The effect of rock composition on cyanobacterial weathering of crystalline basalt and rhyolite.

The weathering of volcanic rocks contributes significantly to the global silicate weathering budget, effecting carbon dioxide drawdown and long-term climate control. The rate of chemical weathering is influenced by the composition of the rock. Rock-dwelling micro-organisms are known to play a role in changing the rate of weathering reactions; however, the influence of rock composition on bio-weathering is unknown. Cyanobacteria are known to be a ubiquitous surface taxon in volcanic rocks. In this study, we used a selection of fast and slow growing cyanobacterial species to compare microbial-mediated weathering of bulk crystalline rocks of basaltic and rhyolitic composition, under batch conditions. Cyanobacterial growth caused an increase in the pH of the medium and an acceleration of rock dissolution compared to the abiotic controls. For example, Anabaena cylindrica increased the linear release rate (R(i)(l)) of Ca, Mg, Si and K from the basalt by more than fivefold (5.21-12.48) and increased the pH of the medium by 1.9 units. Although A. cylindrica enhanced rhyolite weathering, the increase in R(i)(l) was less than threefold (2.04-2.97) and the pH increase was only 0.83 units. The R(i)(l) values obtained with A. cylindrica were at least ninefold greater with the basalt than the rhyolite, whereas in the abiotic controls, the difference was less than fivefold. Factors accounting for the slower rate of rhyolite weathering and lower biomass achieved are likely to include the higher content of quartz, which has a low rate of weathering and lower concentrations of bio-essential elements, such as, Ca, Fe and Mg, which are known to be important in controlling cyanobacterial growth. We show that at conditions where weathering is favoured, biota can enhance the difference between low and high Si-rock weathering. Our data show that cyanobacteria can play a significant role in enhancing rock weathering and likely have done since they evolved on the early Earth.

Enhancing phosphorus removal in constructed wetlands with ochre from mine drainage treatment.

No single end-use has yet been identified that is capable of consuming the projected production of ochre (mainly iron (III) oxides) from mine drainage treatment. However, the high sorption capacity of ochre for phosphorus (up to 26 mg kg(-1)) means that it could be used in constructed wetlands to enhance phosphorus removal. Laboratory batch experiments showed that coarse-grained ochre removes 90% of all phosphorus forms from sewage effluent after 15 minutes of shaking. From a larger-scale experiment, it is estimated that constructed wetlands with an ochre substrate should remove phosphorus from sewage effluent for up to 200-300 years. The suitability of ochre for phosphorus removal is being investigated at the field scale in a wastewater constructed wetland (175 m2 area) in Berwickshire, UK. The hydraulic and treatment performance of the wetland were monitored for 15 months prior to installation at the inlet in November 2003 of a tank containing approximately 1200 kg ochre. Results so far show that improved hydraulic design is required for ochre to increase the mean phosphorus removal efficiency of the system (27 +/- 28%), but potentially toxic metals (Al, Cd, Cr, Cu, Fe, Ni, Pb, Zn) have not been released from the ochre into the wetland outflow.

An IS257-derived hybrid promoter directs transcription of a tetA(K) tetracycline resistance gene in the Staphylococcus aureus chromosomal mec region.

Transcription of the tetA(K) tetracycline resistance determinant encoded by an IS257-flanked cointegrated copy of a pT181-like plasmid, located within the chromosomal mec region of a methicillin-resistant Staphylococcus aureus isolate, has been investigated. The results demonstrated that transcription of tetA(K) in this strain is directed by both an IS257-derived hybrid promoter, which is stronger than the native tetA(K) promoter in the autonomous form of pT181, and a complete outwardly directed promoter identified within one end of IS257. Despite lower gene dosage, the chromosomal configuration was shown to afford a higher level of resistance than that mediated by pT181 in an autonomous multicopy state. Furthermore, competition studies revealed that a strain carrying the chromosomal tetA(K) determinant exhibited a higher level of fitness in the presence of tetracycline but not in its absence. This finding suggests that tetracycline has been a selective factor in the emergence of strains carrying a cointegrated pT181-like plasmid in their chromosomes. The results highlight the potential of IS257 to influence the expression of neighboring genes, a property likely to enhance its capacity to mediate advantageous genetic rearrangements.

HZwint-1, a novel human kinetochore component that interacts with HZW10.

HZwint-1 (Human ZW10 interacting protein-1) was identified in a yeast two hybrid screen for proteins that interact with HZW10. HZwint-1 cDNA encodes a 43 kDa protein predicted to contain an extended coiled-coil domain. Immunofluorescence studies with sera raised against HZwint-1 protein revealed strong kinetochore staining in nocodazole-arrested chromosome spreads. This signal co-localizes at the kinetochore with HZW10, at a position slightly outside of the central part of the centromere as revealed by staining with a CREST serum. The kinetochore localization of HZwint-1 has been confirmed by following GFP fluorescence in HeLa cells transiently transfected with a plasmid encoding a GFP/HZwint-1 fusion protein. In cycling HeLa cells, HZwint-1 localizes to the kinetochore of prophase HeLa cells prior to HZW10 localization, and remains at the kinetochore until late in anaphase. This localization pattern, combined with the two-hybrid results, suggests that HZwint-1 may play a role in targeting HZW10 to the kinetochore at prometaphase. HZwint-1 was also found to localize to neocentromeres and to the active centromere of dicentric chromosomes. HZwint-1 thus appears to associate with all active centromeres, implying that it plays an important role in correct centromere function.

The effect of an insulin releasing agent, BTS 67582, on advanced glycation end product formation in vitro.

BTS 67582 (1,1-dimethyl-2-(2-morpholinophenyl) guanidine fumarate) is an insulin-releasing agent currently in phase II clinical trials. Its effect on advanced glycation end product (AGE) formation was measured in the BSA/D-glucose and L-lysine/glucose-6-phosphate assay systems and Amadori product formation was measured in the BSA/D-glucose assay system, following a 3 week incubation period. In the BSA/D-glucose assay system, 200 mM BTS 67582 caused an approximate 70% inhibition in AGE formation (p<0.001), whilst at 20 mM and 2 mM it caused a marginal inhibition (21%, (p<0.001) and 8% respectively). 200 mM and 20 mM aminoguanidine-HCl inhibited AGE formation by 95% and 69% (p<0.001), respectively, whereas 2 mM aminoguanidine-HCl had no significant effect. Tolbutamide (200 microM) and glibenclamide (100 microM) had significant, but only marginal, effects on AGE formation (16% and 17%, respectively, p<0.01). In the BSA/D-glucose assay system 200 mM BTS 67582 and 200 mM aminoguanidine-HCl retarded Amadori product formation by 88% (p<0.001) and 60% (p<0.01), respectively. BTS 67582 at 20 mM and 2 mM was shown to inhibit Amadori product formation by 67% and 57%, respectively, (p<0.01). In the lysine and glucose-6-phosphate assay system 200 mM BTS 67582 and 200 mM aminoguanidine-HCl were shown to inhibit AGE formation by about 70% and 96% (p<0.001), respectively. Tolbutamide (200 microM) and glibenclamide (100 microM) had no significant effect on AGE formation.

A molecular investigation of the obese phenotype in the Aston University strain of ob/ob mice and the Japanese Kuo Kondo mice.

The aim of this work was to determine the role of the ob gene in the obese phenotype observed in the Aston University Strain of ob/ob mouse and the Japanese Kuo Kondo (KK) mouse. After RT-PCR amplification of the ob RNA, the transcript was cloned into the vector pCR3 and three individual clones from each strain were sequenced. It was confirmed that the Aston University strain of ob/ob mice shared the same C-T mutation found in the Jackson Laboratory C57BL/6J ob/ob strain whereas the Japanese KK mice showed wild-type ob gene expression. This study indicates that the ob mutation has survived unchanged following the separation of the two strains of ob/ob mice, and secondly, that the molecular basis of the obese phenotype in the KK mice is not due to mutations in the ob gene.

The cytochrome P450 4 (CYP4) family.

1. The CYP4 family consists of 11 subfamilies (CYP4A-CYP4M), which encode constitutive and inducible isozymes expressed in both mammals and insects. 2. The CYP4A subfamily encodes several cytochrome P450 enzymes that are capable of hydroxylating the terminal omega-carbon and, to a lesser extent, the (omega-1) position of saturated and unsaturated fatty acids, as well as enzymes active in the omega-hydroxylation of various prostaglandins. 3. The CYP4A1, A2 and A3 genes, the most extensively studied members of the CYP4 family, are expressed constitutively in rat liver and kidney and their expression is induced by a class of chemicals known as peroxisome proliferators, which includes the hypolipidemic drug, clofibrate. 4. Induction of CYP4A expression by clofibrate is due to transcriptional activation, mediated possibly via a peroxisome proliferator activated receptor (PPAR). 5. CYP4A gene expression is hormonally regulated. 6. The CYP4A1-3 genes are expressed constitutively and following induction in pregnant and lactating rats. 7. Translactational and transplacental induction of the CYP4A1-3mRNAs and proteins has been demonstrated. 8. There is a close association between microsomal CYP4A1 induction, peroxisome proliferation and induction of the peroxisomal fatty acid metabolizing system. 9. The CYP4A subfamily may be involved in the metabolism of arachidonic acid leading to the formation of physiologically important metabolites involved in such processes as blood flow in the kidney, cornea and brain.

An investigation of the temporal induction of cytokine mRNAs in LPS-challenged thioglycollate-elicited murine peritoneal macrophages using the reverse transcription polymerase chain reaction.

OBJECTIVE AND DESIGN: A comprehensive study to standardise interleukin (IL)-1 alpha, -1 beta, -6, -10, -12 and tumour necrosis factor alpha (TNF alpha) mRNA detection in murine peritoneal macrophages, using the reverse transcription polymerase chain reaction (RT-PCR) was carried out. SUBJECTS: Thioglycollate-elicited peritoneal exudate cells were harvested from female BALB/c mice and the adherent macrophage fraction isolated for use. TREATMENT: Peritoneal macrophages (1 x 10(6)) were incubated in the presence or absence of lipopolysaccharide (LPS; at a final concentration of 1 microgram/ml) for 0, 1, 2, 3, 4, 5 and 24 h. METHODS: Culture supernatants and cells were harvested at each time point, the secreted cytokine protein levels quantified by sandwich immunoassays and the cytokine mRNA levels assessed by RT-PCR. RESULTS: The IL-6 mRNA was not expressed in detectable amounts in the macrophages, unless challenged with LPS. TNF alpha, IL-1 alpha, IL-1 beta, IL-10 and IL-12 mRNAs were expressed in both stimulated and unstimulated macrophages. The levels of the PCR products and thus mRNAs of all the cytokines increased with LPS stimulation, maximal levels being achieved 3 to 5 h post stimulation. CONCLUSIONS: RT-PCR produced consistent results, indicating that this technique could be used to investigate the effect of biological mediators and novel pharmacological agents on cytokine mRNA levels in macrophages.

Placental transfer of the hypolipidemic drug, clofibrate, induces CYP4A expression in 18.5-day fetal rats.

Rats at day 15.5 of gestation were dosed intraperitoneally with 300 mg.kg-1 of clofibrate for three consecutive days at 24-hr intervals and were culled 24 hr after the final injection. This regime produced maximal induction of the cytochrome P4504A (CYP4A) mRNAs in the maternal liver and kidney and in 18.5-day fetal tissues. The maternal hepatic and renal CYP4A mRNA levels had risen 12- and 2-fold, respectively, above the constitutive levels seen in untreated pregnant rats at an equivalent stage of gestation. Clofibrate was capable of traversing the placenta and modulating the fetal CYP4A mRNA expression as demonstrated by a 3-fold elevation in the mRNA levels in those fetuses explanted from drug-induced mothers, compared with those fetuses removed from untreated mothers. The CYP4A mRNAs were demonstrated in the fetal liver via dot-blot and Northern blot analyses. In addition, low levels of CYP4A mRNA expression were detected in the induced placenta via Northern blot analysis. Western blot analysis revealed that the CYP4A protein levels increased in the maternal liver and in the kidney and fetal livers after exposure to clofibrate. Peroxisome proliferation, a phenomenon associated with induction of CYP4A1 expression in rodents, was demonstrated in both maternal and fetal livers, with the use of light and electron microscopy.

Translactational induction of CYP4A expression in 10.5-day neonatal rats by the hypolipidemic drug clofibrate.

Lactating mothers of 7.5-day neonatal rats were injected intraperitoneally with 500 mg kg-1 clofibrate for 3 consecutive days at 24-hour intervals; 24 hours after the final injection, the maternal cytochrome P450 4A (CYP4A) mRNA levels had risen 14- and 2.5-fold above the constitutive levels of expression seen in the liver and kidney, respectively. Lactational transfer of clofibrate to the suckling 10.5-day litter was demonstrated by the 15- and 5-fold elevation observed in the neonatal hepatic and renal CYP4A mRNAs, respectively, following suckling from drug-induced mothers. A significant decrease in the relative liver weights of these neonatal pups was seen following clofibrate exposure via maternal milk, in total contrast to the normally observed increase in liver/body weight ratios of rats treated with clofibrate. Western blot analysis using a polyclonal goat anti-rat CYP4A1 antibody also demonstrated a rise in the CYP4A protein levels in both the mothers and their litters following maternal clofibrate treatment.

Single photon emission computerised tomography in elderly patients with Alzheimer's disease and multi-infarct dementia. Regional uptake of technetium-labelled HMPAO related to clinical measurements.

Single photon emission computerised tomography (SPECT) was used to measure regional brain uptake of technetium-99m hexamethylpropyleneamine oxine (Tc99m-HMPAO) in elderly patients with Alzheimer's disease (AD), multi-infarct dementia (MID) and normals (n = 20 in each group). Different patterns of uptake were found between groups when cortical uptake was normalised to cerebellar uptake. Reductions occurred in all regions in AD, being most marked in temporal and posterior parietal areas. Significant correlations were found in AD between memory impairment and decreased temporal uptake bilaterally, and between duration of illness and reduced uptake in most brain regions. MID patients showed higher uptake in the anterior parietal region than did the other groups. A variable comparing anterior to posterior uptake significantly discriminated the two patient groups.

Temperamental characteristics of three- to four-year-old boys and girls and child-family interactions.

Mothers interviewed in their homes provided information concerning their child's temperamental characteristics, mild behaviour problems and family interactions. The characteristics active, shy (approach/withdrawal; not settling in) and moody/intense were consistent from 42 to 50 months of age for both sexes. No simple sex differences appeared in these three characteristics. There was little evidence for an association between active and either mild behaviour problems or family interactions. For both girls and boys, moody/intense scores were associated with mild behaviour problems and 'negative' family interactions. However, while shy scores of boys were associated with negative family interactions, shy scores of girls were associated with positive interactions, especially at the later age of 50 months.

An ion exchange chromatographic method for the determination of synaptosomal calcium uptake.

A simple, rapid method for determining depolarization-induced 45Ca influx into synaptosomes is described. Synaptosomes which had been depolarized in the presence of 45Ca were applied to a small column of Chelex-100 resin to separate free Ca2+ from that taken up by the tissue. Approximately 70% of the synaptosomal protein applied to the column was recovered in the initial eluate. The magnitude of 45Ca uptake was dependent on the amount of Ca2+ in the incubation medium and on the KCI concentration. Calcium influx reached a plateau after 90 sec of incubation at 24 degrees C. The Na+ channel activator veratridine also produced a substantial influx of 45Ca, and this effect was blocked by tetrodotoxin. Thus, this ion exchange procedure makes it possible to measure depolarization-induced Ca2+ influx in synaptosomes without subjecting them to high vacuum or centrifugation pressures or to EGTA-containing solutions.

Temperament and relationships.

Temperamental characteristics of preschool children were related to relationships, mothers' mood, and children's feelings towards family members. Active scores were correlated with some problem behaviour and with negative aspects of interactions with mother and father. Active children felt themselves to be independent, while Timid children did not. Timid children had relatively more acute illnesses and worries and fears. Difficult scores were correlated with problem behaviour and with negative aspects of all the types of interaction assessed: mother-child, mother-father, father-child, and sibling-child. Furthermore, the more Difficult the child, the more Anxious, Outwardly Irritable, and Inwardly irritable the mother. In turn, Difficult children expressed relatively few positive and dependency feelings towards their mothers, and more negative feelings towards themselves.

Infant-related influences on birth intervals in rhesus monkeys.

That a mother's relationship with her infant could influence her subsequent reproductive history can be argued as follows. A rhesus monkey infant whose next sibling is to be born in a succeeding birth season could compete with it by trying to postpone the date when its mother next conceives. If the mother preferred a shorter delay than the infant, processes of conflict and negotiation involving the two could show in some of the behavioural interactions constituting their relationship. For example, a mother could try to hasten the onset of her next pregnancy by trying to promote more independent behaviour in her infant, perhaps by rejecting some of its bids for contact. We report here studies on our rhesus monkey colony and other colonies which support this argument.