Thermophilic and mesophilic bacteria in biofilms associated with corrosion in a heat exchanger

Biofilm development on AISI-1020 carbon steel coupons installed at the outlet of a heat exchanger was evaluated at the thirtieth and the sixtieth days of exposure. Water temperature varied between 41 and 60°C. The most probable number technique (MPN) was applied to quantify mesophilic and thermophilic species of aerobic, anaerobic, and sulphate-reducing bacteria (SRB) in planktonic and sessile phases. The results showed predominance of thermophilic aerobic bacteria in both phases, corresponding to 9.5±0.8×10⁶cells/ml in the planktonic phase. In biofilms, maximal aerobic cell concentration, 7.8±0.6×10⁸cells/cm2, was registered at the sixtieth day. An increase in the number of thermophilic anaerobic and SRB with elapsed time was also observed. The results obtained after 60 days were 5.8±0.4×10⁷ and 8.9±0.9×10⁴cells/cm² for anaerobic and sulphate-reducing bacteria, respectively. Scanning electron microscopy showed a varied composition of species in the biofilms and corrosion on the carbon steel surfaces after biofilm removal.     

Molecular cloning and sequence analyses of calcium/calmodulin- dependent protein kinase II from fetal and adult human brain – Sequence analyses of human brain calcium/calmodulin-dependent protein kinase II

The aims of this study were to characterize specific mRNAs and the expression pattern for isoforms of calcium/calmodulin-dependent protein kinase II (CaMKII) in the human brain. We cloned and sequenced the CaMKII and subunit cDNAs, and used them to study the CaMKII expression in human brain. Four distinct isoforms of CAMKII were isolated. Two of them were characterized as CaMKII and subunits. The other two showed similar nucleotide sequences, but one had a 33-bp insertion relative to the subunit, and the other had a 75-bp deletion relative to the subunit. These alterations are located within the variable regions. These two isoforms were characterized as CaMKII _B and _e. Northern blot analysis showed that a 4.4-kb messenger RNA for the isoform and a 3.9-kb messenger RNA for the isoform were expressed in both human fetal and adult brain to different degrees. The results indicate that CaMKII expression is developmentally regulated. The CaMKII isoform expression was confirmed in human fetal and adult brain using RT-PCR with specific primers, which flanked the CaMKII variable regions. The CaMKII , _B, , and _e isoforms were characterized in both human fetal and adult brain.     

A general strategy for the isolation of carbohydrate chains fromN-,O-glycoproteins and its application to human chorionic gonadotrophin

For the structural analysis of the carbohydrate chains ofN-,O-glycoproteins a straightforward strategy was developed based on the cleavage of theN-linked chains with immobilized peptide-N ⁴-(N-acetyl--glucosaminyl) asparagine amidase-F (PN-Gase-F) fromFlavobacterium meningosepticum, followed by alkaline borohydride treatment of the remainingO-glycoprotein material. This methodology was applied to the isolation of the Asn- and Ser-linked carbohydrate chains of human chorionic gonadotrophin. The structures of the isolated oligosaccharides were verified by 500-MHz¹H-NMR spectroscopy. The Asn-linked sugar chains were shown to be: NeuAc2-3Gal1-4GlcNAc1-2Man1-6[NeuAc2-3Gal1-4GlcNAc1-2Man1-3]Man 1-4GlcNAc1-4[Fuc1-6]_0-1GlcNAc and Man1-6[NeuAc2-3Gal1-4GlcNAc1-2Man 1-3]Man1-4GlcNAc1-4GlcNAc. Also some minor constituents occurred. The structures of the Ser-linked oligosaccharides were established in the form of their oligosaccharide-alditols as: NeuAc2-3Gal1-3[NeuAc2-6]GalNAc, NeuAc2-3Gal 1-3GalNAc and NeuAc2-3Gal1-3[NeuAc2-3Gal1-4GlcNAc1-6]GalNAc.Abbreviations hCG human chorionic gonadotrophin - hCG- -subunit - hCG- -subunit - ElA enzyme immunoassay - PNGase-F peptide-N ⁴-(N-acetyl--glucosaminyl)asparagine amidase-F (EC 3.5.1.52) - SDS sodium dodecyl sulphate - GalNAc N-acetylgalactosamine - GlcNAc N-acetylglucosamine - NeuAc N-acetylneuraminic acid - Man mannose - Gal galactose - Fuc fucose     

Lipid profile of rat myelin subfractions

Myelin from adult rat brains was separated on a discontinuous sucrose gradient into three subfractions. Analysis of light, heavy and membrane fraction lipid classes was performed by HPTLC and densitometry while fatty acid composition was determinated by GLC. The more interesting results observed are: i) the membrane fraction resembles in its lipid and fatty acid composition other cell membranes (particulary oligodentrocytes); ii) light and heavy myelin are quite similar between them but the former has a higher content of sphingomyelin, a lower hydroxy/nohhydroxy cerebrosides ratio and a lower content of monoenoic fatty acids than the heavy subfraction. The results obtained could explain the different structures observed in each myelin subfraction since fatty acid composition, hydroxy fatty acids, sphingomyelin and cholesterol play a key role in the stability and structure of membranes.     

Molecular and Functional Studies of the Gamma Subunit of the Sodium Pump

This article reviews our studies of the subunit of the sodium pump. is a member of the FXYD family of small, single transmembrane proteins and is expressed predominantly in the kidney tubule. There are two major variants of which function similarly to bring about two distinct effects, one on K_ATP and the other, on K _K, the affinity of the pump for K ⁺ acting as a competitor of cytoplasmic Na⁺. In this way, is believed to provide a self-regulatory mechanism for maintaining the steady-state activity of the pump in the kidney. Our studies also suggest that K⁺ antagonism of cytoplasmic Na⁺ activation of the pump is relevant not only to the presence of in the kidney, but probably some hitherto undefined factor(s) in other tissues, most notably heart. The interesting possibility that not only but other members of the FXYD family regulate ion transport in a tissue-specific manner is discussed.     

Wall morphogenesis in diatoms: Deposition of silica by cytoplasmic vesicles

Summary Several TEM and SEM techniques were applied to examine developing structures in valves of the centric diatomThalassiosira eccentrica (Ehrenb.) Cleve after cytokinesis. It was possible to confirm that in each stage of the silicification process there is a distinction between a growing zone with a loose assemblage of silica spheres and a compacting zone in an older phase of development. The spherical structure of the silica in the growing zone results from the addition of silica by small cytoplasmic vesicles of about 300 to 400 Å in diameter. The vesicle membrane fuses with the silicalemma and the vesicle content is released into the silica-deposition vesicle. The origin of these vesicles, named STV, is still unknown.     

Optimized Binding of [35S]GTPγS to Gq-Like Proteins Stimulated with Dopamine D1-Like Receptor Agonists

Subtypes of dopamine D₁-like receptors are coupled through the G proteins Gs or Gq to stimulate either adenylate cyclase or phospholipase C signaling cascades. In the present study, we have uncovered the marked enhancement by sodium deoxycholate of D1-like agonist-stimulated [³⁵S]GTPS binding to Gq-like G proteins in brain membranes, and determined the optimal experimental conditions for assessing agonist effects on [³⁵S]GTPS binding in the presence of the detergent. Factors and their optimal levels that were found to significantly enhance the sensitivity and robustness of the agonist-stimulated [³⁵S]GTPS binding reaction include protein concentration at 40 g/ml, cationic concentrations of 120 mM Na⁺, 1.8 mM K⁺, and 20 mM Mg²⁺, a molar guanine nucleotide ratio of 100,000 GDP to [³⁵S]GTPS, the presence of 1 mM deoxycholate, and an overall incubation duration of 30–120 min. Under the optimized conditions, the D₁-like agonist SKF38393 induced potent and highly efficacious (up to 1000%) stimulation of [³⁵S]GTPS binding in membrane preparations from the striatum and other rat brain regions. In striatal membranes incubated with drug for 2 h, immunoprecipitation of the [³⁵S]GTPS-bound proteins with specific G antibodies showed that at least 70% of SKF38393-stimulated [³⁵S]GTPS binding was to Gq. The present reaction parameters are consistent with conditions previously found to support dopaminergic stimulation of phospholipase C-mediated signaling in brain slice preparations. These results imply that different but equally physiologically relevant conditions can be obtained under which subtypes of dopaminergic receptors may couple preferentially to Gs and the adenylate cyclase pathway or to Gq and the phospholipase C pathway.     

TNF-α induced altered signaling mechanism in human neutrophil

In the present study we investigated the TNF- induced signal transduction mechanism in human neutrophil. Exogenously added TNF- affects both PKC activity and its translocation from cytosol to the membrane. Endogenous protein phosphorylation pattern is inhibited in TNF- induced neutrophil in Ca-dependent and Ca-independent manner, including a major 47 and 66 kDa cytosolic proteins, which may be implicated in superoxide anion generation. However TNF- dose dependently enhances the expression of -PKC isotype but not the -PKC. Morphology and cell cytotoxicity are studied in TNF- treated neutrophil to understand the TNF- induced cell death or apoptosis and these experiment is further confirmed by DNA fragmentation analysis. These results clearly demonstrate that TNF- induces cellular death of human neutrophil at least in part by enhanced expression of Ca-independent -PKC. These observations provide an insight towards understanding the function of -PKC in apoptotic pathway.     

Vorläufige Versuche über die Elastizität des Atraktoplasmas

Summary Spindles of nuclei mostly from metaphases of expiants in vitro, Set free according to Dan's method by means of solutions of Duponol (a sodium lauryl sulfonate), have been studied according to their elasticity. The isolated atractoplasm has shown a static modul of elasticityE0.8 to 1.6×10⁶ dynes×cm^–2, and, sometimes, a Poisson number of0.42, that is to say the atractoplasmic gel of the spindles of nuclei is certain elastically. The values of the modul of elongation agree satisfactory with those values recently ascertained by Crick and Hughes with cytoplasm oi explanted cells.     

Hereditary respiration deficiency in Saccharomycodes ludwigii

Saccharomycodes ludwigii, supposed to be petite-negative, gave rise to respiration-deficient mutants when acriflavine and ultraviolet irradiation, respectively, were applied to this yeast, strain IFO 1194. The frequency of such mutants was very low as compared with that in Saccharomyces cerevisiae and other petite-positive yeasts. Cytochrome composition was characterized by spectrophotometry at the temperature of liquid nitrogen. The respiratory mutants examined contained cytochrome c unaltered in quality and quantity. Cytochrome b was often present only in small amounts though never absent, while cytochrome a+a₃ was either present or absent. The respiratory mutants could form zygotes after conjugation with a wild-type culture of opposite mating type ( vs. a). The hybridization and segregation analysis of spore tetrads showed the inheritance of respiratory mutant character to be either Mendelian or non-Mendelian and similar to that of pet (nuclear) and rho^- (cytoplasmic) mutants, respectively, in Saccharomyces cerevisiae.     

Antlers as biomonitors of environmental pollution by lead and fluoride: A review

Antlers are periodically cast and re-grown cranial appendages of deer. Both endochondral and intramembranous ossification are involved in the formation of antler bone. After velvet shedding, antlers are completely bony structures which are referred to as hard antlers. Growing antlers accumulate substances with an affinity to mineralized tissues. Among these substances are lead and fluoride. Due to the seasonally limited life span of antlers, the concentrations of lead and fluoride in hard antlers reflect uptake by the forming bone during a defined, species-specific period of some months. Antlers can thus be viewed as naturally standardized environmental samples that are well suited as biomonitors of environmental pollution by bone-seeking contaminants. Because hard antlers are collected by hunters as trophies and kept in private or public collections, material for study can be obtained rather easily. So far, lead concentrations in hard antlers have been reported only from Europe, whereas data for fluoride are available from both Europe and North America. Some studies compared contaminant concentrations in antlers from different regions, while others analyzed time-trends in contaminant levels in antlers from a single region. Using the latter approach, a pronounced drop of lead concentrations during recent decades has been reported for antlers from various European countries. This indicates a marked decline of environmental lead levels that can be attributed mainly to the phase-out of leaded gasoline and, in addition, to a reduction of lead emissions also from stationary sources. In Germany, a corresponding drop has also been recorded for fluoride concentrations in antlers, which is attributed to a decline of fluoride emissions from stationary sources due to improved emission-control measures. There is some evidence that exposure to higher levels of fluoride may lead to an impaired antler mineralization. Using antlers as biomonitors has been shown to be an efficient method for assessing environmental pollution by lead and fluoride at low cost. Further studies using this now well established approach are therefore encouraged.This revised version was published online in May 2005. The following errors were eliminated:1. In the chapter Antler growth and mineralization in the last sentence of the first paragraph a slash was missing between the words reindeer and caribou; 2. In the second paragraph of the same chapter the word reiudees was corrected to reindeer; 3. In the chapter Accumulation of lead and fluoride in growing antlers the word lightest was corrected to highest.     

Photosynthetic potential of phytoplankton in the deep water of Lake Baikal,Russia

We examined the photosynthetic activity of the phytoplankton community collected from the surface to a depth of 1000m in the south basin of Lake Baikal. Experiments were conducted in June (mixing period) and August (stratified period). The carbon fixation rate was measured by the use of a ¹³C tracer after the incubation of samples under light conditions in the upper water column. Photosynthetic fixation of ¹³C was detected for samples collected from a depth of 500m, indicating the viability of phytoplankton in deep water. The community composition was dominated by Bacillariophyceae in deep water. The finding of lower activity at a depth of 200m than that at a depth of 500m in August suggests that the spring diatom bloom could be a significant source of viable populations at a depth of 500m. Photosynthetic activity was not detected in samples collected at a depth of 1000m.     

Effects of aquatic macrophytes on water quality and phytoplankton communities in shallow lakes

We investigated aquatic macrophytes, water quality, and phytoplankton biomass and species composition in three shallow lakes with different levels of vegetation cover and nutrient concentration in Kushiro Moor, during August 2000. Trapa japonica can live in a wide range of nutrient levels. This species forms an environment with a steeper extinction of light, higher concentrations of dissolved organic carbon (DOC), lower concentrations of dissolved oxygen (DO) near the bottom, and lower concentrations of nitrate+nitrite and soluble reactive phosphorus (SRP) than other vegetation types. The pH was much higher in a Polygonum amphibium community, and the DO near the bottom did not decrease compared to a T.japonica community in the summer. The relationship between chlorophyll a and the limiting nutrient (total phosphorus (TP) when total nitrogen (TN):TPis 10 and TN/10 when TN:TP is <10) significantly differed between lakes with and without submerged vegetation. The chlorophyll a concentrations at a given nutrient level were significantly lower in water with submerged macrophytes than in water without them. Correspondence analysis showed that the difference in phytoplankton community structure across sites was largely due to the presence or absence of submerged macrophytes, and the ordination of phytoplankton species in the lakes with submerged macrophytes is best explained by environmental gradients of TN, chlorophyll, pH and SRP.     

Structure of theEscherichia coli ATP synthase and role of the γ and ε subunits in coupling catalytic site and proton channeling functions

The structure of theEscherichia coli ATP synthase has been studied by electron microscopy and a model developed in which the and subunits of the F₁ part are arranged hexagonally (in top view) alternating with one another and surrounding a central cavity of around 35 Å at its widest point. The and subunits are interdigitated in side view for around 60 Å of the 90 Å length of the molecule. The F₁ narrows and has three-fold symmetry at the end furthest from the F₀ part. The F₁ is linked to F₀ by a stalk approximately 45 Å long and 25–30 Å in diameter. The F₀ part is mostly buried in the lipid bilayer. The subunit provides a domain that extends into the central cavity of the F₁ part. The and subunits are in a different conformation when ATP+Mg²⁺ are present in catalytic sites than when ATP+EDTA are present. This is consistent with these two small subunits switching conformations as a function of whether or not phosphate is bound to the enzyme at the position of the phosphate of ATP. We suggest that this switching is the key to the coupling of catalytic site events with proton translocation in the F₀ part of the complex.     

Proliferation and germination of somatic embryos from embryogenic suspension cultures in <Emphasis Type="Italic">Phœnix dactylifera</Emphasis>

The present study demonstrates a procedure for the rapid development of a high number of somatic embryos from embryogenic suspension culture. This method might be efficient for mass propagation of Phnix dactylifera L. Embryogenic callus placed in liquid medium with 10^–5M ABA yielded an average 72 embryos per 100ml of culture medium within 2months, while those placed on solid medium yielded an average of 33, 20 and 16 embryos per 100ml of culture medium respectively for 10^–7, 10^–6 and 10^–5 M ABA after 4months. The combination of 2,4-DIchlorophenoxyacetic acid (2,4-D) (4.5×10^–7M), glutamine (6.7×10^–4M), and ABA (10^–5M) (L8 liquid medium) showed a beneficial effect on somatic embryos production compared to 2,4-D and glutamine alone, while this combination significantly (p<0.05) increased the accumulation of storage proteins (144 and 138mgg^–1 DW respectively for Jihel and Bousthami noir cultivars) in somatic embryos. The somatic embryos which underwent maturation on medium containing only 4.5×10^–7M 2,4-D and 10^–5M ABA (L6 liquid medium) accumulated more sugars (292 and 265mgg^–1 DW respectively for Jihel and Bousthami noir) than those matured on any other liquid medium. Histological studies revealed that somatic embryos (developed in L6 and L8 liquid media) accumulated less reserve compounds (proteins and sugars) than zygotic embryos. The addition of activated charcoal (0.25 and 0.5gl^–1) and phytagel^® (2.5gl^–1) to the germination medium may be useful for enhancing the germination of Phnix dactyliferasomatic embryos.     

Multiple pathways for amino acid transport inMytilus gill

Summary The epidermal tissues of marine mussels can accumulate amino acids from surrounding sea water. In the present study, gill tissue isolated from the California coastal mussel,Mytilus californianus, was used in conjunction with intact, actively pumping mussels to study epidermal transport processes. There appeared to be at least four pathways for this uptake: i) a -neutral pathway which transports taurine; ii) an -acidic pathway specific for substrates such as aspartate; iii) an -neutral pathway having a general specificity for this class of compound, but which also accepts the basic amino acid, lysine; and iv) a second -neutral pathway, also of broad specificity, capable of accepting the imino acid, proline, as a substrate. Replacement of Na in sea water with choline reduced uptake of leucine, taurine, aspartate, and proline by more than 95%, and reduced lysine uptake by 75%, suggesting that Na-independent pathways play no significant role in epidermal transport in the gill. Isolated gill tissue was used to estimate the maximum transport capacities (J _max's) of the pathways, which ranged from approximately 5 to 25 mol/(g·hr). Apparent Michaelis constants (K _t ^*'s) of the epidermal transporters were estimated using a convection-diffusion model introduced previously (Wright and Secomb, Am J Physiol 247:R346–R355, 1984). TheseK _t ^*'s ranged from 1 to 5 M. The characteristics of the epidermal transporters are such that they can play a significant role in both animal nutrition and in the reacumulation of endogenous amino acids lost from surface cells through passive diffusion.Abbreviation ASW artificial sea water     

The cytology of phytophthora infestans

Summary Phytophthora infestans has three kinds of somatic nuclei: an oval shaped nucleus (approx. 3.1×2.7 ) which stains diffusely except for a crescent shaped Feulgen positive cap which stains intensely; a granular nucleus whose contents are organized into a fairly constant number of stained bodies, and, a deeply staining condensed nucleus. The capped nucleus is thought to be metabolic or resting and the granular nucleus is thought to be dividing as it is most commonly found in hyphal tips. Attenuated forms of all three kinds of nuclei are found.Nuclear division is mitotic and intranuclear. Eight—ten chromosomes are seen at metaphase.Sporangia have a mean of 6.3 nuclei which is constant for age and strain of culture. Sporangia become multinucleate as a result of nuclear migration and not by division in the developing sporangium. Zoospores are usually uninucleate.The nuclear cap is persistent throughout nuclear division when it also divides. It is associated with flagella production and nuclear migration and has some of the properties of a blepharoplast.     

Somatic embryogenesis from cultured leaf segments of Zea mays

Basal leaf segments of 3 to 4 week old maize (Zea mays L.) seedlings plated on SH medium with 30 M dicamba produced embryogenic callus and/or somatic embryos. Histological evidence showed that some of the embryos arose directly from the explant. When leaf segments with embryos were transferred to MS medium with 1.0 M NAA, 1.0 M IAA, 2.0 M 2iP, and 60 g/l sucrose, the embryos germinated and the resulting seedlings could be established in culture tubes. These responses were obtained from three inbred lines, CHI31, S615, and S7.Abbreviations SH Schenk and Hildebrandt (1972) medium - MS Murashige and Skoog (1962) medium - dicamba 3,6-dichloro-o-anisic acid - IAA indole-3-acetic acid - NAA -naphthaleneacetic acid - 2iP 2-isopentyladenine     

Subcellular localisation of some acid hydrolases Triturus cristatus spermatozoa

Summary The histochemical localisation of acid -glycerophosphatase has been studied, by optical and electron microscopy, in the sperm cells of the newt. The enzymatic activity is restricted to some of the acrosomal structures, other parts of the cell being unreactive.The subcellular localisation of this enzyme, and of other four acid hydrolases (Proteinase, DNase, RNase and -glucuronidase) has been studied using biochemical techniques. The sedimentability of the before mentioned hydrolases with the acrosomes detached from the entire cells and their solubilisation by Triton X-100, have been examined.The evidence obtained was discussed in relation to the hypothesis of a lysosomal-like nature of the acrosome.     

Protoplasmic streaming in the giant unicellular green algaAcetabularia mediterranea

Summary Protoplasmic Streaming inAcetabularia mediteranea has been studied by microcinematography in 1. germinating zygotes, 2. germlings before the differentiation of rhizoids and apices, 3. young cells with rhizoids and apices, 4. vegetative cells-several centimeters in length, 5. cells with a maximum sized cap, containing secondary nuclei, and 6. cells after cyst formation. Intracellular transport is found to occur at a network-system of thin filaments and at a different system of headed streaming bands. At the network of filaments chloroplasts are found to move at a velocity of 1–2 m/sec. Headed streaming bands move along the filaments and may lead without interruption from the rhizoid to the apex of the cell andvice versa. The front zone of the streaming bands is occupied by a leading cytoplasmic head-structure. Small vesicles, polyphosphate granula and secondary nuclei are the predominant moving structures in headed streaming bands. The velocity of these particles is found to be 3–11 m/sec. The filament system is found during all developmental stages. Headed streaming bands are undetectable in germinating zygotes and develop from small cytoplasmic droplets in germlings to broad heavily loaded bands in the huge vegetative cell.Transport of secondary nuclei by headed streaming bands is not observed during mitotic divisions and after cyst formation, though moving bands are still present for several weeks after cyst formation.