Front-face fluorescence measurement of photosensitizers and lipid oxidation products during the photooxidation of butter

This paper shows that fluorescence spectroscopy can measure both degradation of photosensitizers and formation of lipid oxidation products in light-exposed butter. The photosensitizers were already notably degraded after 4 h of light exposure, whereas fluorescent lipid oxidation products were detected after 5 d. The fluorescence measurements were highly correlated with sensory assessments of acidic and rancid flavor. Photosensitizer degradation is therefore a promising indirect indicator of the onset of lipid oxidation in butter. Sensory analysis and measurement of peroxide value showed that the level of lipid oxidation was significantly higher for butter stored in air compared with butter stored in nitrogen (N₂). This might be explained by the formation of singlet oxygen from direct photooxidation and type II photosensitized oxidation. Addition of the singlet oxygen quencher β-carotene reduced the rancid flavor intensity in the air and N₂ packages from 9.0 to 4.9 and from 6.5 to 4.7, respectively. Results indicate that lipid oxidation in the butter stored in N₂ was mainly caused by type I photosensitized reactions, because addition of β-carotene had little effect on the rancid flavor intensity.     

Cluster analysis and disease mapping--why, when, and how? A step by step guide

Growing public awareness of environmental hazards has led to an increased demand for public health authorities to investigate geographical clustering of diseases. Although such cluster analysis is nearly always ineffective in identifying causes of disease, it often has to be used to address public concern about environmental hazards. Interpreting the resulting data is not straightforward, however, and this paper presents a guide for the non-specialist. The pitfalls include the fact that cluster analyses are usually done post hoc, and not as a result of a prior hypothesis. This is particularly true for investigations prompted by reported clusters, which have the inherent danger of overestimating the disease rate through "boundary shrinkage" of the population from which the cases are assumed to have arisen. In disease surveillance the problem of making multiple comparisons can be overcome by testing for clustering and autocorrelation. When rates of disease are illustrated in disease maps undue focus on areas where random fluctuation is greatest can be minimised by smoothing techniques. Despite the fact that cluster analyses rarely prove fruitful in identifying causation, they may-like single case reports-have the potential to generate new knowledge.     

Regulation in the heart field of zebrafish

In many vertebrates, removal of early embryonic heart precursors can be repaired, leaving the heart and embryo without visible deficit. One possibility is that this 'regulation' involves a cell fate switch whereby cells, perhaps in regions surrounding normal progenitors, are redirected to the heart cell fate. However, the lineage and spatial relationships between cells that are normal heart progenitors and those that can assume that role after injury are not known, nor are their molecular distinctions. We have adapted a laser-activated technique to label single or small patches of cells in the lateral plate mesoderm of the zebrafish and to track their subsequent lineage. We find that the heart precursor cells are clustered in a region adjacent to the prechordal plate, just anterior to the notochord tip. Complete unilateral ablation of all heart precursors with a laser does not disrupt heart development, if performed before the 18-somite stage. By combining extirpation of the heart precursors with cell labeling, we find that cells anterior to the normal cardiogenic compartments constitute the source of regulatory cells that compensate for the loss of the progenitors. One of the earliest embryonic markers of the premyocardial cells is the divergent homeodomain gene, Nkx2.5. Interestingly, normal cardiogenic progenitors derive from only the anterior half of the Nkx2.5-expressing region in the lateral plate mesoderm. The posterior half, adjacent to the notochord, does not include cardiac progenitors and the posterior Nkx2.5-expressing cells do not contribute to the heart, even after ablation of the normal cardiogenic region. The cells that can acquire a cardiac cell fate after injury to the normal progenitors also reside near the prechordal plate, but anterior to the Nkx2.5-expressing domain. Normally they give rise to head mesenchyme. They share with cardiac progenitors early expression of GATA 4. The location of the different elements of the cardiac field, and their response to injury, suggests that the prechordal plate supports and/or the notochord suppresses the cardiac fate.     

Protein kinase C in platelets of depressed patients

Though described in 1769, the etiology of Zenker's diverticulum remains unclear. Various primary esophageal motor disorders have been proposed, but no consistent manometric pattern or anatomic etiology has been uniformly recognized. An association with clinical neurologic disease at our institution prompted a review of 12 cases of Zenker's diverticulum in patients over 60 years of age, treated in the last 8 years. Nine patients (75%) underwent cricopharyngeus myotomy and diverticulectomy, with uniformly good results. Ten patients (83%) had an associated neurologic disorder, substantiated by cranial CT or MRI, in most cases. A wide range of neurologic problems were identified, but a strong trend toward brainstem or basilar lesions was present. As expected, the etiology of the neurologic abnormality in most patients in this group was cerebrovascular disease, but two patients had peripheral neuropathies. We suggest that the etiology of Zenker's diverticulum in the elderly may be neurologic in origin. Esophageal motor disorders, including incomplete upper esophageal sphincter opening and increased hypopharyngeal pressures, which may result in Zenker's diverticulum, may be a manifestation of central or peripheral neurologic disease in the elderly.     

Physical and antibacterial properties of edible films formulated with apple skin polyphenols

Fruit and vegetable skins have polyphenolic compounds, terpenes, and phenols with antimicrobial and antioxidant activity. These flavoring plant essential oil components are generally regarded as safe. Edible films made from fruits or vegetables containing apple skin polyphenols have the potential to be used commercially to protect food against contamination by pathogenic bacteria. The main objective of this study was to evaluate physical properties as well as antimicrobial activities against Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella enterica of apple skin polyphenols at 0% to 10% (w/w) concentrations in apple puree film-forming solutions formulated into edible films. Commercial apple skin polyphenol powder had a water activity of 0.44 and high total soluble phenolic compounds and antioxidant capacity (995.3 mg chlorogenic acid/100 g and 14.4 mg Trolox/g, respectively). Antimicrobial activities of edible film containing apple skin polyphenols were determined by the overlay method. Apple edible film with apple skin polyphenols was highly effective against L. monocytogenes. The minimum concentration need to inactive L. monocytogenes was 1.5%. However, apple skin polyphenols did not show any antimicrobial effect against E. coli O157:H7 and S. enterica even at 10% level. The presence of apple skin polyphenols reduced water vapor permeability of films. Apple skin polyphenols increased elongation of films and darkened the color of films. The results of the present study show that apple skin polyphenols can be used to prepare apple-based antimicrobial edible films with good physical properties for food applications by direct contact.     

Impact on the photothermal emission from single wall nanotubes by some alkali halide salts

We demonstrate that alkali-halide salts, particularly potassium bromide, can reduce the photothermal emission (PTE) from single walled carbon nanotubes (SWNT). PTE is a prominent spectral feature in Raman spectroscopy when a near infrared laser is used to analyze a dark colored sample. We subsequently show that trapping salts inside SWNT and coating SWNT with the salt has a more pronounced impact on not only reducing PTE, but also enhancing the intensity of the Raman spectral features. The effect, which we have called nanotube enhanced Raman spectroscopy (NERS), has differences and similarities to the widely studied surface enhanced Raman spectroscopy (SERS).     

The influence of different zeolitic supports on hydrogen production and waste degradation

The photocatalyst composition affects the chemical–physical properties and directly impacts photocatalytic activity, both in the hydrogen production and degradation of organic contaminants. In this work, the influence of zeolitic structures NaA, NaY, and ZSM-5 combined with a 10% active phase, TiO₂ catalyst doped with 1% copper, and cobalt cocatalysts was tested to mineralize the reactive blue dye (CI250) and to produce hydrogen by photocatalysis under ultraviolet radiation. The band gap energy was affected mainly by the cocatalyst, while the Brunauer-Emmett-Teller method (BET) area was affected by the zeolite structure as well as the X-ray diffraction (XRD). The most active catalyst was the Cu@TiO₂/NaY, which promoted a hydrogen production rate of 240 μmolH₂gcat⁻¹ using 10% ethanol (v/v) aqueous solution as a sacrificial agent and mineralization of 53% of the organic dye, followed by the catalysts impregnated on ZSM-5 zeolites, which had discolouration up to 50% and hydrogen evolution of 92.6 and 109.7 μmolH₂gcat⁻¹ for the catalyst doped with Cu and Co, respectively.     

Epitope mapping using mRNA display and a unidirectional nested deletion library

In vitro selection targeting an anti-polyhistidine monoclonal antibody was performed using mRNA display with a random, unconstrained 27-mer peptide library. After six rounds of selection, epitope-like peptides were identified that contain two to five consecutive, internal histidines and are biased for arginine residues, without any other identifiable consensus. The epitope was further refined by constructing a high-complexity, unidirectional fragment library from the final selection pool. Selection by mRNA display minimized the dominant peptide from the original selection to a 15-residue functional sequence (peptide Cmin: RHDAGDHHHHHGVRQ; K(D) = 38 nM). Other peptides recovered from the fragment library selection revealed a separate consensus motif (ARRXA) C-terminal to the histidine track. Kinetics measurements made by surface plasmon resonance, using purified Fab (antigen-binding fragment) to prevent avidity effects, demonstrate that the selected peptides bind with 10- to 75-fold higher affinities than a hexahistidine peptide. The highest affinity peptides (K(D) approximately 10 nM) encode both a short histidine track and the ARRXA motif, suggesting that the motif and other flanking residues make important contacts adjacent to the core polyhistidine-binding site and can contribute >2.5 kcal/mol of binding free energy. The fragment library construction methodology described here is applicable to the development of high-complexity protein or cDNA expression libraries for the identification of protein-protein interaction domains.     

Chemical synthesis of a dual branched malto-decaose: a potential substrate for alpha-amylases

A convergent block strategy for general use in efficient synthesis of complex alpha-(1-->4)- and alpha-(1-->6)-malto-oligosaccharides is demonstrated with the first chemical synthesis of a malto-oligosaccharide, the decasaccharide 6,6'-bis(alpha-maltosyl)-maltohexaose, with two branch points. Using this chemically defined branched oligosaccharide as a substrate, the cleavage pattern of seven different alpha-amylases were investigated. Alpha-amylases from human saliva, porcine pancreas, barley alpha-amylase 2 and recombinant barley alpha-amylase 1 all hydrolysed the decasaccharide selectively. This resulted in a branched hexasaccharide and a branched tetrasaccharide. Alpha-amylases from Asperagillus oryzae, Bacillus licheniformis and Bacillus sp. cleaved the decasaccharide at two distinct sites, either producing two branched pentasaccharides, or a branched hexasaccharide and a branched tetrasaccharide. In addition, the enzymes were tested on the single-branched octasaccharide 6-alpha-maltosyl-maltohexaose, which was prepared from 6,6'-bis(alpha-maltosyl)-maltohexaose by treatment with malt limit dextrinase. A similar cleavage pattern to that found for the corresponding linear malto-oligosaccharide substrate was observed.