and folic acid (FA) as doping agents by green reactive electrospinning. Chemical and physical structures, surface morphology and electrical properties were investigated. Effects of matrix/partner polymer ratios, doping agents, absorption time of NaOH, and temperature on electrical parameters of NFCs were evaluated. The presence of FA and increasing dextran fraction in NFCs resulted in reducing fiber diameter and improving diameter distribution. High complexing behaviors of matrix/partner polymer chains, organoclay, FA, and NaOH significantly improved conductivity parameters, especially 5-min of absorption time (≈10-2-10-3Sm-1). The conductivity of the samples decreased with increasing temperature. NFCs fabricated for the first time are promising candidates for various biomedical, electrochemical and electronic applications as electro-active platforms.

Folic acid conjugated and Ag-carrying organoclay nanoparticles and their response to L929 fibroblast and DLD-1 cancer cells.

This work presents the synthesis and characterisation of intercalated nanocomposites (NCs) from dispersed water solution blends of octadecyl amine-montmorillonite (ODA-MMT) (NC-0), folic acid (FA) conjugated ODA-MMT (NC-1) and Ag-MMT clay as a stable silver carrying agent (NC-2). The composition, chemical/physical and morphology of NCs with in situ intercalating nanostructures were investigated. Effect of organoclay, FA and Ag-MMT on L929 fibroblast (control), human colon carcinoma (DLD-1) cell lines, and the cytotoxicity, apoptosis and necrosis degree were estimated via WST-1/hemocytometric, double staining (as a ribonuclease A enzyme based method) and fluorescence microscopy methods in a dose-dependent manner. The mentioned cell lines integrated with NCs resulted in remarkable change in both morphology and nuclei of DLD-1 and fibroblast cells by apoptosis analysis. The number of necrotic cells were remarkably increased, as the toxic effects of nanocomposite nanoparticles were applied to both cell lines. Finally, the molecular mechanism of anticancer action of functionalised organoclays was elucidated.

Fabrication and characterization of PVA/ODA-MMT-poly(MA-alt-1-octadecene)-g-graphene oxide e-spun nanofiber electrolytes and their response to bone cancer cells.

This work presents a new approach to fabrication and characterization of novel polymer nanofiber electrolytes from intercalated PVA/ODA-MMT nanocomposite as a matrix polymer and encapsulated graphene oxide (GO) nanosheets with amphiphilic reactive copolymer as partner polymers using electrospinning method. The chemical and physical structures, surface morphology, thermal behaviors and electric conductivity of nanocomposites and nanofibers were investigated using analyses methods including FTIR, XRD, SEM, DSC-TGA and conductivity analysis. Significant improvements in nanofiber morphology and size distribution were observed when GO and reactive organoclay were incorporated as reinforcement fillers into various matrix/partner solution blends. The structural factors of matrix-partner polymer nanocomposite particles with higher zeta-potential play important roles in both chemical and physical interfacial interactions and phase separation processing and also lead to the formation of nanofibers with unique surface morphologies and good conductivities. The cytotoxic, necrotic and apoptotic effects of chosen nanofibers on osteocarcinoma cells were also investigated. These multifunctional, self-assembled, nanofibrous surfaces can serve as semi-conductive and bioactive platforms in various electrochemical and bio-engineering processes, as well as reactive matrices used for the immobilization of various biopolymer precursors.

Fabrication and characterization of novel starch-grafted poly l-lactic acid/montmorillonite organoclay nanocomposites.

In this work, poly(L-lactic acid)-g-starch layered silicate nanocomposites (NCs) (PLLA-g-starch/MMT) were fabricated by intercalative bulk graft copolymerization of LA with starch, in the presence of either stannous octoate acting as a catalyst or LA-MMT organoclay acting as a cocatalyst-nanofiller. This procedure was performed inside a custom vacuum micro-reactor. To better understand the graft copolymerization mechanism, in situ processing types, interfacial interactions and nanostructure formation of PLLA-g-starch/MMT NCs, methods such as FT-IR, XRD, (1)H NMR, (13)C CP/MAS-NMR, DSC/TGA, TEM and SEM were utilized. The morphology and thermal behaviors of nanocomposites were found to be strongly dependent on the loading mass fraction of LA-MMT within the nanocomposite structure and the type of in situ processing such as interfacial, physical and chemical interactions. Preintercalated LA-MMT organoclay exhibited dual functions. It demonstrated the ability to act as a catalyst, essentially accelerating in situ graft copolymerization via esterification of LA carboxyl groups with hydroxyl groups of starch macromolecules, whilst also acting as a nananofiller-compatibilizer.

Novel multifunctional colloidal carbohydrate nanofiber electrolytes with excellent conductivity and responses to bone cancer cells.

This work presents a new approach to fabricating novel polymer nanofiber composites (NFCs) from water solution blends of PVA (hydrolyzed 89%)/ODA-MMT and Na-CMC/ODA-MMT nanocomposites as well as their folic acid (FA) incorporated modifications (NC-3-FA and NC-4-FA) through green electrospinning nanotechnology. The chemical and physical structures and surface morphology of the nanofiber composites were confirmed. Significant improvements in nanofiber morphology and size distribution of the NFC-3-FA and NFC-4-FA nanofibers with lower average means 110 and 113nm compared with those of NFC-1/NFC-2 nanofibers (270 and 323nm) were observed. The structural elements of polymer NFCs, particularly loaded partner NC-2, plays an important role in chemical and physical interfacial interactions, phase separation processing and enables the formation of nanofibers with unique morphology and excellent conductivity (NFC-3-FA 3.25×10(-9)S/cm and NFC-4-FA 8.33×10(-4)S/cm). This is attributed to the higher surface contact areas and multifunctional self-assembled supramacromolecular nanostructures of amorphous colloidal electrolytes. The anticancer activity of FA-containing nanofibers against osteocarcinoma cells were evaluated by cytotoxicity, apoptotic and necrotic analysis methods.

Controlled graft copolymerization of lactic acid onto starch in a supercritical carbon dioxide medium.

This work presents a new approach for the synthesis of a starch-g-poly L-lactic acid (St-g-PLA) copolymer via the graft copolymerization of LA onto starch using stannous 2-ethyl hexanoate (Sn(Oct)2) as a catalyst in a supercritical carbon dioxide (scCO2) medium. The effects of several process parameters, including the pressure, temperature, scCO2 flow rate and reaction time, on the polymerization yield and grafting degree were studied. Amorphous graft St-g-PLA copolymers with increased thermal stability and processability were produced with a high efficiency. The maximum grafting degree (i.e., 52% PLA) was achieved with the following reaction conditions: 6h, 100°C, 200 bar and a 1:3 (w/w) ratio of St/LA. It was concluded that these low cost biobased graft biopolymers are potential candidates for several environment-friendly applications.

Impact of MBR cleaning and breaching on passage of selected microorganisms and subsequent inactivation by free chlorine.

Membrane bioreactors (MBRs) produce better quality effluent compared to conventional treatment processes but they are still subjected to the same disinfection requirements as conventional processes by many regulatory agencies. A research study consisting of bench-, pilot- and full-scale studies was conducted to characterize effluents produced from an MBR system operating under routine and challenged conditions and to assess the disinfection requirements for these effluents. Membrane cleaning did not seem to pose a substantial risk with respect to passage of target microorganisms; however, the membrane under breached conditions (turbidity > 0.5 NTU) resulted in an increase in a total coliform bacterial concentration up to 8500 CFU/100 mL. Adenoviruses were always detected in MBR filtrate samples by PCR (method detection limit of 10(3) genome copies per 25 µL reaction) irrespective of the membrane cleaning or breaching status. Passage of MS-2 bacteriophage through a breached membrane was lower compared to total coliform bacteria potentially due to their lower densities in the mixed liquor. Despite an increase in microbial concentration, a free chlorine CT of 30 mg-min/L was sufficient to achieve greater than 5-log removal of seeded MS-2 bacteriophage and removal of total coliform bacteria at or below the method detection limit (2 CFU/100 mL) for samples with a filtrate turbidity of 1.0 NTU. If such lower CT were to be employed, a significant decrease in plant footprint and operational costs could be realized.

Characterization of effluent water qualities from satellite membrane bioreactor facilities.

Membrane bioreactors (MBRs) are often a preferred treatment technology for satellite water recycling facilities since they produce consistent effluent water quality with a small footprint and require little or no supervision. While the water quality produced from centralized MBRs has been widely reported, there is no study in the literature addressing the effluent quality from a broad range of satellite facilities. Thus, a study was conducted to characterize effluent water qualities produced by satellite MBRs with respect to organic, inorganic, physical and microbial parameters. Results from sampling 38 satellite MBR facilities across the U.S. demonstrated that 90% of these facilities produced nitrified (NH4-N <0.4 mg/L-N) effluents that have low organic carbon (TOC <8.1 mg/L), turbidities of <0.7 NTU, total coliform bacterial concentrations <100 CFU/100 mL and indigenous MS-2 bacteriophage concentrations <21 PFU/100 mL. Multiple sampling events from selected satellite facilities demonstrated process capability to consistently produce effluent with low concentrations of ammonia, TOC and turbidity. UV-254 transmittance values varied substantially during multiple sampling events indicating a need for attention in designing downstream UV disinfection systems. Although enteroviruses, rotaviruses and hepatitis A viruses (HAV) were absent in all samples, adenoviruses were detected in effluents of all nine MBR facilities sampled. The presence of Giardia cysts in filtrate samples of two of nine MBR facilities sampled demonstrated the need for an appropriate disinfection process at these facilities.

Occurrence and removal of microbial indicators from municipal wastewaters by nine different MBR systems.

Nine different membrane bioreactor (MBR) systems with different process configurations (submerged and external), membrane geometries (hollow-fiber, flat-sheet, and tubular), membrane materials (polyethersulfone (PES), polyvinylidene fluoride (PVDF), and polytetrafluoroethylene (PTFE)) and membrane nominal pore sizes (0.03-0.2 µm) were evaluated to assess the impact of influent microbial concentration, membrane pore size and membrane material and geometries on removal of microbial indicators by MBR technology. The log removal values (LRVs) for microbial indicators increased as the influent concentrations increased. Among the wide range of MBR systems evaluated, the total and fecal coliform bacteria and indigenous MS-2 coliphage were detected in 32, 9 and 15% of the samples, respectively; the 50th percentile LRVs were measured at 6.6, 5.9 and 4.5 logs, respectively. The nominal pore sizes of the membranes, membrane materials and geometries did not show a strong correlation with the LRVs.

Bioengineering functional copolymers. XXI. Synthesis of a novel end carboxyl-trithiocarbonate functionalized poly(maleic anhydride) and its interaction with cancer cells.

A novel carboxyl-trithiocarbonate functionalized polymer with a highly selective antitumor activity was synthesized by a reversible addition-fragmentation chain transfer (RAFT) polymerization of maleic anhydride (MA) with benzoyl peroxide as an initiator and S-1-dodecyl-S-(α,α'-dimethyl-α"-acetic acid)trithiocarbonate as a RAFT agent with the aim to design and synthesize an effective anticancer agent with minimum side effects. The structure, molecular weights and composition of synthesized polymers were investigated by (1)H ((13)C) NMR, MALDI-TOF-MS and GPC analyzes. It was demonstrated that RAFT polymerization of MA was accompanied by a partially controlled decarboxylation of anhydride units and the formation of conjugated double bond fragments in backbone macromolecular chains. The mechanism of interaction of pristine RAFT agent and PMA-RAFT polymer with cancer (HeLa human cervix carcinoma) and normal (L929 Fibroblast) cells was investigated by using a combination of chemical, biochemical, statistical, spectroscopic (SEM and fluorescence inverted microscope) and real-time analysis (RTCA) methods. PMA-RAFT exhibited higher and selective cytotoxicity, apoptotic and necrotic effects toward HeLa cells at relatively low concentrations (around 7.5-75 µg mL(-1), IC(50) = 11.183 µg mL(-1)) and toward Fibroblast cells at high concentrations (IC(50) > 100 µg mL(-1)). The observed highly selective antitumor activity render PMA-RAFT polymers as promising candidates for the utilization in cancer chemotherapy.

Morphology and innervation of the vestibular lagena in pigeons.

The morphological characteristics of the pigeon lagena were examined using histology, scanning electron microscopy, and biotinylated dextran amine (BDA) neural tracers. The lagena epithelium was observed to lie partially in a parasagittal plane, but was also U-shaped with orthogonal (lateral) directed tips. Hair cell planar polarities were oriented away from a central reversal line that ran nearly the length of the epithelium. Similar to the vertebrate utricle and saccule, three afferent classes were observed based upon their terminal innervation pattern, which include calyx, dimorph, and bouton fibers. Calyx and dimorph afferents innervated the striola region of the lagena, whereas bouton afferents innervated the extrastriola and a small region of the central striola known as the type II band. Calyx units had large calyceal terminal structures that innervated only type I hair cells. Dimorph afferents innervated both type I and II hair cells, with calyx and bouton terminals. Bouton afferents had the largest most complex innervation patterns and the greatest terminal areas contacting many hair cells.

Functional copolymer/organo-MMT nanoarchitectures. VI. Synthesis and characterization of novel nanocomposites by interlamellar controlled/living radical copolymerization via preintercalated RAFT-agent/organoclay complexes.

We have developed a new approach for the synthesis of polymer nanocomposites using a bifunctional reversible addition-fragmentation chain transfer (RAFT) agent, two types of organo-montmorillonites, such as a non-reactive dimethyldodecyl ammonium (DMDA)-MMT and a reactive octadecylamine (ODA)-MMT organoclays, and a radical initiator. The method includes the following stages: (1) synthesis of RAFT intercalated O-MMTs by a physical or chemical interaction of the RAFT agent having two pendant carboxylic groups [S,S-bis(alpha,alpha'-dimethyl-alpha"-acetic acid)trithiocarbonate] with surface alkyl amines of O-MMT containing tertiary ammonium cation or primary amine groups through strong H-bonding and complexing/amidization reactions, respectively, and (2) utilization of these well-dispersed and intercalated RAFT ... O-MMT complexes and their amide derivatives as new modified RAFT agents in radical-initiated interlamellar controlled/living copolymerization of itaconic acid (IA)-n-butylmethacrylate (BMA) monomer pair. The structure and compositions of the synthesized RAFT ... O-MMT complexes and functional copolymer/O-MMT hybrids were confirmed by FTIR, XRD, thermal (DSC-TGA), SEM and TEM morphology analyses. It was demonstrated that the degree of interaction/exfoliation, morphology and thermal behavior of nanocomposites significantly depended on the type of organoclay and in situ interaction, as well as on the content of flexible butyl-ester linkages as a internal plasticizer. The results of the comparative analysis of the nanocomposites structure-composition-property relations show that the functional copolymer-organoclay hybrids prepared with reactive RAFT ... ODA-MMT complex and containing a combination of partially intercalated and predominantly exfoliated nano-structures exhibit relatively higher thermal stability and fine dispersed morphology. These effects were explained by in situ interfacial chemical reactions through amidization of RAFT with surface alkyl amine of MMT clay in interlamellar copolymerization. This simple and versatile method can be applied to a wide range of functional monomer/comonomer systems and mono- and bifunctional RAFT compounds for preparation new generation of nanomaterials.

Spatial and temporal characteristics of vestibular convergence.

In all species studied, afferents from semicircular canals and otolith organs converge on central neurons in the brainstem. However, the spatial and temporal relationships between converging inputs and how these contribute to vestibular behaviors is not well understood. In the current study, we used discrete rotational and translational motion stimuli to characterize canal- and otolith-driven response components of convergent non-eye movement (NEM) neurons in the vestibular nuclear complex of alert pigeons. When compared to afferent responses, convergent canal signals had similar gain and phase ranges but exhibited greater spatial variability in their axes of preferred rotation. Convergent otolith signals also had similar mean gain and phase values to the afferent population but were spatially well-matched with the corresponding canal signals, cell-by-cell. However, neither response component alone nor a simple linear combination of these components was sufficient to predict actual net responses during combined canal-otolith stimulation. We discuss these findings in the context of previous studies of pigeon vestibular behaviors, and we compare our findings to similar studies in other species.

Bioengineering functional copolymers. XIV. Synthesis and interaction of poly(N-isopropylacrylamide-co-3,4-dihydro-2H-pyran-alt-maleic anhydride)s with SCLC cancer cells.

Novel antitumor active functional polymers with supramacromolecular structures were synthesized by a complex-radical terpolymerization of N-isopropylacrylamide (NIPAm), 3,4-dihydro-2H-pyran (DHP), and maleic anhydride (MA) with 2,2'-azoisobisbutyronitrile as a radical initiator in 1,4-dioxane at 65°C under nitrogen atmosphere. The structure and composition of terpolymers were investigated by (1)H ((13)C) NMR spectroscopy. Interaction of terpolymers with human lung small cell carcinoma (SCLC) were investigated by using different methods such as cytotoxicity, statistical, apoptotic and necrotic cell indexes, double staining and caspase-3 immunostaining, light and fluorescence inverted microscopy analyses. Investigations into structure, composition, and antitumor activity relationships revealed that terpolymers containing a combination of ionisable amide-pyran linkages and H-bonded carboxylic groups exhibited higher cytotoxicity. It was observed that terpolymer with nearly alternating structure provides a maximum concentration of ionisable and H-bonded antitumor sites, and therefore, exhibits higher in vitro cytotoxicity, apoptotic and necrotic effects towards SCLC cancer cells.

Novel compound from Trachyspermum ammi (Ajowan caraway) seeds with antibiofilm and antiadherence activities against Streptococcus mutans: a potential chemotherapeutic agent against dental caries.

AIM: The objective of this study was to isolate and characterize the active compound from Trachyspermum ammi seeds, exhibiting antibiofilm activity against Streptococcus mutans, a major causal organism of dental caries. METHODS AND RESULTS: Purification of the active compound from the seeds was performed by silica gel chromatography, and spectroscopic methods (FTIR, NMR and MS) were employed for its identification and structure determination. Antibiofilm and antiadherence activities of the active compound against S. mutans were analysed. Confocal microscopy was performed to visualize the effect of the compound on biofilm structure of S. mutans. Around 50% reduction was observed in adherence at 39·06 µg ml(-1) and in biofilm at 78·13 µg ml(-1) . It was found effective against adherent cells of S. mutans, reduced water-insoluble glucan synthesis and inhibited the reduction in pH. Confocal microscopy revealed scattered cells at sub-MIC concentration of the compound, resulting in distorted biofilm architecture in contrast to clustered cells seen in control. CONCLUSION: This study revealed a novel compound, a naphthalene derivative, isolated first time from T. ammi seeds with antibiofilm activity against S. mutans. SIGNIFICANCE AND IMPACT OF THE STUDY: Trachyspermum ammi represents an interesting source of a novel compound, (4aS, 5R, 8aS) 5, 8a-di-1-propyl-octahydronaphthalen-1-(2H)-one, with a great potential to be used as a therapeutic agent against dental caries.

Peak flux performance and microbial removal by selected membrane bioreactor systems.

A pilot study was conducted over a period of 18 months at the Point Loma Wastewater Treatment Plant (PLWWTP) in San Diego, CA to evaluate the operational and water quality performance of six selected membrane bioreactor (MBR) systems at average and peak flux operation. Each of these systems was operated at peak flux for 4 h a day for six consecutive days to assess peak flux performance. Virus seeding studies were also conducted during peak flux operation to assess the capability of these systems to reject MS-2 coliphage. When operating at steady state, these MBR systems achieved an effluent BOD concentration of <2 mg/L and a turbidity of <0.1 NTU. Peak flux for the MBR systems ranged from 56 to 76 L/m2/h (liters per square meter per hour) with peaking factors in the range of 1.5-3.2. When switching from average to peak flux operation, a reversible drop of 22-32% in temperature-corrected permeability was observed for all submerged MBR systems. The percent drop in permeability increased as MLSS concentration in the membrane tank increased from 11,100 mg/L to 15,300 mg/L and was observed to be highest for the system operating at highest MLSS concentration. Such trends were not observed with an external MBR system. Each MBR system was able to sustain a 4-h-a-day peak flow for six consecutive days with only moderate membrane fouling. The membrane fouling was quantified by measuring the drop in temperature-corrected permeability. This drop ranged from 13 to 33% over six days for different submerged MBR systems. The MBR systems achieved microbial removal in the range of 5.8-6.9 logs for total coliform bacteria, >5.5 to >6.0 logs for fecal coliform bacteria and 2.6 to >3.4 logs for indigenous MS-2 coliphages. When operating at peak flux, seeded MS-2 coliphage removal ranged from 1.0 to 4.4 logs, respectively. The higher log removal values (LRVs) for indigenous MS-2 coliphage among different MBR systems were probably the result of particle association of indigenous coliphage. Differences in membrane pore size (0.04-0.2 microm) amongst the MBR systems evaluated did not have a substantial impact on indigenous MS-2 coliphage removal, but seeded MS-2 coliphage removal varied among the different MBR systems.

Preparation and properties of invertase immobilized on a poly(maleic anhydride-hexen-1) membrane.

Poly(maleic anhydride-alt-hexen-1)(poly(MA-alt-H-1)) has been synthesized by radical polymerization and characterized by DSC, FT-IR, acid number determination, viscometric and NMR methods. Data showed that the co-polymer is an alternating co-polymer whose composition does not depend on the monomer feed composition. Invertase was immobilized onto a poly(MA-alt-H-1) membrane via glutaraldehyde and bovine serum albumin. The Km value of poly(MA-alt-H-1)-invertase was approximately 4.4-fold higher than the free enzyme, indicating decreased affinity by the invertase for its substrate (sucrose), whereas Vmax was lower for the immobilized invertase. Immobilization improved the pH stability of the enzyme, as well as its temperature stability. Immobilized samples obtained were stable and could be used many times over a period of 2 months without considerable activity loss.

Bioengineering Functional Copolymers. IX. Poly[(maleic anhydride-co-hexene-1)-g-poly(ethylene oxide)].

Amphiphilic bioengineering copolymers having a combination of hydrophilic/hydrophobic linkages and polyelectrolyte behavior, along with an ability to interact with biomacromolecules, in particular with the invertase enzyme, have been synthesized by (a) complex-radical copolymerization of maleic anhydride (MA, the acceptor) and hexene-1 (H-1, the donor) monomers with benzoyl peroxide as the initiator in 1,4-dioxane at 65 degrees C under high-conversion conditions and (b) subsequent grafting (polyesterification) of synthesized poly(MA-alt-H-1) with alpha-methoxy-omega-hydroxy-poly(ethylene oxide) (PEO). Copolymerizations were also carried out in the steady state, in order to essentially reduce the effect of copolymer composition drift. The values of the monomer reactivity ratios (r(1) and r(2)) determined by using the known terminal models of Fineman-Ross (FR) and Kelen-Tüdös (KT), as well as by nonlinear regression (NLR) analysis, are: r(1) = 0.16 and r(2) = 0.30 (FR), r(1) = 0.14 and r(2) = 0.27 (KT), and r(1) = 0.15 and r(2) = 0.29 (NLR), respectively. All the copolymers and graft copolymers were characterized by FTIR spectroscopy, (1)H{(13)C} NMR spectroscopy, viscometric measurements, and chemical (acid number), thermal (DSC and TGA), and X-ray diffraction analyses. Unlike poly(MA-alt-H-1)s, PEO macrobranched graft copolymers exhibit expressed polyelectrolyte and swelling behavior in diluted and concentrated dioxane solutions, respectively. The copolymer and its PEO hyperbranched derivatives can be used as carriers for enzyme immobilization.

Emx2 and Pax6 function in cooperation with Otx2 and Otx1 to develop caudal forebrain primordium that includes future archipallium.

One of the central issues in developmental neurobiology is how the forebrain is organized ontogenetically. The traditional view is that the anterior neuroectoderm first develops into mesencephalic and prosencephalic vesicles; the latter vesicle subsequently develops into the diencephalon and secondary prosencephalon, of which dorsal parts protrude to generate the telencephalon. The diencephalon yields the pretectum, thalamus, and prethalamus, and the telencephalon produces the archipallium, neopallium, and ganglionic eminences. By identifying cell descendants that once expressed Emx2 with use of the Cre knock-in mutant into the Emx2 locus and analyzing phenotypes of double mutants between Emx2 and Otx2/Otx1 and between Emx2 and Pax6, we propose that at the 3-6 somite stage, the anterior neuroectoderm develops into three primordia: midbrain, caudal forebrain, and rostral forebrain. The caudal forebrain primordium generates not only the pretectum, thalamus, and prethalamus but also the archipallium, cortical hem, choroid plexus, choroidal roof, and eminentia thalami. The primordium corresponds to the Emx2- or Pax6-positive region at the 3-6 somite stage that most probably does not include the future neopallium or commissural plate. Otx2 and Otx1 that are expressed in the entire future forebrain and midbrain cooperate with this Emx2 and Pax6 expression in the development of the caudal forebrain primordium; Emx2 and Pax6 functions are redundant. In the embryonic day 9.5 Emx2-/-Pax6-/- double mutant, the caudal forebrain remained unspecified and subsequently transformed into tectum in a mirror image of the endogenous one.