Tapping into cyanobacteria electron transfer for higher exoelectrogenic activity by imposing iron limited growth.

The exoelectrogenic capacity of the cyanobacterium Synechococcus elongatus PCC7942 was studied in iron limited growth in order to establish conditions favouring extracellular electron transfer in cyanobacteria for photo-bioelectricity generation. Investigation into extracellular reduction of ferricyanide by Synechococcus elongatus PCC7942 demonstrated enhanced capability for the iron limited conditions in comparison to the iron sufficient conditions. Furtheremore, the significance of pH showed that higher rates of ferricyanide reduction occurred at pH 7, with a 2.7-fold increase with respect to pH 9.5 for iron sufficient cultures and 24-fold increase for iron limited cultures. The strategy presented induced exoelectrogenesis driven mainly by photosynthesis and an estimated redirection of the 28% of electrons from photosynthetic activity was achieved by the iron limited conditions. In addition, ferricyanide reduction in the dark by iron limited cultures also presented a significant improvement, with a 6-fold increase in comparison to iron sufficient cultures. Synechococcus elongatus PCC7942 ferricyanide reduction rates are unprecedented for cyanobacteria and they are comparable to those of microalgae. The redox activity of biofilms directly on ITO-coated glass, in the absence of any artificial mediator, was also enhanced under the iron limited conditions, implying that iron limitation increased exoelectrogenesis at the outer membrane level. Cyclic voltammetry of Synechococcus elongatus PCC7942 biofilms on ITO-coated glass showed a midpoint potential around 0.22 V vs. Ag/AgCl and iron limited biofilms had the capability to sustain currents in a saturated-like fashion. The present work proposes an iron related exoelectrogenic capacity of Synechococcus elongatus PCC7942 and sets a starting point for the study of this strain in order to improve photo-bioelectricity and dark-bioelectricity generation by cyanobacteria, including more sustainable mediatorless systems.

Evaluation of pre-PCR processing approaches for enumeration of Salmonella enterica in naturally contaminated animal feed.

AIMS: Three pre-PCR processing strategies for the detection and/or quantification of Salmonella in naturally contaminated soya bean meal were evaluated. METHODS AND RESULTS: Methods included: (i) flotation-qPCR [enumeration of intact Salmonella cells prior to quantitative PCR (qPCR)], (ii) MPN-PCR (modified most probable number method combined with qPCR) and (iii) qualitative culture enrichment PCR. The limit of quantification was 1·8 × 10(2) CFU g(-1) (flotation-qPCR) and 0·02 MPN g(-1) (MPN-PCR). Fifteen naturally contaminated Salmonella positive soya bean meal samples from one lot were analysed in parallel with the three methods, using 2·5, 50 and 25 g of feed, respectively, resulting in detection of Salmonella in 6, 15 and 9 bags. Enumeration resulted in 1·8 × 10(2) -7·8 × 10(3) CFU g(-1) (flotation-qPCR) and 0·024 to >5·2 MPN g(-1) (MPN-PCR). CONCLUSIONS: Except for differences in methodology, results obtained with the three techniques could be due to the presence of nonculturable Salmonella and/or a heterogeneous distribution of Salmonella in the material. SIGNIFICANCE AND IMPACT OF THE STUDY: The evaluated methods provide different possibilities to assess the prevalence of Salmonella in feed, together with the numbers of culturable, as well as nonculturable cells, and can be applied to generate data to allow more accurate quantitative microbial risk assessment for Salmonella in the feed chain.

The initial steps of biogenesis of cyanobacterial photosystems occur in plasma membranes.

During oxygenic photosynthesis in cyanobacteria and chloroplasts of plants and eukaryotic algae, conversion of light energy to biologically useful chemical energy occurs in the specialized thylakoid membranes. Light-induced charge separation at the reaction centers of photosystems I and II, two multisubunit pigment-protein complexes in the thylakoid membranes, energetically drive sequential photosynthetic electron transfer reactions in this membrane system. In general, in the prokaryotic cyanobacterial cells, the thylakoid membrane is distinctly different from the plasma membrane. We have recently developed a two-dimensional separation procedure to purify thylakoid and plasma membranes from the genetically widely studied cyanobacterium Synechocystis sp. PCC 6803. Immunoblotting analysis demonstrated that the purified plasma membrane contained a number of protein components closely associated with the reaction centers of both photosystems. Moreover, these proteins were assembled in the plasma membrane as chlorophyll-containing multiprotein complexes, as evidenced from nondenaturing green gel and low-temperature fluorescence spectroscopy data. Furthermore, electron paramagnetic resonance spectroscopic analysis showed that in the partially assembled photosystem I core complex in the plasma membrane, the P700 reaction center was capable of undergoing light-induced charge separation. Based on these data, we propose that the plasma membrane, and not the thylakoid membrane, is the site for a number of the early steps of biogenesis of the photosynthetic reaction center complexes in these cyanobacterial cells.

Proteomics of Synechocystis sp. strain PCC 6803. Identification of periplasmic proteins in cells grown at low and high salt concentrations.

Periplasmic proteins isolated by cold osmotic shock of Synechocystis sp. PCC 6803 cells were identified using 2D PAGE, MS and genome analysis. Most of the periplasmic proteins represent 'hypothetical proteins' with unknown function. A number of proteases of different specificity, and several enzymes involved in cell wall biosynthesis were also found. In salt-adapted cells, six proteins were greatly enhanced and three proteins were newly induced. Most of the salt-enhanced proteins are involved in the alteration of cell wall structure of salt-adapted cells. The precursors of all 57 periplasmic proteins identified have a signal peptide; 47 of them contain a typical Sec-dependent signal peptide, whereas 10 contain a putative twin-arginine signal peptide.

Effect of varying fixture width on stress and strain distribution associated with an implant stack system.

The purpose of this investigation was to evaluate the dissipation of a force applied to an assembled stack of implant components. The stack consisted of a 10-mm threaded implant, a screw-retained abutment and a screw-retained gold crown. The dissipation of force was analyzed in relation to varying the implant diameter with and without a concomitant change in abutment diameter. Two experimental groups were evaluated. The first group consisted of 25 titanium screw-form implants (Implant Innovations, Inc.). These implants measured 10 mm in length and 3.25 mm, 3.75 mm, 4.0 mm, 5.0 mm, and 6.0 mm in diameter. The second group included 15 titanium screw-form implants (Nobel Biocare, Inc.) measuring 10 mm in length and 3.75 mm, 4.0 mm, and 5.0 mm in diameter. All implants were embedded in standardized photoelastic resin blocks. Points of interest were marked on each block using standardized templates to ensure consistency. Implants were restored using system-specific conical abutments and standardized single-unit restorations. A strain gauge was affixed to each abutment, and an eccentric load of 176 N was applied to the restoration. Periimplant stresses were measured using photoelastic analysis. Abutment strain was determined using an electronic strain indicator. Data were collated and compared using ANOVA and the Duncan multiple range statistical tests. When stress was analyzed at points on the resin-implant interface or a fixed distance from the interface, stress tended to decrease from the 5-mm-wide implant to the 6-mm-wide implant. Stress in relation to the 3.25-mm, 3.75-mm, and 4.0-mm implant was not as well defined, indicating the possibility that some deformation of implants was occurring. Increased abutment width resulted in decreased abutment strain. Therefore, using a wider abutment may be helpful in preventing preload reduction in clinical applications. This may reduce the incidence of loosening and fracture of abutment and restoration screws.

Quantification of leucite concentration using X-ray diffraction.

OBJECTIVES: This study sought to evaluate the efficacy of using an X-ray diffractometer for the determination of leucite in Finesse, Ceramco II, and IPS Empress porcelains. METHODS: An internal standard, copper, was used. Two quantification methods are presented: (1) the generation of a calibration curve using peak height ratios; and (2) the generation of a calibration curve using peak area ratio. RESULTS: The leucite concentration obtained from the peak height versus concentration calibration curve was observed to be statistically different from leucite concentration obtained from the peak area versus concentration calibration curve. Other information obtained from X-ray powder diffraction include the lattice parameters and volume of the unit leucite cell. SIGNIFICANCE: The leucite contained in the dental porcelains (Finesse, Ceramco II, and Empress) has expanded a-lattice spacings and contracted c-lattice spacings relative to standard leucite. These changes in the lattice parameters resulted in a net expansion of the leucite cell volume.

Subcellular localization of the BtpA protein in the cyanobacterium Synechocystis sp. PCC 6803.

Photosystem I is a large pigment-protein complex embedded in the thylakoid membranes of chloroplasts and cyanobacteria. In the cyanobacterium Synechocystis sp. PCC 6803, the btpA gene encodes a 30-kDa polypeptide. Mutations in this gene significantly affect accumulation of the reaction center proteins of photosystem I in Synechocystis 6803 [Bartsevich, V. V. & Pakrasi, H. B. (1997) J. Biol. Chem. 272, 6372-6378]. We describe here the intracellular localization of the BtpA protein. Immunolocalization in Synechocystis 6803 cells demonstrated that the BtpA protein is tightly associated with the thylakoid membranes. Phase fractionation in the detergent Triton X-114 indicated that BtpA is a peripheral membrane protein. To determine which surface of the thylakoid membrane BtpA is exposed to, we used a two-phase polymer partitioning technique to develop a novel method to isolate inside-out and right-side-out thylakoid vesicles from Synechocystis 6803. Treatments of such vesicles with different salts and protease showed that the BtpA protein is an extrinsic membrane protein which is exposed to the cytoplasmic face of the thylakoid membrane.

2D-isolation of pure plasma and thylakoid membranes from the cyanobacterium Synechocystis sp. PCC 6803.

Aqueous polymer two-phase partitioning in combination with sucrose density centrifugation offered, for the first time, a 2D-separation method for the isolation of pure plasma and thylakoid membranes from the cyanobacterium Synechocystis 6803 without any cross-contaminations. The purity of the membrane fractions was verified by immunoblot analysis using antibodies against membrane-specific marker proteins. As an initiation of a proteomics project, two prominent proteins, which were observed only in the plasma membrane (Slr1513, a hypothetical protein, and HofG, a general secretion pathway protein), or in the thylakoid membrane (PsaE, a photosystem I protein, and NdhH, a subunit of NADH dehydrogenase), were identified.

Do combinations of 1 mg estradiol and low doses of NETA effectively control menopausal symptoms?

OBJECTIVES: This study compared two continuous-combined hormone replacement therapy (HRT) formulations, 1 mg estradiol (E2)/0.25 mg norethisterone acetate (NETA) and 1 mg E2/0.5 mg NETA, with placebo, with regard to the efficacy for vasomotor symptom relief in menopausal women. METHODS: A total of 119 women aged 45-61 years with moderate and severe hot flushes and with amenorrhea for at least 3 months were randomly assigned to 12 weeks' treatment with 1 mg E2/0.25 mg NETA, 1 mg E2/0.5 mg NETA or placebo. The number and severity of hot flushes, as well as any vaginal bleeding, were recorded on a daily basis. The Kupperman Menopausal Index, Greene Climacteric Scale and visual analog scales for various symptoms were assessed before and after treatment. Subpopulation analysis according to menopausal status was performed. RESULTS: Both combinations significantly reduced the number and severity of hot flushes, compared to placebo. A reduction of approximately 85% in vasomotor symptomatology occurred in the two combination groups by week 4 of treatment, and this was further diminished throughout the study to approximately 97% reduction by week 12. At the end of the study, 85% of the women receiving 1 mg E2/0.5 mg NETA and 71% of the women receiving 1 mg E2/0.25 mg NETA were considered to the clinically adequate responders to treatment. Both combinations were associated with significant improvements, compared to placebo, in visual analog scales for overall general condition, Kupperman Menopausal Index, and Greene Climacteric vasomotor and psychological subscales. While both combinations resulted in similar bleeding profiles in postmenopausal women, the combination of 1 mg E2/0.5 mg NETA resulted in the lowest incidence of bleeding in late perimenopausal women. CONCLUSIONS: The combinations of 1 mg E2/0.25 mg NETA and 1 mg E2/0.5 mg NETA rapidly relieve vasomotor symptoms and are efficacious in the majority of menopausal women, including those with severe hot flushes.

An investigation of new metal framework design for metal ceramic restorations.

STATEMENT OF PROBLEM: Metal ceramic restorations have been implicated in the discoloration of associated gingival tissues. Attempts to remedy this by altering the design of the metal frameworks for such restorations may lead to unacceptable decreases in fracture resistance. PURPOSE: This study evaluated a new metal framework design for metal-ceramic restorations. MATERIAL AND METHODS: Twenty artificial crowns were fabricated with various degrees of facial metal reduction; 0, 1, 2, and 3 mm. The study was conducted in two parts. The first part evaluated changes in light transmission into adjacent root tissue. A light box was fabricated so sample crowns could be illuminated on a mounted natural tooth. The root of the tooth remained outside the light box, and the light transmitted through the crowns into root tissue was measured with a light meter. The second part of the study evaluated changes in fracture strength. The sample crowns were subjected to a vertical load until fracture with use of an Instron machine at a crosshead speed of 1 mm per minute. The load at fracture was recorded. RESULTS: Results indicated a statistically significant increase in light transmission with 1 mm framework reduction or greater, and fracture strengths did not decrease with up to 1 mm of framework reduction. A 1 mm facial axial reduction of the metal framework may be indicated for anterior metal-ceramic restorations.

A rapid sample preparation method for PCR detection of food pathogens based on buoyant density centrifugation.

The use of buoyant density centrifugation (BDC) to prepare samples for PCR analysis of food pathogens is described. Blue cheese and milk homogenates were inoculated with Shigella flexneri and layered on top of Percoll media. After BDC, the food homogenates remained in the upper part of the centrifuge tube, separated from the bacteria, which retained viability and were concentrated below the lighter Percoll layer. PCR inhibitors stayed in the homogenate and PCR analyses of treated samples consistently detected 10(4) cfu g-1 of blue cheese and 500 cfu ml-1 of milk, respectively. Differences in the density of live and killed Sh. flexneri and Yersinia enterocolitica were detected by BDC but were dependent on the mechanism of killing.

Directly placed esthetic restorative materials--the continuum.

Because new restorative materials have had little clinical testing, it is difficult to make specific material recommendations for the esthetic restoration of carious teeth. Although fluoride-releasing materials have long been used successfully to restore carious teeth, little clinical documentation has been presented to support their use to inhibit recurrent caries, and their use as an effective restorative material may be questioned. Glass ionomers, compomers, and resin-modified glass ionomers are esthetic fluoride-releasing materials designed to restore teeth by bonding to tooth structure. This article describes the continuum of directly placed esthetic dental restorative materials, the efficacy of amalgam replacement restorative materials, and the role that fluoride-releasing materials may play in the inhibition of recurrent caries in vitro and in vivo.

Gypsum compatibility of antimicrobial alginates after spray disinfection.

PURPOSE: This investigation examined the gypsum compatibility of two antimicrobial alginates after spray disinfection. Subjective compatibility evaluations were compared with objective quantitative profilometer readings of gypsum cast surface roughness. MATERIALS AND METHODS: COE Hydrophilic Gel Alginate, Jeltrate Plus, Antimicrobial Alginate, and their nonantimicrobial counterparts, Coe Alginate and Jeltrate Plus, were used in this study. After spray disinfection with water (control), Alcide LD, Biocide, OMC II, and 0.5% NaOCI, impressions of the American National Standards Institute/American Dental Association (ANSI/ADA) specification no. 18 detail reproduction die and impressions made simultaneously of a smooth glass die were cast in Microstone, Silky-Rock, and Die-Keen. Five specimens were made for each alginate/disinfectant/gypsum combination for a total of 300 samples each for both the subjective and objective analyses. For the subjective analysis of gypsum compatibility, the specimens made from the ANSI/ADA specification no. 18 test die were evaluated by using a 1-to-4 visual rating system at magnification x12. For the objective analysis, the arithmetic average surface roughness of each specimen made from the smooth glass die was recorded three times with a 200-mg skidless stylus instruments. RESULTS: The results of the ANSI/ADA specification no. 18 testing for gypsum compatibility showed that 11 of 60 possible combinations did not pass the test. All impressions made with nonantimicrobial COE Alginate passed the test regardless of the disinfectant/gypsum combination. The results of the three-factor analysis of variance for the subjective and objective analyses showed significant interactions between alginates, disinfectants, and stones at the P < .05 level. To further delineate these differences, unpaired t tests (P < .05) within brands for each disinfectant/gypsum combination were performed. CONCLUSIONS: The addition of 1% chlorhexidine diacetate to COE Hydrophilic Gel Alginate has decreased its compatibility with the dental stones and disinfectants tested when compared with its nonantimicrobial counterpart. In terms of gypsum compatibility, the nonantimicrobial COE Alginate was compatible with all disinfectant and gypsum combinations tested. The addition of 1.70% didecyldimethyl ammonium chloride to Jeltrate Plus Antimicrobial Alginate has increased its compatibility with all the dental stones tested. A strong positive correlation (r = 0.7398) was found between visual gypsum compatibility evaluation scores and surface roughness of gypsum casts.

Subfractional analysis of cyanobacterial membranes and isolation of plasma membranes by aqueous polymer two-phase partitioning.

The present work demonstrates that partition in aqueous polymer two-phase systems offers a rapid method for separation and isolation of thylakoid, plasma, and outer membranes from cyanobacteria. Pure plasma membranes from Phormidium laminosum can be isolated by this method within 3 h, starting with total membranes obtained by French press treatment of the cyanobacterial cells. The isolated plasma membranes have a broad density profile, giving rise to three subpopulations. The main fraction has the same density as the abundant thylakoid membranes. This fraction has not been resolved in previous separations based on sucrose gradient centrifugation, which is the only method previously used for isolation of cyanobacterial plasma membranes. Another advantage of the aqueous polymer two-phase system is that it can handle large quantities of starting material, which is essential to obtain a satisfactory yield since plasma membranes constitute only a very small fraction of the total membrane content in a cyanobacterial cell. The isolation procedure results in a pure plasma membrane preparation with retained cytochrome c oxidase activity. The results also point to the possibility of a lateral heterogeneity in the organization of the cyanobacterial plasma membrane.

Mercury in solution following exposure of various amalgams to carbamide peroxides.

Carbamide peroxide (CP) is an easily administered material for whitening teeth. Although toxicological research on CP alone has revealed no adverse health effects, possible oxidation and release of mercury from amalgams have not previously been investigated. This research evaluated the quantitative release of mercury from amalgams into solution by CP. CP preparations can generally be divided into two classes based on the presence or absence of carbopol, an oxygen-releasing inhibitor. Rembrandt (R), a 10% CP with carbopol and White and Brite (WB), a 10% CP without carbopol were used in this study. Four different types of amalgams [Dispersalloy (D), Sybraloy (S), Tytin (T) and Valiant Ph.D. (V)] were selected. Uniform samples of the four amalgams were prepared and stored at 37 degrees C for 1 week. Vials of saline (10 ml), R and WB were prepared. R and WB were mixed with saline to a 50:50 solution to reduce viscosity and facilitate stirring. Magnetic teflon coated stir bars were placed in all vials, and one amalgam specimen was placed in each non-control vial. After being stirred for 8 hours, solutions were analyzed for elemental mercury content using a Jerome Gold Film Mercury Analyzer. All background mercury levels were zero, but following the experiment there were significantly higher amounts of mercury in the CP solutions as compared to the 100% saline solutions. These results suggest there is an active oxidation of the amalgam releasing mercury ions into solution.

Fibronectin and basement membrane components in renal amyloid deposits in patients with primary and secondary amyloidosis.

Kidney biopsies from one patient with primary (AL) and three with secondary (AA) amyloidosis were used for an ultrastructural study of the collocalization of basement membrane proteins and the extracellular matrix protein fibronectin within amyloid deposits. Antibodies against amyloid P component, laminin, and heparan sulphate proteoglycan core protein all reacted with the basement membranes and the amyloid depositions in AA and AL amyloidosis. Monoclonal and polyclonal antibodies against collagen type IV reacted only with the basement membranes. Anti-fibronectin reaction was found in association with the basement membranes in all four cases, while labelling of amyloid depositions was found only in one of the AA amyloid cases and in the AL amyloid depositions. It is concluded that basement membrane components may be of importance for the formation of amyloid fibrils.

Genital human papilloma virus infection in Oslo studied by dot blot DNA hybridization and the polymerase chain reaction.

Samples from patients with genital condyloma acuminata or with cervical condylomas and/or dysplasia and from women without cytological/clinical evidence of cervical affection were examined by dot blot DNA hybridization or the polymerase chain reaction (PCR). The PCR was much more sensitive than dot blot, more than doubling the human papilloma virus (HPV) findings. HPV DNA, mainly HPV 6/11, was detected in 18 of 19 biopsies of condyloma acuminata, whereas HPV 16 was most frequently detected in the 21 cervices (76%) with condyloma and/or dysplasia. HPV 16 was detected in eight of 103 cervical smears with no signs of infection. The prevalence of HPV 16 in cervical samples was somewhat higher than expected. This suggests that, in Oslo, HPV 16 is a common HPV type in women with cytologically normal cervices. HPV 18 was relatively rare and was detected only in combination with other HPVs.

Inhibition of lipid peroxidation by ubiquinol in submitochondrial particles in the absence of vitamin E.

The relationship between the antioxidant effects of reduced coenzyme Q10 (ubiquinol, UQH2) and vitamin E (alpha-tocopherol) was investigated in beef heart submitochondrial particles in which lipid peroxidation was initiated by incubation with ascorbate + ADP-Fe3+. These effects were examined after extraction of coenzyme Q10 (UQ-10) and vitamin E from the particles and reincorporation of the same components alone or in combination. The results show that UQH2 efficiently inhibits lipid peroxidation even when vitamin E is absent. It is concluded that UQH2 can inhibit lipid peroxidation directly, without the mediation of vitamin E.