Synaptic vesicle transport and synaptic membrane transporter sites in excitatory amino acid nerve terminals in Alzheimer disease

The apparent L-[H-3]glutamate uptake rate (v') was measured in synaptic vesicles isolated from cerebral cortex synaptosomes prepared from autopsied Alzheimer and non-Alzheimer dementia cases, and age-matched controls. The initial synaptosome preparations exhibited similar densities of D-[H-3]aspartate membrane binding sites (B-MAX values) in the three groups. In control brain the temporal cortex D-[H-3]aspartate B-MAX was 132% of that in motor cortex, parallel with the L- [H-3]glutamate v' values (temporal = 139% of motor; NS). Unlike D- [H-3]aspartate B-MAX values, L- [H-3]glutamate v' values were markedly and selectively lower in Alzheimer brain preparations than in controls, particularly in temporal cortex. The difference could not be attributed to differential effects of autopsy interval or age at death. Non-Alzheimer dementia cases resembled controls. The selective loss of vesicular glutamate transport is consistent with a dysfunction in the recycling of transmitter glutamate.

Growth hormone receptor and insulin-like growth factor-I immunoreactivity in osteoclast-like cells during tooth eruption in the toothless (Osteopetrotic) rat following treatment with colony-stimulating factor-1

In the toothless (tl/tl) osteopetrotic rat, teeth form but fail to erupt. Treatment of tl/tl rats with colony-stimulating factor-1 (CSF-1) activates bone resorption by osteoclasts, permits tooth eruption, and up-regulates the immunoreactivity of bone marrow mononuclear cells to growth hormone receptor (GHr) and insulin-like growth factor (IGF)-I. This study examined the distribution of tartrate-resistant acid phosphatase (TRAP) and immunoreactivity for GHr and IGF-I in osteoclast-like cells located on the alveolar bone margin, adjacent to the lower first molar crown, in 14-day-old normal and tl/tl rats, following treatment with CSF-1. Osteoclast-like cells demonstrated a positive reaction for TRAP, GHr, and IGF-I in all groups. However, in tl/tl tissue, osteoclast-like cells were generally negative for GHr. There was no significant difference in the total number of TRAP, GHr, and IGF-I-positive osteoclast-like cells on the adjacent bone margin in normal, normal treated with CSF-1, and tl/tl rats. CSF-1 treatment of the tl/tl rat significantly increased the total number of osteoclast-like cells, which were positive for TRAP (p < 0.001), GHr (p < 0.05) and IGF-I (P < 0.01).

Lactobacillus ruminis is a predominant lactic acid producing bacterium in the caecum and rectum of the pig

Aims: To identify the predominant lactic acid producing bacteria in the small intestine, caecum and the rectum of the healthy pig. Methods and Results: Samples obtained from the large intestine of healthy pigs post-mortem were cultured using a modified agar-MRS medium in roll tubes. Thirteen isolates were selected on the basis of their morphological characteristics and Gram stain reaction for gene sequencing. These isolates were characterized by DNA sequence analysis of 16S rDNA. Eight isolates were identified as Lactobacillus ruminis , two as Enterococcus faecium , one as Mitsuokella multiacidus and two as Escherichia coli . Conclusion: This is the first report of Lact. ruminis as the dominant lactic acid bacteria in the large intestine of the pig. Significance and Impact of the Study: The results suggest that Lact. ruminis is a dominant bacterium in the large intestine of the healthy pig. Future work should focus on the role of this bacterium in relation to the physiological function of the intestine and the health of the animal.

Part 2. Carcass composition and fatty acid profiles of adipose tissue of male goats: effects of genotype and liveweight at slaughter

The dissected carcass composition and fatty acid profiles of intermuscular fat from 110 male goat kids from six genotypes i.e. Boer x Angora (BA), Boer x Feral (BF), Boer x Saanen (BS), Feral x Feral (1717), Saanen x Angora (SA) and Saanen x Feral (SF) and two slaughter weight groups i.e. Capretto and Chevon (liveweight at slaughter 14-22 and 30-35 kg, respectively) were compared. Carcass tissue distribution for various genotypes was: muscle (63-66%), fat (10-13%) and bone (21-24%). Genotype significantly (P < 0.05) influenced the carcass composition; BA and FF carcasses had significantly higher muscle to bone ratio, while carcasses from BS kids were leaner compared to other genotypes. However, the two slaughter weight groups did not differ significantly (P > 0.05) in terms of carcass composition, when compared at the same carcass weight. In the present study, significant (P < 0.01) correlations were observed between percentage of muscle, fat and bone in most of the primal cuts and that in the carcass side. The main saturated fatty acids (SFAs) identified were palmitic (16:0) and stearic acid (18:0), while oleic acid (18: 1, omega9) was the main unsaturated fatty acid (UFA) in the intermuscular fat from goat kids. There were significant (P < 0.05) differences between genotypes in the proportions of individual fatty acids. Adipose tissue from BS kids had significantly higher UFAs (mainly oleic acid) and thus had a significantly lower melting point compared to other genotypes. There were significantly higher proportions of palmitic acid (35%) in the adipose tissue from Capretto kids compared to that from Chevon kids (22%). The concentration of UFAs increased in the adipose tissue from Capretto to Chevon carcasses. (C) 2003 Elsevier Science B.V. All rights reserved.

Nitrogen cycling in degraded Leucaena leucocephala-Brachiaria decumbens pastures on an acid infertile soil in south-east Queensland, Australia

A grazing trial was conducted to quantify N cycling in degraded Leucaena leucocephala (leucaena)-Brachiaria decumbens (signal grass) pastures grown on an acid, infertile, podzolic soil in south-east Queensland. Nitrogen accumulation and cycling in leucaena-signal grass pastures were evaluated for 9 weeks until all of the leucaena on offer (mean 600 kg edible dry matter (EDM)/ha, 28% of total pasture EDM) was consumed. Nitrogen pools in the grass, leucaena, soil, cattle liveweight, faeces and urine were estimated. The podzolic soil (pH 4.8-5.9) was found to be deficient in P, Ca and K. Leucaena leaf tissues contained deficient levels of N, P and Ca. Grass tissues were deficient in N and P. Grazing was found to cycle 65% of N on offer in pasture herbage. However, due to the effect of the plant nutrient imbalances described above, biological N fixation by leucaena contributed only 15 kg/ha N to the pasture system over the 9-month regrowth period, of which 13 kg/ha N was cycled. Cattle retained 1.8 kg/ha N (8% of total N consumed) in body tissue and the remainder was excreted in dung and urine in approximately equal proportions. Mineral soil N concentrations did not change significantly (-3.5 kg/ha N) over the trial period. The ramifications of grazing and fertiliser management strategies, and implications for pasture rundown and sustainability are discussed.

Diagnosing the cause of proteolysis in UHT milk

Proteolysis of UHT milk during storage at room temperature is a major factor limiting its shelf-life through changes in its flavour and texture. The latter is characterised by increases in viscosity leading in some cases to gel formation. The enzymes responsible for the proteolysis are the native milk alkaline proteinase, plasmin, and heat-stable, extracellular bacterial proteinases produced by psychrotrophic bacterial contaminants in the milk prior to heat processing. These proteinases react differently with the milk proteins and produce different peptides in the UHT milk. In order to differentiate these peptide products, reversed-phase HPLC and the fluorescamine method were used to analyse the peptides soluble in 12% trichloroacetic acid (TCA) and those soluble at pH 4.6. The TCA filtrate showed substantial peptide peaks only if the milk was contaminated by bacterial proteinase, while the pH 4.6 filtrate showed peptide peaks when either or both bacterial and native milk proteinases caused the proteolysis. Results from the fluorescamine test were in accordance with the HPLC results whereby the TCA filtrate exhibited significant proteolysis values only when bacterial proteinases were present, but the pH 4.6 filtrates showed significant values when the milk contained either or both types of proteinase. A procedure based on these analyses is proposed as a diagnostic test for determining which type of proteinase-milk plasmin, bacterial proteinase, or both-is responsible for proteolysis in UHT milk. (C) 2003 Swiss Society of Food Science and Technology. Published by Elsevier Science Ltd. All rights reserved.

Effect of storage time and conditions on the cotyledon cell wall of the adzuki bean (Vigna angularis)

Two varieties of adzuki grown in Australia, Bloodwood and Erimo, were stored for up to 6 months at three temperatures (10, 20 and 30 degreesC), and two relative humidities (RH; 40 and 65%). The amount of cell wall material increased with time under all storage conditions. This increase was greatest at 30 degreesC and 40% RH. Storage time and conditions did not affect the total pectin levels in the cell wall. Erimo constantly exhibited a higher total pectin level than Bloodwood. The Bloodwood soluble pectin, Ca++ and Mg++ and Erimo Ca++ in the cell wall remained stable during storage, while the Erimo soluble pectin and Mg++ exhibited a slight decrease at 20 and 30 degreesC after 3 months of storage. (C) 2002 Elsevier Science Ltd. All rights reserved.

Flavonoids, phenolic acids and abscisic acid in Australian and New Zealand Leptospermum honeys

Flavonoids, phenolic acids and abscisic acid of Australian and New Zealand Leptospermum honeys were analyzed by HPLC. Fifteen flavonoids were isolated in Australian jelly bush honey (Leptospermum polygalifolium), with an average content of 2.22 mg/100 g honey. Myricetin (3,5,7,3',4',5'-hexahydroxyflavone), luteolin (5,7,3',4'-tetrahydroxyflavone) and tricetin (5,7,3',4',5'-pentahydroxyflavone) were the main flavonoids identified. The mean content of total phenolic acids in jelly bush honey was 5.14 mg/100 g honey, with gallic and coumaric acids as the potential phenolic acids. Abscisic acid was quantified as twice the amount (11.6 mg/100 g honey) of the phenolic acids in this honey. The flavonoid profile mainly consisted of quercetin (3,5,7,3',4'-pentahydroxyflavone), isorhamnetin (3,5,7,4'-tetrahydroxyflavone 3'-methyl ethyl), chrysin (5,7-dihydroxyflavone), luteolin and an unknown flavanone in New Zealand manuka (Leptospermum scoparium) honey with an average content of total flavonoids of 3.06 mg/100 g honey. The content of total phenolic acids was up to 14.0 mg/100 g honey, with gallic acid as the main component. A substantial quantity (32.8 mg/100 g honey) of abscisic acid was present in manuka honey. These results showed that flavonoids and phenolic acids could be used for authenticating honey floral origins, and abscisic acid may aid in this authentication. (C) 2002 Published by Elsevier Science Ltd.

Surface stickiness of drops of carbohydrate and organic acid solutions during convective drying: Experiments and modeling

Drying kinetics of low molecular weight sugars such as fructose, glucose, sucrose and organic acid such as citric acid and high molecular weight carbohydrate such as maltodextrin (DE 6) were determined experimentally using single drop drying experiments as well as predicted numerically by solving the mass and heat transfer equations. The predicted moisture and temperature histories agreed with the experimental ones within 6% average relative (absolute) error and average difference of +/- 1degreesC, respectively. The stickiness histories of these drops were determined experimentally and predicted numerically based on the glass transition temperature (T-g) of surface layer. The model predicted the experimental observations with good accuracy. A nonsticky regime for these materials during spray drying is proposed by simulating a drop, initially 120 mum in diameter, in a spray drying environment.

Preparation and characterisation of biodegradable starch-based nanocomposite materials

Different formulations of biodegradable starch-polyester blend nanocomposite materials have been film blown on a pilot scale film blowing tower. The physical properties of different films have been examined by thermal and mechanical analysis and X-ray diffraction. The results show that the addition of an organoclay (from 0 to 5 wt%) significantly improves both the processing and tensile properties over the original starch blends. Wide angle X-ray diffraction (WAXD) results indicate that the best results were obtained for 30wt% starch blends, and the level of delamination depends on the ratio of starch to polyester and amount of organoclay added. The crystallisation temperature of the nanocomposite blends is significantly lower than the base blend. This is probably due to the platelets inhibiting order, and hence crystallisation, of the starch and polyester. The mechanical and thermal properties of the blends are also sensitive to the way the clay particles are dispersed. (C) 2003 Society of Chemical Industry.

Biodegradation and ecotoxicity evaluation of a bionolle and starch blend and its degradation products in compost

A polymer based on a blend of starch and Bionolle(TM) has been prepared and tested for biodegradation in compost. The polymer was completely mineralised to carbon dioxide in 45 days. The potential toxicity of the polymer was tested against the earthworm Eisenia fetida using a modification of the American Standard for Testing Materials E1976-97. The earthworms were exposed to 30 g of the polymer for 28 days and changes in weight recorded. In addition, the polymer was firstly degraded by the compost and the worms exposed to the breakdown products for 28 days. Differences in weight were also recorded. In each case the production of juveniles was noted and all earthworms were examined for pathology. The results obtained were processed statistically using a t-test. The number of juveniles, produced from the breakdown products, was highly significant (P < 0.001) when compared to the earthworms added to the intact polymer. There was a definitely significant difference (P < 0.01, t = 3.25) in change in weight between the earthworms that were exposed to the polymer directly and those that were exposed to the breakdown products. There was no indication of any pathology of any earthworms. The polymer is considered safe for this species. (C) 2002 Elsevier Science Ltd. All rights reserved.

Radical formation on gamma-radiolysis of poly [(methacrylic acid)-co-acrylonitri

The radicals formed on gamma-radiolysis of a series of copolymers of methacrylic acid and acrylonitrile have been investigated by ESR spectroscopy. This series of copolymers spanned the full composition range and the study was carried out at 77 K and ambient temperature. The radicals formed in the copolymers at 77 and 303 K were found to be similar to those found in the two homopolymers, but in the intermediate composition range the presence of acrylonitrile propagation radicals was also detected. These radicals were not observed to be formed in significant quantities on the radiolysis of polyacrylonitrile. They are believed to result from a scission of the main chain at methacrylic acid/acrylonitrile diad sequences following loss of the methacrylic acid carboxyl group. At 77 K, the copolymers with high methacrylic acid contents were found to be more sensitive to radical formation than the methacrylic acid homopolymer, but this enhanced sensitivity was not evident at ambient temperature, where the G-values for radical formation for the copolymers were slightly less than the values for the homopolymers. (C) 2003 Society of Chemical Industry.

Ácidos húmicos extraídos do lodo de esgoto sanitário e seus efeitos em plantas

O lodo de esgoto possui alto teor de matéria orgânica porém, também estão presentes diferentes poluentes e patógenos. Desta forma, neste trabalho foi extraído ácido húmico (AH) o qual é um composto resultante do fracionamento de substâncias húmicas que compreendem um grupo de compostos de carbono gerados na decomposição de resíduos orgânicos que sofrem ressíntese formando o húmus. O ácido húmico promove diversos benefícios nos vegetais, como crescimento, elongação celular, tolerância a estresses e aumento da permeabilidade da membrana plasmática. Com base nestas características, o presente trabalho teve como objetivo avaliar os efeitos citogenéticos (ensaio Allium cepa), fisiológicos, anatômicos e bioquímicos do ácido húmico do lodo de esgoto sanitário. Foi realizada caracterização elementar do material para a definição das doses. Após 20 dias de tratamento, foram realizadas coletas do material e, posteriormente, analisadas. A caracterização química do AH indicou-o como bom condicionador para culturas apresentando elevadas taxas de C, H e N. Não foi observado efeito de toxicidade, citotoxicidade, genotoxicidade e mutagenicidade do AH. Foi verificado aumento expressivo de todos os pigmentos fotossintéticos vegetais nas concentrações mais altas (2 mM C L-1 e 4 mM C L-1). Houve aumento da expressão da ATPAse em todos os tratamentos e das enzimas do estresse oxidativo (CAT, SOD, APX, GST) em diferentes concentrações. A integração destas análises permitiu concluir que o ácido húmico do lodo de esgoto pode ser utilizado como adubo orgânico, pois foi observado benefícios no vegetal tais como maior crescimento vegetal e aumento da atividade nas enzimas do estresse oxidativo.

Electrospun silk-elastin-like fiber mats for tissue engineering applications

Protein-based polymers are present in a wide variety of organisms fulfilling structural and mechanical roles. Advances in protein engineering and recombinant DNA technology allow the design and production of recombinant protein-based polymers (rPBPs) with an absolute control of its composition. Although the application of recombinant proteins as biomaterials is still an emerging technology, the possibilities are limitless and far superior to natural or synthetic materials, as the complexity of the structural design can be fully customized. In this work, we report the electrospinning of two new genetically engineered silk-elastin-like proteins (SELPs) consisting of alternate silk- and elastin-like blocks. Electrospinning was performed with formic acid and aqueous solutions at different concentrations without addition of further agents. The size and morphology of the electrospun structures was characterized by scanning electron microscopy showing to be dependent of concentration and solvent used. Treatment with air saturated with methanol was employed to stabilize the structure and promote water insolubility through a time-dependent conversion of random coils into β-sheets (FTIR). The resultant methanol-treated electrospun mats were characterized for swelling degree (570-720%), water vapour transmission rate (1083 g/m2/day) and mechanical properties (modulus of elasticity of ~126 MPa). Furthermore, the methanol-treated SELP fiber mats showed no cytotoxicity and were able to support adhesion and proliferation of normal human skin fibroblasts. Adhesion was characterized by a filopodia-mediated mechanism. These results demonstrate that SELP fiber mats can provide promising solutions for the development of novel biomaterials suitable for tissue engineering applications.