Dry surface biofilms in the food processing industry: An overview on surface characteristics, adhesion and biofilm formation, detection of biofilms, and dry sanitization methods.

Bacterial biofilm formation in low moisture food processing (LMF) plants is related to matters of food safety, production efficiency, economic loss, and reduced consumer trust. Dry surfaces may appear dry to the naked eye, however, it is common to find a coverage of thin liquid films and microdroplets, known as microscopic surface wetness (MSW). The MSW may favor dry surface biofilm (DSB) formation. DSB formation is similar in other industries, it occurs through the processes of adhesion, production of extracellular polymeric substances, development of microcolonies and maturation, it is mediated by a quorum sensing (QS) system and is followed by dispersal, leading to disaggregation. Species that survive on dry surfaces develop tolerance to different stresses. DSB are recalcitrant and contribute to higher resistance to sanitation, becoming potential sources of contamination, related to the spoilage of processed products and foodborne disease outbreaks. In LMF industries, sanitization is performed using physical methods without the presence of water. Although alternative dry sanitizing methods can be efficiently used, additional studies are still required to develop and assess the effect of emerging technologies, and to propose possible combinations with traditional methods to enhance their effects on the sanitization process. Overall, more information about the different technologies can help to find the most appropriate method/s, contributing to the development of new sanitization protocols. Thus, this review aimed to identify the main characteristics and challenges of biofilm management in low moisture food industries, and summarizes the mechanisms of action of different dry sanitizing methods (alcohol, hot air, UV-C light, pulsed light, gaseous ozone, and cold plasma) and their effects on microbial metabolism.

Determination of Hydrogen Bond Structure in Water versus Aprotic Environments To Test the Relationship Between Length and Stability.

Hydrogen bonds profoundly influence the architecture and activity of biological macromolecules. Deep appreciation of hydrogen bond contributions to biomolecular function thus requires a detailed understanding of hydrogen bond structure and energetics and the relationship between these properties. Hydrogen bond formation energies (ΔGf) are enormously more favorable in aprotic solvents than in water, and two classes of contributing factors have been proposed to explain this energetic difference, focusing respectively on the isolated and hydrogen-bonded species: (I) water stabilizes the dissociated donor and acceptor groups much better than aprotic solvents, thereby reducing the driving force for hydrogen bond formation; and (II) water lengthens hydrogen bonds compared to aprotic environments, thereby decreasing the potential energy within the hydrogen bond. Each model has been proposed to provide a dominant contribution to ΔGf, but incisive tests that distinguish the importance of these contributions are lacking. Here we directly test the structural basis of model II. Neutron crystallography, NMR spectroscopy, and quantum mechanical calculations demonstrate that O-H···O hydrogen bonds in crystals, chloroform, acetone, and water have nearly identical lengths and very similar potential energy surfaces despite ΔGf differences >8 kcal/mol across these solvents. These results rule out a substantial contribution from solvent-dependent differences in hydrogen bond structure and potential energy after association (model II) and thus support the conclusion that differences in hydrogen bond ΔGf are predominantly determined by solvent interactions with the dissociated groups (model I). These findings advance our understanding of universal hydrogen-bonding interactions and have important implications for biology and engineering.

Genetic variability of isoflavones in the USDA red clover core collection

Red clover is one of the most utilized forage in agriculture and contains many of the isoflavones known for their human health benefits. The objectives of this study were: i) to quantify, using HPLC analysis, isoflavones in 77 accessions from the USDA core collection and a Brazilian line; ii) to verify possible relationships depending on their origin, improvement status or maturity type and; iii) to verify the seasonal variation. The isoflavone mean contents were 29.27 µg g-1 of dry material for daidzein, 163.69 µg g-1 for genistein, 11353.29 µg g-1 for formononetin and 6568.8 µg g-1 for biochanin A. Clustering was mainly influenced by the total amount of isoflavones and partially due to maturity type, improvement status and geographic origin. The seasonal evaluation demonstrated an increase of concentration during winter, and decrease during spring. These results highlighted accessions that can be used to develop new varieties with low or high isoflavones concentration.

Structural, theoretical and spectroscopic studies of the dichloride hexahydrate cube [Cl2(H2O)6]2-.

The structure of the dichloride hexahydrate cube, [Cl(2)(H(2)O)(6)](2-), as a salt with the tris(diisopropylamino)cyclopropenium cation, [C(3)(N(i)Pr(2))(3)](+), has been determined by low-temperature X-ray and neutron-diffraction studies. H atoms not involved in O-H···Cl bonding are disordered over two 0.5 occupancy sites around the O(6) ring. Calculations of the dianionic cube in the gas phase show remarkably good agreement with the solid-state structures with the exception of short O-H bond distances around the O(6) ring that suggests the involvement of a dynamic process. The cluster was also characterised by single-crystal infrared spectroscopy, and vibrational wavenumbers were found to be in good agreement with hydrogen bonding distances. Dibromide and difluoride hexahydrates were also studied theoretically, and O···O distances were found to decrease in the order difluoride > dichloride > dibromide > (H(2)O)(6) and as O···O···O angles increased towards an almost planar ring in (H(2)O)(6). NMR spectra of a chloroform solution of the hydrated salt at -25 °C is consistent with cluster formation.

Revisiting the polyoxometalate-based late-transition-metal-oxo complexes: the "oxo wall" stands.

Terminal oxo complexes of the late transition metals Pt, Pd, and Au have been reported by us in Science and Journal of the American Chemical Society. Despite thoroughness in characterizing these complexes (multiple independent structural methods and up to 17 analytical methods in one case), we have continued to study these structures. Initial work on these systems was motivated by structural data from X-ray crystallography and neutron diffraction and (17)O and (31)P NMR signatures which all indicated differences from all previously published compounds. With significant new data, we now revisit these studies. New X-ray crystal structures of previously reported complexes K(14)[P(2)W(19)O(69)(OH(2))] and "K(10)Na(3)[Pd(IV)(O)(OH)WO(OH(2))(PW(9)O(34))(2)]" and a closer examination of these structures are provided. Also presented are the (17)O NMR spectrum of an (17)O-enriched sample of [PW(11)O(39)](7-) and a careful combined (31)P NMR-titration study of the previously reported "K(7)H(2)[Au(O)(OH(2))P(2)W(20)O(70)(OH(2))(2)]." These and considerable other data collectively indicate that previously assigned terminal Pt-oxo and Au-oxo complexes are in fact cocrystals of the all-tungsten structural analogues with noble metal cations, while the Pd-oxo complex is a disordered Pd(II)-substituted polyoxometalate. The neutron diffraction data have been re-analyzed, and new refinements are fully consistent with the all-tungsten formulations of the Pt-oxo and Au-oxo polyoxometalate species.

First record of culex (Culex) brethesi (Dyar) (Diptera: Culicidae) in Rio Grande do Sul State, Brazil.

This is the first record of Culex (Culex) brethesi (Dyar) in Rio Grande do Sul state, Brazil. The species was identified from specimens collected in a sand bar vegetation with the aid of a Nasci's trap, during an expedition for surveillance of the West Nile Virus in July of 2006, in the city of Mostardas, Rio Grande do Sul, Brazil.

Development and loss of ferromagnetism controlled by the interplay of Ge concentration and Mn vacancies in structurally modulated Y4Mn(1-x)Ga(12-y)Ge(y).

The cubic intermetallic phase Y(4)Mn(1-x)Ga(12-y)Ge(y) (x = 0-0.26, y = 0-4.0) has been isolated from a molten gallium flux reaction. It presents a rare example of a system where ferromagnetism can be induced by controlling the vacancies of the magnetic centers. The Y(4)PdGa(12) type crystal structure is made up of a corner-sharing octahedral network of Ga and Ge atoms with Mn atoms at the centers of half the octahedra and Y atoms in the voids. At the highest Ge concentration, y = 4.0, the Mn site is nearly fully occupied, x = 0.05, and the samples are paramagnetic. At a lower Ge concentration, y = 1.0, Mn deficiency develops with x = 0.10. Surprisingly, strong ferromagnetism is observed with T(c) = 223 K. When Ge is excluded, y = 0, Mn is substantially deficient at x = 0.26 and ferromagnetism is maintained with a T(c) of approximately 160 K. In addition, a 6-fold modulated superstructure appears owing to an ordered slab-like segregation of Mn atoms and vacancies. Corresponding bond distortions propagate throughout the octahedral Ga network. Structure-property relationships are examined with X-ray and neutron diffraction, magnetic susceptibility, and electrical resistivity measurements.

Chloro and azido diruthenium complexes bearing electron-rich N,N',N''-triphenylguanidinate ligands.

The reaction of Ru(2)(OAc)(4)Cl with N,N',N''-triphenylguanidine (HTPG) produces one of two different compounds depending on the reaction conditions. In acetone in the presence of triethyl amine, the reaction produces tri-substituted Ru(2)(TPG)(3)(OAc)Cl, and in refluxing xylene, the tetra-substituted Ru(2)(TPG)(4)Cl is produced. Both of these new complexes can be cleanly converted into their corresponding azido analogues by reaction with sodium azide in methanol. The X-ray crystal structures of Ru(2)(TPG)(3)(OAc)Cl, Ru(2)(TPG)(3)(OAc)N(3), and Ru(2)(TPG)(4)Cl are presented, along with magnetic, electrochemical, and spectral measurements for each compound. Studies in solution show that, in contrast to Ru(2)(TPG)(3)(OAc)Cl, Ru(2)(TPG)(4)Cl is sterically hindered at the axial positions, and readily dissociates a chloride ion at high ionic strength. Equilibrium constants for chloride association and dissociation have been estimated. Mass spectrometric data suggest that the two azido complexes are precursors to new diruthenium nitrido species.

Biotin-avidin sandwich elisa with specific human isotypes IgG1 and IgG4 for Culicidae mosquito blood meal identification from an epizootic yellow fever area in Brazil

With a view toward investigating the feeding behavior of Culicidae mosquitoes from an area of epizootic yellow fever transmission in the municipalities of Garruchos and Santo Antônio das Missões, Rio Grande do Sul State, Brazil, specimens were collected by aspiration from September 2005 to April 2007. The engorged females were submitted to blood meal identification by enzyme-linked immunosorbent assay (ELISA). A total of 142 blood-engorged samples were examined for human or monkey blood through species-specific IgG. Additional tests for specificity utilizing isotypes IgG1 and IgG4 of human monoclonal antibodies showed that only anti-human IgG1 was effective in recognizing blood meals of human origin. The results indicated a significant difference (p = 0.027) in detection patterns in samples of Haemagogus leucocelaenus recorded from human blood meals at Santo Antônio das Missões, which suggests some degree of exposure, since it was an area where epizootic outbreaks have been reported.

Variable temperature neutron diffraction and X-ray charge density studies of tetraacetylethane.

Single crystal neutron diffraction data have been collected on a sample of enolized 3,4-diacetyl-2,5-hexanedione (tetraacetylethane, TAE) at five temperatures between 20 and 298 K to characterize the temperature-dependent behavior of the short, strong, intramolecular hydrogen bond. Upon decreasing the temperature from 298 K to 20 K, the O2-H1 distance decreases from 1.171(11) to 1.081(2) A and the O1...H1 distance increases from 1.327(10) to 1.416(6) A. The convergence of the C-O bond lengths from inequivalent distances at low temperature to identical values (1.285(4) A) at 298 K is consistent with a resonance-assisted hydrogen bond. However, a rigid bond analysis indicates that the structure at 298 K is disordered. The disorder vanishes at lower temperatures. Short intermolecular C-H...O contacts may be responsible for the ordering at low temperature. The intramolecular O...O distance (2.432 +/- 0.006 A) does not change with temperature. X-ray data at 20 K were measured to analyze the charge density and to gain additional insight into the nature of the strong hydrogen bond. Quantum mechanical calculations demonstrate that periodic boundary conditions provide significant enhancement over gas phase models in that superior agreement with the experimental structure is achieved when applying periodicity. One-dimensional potential energy calculations followed by quantum treatment of the proton reproduce the location of the proton nearer to the O2 site reasonably well, although they overestimate the O-H distance at low temperatures. The choice of the single-point energy calculation strategy for the proton potential is justified by the fact that the proton is preferably located nearer to O2 rather than being equally distant to O1 and O2 or evenly distributed (disordered) between them.

Sigma-borane complexes of iridium: synthesis and structural characterization.

Reaction of NaBH4 with (tBuPOCOP)IrHCl affords the previously reported complex (tBuPOCOP)IrH2(BH3) (1) (tBuPOCOP = kappa(3)-C6H3-1,3-[OP(tBu)2]2). The structure of 1 determined from neutron diffraction data contains a B-H sigma-bond to iridium with an elongated B-H bond distance of 1.45(5) A. Compound 1 crystallizes in the space group P1 (Z = 2) with a = 8.262 (5) A, b = 12.264 (5) A, c = 13.394 (4) A, and V = 1256.2 (1) A(3) (30 K). Complex 1 can also be prepared by reaction of BH3 x THF with (tBuPOCOP)IrH2. Reaction of (tBuPOCOP)IrH2 with pinacol borane gave initially complex 2, which is assigned a structure analogous to that of 1 based on spectroscopic measurements. Complex 2 evolves H2 at room temperature leading to the borane complex 3, which is formed cleanly when 2 is subjected to dynamic vacuum. The structure of 3 has been determined by X-ray diffraction and consists of the (tBuPOCOP)Ir core with a sigma-bound pinacol borane ligand in an approximately square planar complex. Compound 3 crystallizes in the space group C2/c (Z = 4) with a = 41.2238 (2) A, b = 11.1233 (2) A, c = 14.6122 (3) A, and V = 6700.21 (19) A(3) (130 K). Reaction of (tBuPOCOP)IrH2 with 9-borobicyclononane (9-BBN) affords complex 4. Complex 4 displays (1)H NMR resonances analogous to 1 and exists in equilibrium with (tBuPOCOP)IrH2 in THF solutions.

Hydrogen bonding effects on the electronic configuration of five-coordinate high-spin iron(II) porphyrinates.

The characterization of a new five-coordinate derivative of (2-methylimidazole)(tetraphenylporphinato)iron(II) provides new and unique information about the effects of forming a hydrogen bond to the coordinated imidazole on the geometric and electronic structure of iron in these species. The complex studied has two crystallographically distinct iron sites; one site has an axial imidazole ligand modified by an external hydrogen bond, and the other site has an axial imidazole ligand with no external interactions. The iron atoms at the two sites have distinct geometric features, as revealed in their molecular structures, and distinct electronic structures, as shown by Mössbauer spectroscopy, although both are high spin (S = 2). The molecule with the external hydrogen bond has longer equatorial Fe-N(p) bonds, a larger displacement of the iron atom out of the porphyrin plane, and a shorter axial bond compared to its counterpart with no hydrogen bonding. The Mössbauer features are distinct for the two sites, with differing quadrupole splitting and isomer shift values and probably differing signs for the quadrupole splitting as shown by variable-temperature measurements in applied magnetic field. These features are consistent with a significant change in the nature of the doubly populated d orbital and are all in the direction of the dichotomy displayed by related imidazole and imidazolate species where deprotonation leads to major differences. The results points out the possible effects of strong hydrogen bonding in heme proteins.

X-ray and neutron diffraction studies of water-encapsulated inside potassium aryloxide aggregates: complementary host-guest stabilization of mono- and dihydrated cages.

The controlled hydrolysis of potassium 2-tert-butylphenoxide or 2-isopropylphenoxide leads to the unexpected encapsulation of the water inside K6O6 hexameric drum aggregates. Encapsulation of the neutral molecules is enabled in these instances through the formation of strong hydrogen bonds and dative interactions between the host and guest.

Terminal gold-oxo complexes.

In contradiction to current bonding paradigms, two terminal Au-oxo molecular complexes have been synthesized by reaction of AuCl3 with metal oxide-cluster ligands that model redox-active metal oxide surfaces. Use of K10[alpha2-P2W17O61].20H2O and K2WO4 (forming the [A-PW9O34]9- ligand in situ) produces K15H2[Au(O)(OH2)P2W18O68].25H2O (1); use of K10[P2W20O70(OH2)2].22H2O (3) produces K7H2[Au(O)(OH2)P2W20O70(OH2)2].27H2O (2). Complex 1 crystallizes in orthorhombic Fddd, with a=28.594(4) A, b=31.866(4) A, c=38.241(5) A, V=34844(7) A3, Z=16 (final R=0.0540), and complex 2 crystallizes in hexagonal P6(3)/mmc, with a=16.1730(9) A, b=16.1730(9) A, c=19.7659(15) A, V=4477.4(5) A3, Z=2 (final R=0.0634). The polyanion unit in 1 is disorder-free. Very short (approximately 1.76 A) Au-oxo distances are established by both X-ray and 30 K neutron diffraction studies, and the latter confirms oxo and trans aqua (H2O) ligands on Au. Seven findings clarify that Au and not W is present in the Au-oxo position in 1 and 2. Five lines of evidence are consistent with the presence of d8 Au(III) centers that are stabilized by the flanking polytungstate ligands in both 1 and 2: redox titrations, electrochemical measurements, 17 K optical spectra, Au L2 edge X-ray absorption spectroscopy, and Au-oxo bond distances. Variable-temperature magnetic susceptibility data for crystalline 1 and 2 establish that both solids are diamagnetic, and 31P and 17O NMR spectroscopy confirm that both remain diamagnetic in solution. Both complexes have been further characterized by FT-IR, thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), and other techniques.

A lipid-soluble iron chelator alters cell cycle regulatory protein binding in breast cancer cells compared to normal breast cells.

The lipid-soluble iron chelator desferri-exochelin (D-Exo) causes reversible cell cycle arrest in normal human mammary epithelial cells (NHMEC) but triggers apoptotic cell death in human breast cancer cells. We studied the effects of iron chelation with D-Exo on cell cycle regulatory proteins in cultures of NHMEC and MCF-7 breast cancer cells. In co-immunoprecipitation studies, D-Exo inhibited binding of cyclins A and E to cyclin dependent kinase 2 (CDK2) in NHMEC, but in MCF-7 cells binding of these cyclins to CDK2 was enhanced. D-Exo treatment markedly increased expression of p53 and increased binding of p21 to CDK2 in the MCF-7 cells but not in NHMEC. Therefore differences in effects of iron chelation on cell cycle protein binding in cancer cells compared to normal cells may trigger apoptosis in cancer cells while normal breast cells are spared.

Synthesis, structure, and properties of BaAl2Si2.

Single crystals of BaAl2Si2 were grown from an Al molten flux and characterized using single-crystal X-ray diffraction at 10 and 90 K and neutron diffraction at room temperature. BaAl2Si2 crystallizes with the alpha-BaCu2S2 structure type (Pnma), is isostructural with alpha-BaAl2Ge2, and is an open 3D framework compound, where Al and Si form a covalent cagelike network with Ba2+ cations residing in the cages. BaAl2Si2 has a unit cell of a=10.070(3) A, b=4.234(1) A, and c=10.866(3) A, as determined by room-temperature single-crystal neutron diffraction (R1=0.0533, wR2=0.1034). The structure as determined by single-crystal neutron and X-ray diffraction (10 and 90 K) indicates that BaAl2Si2 (Pnma) is strictly isostructural to other (alpha)-BaCu2S2-type structures, requiring site specificity for Al and Si. Unlike BaAl2Ge2, no evidence for an alpha to beta (BaZn2P2-type, I4/mmm) phase transition was observed. This compound shows metallic electronic resistivity and Pauli paramagnetic behavior.