High-latitude platform carbonate deposition constitutes a climate conundrum at the terminal Mesoproterozoic.

During the Mesoproterozoic Era, 1600 to 1000 million years ago, global climate was warm with very little evidence of glaciation. Substantial greenhouse warming would have been required to sustain this ice-free state given 5-18% lower solar luminosity. Paleomagnetic data reported here place voluminous ca. 1.2 Ga shallow marine carbonate deposits from India at an unexpectedly high latitude of around 70° from the equator. Previous studies noted high latitudes, but their implication was never considered. Here, we evaluate the temporal-latitudinal distribution of neritic carbonate deposits across the Proterozoic and identify similar deposits from North China that together with those from India are seemingly unique to the late Mesoproterozoic. A uniformitarian interpretation implies that this is cold-water carbonate deposition, but facies similarity with low-latitude neritic deposits rather suggests a hotter climate and elevated polar ocean temperatures of 15-20° or higher. This interpretation represents a climate conundrum that would require much greater greenhouse warming than documented for the Mesoproterozoic.

Elastohydrodynamic lubricant flow with nanoparticle tracking.

Lubricants operating in elastohydrodynamic (EHD) contacts exhibit local variations in rheological properties when the contact pressure rises. Direct evidence of this behaviour has only been obtained by examining through-thickness velocity profiles U(z) of lubricants in a contact using luminescence-based imaging velocimetry. In the present study, nanoparticles (NPs) are added to polybutene (PB) as tracers to investigate the effect of pressure on the flow of PB in an EHD contact. By tracking NPs in the contact, particle velocity distributions f(U) under various pressures are obtained and found to be pressure dependent. Results show quantitatively that f(U) and U(z) are correlated and thus confirm that U(z) of PB changes from Couette flow to partial plug flow above a critical pressure. This confirmation highlights the complexity of lubricant rheology in a high pressure contact.

Comparing the molecular and global rheology of a fluid under high pressures.

The viscosity of liquids is a strong function of pressure. While viscosity is relatively easy to measure at low pressure, high-pressure rheology presents significant experimental challenges. As a result, rheological models are often used to extrapolate viscosity from low pressure measurements to higher pressures. Techniques to obtain data over a wide range of pressures and shear rates, as well as understanding the validity and limitations of methods to fill the gaps in the available data, are therefore of crucial practical and theoretical importance. This work examines the viscosity of polyalphaolefin (PAO) by combining average global area averaged measurements at high pressure and local molecular viscosity measurements at moderate pressures. Viscosities spanning five orders of magnitude are examined at pressures up to 720 MPa. High pressure results were obtained with friction measurements where the fluid is sheared between two surfaces in a loaded point contact. The local molecular microviscosity at medium and low pressures was measured by applying a technique based on fluorescence anisotropy, which probes the rotational motion of dye molecules in a nanoscale film under shear. Both sets of measurements are taken in the same configuration, an elastohydrodynamic (EHD) contact. This is the first set of quantitative local viscosity measurements that have been verified against both friction and high pressure rheometry measurements. Commonly used rheological models were compared to experimental results. Our work shows that fluorescence anisotropy and friction measurements can be used to determine the viscosity of liquids over a wide range of conditions from a single experimental setup. The results obtained match results from low- and high-pressure rheometry for PAO. The importance of correcting friction data for pressure non-uniformity, temperature and shear thinning is also highlighted.

Diagnostic accuracy of colour Doppler ultrasonography, CT angiography and blood-pool-enhanced MR angiography in assessing carotid stenosis: a comparative study with DSA in 170 patients.

PURPOSE: This study was undertaken to prospectively evaluate the diagnostic performance of colour Doppler ultrasonography (CDUS), first-pass (FP) and steady-state (SS) contrast-enhanced magnetic resonance angiography (MRA) and computed tomography angiography (CTA) of the carotid arteries using digital subtraction angiography (DSA) as the reference standard. MATERIALS AND METHODS: A total of 170 patients with previous cerebrovascular events and suspected carotid artery stenoses underwent CDUS, blood-pool MRA, CTA and DSA. Accuracy, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were calculated for CDUS, FP MRA, SS MRA and CTA. The McNemar and Wilcoxon tests and receiver operating characteristic (ROC) curve analysis were used to determine significant differences (p<0.05) between the diagnostic performances of the four modalities, and the degree of stenosis was compared using linear regression. RESULTS: A total of 336 carotid bifurcations were studied. The area under the curve (AUC) for degree of stenosis was: CDUS 0.85±0.02, FP MRA 0.982±0.005, SS MRA 0.994±0.002 and CTA 0.997±0.001. AUC analysis showed no statistically significant difference between CTA and MRA (p=0.0174) and a statistically significant difference between CDUS and the other techniques (p<0.001). Plaque morphology analysis showed no significant difference between CTA and SS MRA; a significant difference was seen between CTA and SS MRA versus FP MRA (p=0.04) and CDUS (p=0.038). Plaque ulceration analysis showed a statistically significant difference between MRA and CTA (0.04< p<0.046) versus CDUS (p=0.019). CONCLUSIONS: CTA is the most accurate technique for evaluating carotid stenoses, with a slightly better performance than MRA (97% vs. 95% for SS MRA and 92% for FP MRA) and a greater accuracy than CDUS (97% vs. 76%). Blood-pool contrast-enhanced SS sequences offer improved evaluation of degree of stenosis and plaque morphology with accuracy substantially identical to CTA.

[Current treatment strategies for sigmoid diverticulitis]. / Nouveautés dans la stratégie thérapeutique de la diverticulite sigmoïdienne.

Treatment of colonic diverticular disease has evolved over the past years. Most episodes are simple and can be successfully treated with antibiotics alone. For complicated diverticulitis, a strong trend is developing towards less invasive therapies including interventional radiology and laparoscopic lavage in an effort to avoid the morbidity and discomfort of a diverting colostomy. Based on a better understanding of the natural history of the disease, the indication to prophylactic colectomy after a few episodes of simple diverticulitis has been seriously challenged. For those patients who need a colectomy, single port laparoscopy, NOTES and transanal specimen extraction are being proposed. However larger studies are needed to confirm the hypothetical advantages of these evolving techniques.

c-Jun N-terminal kinase binding domain-dependent phosphorylation of mitogen-activated protein kinase kinase 4 and mitogen-activated protein kinase kinase 7 and balancing cross-talk between c-Jun N-terminal kinase and extracellular signal-regulated kinase pathways in cortical neurons.

The c-Jun N-terminal kinase (JNK) is a mitogen-activated protein kinase (MAPK) activated by stress-signals and involved in many different diseases. Previous results proved the powerful effect of the cell permeable peptide inhibitor d-JNKI1 (d-retro-inverso form of c-Jun N-terminal kinase-inhibitor) against neuronal death in CNS diseases, but the precise features of this neuroprotection remain unclear. We here performed cell-free and in vitro experiments for a deeper characterization of d-JNKI1 features in physiological conditions. This peptide works by preventing JNK interaction with its c-Jun N-terminal kinase-binding domain (JBD) dependent targets. We here focused on the two JNK upstream MAPKKs, mitogen-activated protein kinase kinase 4 (MKK4) and mitogen-activated protein kinase kinase 7 (MKK7), because they contain a JBD homology domain. We proved that d-JNKI1 prevents MKK4 and MKK7 activity in cell-free and in vitro experiments: these MAPKK could be considered not only activators but also substrates of JNK. This means that d-JNKI1 can interrupt downstream but also upstream events along the JNK cascade, highlighting a new remarkable feature of this peptide. We also showed the lack of any direct effect of the peptide on p38, MEK1, and extracellular signal-regulated kinase (ERK) in cell free, while in rat primary cortical neurons JNK inhibition activates the MEK1-ERK-Ets1/c-Fos cascade. JNK inhibition induces a compensatory effect and leads to ERK activation via MEK1, resulting in an activation of the survival pathway-(MEK1/ERK) as a consequence of the death pathway-(JNK) inhibition. This study should hold as an important step to clarify the strong neuroprotective effect of d-JNKI1.

Molecular epidemiology of Mycobacterium bovis isolates from free-ranging wildlife in South African game reserves.

Bovine tuberculosis is endemic in African buffalo and a number of other wildlife species in the Kruger National Park (KNP) and Hluhluwe-iMfolozi Park (HiP) in South Africa. It was thought that the infection had been introduced into the KNP ecosystem through direct contact between cattle and buffalo, a hypothesis which was confirmed in this study by IS6110 and PGRS restriction fragment length polymorphism (RFLP) typing. The molecular characterisation of 189 Mycobacterium bovis isolates from nine wildlife species in the HiP, including three smaller associated parks, and the Kruger National Park with adjacent areas showed that the respective epidemics were each caused by an infiltration of a single M. bovis genotype. The two M. bovis strains had different genetic profiles, as demonstrated by hybridisation with the IS6110 and PGRS RFLP probes, as well as with regard to evidence of evolutionary changes to the IS profile. While the M. bovis type in HiP was transmitted between buffaloes and to at least baboon, bushpig and lion without obvious genetic changes in the RFLP patterns, in the KNP a dominant strain was represented in 73% of the M. bovis isolates, whilst the remaining 27% were variants of this strain. No species-specific variants were observed, except for one IS6110 type which was found only in a group of five epidemiologically related greater kudu. This finding was attributed to species-specific behaviour patterns rather than an advanced host-pathogen interaction.

High Mycobacterium bovis genetic diversity in a low prevalence setting.

The genetic diversity among South African Mycobacterium bovis isolates from cattle was determined by genetic fingerprinting. The restriction fragment length polymorphism (RFLP) markers IS6110 and polymorphic GC-rich sequence (PGRS) as well as spoligotyping and determination of variable number of tandem repeats (VNTR) were used to characterize sub samples of 91 M. bovis field isolates. PGRS RFLP was the single most discriminatory method and combinations of typing methods, which included IS6110 and/or PGRS had the highest discriminatory power, able to reveal 29 distinct genotypes among 35 farms with no epidemiological link. Three of the farms were co-infected with two genetically unrelated strains. In contrast to reports from European and also other colonised countries on the African continent our findings are suggestive of a high genetic diversity of M. bovis in South Africa's cattle population, implying a variety of unrelated ancestor strains. Despite effective intervention through test-and-slaughter campaigns no indication of a 'founder effect' was apparent in the panel of isolates derived from all infected provinces.

Feeding strategies for E. coli fermentations demanding an enriched environment.

The addition of a carbon nutrient feed to a fed-batch cultivation is often not enough to obtain satisfactory growth and/or production. In some cases, an additional feed with for example supplementary amino acids or complex media is required. This work presents the development of feeding strategies where more than one feed is required and the knowledge of the growth requirements is low. Simulations and cultivations with E. coli are shown using the proposed feed controllers which are based on a probing control concept. The strategies work well and they can be used to shorten the process development phase considerably.

Adhesion of Lactobacillus plantarum 423 and Lactobacillus salivarius 241 to the intestinal tract of piglets, as recorded with fluorescent in situ hybridization (FISH), and production of plantaricin 423 by cells colonized to the ileum.

AIMS: To determine which intestinal section of pre and postweaned piglets are colonized by Lactobacillus plantarum 423 and Lactobacillus salivarius 241, and follow production of plantaricin 423 in a gastro-intestinal model. METHODS AND RESULTS: Lactobacillus plantarum 423 and Lact. salivarius 241, single or in combination, were administered to 1-, 14- and 28-day-old (postweaned) piglets. According to results obtained by fluorescent in situ hybridization (FISH), Lact. plantarum 423 adhered strongly to the ileum and posterior colon and Lact. salivarius 241 to the duodenum in preweaned piglets. High numbers of strain 241 were recorded in the duodenum and posterior colon of postweaned piglets, whereas strain 423 remained localized to the ileum. Lowering in Enterococcus faecalis cell numbers were recorded when preweaned piglets were challenged with strain 241. Plantaricin 423 was produced for 96 h in the ileum section of a gastro-intestinal model. CONCLUSIONS: Lactobacillus plantarum 423 and Lact. salivarius 241 adhere to different sections of the intestinal tract, depending on the piglet's age. Ent. faecalis were inhibited in vivo, probably by plantaricin 423. SIGNIFICANCE AND IMPACT OF THE STUDY: Fluorescent in situ hybridization proved valuable in the detection of probiotic bacteria adhered to the intestine. This is the first report of bacteriocin production in a model simulating the porcine gastro-intestinal tract.

A cultivation technique for E. coli fed-batch cultivations operating close to the maximum oxygen transfer capacity of the reactor.

A cultivation strategy combining the advantages of temperature-limited fed-batch and probing feeding control is presented. The technique was evaluated in fed-batch cultivations with E. coli BL21(DE3) producing xylanase in a 3 liter bioreactor. A 20% increase in cell mass was achieved and the usual decrease in specific enzyme activity normally observed during the late production phase was diminished with the new technique. The method was further tested by growing E. coli W3110 in a larger bioreactor (50 l). It is a suitable cultivation technique when the O2 transfer capacity of the reactor is reached and it is desired to continue to produce the recombinant protein.

Probing control of glucose feeding in Vibrio cholerae cultivations.

Infection with Vibrio cholerae is a significant problem in many developing countries. Cultivation of V. cholerae is used in production of cholera toxin B subunit, which is a component in a cholera vaccine. Fed-batch cultivations with V. cholerae in defined media have been conducted and reproducible results were obtained. A probing feeding strategy developed by Akesson for Escherichia coli cultivations has been tested. The strategy is working as well for V. cholerae as for E. coli in minimizing the amount of acetic acid formed and avoiding anaerobic conditions. At 2 h after the feed start most of the acetic acid accumulated during the batch phase is consumed. The resulting feed rate tends to be the highest possible with respect to the constraints from cell metabolism and mass transfer, thus maximizing productivity in terms of biomass. A cell dry weight of 20-23 g/l is obtained after 12 h of feeding.

Production of heterologous thermostable glycoside hydrolases and the presence of host-cell proteases in substrate limited fed-batch cultures of Escherichia coli BL21(DE3).

Metabolic stress is a phenomenon often discussed in conjunction with recombinant protein production in Escherichia coli. This investigation shows how heterologous protein production and the presence of host cell proteases is related to: (1) Isopropyl-beta- D-thiogalactopyranoside (IPTG) induction, (2) cell-mass concentration at the time of induction, and (3) the presence of metabolites (glutamic acid or those from tryptone soy broth) during the post-induction phase of high cell density fed-batch cultivations. Two thermostable xylanase variants and one thermostable cellulase, all originating from Rhodothermus marinus, were expressed in E. coli strain BL21 (DE3). A three-fold difference in the specific activity of both xylanase variants [between 7,000 and 21,000 U/(g cell dry weight)], was observed under the different conditions tested. Upon induction at high cell-mass concentrations employing a nutrient feed devoid of the metabolites above, the specific activity of the xylanase variants, was initially higher but decreased 2-3 h into the post-induction phase and simultaneously protease activity was detected. Furthermore, protease activity was detected in all induced cultivations employing this nutrient feed, but was undetected in uninduced control cultivations (final cell-mass concentration of 40 g/l(-1)), as well as in induced cultivations employing metabolite-supplemented nutrient feeds. By contrast, maximum specific cellulase activity [between 700 and 900 U/(g cell dry weight)] remained relatively unaffected in all cases. The results demonstrate that detectable host cell proteases was not the primary reason for the decrease in post-induction activity observed under certain conditions, and possible causes for the differing production levels of heterologous proteins are discussed.

Characterization of south african isolates of Salmonella enteritidis by phage typing, numerical analysis of RAPD-PCR banding patterns and plasmid profiles.

Eleven of the 33 strains of Salmonella enteritidis (S.E.) included in this study belonged to phage type 34. Six strains belonged to phage type 14, six strains to phage type 4 and four strains to phage type 7. The remaining six strains belonged to phage types 35, 1, 24var (a variation of phage type 24), 9a, 1b and an unknown phage type. The majority of S.E. phage type 34 strains (eight of the 11) grouped at R2 > or =0.45 into one RAPD-PCR cluster with two strains of phage types 4, a strain of phage type 24var and a strain of phage type 9a, indicating that they consist of a genetically heterogeneous collection of strains. Two of the remaining three phage type 34 strains grouped into two different clusters, well separated from the other phage type 34 strains. One strain of phage type 34 was genetically diverse and did not cluster with any of the strains included in this study. Three of the phage type 14 strains grouped into cluster 11 at R2 > or =0.72, suggesting that they are genetically closely related. However, the remaining three strains of phage type 14 grouped into two separate clusters. Strains of phage types 7, 35, and 1 grouped in one cluster at R2 > or = 0.55. Our results clearly indicated that S.E. strains of the same phage type are not always genetically related. On the other hand, strains of a high genetic relatedness classified as different phage types. No specific plasmid profile could be linked to any of the phage types. Based on results obtained by LD50 virulence tests, strains containing the 38 MDa plasmid are more virulent compared to strains which do not contain the plasmid.

Phage types of Salmonella enterica serovar Enteritidis isolated in South Africa from 1991-1995.

A total of 615 strains of Salmonella enterica serovar Enteritidis (SE), received from 1991-1995 at the Onderstepoort Veterinary Institute (OVI), were phage typed. Most SE isolates (54,7%) originated from poultry followed by humans (28,5 %) and poultry eggs (9,6 %). Phage type 34 was the most prevalent (40,5 %) of all isolates, followed by phage type 4 (33,8 %). Other phage types identified were 1, lb, 4a, 7, 7a, 9a, 14, 24, 24var and 35 (in total 2,4% of isolates). Most isolates of SE were received from the Western Cape Province (47,4%) and Gauteng (22,3%). In poultry phage type 4 was dominant, but in humans, eggs, goats, ducks, sheep, pigs and rabbits, phage type 34 was the dominant type. It appeared as if the poultry-associated epidemic of SE in South Africa that occurred from 1991-1995 originated in the Western Cape Province during 1991 amongst poultry and then spread from there to humans and eggs and then to the rest of the country, where it emerged during 1993. Results indicate that phage type 34 was the dominant phage type from 1991-1993, but during 1994-1995 its presence declined. During this latter period the presence of phage type 4 increased. This may suggest that two smaller epidemics consisting of the two different phage types might have been responsible for the epidemic that occurred from 1991-1995.

Odontoma in an African elephant (Loxodonta africana).

The first known case of an odontoma in an African elephant (Loxodonta africana) is described. The tumour was fused with the coronal cementum of the sixth right mandibular molar tooth, thus preventing its eruption.