The neural bases of the short-term storage of verbal information are anatomically variable across individuals

What are the precise brain regions supporting the short-term retention of verbal information? A previous functional magnetic resonance imaging (fMRI) study suggested that they may be topographically variable across individuals, occurring, in most, in regions posterior to prefrontal cortex (PFC), and that detection of these regions may be best suited to a single-subject (SS) approach to fMRI analysis (Feredoes and Postle, 2007). In contrast, other studies using spatially normalized group-averaged (SNGA) analyses have localized storage-related activity to PFC. To evaluate the necessity of the regions identified by these two methods, we applied repetitive transcranial magnetic stimulation (rTMS) to SS- and SNGA-identified regions throughout the retention period of a delayed letter-recognition task. Results indicated that rTMS targeting SS analysis-identified regions of left perisylvian and sensorimotor cortex impaired performance, whereas rTMS targeting the SNGA-identified region of left caudal PFC had no effect on performance. Our results support the view that the short-term retention of verbal information can be supported by regions associated with acoustic, lexical, phonological, and speech-based representation of information. They also suggest that the brain bases of some cognitive functions may be better detected by SS than by SNGA approaches to fMRI data analysis.

Hydrogen-bonded complexes and blends of poly(acrylic acid) and methylcellulose: nanoparticles and mucoadhesive films for Ocular Delivery of Riboflavin

Poly(acrylic acid) (PAA) and methylcellulose (MC) are able to form hydrogen-bonded interpolymer complexes (IPCs) in aqueous solutions. In this study, the complexation between PAA andMC is explored in dilute aqueous solutions under acidic conditions. The formation of stable nanoparticles is established,whose size and colloidal stability are greatly dependent on solution pH and polymers ratio in the mixture. Poly(acrylic acid) and methylcellulose are also used to prepare polymeric films by casting from aqueous solutions. It is established that uniform films can be prepared by casting from polymer mixture solutions at pH 3.4–4.5. At lower pHs (pH<3.0) the films have inhomogeneous morphology resulting from strong interpolymer complexation and precipitation of polycomplexes, whereas at higher pHs (pH 8.3) the polymers form fully immiscible blends because of the lack of interpolymer hydrogen-bonding. The PAA/MC films cast at pH 4 are shown to be non-irritant to mucosal surfaces. These films provide a platform for ocular formulation of riboflavin, a drug used for corneal crosslinking in the treatment of keratoconus. An in vitro release of riboflavin as well as an in vivo retention of the films on corneal surfaces can be controlled by adjusting PAA/MC ratio in the formulations.

Enhancing the crystallization and orientation of electrospinning poly (lactic acid) (PLLA) by combining with additives

PLLA is a thermoplastic biopolymer and can be used in industrial applications for medical and filtration applications. The brittleness of PLLA is attributed to slow crystallization rates and its glass transition temperature (Tg) is high (60 °C); for this reason, its applications are limited. The orientation, morphology, and crystal structure of the electrospun fibers was investigated by SEM, POM, DSC, FTIR, XRD, and SAXS. Combining with additives leads to a large decrease of fiber diameter, viscosity, and changes of fiber morphology and crystal structure compared to pure PLLA. DSC showed that the Tg of PLLA decreased about 15 °C and there was no change in relaxation enthalpy by the addition of plasticizer. FT-IR indicate a strong interaction between PLLA and additives; a new band appears in the PLLA blend at 1,756 cm−1 at room temperature as a crystalline band without any annealing. In addition, WAXD indicated that the intensities of the two peaks at (200/110) and (203) increased for the blend at room temperature without any annealing in comparison with PLLA; this means that PHB crystallizes in the amorphous region of PLLA. The POM experiments agree with the results from DSC, FTIR, and WAXS measurements, confirming that adding PHB results in an increase in the number of nuclei with much smaller spherulites and enhances the crystallization behavior of this material, thereby improving its potential for applications.

An intronic G run within HIV-1 intron 2 is critical for splicing regulation of vif mRNA

Within target T lymphocytes, human immunodeficiency virus type I (HIV-1) encounters the retroviral restriction factor APOBEC3G (apolipoprotein B mRNA-editing enzyme, catalytic polypeptide-like 3G; A3G), which is counteracted by the HIV-1 accessory protein Vif. Vif is encoded by intron-containing viral RNAs that are generated by splicing at 3' splice site (3'ss) A1 but lack splicing at 5'ss D2, which results in the retention of a large downstream intron. Hence, the extents of activation of 3'ss A1 and repression of D2, respectively, determine the levels of vif mRNA and thus the ability to evade A3G-mediated antiviral effects. The use of 3'ss A1 can be enhanced or repressed by splicing regulatory elements that control the recognition of downstream 5'ss D2. Here we show that an intronic G run (G(I2)-1) represses the use of a second 5'ss, termed D2b, that is embedded within intron 2 and, as determined by RNA deep-sequencing analysis, is normally inefficiently used. Mutations of G(I2)-1 and activation of D2b led to the generation of transcripts coding for Gp41 and Rev protein isoforms but primarily led to considerable upregulation of vif mRNA expression. We further demonstrate, however, that higher levels of Vif protein are actually detrimental to viral replication in A3G-expressing T cell lines but not in A3G-deficient cells. These observations suggest that an appropriate ratio of Vif-to-A3G protein levels is required for optimal virus replication and that part of Vif level regulation is effected by the novel G run identified here.

A review of biochars’ potential role in the remediation, revegetation and restoration of contaminated soils

Biochars are biological residues combusted under low oxygen conditions, resulting in a porous, low density carbon rich material. Their large surface areas and cation exchange capacities, determined to a large extent by source materials and pyrolysis temperatures, enables enhanced sorption of both organic and inorganic contaminants to their surfaces, reducing pollutant mobility when amending contaminated soils. Liming effects or release of carbon into soil solution may increase arsenic mobility, whilst low capital but enhanced retention of plant nutrients can restrict revegetation on degraded soils amended only with biochars; the combination of composts, manures and other amendments with biochars could be their most effective deployment to soils requiring stabilisation by revegetation. Specific mechanisms of contaminant-biochar retention and release over time and the environmental impact of biochar amendments on soil organisms remain somewhat unclear but must be investigated to ensure that the management of environmental pollution coincides with ecological sustainability.

Targeted activation of toll-like receptors: conjugation of a toll-like receptor 7 agonist to a monoclonal antibody maintains antigen binding and specificity

Therapeutic activation of Toll-like receptors (TLR) has potential for cancer immunotherapy, for augmenting the activity of anti-tumor monoclonal antibodies (mAbs), and for improved vaccine adjuvants. A previous attempt to specifically target TLR agonists to dendritic cells (DC) using mAbs failed because conjugation led to non-specific binding and mAbs lost specificity. We demonstrate here for the first time the successful conjugation of a small molecule TLR7 agonist to an anti-tumour mAb (the anti-hCD 20 rituximab) without compromising antigen specificity. The TLR7 agonist UC-1V150 was conjugated to rituximab using two conjugation methods and yield, molecular substitution ratio, retention of TLR7 activity and specificity of antigen binding were compared. Both conjugation methods produced rituximab-UC-1V150 conjugates with UC-1V150 : rituximab ratio ranging from 1:1 to 3:1 with drug loading quantified by UV spectroscopy and drug substitution ratio verified by MALDI TOF mass spectroscopy. The yield of purified conjugates varied with conjugation method, and dropped as low as 31% using a method previously described for conjugating UC-1V150 to proteins, where a bifunctional crosslinker was firstly reacted with rituximab, and secondly to the TLR7 agonist. We therefore developed a direct conjugation method by producing an amine-reactive UV active version of UC-1V150, termed NHS:UC-1V150. Direct conjugation with NHS:UC-1V150 was quick and simple and gave improved conjugate yields of 65-78%. Rituximab-UC-1V150 conjugates had the expected pro-inflammatory activity in vitro (EC50 28-53 nM) with a significantly increased activity over unconjugated UC-1V150 (EC50 547 nM). Antigen binding and specificity of the rituxuimab-UC-1V150 conjugates was retained, and after incubation with human peripheral blood leukocytes, all conjugates bound strongly only to CD20-expressing B cells whilst no non-specific binding to CD20-negative cells was observed. Selective targeting of Toll-like receptor activation directly within tumors or to DC is now feasible.

Indoor microclimate in a South Africa School: impact of indoor environmental factors

Demand for good indoor air quality is increasing as people recorgnise the risks to their health and productivity from indoor pollutants. There is a tendency to reduce ventilation rates to ensure energy conservation in buildings; in this instance schools. However, evidence reviewed shows that this can be detrimental to health and wellbeing in schools because of the learner density within a small area (1.8 - 2.4m2/person); eventually indicating that carbon dioxide (CO2) levels can rise to very high levels in classroom occupancy periods. A preliminary study to investigate the impact of indoor environmental parameters has been performed in a secondary school classroom in Pretoria, South Africa. Factors monitored include temperature, relative humidity, lighting, air velocities and CO2 concentrations. From the results low air velocities are recorded (i.e. 0.1-0.3m/s) impacting on the retention of CO2 build-up in the classroom. Results presented in this paper are the initial findings of ongoing research.

Self-assembly of telechelic tyrosine end-capped PEO and poly(alanine) polymers in aqueous solution

The self-assembly in aqueous solution of three novel telechelic conjugates comprising a central hydrophilic polymer and short (trimeric or pentameric) tyrosine end-caps has been investigated. Two of the conjugates have a central poly(oxyethylene) (polyethylene oxide, PEO) central block with different molar masses. The other conjugate has a central poly(l-alanine) (PAla) sequence in a purely amino-acid based conjugate. All three conjugates self-assemble into β-sheet based fibrillar structures, although the fibrillar morphology revealed by cryogenic-TEM is distinct for the three polymers—in particular the Tyr5-PEO6k-Tyr5 forms a population of short straight fibrils in contrast to the more diffuse fibril aggregates observed for Tyr5-PEO2k-Tyr5 and Tyr3-PAla-Tyr3. Hydrogel formation was not observed for these samples (in contrast to prior work on related systems) up to quite high concentrations, showing that it is possible to prepare solutions of peptide–polymer-peptide conjugates with hydrophobic end-caps without conformational constraints associated with hydrogelation. The Tyr5-PEO6k-Tyr5 shows significant PEO crystallization upon drying in contrast to the Tyr5-PEO2k-Tyr5 conjugate. Our findings point to the remarkable ability of short hydrophobic peptide end groups to modulate the self-assembly properties of polymers in solution in model peptide-capped “associative polymers”. Retention of fluidity at high conjugate concentration may be valuable in potential future applications of these conjugates as bioresponsive or biocompatible materials, for example exploiting the enzyme-responsiveness of the tyrosine end-groups

High hydrostatic pressure blanching of baby spinach (Spinacia oleracea L.)

Given the high susceptibility of baby spinach leaves to thermal processing, the use of high hydrostatic pressure (HHP) is explored as a non-thermal blanching method. The effects of HHP were compared with thermal blanching by following residual activity of polyphenol oxidases and peroxidases, colour retention, chlorophyll and carotenoids content, antioxidant capacity and total polyphenols content. Spinach subjected to 700 MPa at 20 ºC for 15 min represented the best treatment among the conditions studied due to its balanced effect on target enzymes and quality indices. The latter treatment reduced enzyme activities of polyphenol oxidases and peroxidases by 86.4 and 76.7 %, respectively. Furthermore, leaves did not present changes in colour and an increase by 13.6 % and 15.6 % was found in chlorophyll and carotenoids content, respectively; regarding phytochemical compounds, retentions of 28.2 % of antioxidant capacity and 77.1 % of polyphenols content were found. Results demonstrated that HHP (700 MPa) at room temperature, when compared with thermal treatments, presented better retention of polyphenols, not significantly different chlorophyll and carotenoids content and no perceptible differences in the instrumental colour evaluated through ΔE value; therefore, it can be considered a realistic practical alternative to the widely used thermal blanching.

Effect of monensin on mineral balance in growing ruminants reared under different environmental temperatures

To test the effect of monensin on the mineral balance of growing cattle under different environmental temperatures, 24 male steers were assigned in a 2 x 2 factorial arrangement, contrasting 0 and 85 mg monensin/animal per day at 24.3 and 33.2 degrees C (environmental temperatures). Monensin effect was directly modulated by the environmental temperature: it increased apparent retentions of P (P=0.066), Na (P=0.005) and K (P=0.003), at the higher temperature and decreased these apparent retentions at the lower temperature, as compared with non-supplemented animals. Monensin increased fecal Ca (P=0.037), and urinary P (P=0.002), Na (P=0.003), K (P=0.014), Mg (P=0.051) and Zn (P=0.091), with higher concentrations of these minerals in animals held at 24.3 degrees C and lower concentrations in those at 33.2 degrees C, as compared with non-supplemented animals. Monensin decreased serum Mg (P=0.001) and increased serum Zn (P=0.071) in animals at 33.2 degrees C and increased serum Mg and decreased serum Zn at 24.3 degrees C. Irrespective of temperature, monensin increased both apparent absorption (P=0.058) and apparent retention (P=0.093) of P, and also urine Cu (P=0.085). Environmental temperature modulated monensin effects on mineral balance. Monensin increased apparent retention of several minerals in animals under heat stress. (C) 2007 Elsevier B.V. All rights reserved.

Garlic powder and wheat bran as fillers: Their effect on the physicochemical properties of edible biocomposites

Biocomposites with two different fillers, garlic and wheat bran, were studied. They were based on cassava starch and contained glycerol as a plasticizer and potassium sorbate as an antimicrobial agent and were characterized by scanning electron microscopy (SEM), differential scanning calorimetry (DSC) and infrared spectroscopy (IR). The mechanical performance at room and lower temperatures was also studied. SEM micrographies of fractured surfaces of the wheat bran composite films showed some ruptured particles of fiber while fibrils of garlic on the order of nanometers were observed when garlic composite films were studied. Mechanical tests, at room temperature, showed that the addition of wheat bran led to an increment in the storage modulus (E`) and hardening and a decrease in Tan delta, while the garlic composite showed a diminishing in the E` and hardening and did not produce significant changes in Tan delta values when compared with systems without fillers (matrix). In the range between -90 degrees C and 20 degrees C. all the materials studied presented two peaks in the Tan delta curve. In the case of the wheat bran composite, both relaxation peaks shifted slightly to higher temperatures, broadened and diminished their intensity when compared with those of the matrix; however garlic composite showed a similar behavior to the matrix. DSC thermograms of aqueous systems showed a slight shift of gelatinization temperature (T(gelatinization)) to higher values when the fillers were present. Thermograms of films showed that both, garlic and wheat bran composites, had a lower melting point than the matrix. IR data indicated that interaction between starch and fillers determined an increase in the availability of hydroxyl groups to be involved in a dynamic exchange with water. (C) 2010 Elsevier B.V. All rights reserved.

Reduction of oil uptake in deep fat fried chicken nuggets using edible coatings based on cassava starch and methylcellulose

The aim of this study was to evaluate the effect of edible coatings based on methylcellulose (MC) and cassava starch (CS) to reduce oil uptake and improve water retention of chicken nuggets during deep fat frying. Edible coatings were prepared with I g of MC/100 g solution and 4 g of CS/100 g solution, with 25 or 55 g glycerol/100 g biopolymer. These solutions were applied to nugget samples before battering. Pre-fried and fried nuggets were analyzed to determine lipid and water contents. Color and texture were also measured in the fried nuggets. In general, there was no effect of the two concentrations of plasticizer of either of the biopolymers on the water retention of whole nuggets. But, higher oil uptake reduction, and consequently, lower lipid content was observed on nuggets coated with CS and 25% plasticizer. The coated samples were darker and had a brighter yellow color when compared with the control. There was also a significant decrease in the shearing force of the fried coated samples, indicating reduced hardness of these samples.

Paleoamerican Morphology in the Context of European and East Asian Late Pleistocene Variation: Implications for Human Dispersion Into the New World

Early American crania show a different morphological pattern from the one shared by late Native Americans. Although the origin of the diachronic morphological diversity seen on the continents is still debated, the distinct morphology of early Americans is well documented and widely dispersed. This morphology has been described extensively for South America, where larger samples are available. Here we test the hypotheses that the morphology of Early Americans results from retention of the morphological pattern of Late Pleistocene modern humans and that the occupation of the New World precedes the morphological differentiation that gave rise to recent Eurasian and American morphology. We compare Early American samples with European Upper Paleolithic skulls, the East Asian Zhoukoudian Upper Cave specimens and a series of 20 modern human reference crania. Canonical Analysis and Minimum Spanning Tree were used to assess the morphological affinities among the series, while Mantel and Dow-Cheverud tests based on Mahalanobis Squared Distances were used to test different evolutionary scenarios. Our results show strong morphological affinities among the early series irrespective of geographical origin, which together with the matrix analyses results favor the scenario of a late morphological differentiation of modern humans. We conclude that the geographic differentiation of modern human morphology is a late phenomenon that occurred after the initial settlement of the Americas. Am J Phys Anthropol 144:442-453, 2011. (c) 2010 Wiley-Liss, Inc.

Peptidomic Analysis of Human Cell Lines

Peptides have been proposed to function in intracellular signaling within the cytosol. Although cytosolic peptides are considered to be highly unstable, a large number of peptides have been detected in mouse brain and other biological samples. In the present study, we evaluated the peptidome of three diverse cell lines: SH-SY5Y, MCF7, and HEIC293 cells. A comparison of the peptidomes revealed considerable overlap in the identity of the peptides found in each cell line. The majority of the observed peptides are not derived from the most abundant or least stable proteins in the cell, and approximately half of the cellular peptides correspond to the N- or C- termini of the precursor proteins. Cleavage site analysis revealed a preference for hydrophobic residues in the PI position. Quantitative peptidomic analysis indicated that the levels of most cellular peptides are not altered in response to elevated intracellular calcium, suggesting that calpain is not responsible for their production. The similarity of the peptidomes of the three cell lines and the lack of correlation with the predicted cellular degradome implies the selective formation or retention of these peptides, consistent with the hypothesis that they are functional in the cells.

Sequence diversity and evolutionary dynamics of the dimorphic antigen merozoite surface protein-6 and other Msp genes of Plasmodium falciparum

Immune evasion by Plasmodium falciparum is favored by extensive allelic diversity of surface antigens. Some of them, most notably the vaccine-candidate merozoite surface protein (MSP)-1, exhibit a poorly understood pattern of allelic dimorphism, in which all observed alleles group into two highly diverged allelic families with few or no inter-family recombinants. Here we describe contrasting levels and patterns of sequence diversity in genes encoding three MSP-1-associated surface antigens of P. falciparum, ranging from an ancient allelic dimorphism in the Msp-6 gene to a near lack of allelic divergence in Msp-9 to a more classical multi-allele polymorphism in Msp-7 Other members of the Msp-7 gene family exhibit very little polymorphism in non-repetitive regions. A comparison of P. falciparum Msp-6 sequences to an orthologous sequence from P. reichenowi provided evidence for distinct evolutionary histories of the 5` and 3` segments of the dimorphic region in PfMsp-6, consistent with one dimorphic lineage having arisen from recombination between now-extinct ancestral alleles. In addition. we uncovered two surprising patterns of evolution in repetitive sequence. Firsts in Msp-6, large deletions are associated with (nearly) identical sequence motifs at their borders. Second, a comparison of PfMsp-9 with the P. reichenowi ortholog indicated retention of a significant inter-unit diversity within an 18-base pair repeat within the coding region of P. falciparum, but homogenization in P. reichenowi. (C) 2009 Elsevier B.V. All rights reserved.