Thermal Marangoni trapping driven by laser absorption in evaporating droplets for particle deposition.

Controlling the deposition of particles is of great importance in many applications. In this work, we study particle deposition driven by Marangoni flows, triggered by laser absorption inside an evaporating droplet. When the laser is turned on, thermal gradients are generated and produce a toroidal Marangoni flow that concentrates the particles around the laser beam and ultimately controls the final deposition. We experimentally characterize the radius of the Marangoni flows as a function of the laser parameters. Counter-intuitively, the radius of the Marangoni region appears to remain constant and is not proportional to the thickness of the drop which decreases due to evaporation. We develop a model to predict the size of the Marangoni region that combines evaporative flows and laser-induced Marangoni flows. The experimental data are in good agreement with the predictions, allowing us to estimate the particle overconcentration factor resulting from the laser heating effects. The addition of surfactants to the solution allows the coupling of solutal Marangoni flows with thermal ones to achieve a final micron-scale deposit located at the laser spot. These results pave the way for new methods with high tunability provided by spatio-temporal light control for surface patterning applications.

Pyrosequencing as a tool for the detection of Phytophthora species: error rate and risk of false Molecular Operational Taxonomic Units.

AIMS: To evaluate the accuracy of pyrosequencing for the description of Phytophthora communities in terms of taxa identification and risk of assignment for false Molecular Operational Taxonomic Units (MOTUs). METHODS AND RESULTS: Pyrosequencing of Internal Transcribed Spacer 1 (ITS1) amplicons was used to describe the structure of a DNA mixture comprising eight Phytophthora spp. and Pythium vexans. Pyrosequencing resulted in 16 965 reads, detecting all species in the template DNA mixture. Reducing the ITS1 sequence identity threshold resulted in a decrease in numbers of unmatched reads but a concomitant increase in the numbers of false MOTUs. The total error rate was 0·63% and comprised mainly mismatches (0·25%) CONCLUSIONS: Pyrosequencing of ITS1 region is an efficient and accurate technique for the detection and identification of Phytophthora spp. in environmental samples. However, the risk of allocating false MOTUs, even when demonstrated to be low, may require additional validation with alternative detection methods. SIGNIFICANCE AND IMPACT OF THE STUDY: Phytophthora spp. are considered among the most destructive groups of invasive plant pathogens, affecting thousands of cultivated and wild plants worldwide. Simultaneous early detection of Phytophthora complexes in environmental samples offers an unique opportunity for the interception of known and unknown species along pathways of introduction, along with the identification of these organisms in invaded environments.

A long term feed supplementation based on phosphate binders in Feline Chronic Kidney Disease.

Chronic kidney disease (CKD) is a very common disorder in elderly cats. A proper renal diet represents the most efficient therapeutic intervention to improve survival and life quality in feline patients with 3 and 4 International Renal Interest Society (IRIS) stages. Twenty cats were selected in this study. Ten were administered the dietary supplementation for 360 days and the other ten, whose owners did not give consent for any supplemental therapies apart from the renal diet, were selected from a clinical database and used as control group. The present study is a long term study (360 days) aiming to evaluate the efficacy and palatability of a dietary supplementation containing calcium carbonate, calcium-lactate gluconate, chitosan and sodium bicarbonate in cats diagnosed with 3 and 4 IRIS stages of CKD. The owners were asked to fill in questionnaires to get information on the cat's appetite, the palatability of the given supplement, the presence of vomit and/or diarrhoea, general health and vitality. Hematochemical, biochemical and urinary analyses were performed on day 0, 30, 60, 90, 120, 150,180 and 360. GraphPad Prism® software was used to perform statistical analysis. Our study shows that the given dietary supplement reduced serum phosphorus and increased serum bicarbonate values in cats with CKD. In turn, this supplement could be used as a support therapy in cats with advanced CKD improving their clinical conditions without any adverse reaction. Finally, it is important to underline that all the animals completed the study and the owners reported a good palatability of the feed supplement.

Glucose-6-phosphatase mRNA and activity are increased to the same extent in kidney and liver of diabetic rats.

Using Northern blot with a specific glucose-6-phosphatase (Glc6Pase) cDNA probe and enzymatic activity determination, we studied the effect of streptozotocin-induced diabetes on Glc6Pase in rat gluconeogenic tissues. The Glc6Pase mRNA abundance was increased four to five times in both the liver and kidney of diabetic rats. This was correlated with a concomitant 130% increase in Glc6Pase catalytic subunit in both tissues. The elevated level of Glc6Pase mRNA was significantly corrected in both the liver and kidney of diabetic rats after a 12-h insulin treatment. We also studied Glc6Pase mRNA and activity in gluconeogenic tissues during the fed-fasted and fasted-refed transitions in normal rats. In the liver, the abundance of Glc6Pase mRNA was sharply increased about four times after 24 or 48 h of fasting. In the kidney, the Glc6Pase mRNA level was gradually increased some three and five times after 24 and 48 h of fasting, respectively. The increase of Glc6Pase mRNA in both organs was matched with a doubling of the activity of Glc6Pase catalytic subunit: rapid in the liver and gradual in the kidney. The liver Glc6Pase mRNA abundance in 48-h fasted rats was acutely and importantly decreased upon refeeding. The kidney Glc6Pase mRNA level was also significantly lowered under these conditions, albeit less rapidly. These data demonstrate that efficient control of Glc6Pase takes place in both gluconeogenic organs at the pretranslational level and suggest that insulin might play an important role in this control. In addition, using reverse transcription-polymerase chain reaction and Northern blot, we report that Glc6Pase mRNA is not detectable in several other tissues previously assumed to express the enzyme.

Development and regulation of glucose-6-phosphatase gene expression in rat liver, intestine, and kidney: in vivo and in vitro studies in cultured fetal hepatocytes.

The mRNA and the activity of glucose-6-phosphatase (Glc-6-Pase) were present in the liver, kidney, and small intestine of 15-day-old suckling rats, but were absent from the stomach, colon, lung, white and brown adipose tissues, muscle, heart, brain, and spleen. The mRNA encoding Glc-6-Pase was present in the liver of 21-day-old fetal rats and increased markedly immediately after birth. From 5 days after birth to the end of the suckling period, it returned to 50% of the level found in the liver of 48-h starved adult rats. When rats were weaned at 21 days onto a high-carbohydrate, low-fat (HCLF) diet, the concentration of liver Glc-6-Pase mRNA was markedly increased. In the fetal rat jejunum, the activity and mRNA of Glc-6-Pase were very low. It increased during the 5 days after birth and then declined to reach very low levels. Neither mRNA nor activity of Glc-6-Pase was present in the fetal kidney. They appeared and increased slowly during the suckling period to reach maximal levels 15 days after birth and then remained constant. Weaning onto the HCLF diet did not change the Glc-6-Pase gene expression, neither in the jejunum nor in the kidney. The regulation of Glc-6-Pase gene expression by hormones and nutrients was studied in cultured hepatocytes from 20-day-old rat fetuses. Bt2cAMP stimulated the Glc-6-Pase gene expression in a dose-dependent manner. This probably resulted from an increased gene transcription since the half-life of the transcript was not affected by dibutyryl cAMP (Bt2cAMP). The Bt2cAMP-induced Glc-6-Pase mRNA accumulation was antagonized by insulin in a dose-dependent manner. Long-chain fatty acids (LCFAs), but not medium-chain fatty acids, induced the accumulation of Glc-6-Pase mRNA and the stabilization of the transcript. The peroxisome proliferator, clofibrate, induced a threefold increase in Glc-6-Pase mRNA concentration. Both stimulation of Glc-6-Pase mRNA by LCFAs and clofibrate were inhibited by insulin. Increasing concentrations of glucose (from 0 to 20 mmol/l) did not affect the Bt2cAMP-induced Glc-6-Pase gene expression. By contrast, high glucose concentration (25 mmol/l) markedly induced the Glc-6-Pase gene expression in fed adult rat hepatocytes. The difference in the response to glucose between fetal and adult rat hepatocytes is discussed. We conclude that the rapid increase in hepatic Glc-6-Pase mRNA levels that accompanies the fetal-to-neonatal transition in the rat is triggered by the reciprocal change in circulating insulin and LCFA concentrations, coupled to the rise in liver cAMP concentration.

Reshaping ethnography: contemporary postpositivist possibilities.

Following Leinginger's introduction of ethnography into the field of nursing research, numerous descriptive and interpretive studies of health care beliefs and practices have been conducted. The resultant data have been translated into recommendations relative to the areas of nursing education, administration and clinical practice in an effort to ensure that the identified cultural needs are recognized and met. In this paper the discourses that inform such work are explored. Its practices and emergent dilemmas are reassessed in the light of an emerging body of work that challenges its foundational assumptions. Linked under the umbrella of 'postpositivist ethnography', such work recognizes the research area as a social and political field of which the researcher is an integral part. Hence, as an informed subject, the researcher, like the informants, is seen to be implicated in the generation of data. She, or he, is not charged with occupying the opposing roles of objective researcher and subjective participant, nor with reporting 'the truth' as told by informants. This emergent tradition is not without its dilemmas and of particular concern is the issue of authority; that is, whose voice constructs the text? As nursing academics grapple with questions regarding the nature of the knowledge that informs their discipline, it is imperative that they critique potentially fruitful research practices before they appropriate them. Failure to do so may lead them to unwittingly generate knowledge that is inimical to their particular quest. The 'new ethnography' discussed in this paper, offers academics and others interested in the generation of knowledge, not only a methodology that invites the possibility of opening up previously hidden areas of practice, but one that actively involves the researcher in challenging her taken-for-granted assumptions.

The nurse educator as teacher: exploring the construction of the 'reluctant instructor'.

This paper explores the discursive construction of the category of teacher identified as the 'reluctant instructor'. Data are drawn from my doctoral study, an ethnography of a school of nursing located in the higher education sector in which I was employed. This study explored the question: What shapes nurse educators and what do they, in turn, shape? Participants included nurse educators who taught a pre-registration programme. Data were gathered through interview and observations of classroom teaching. Analysis focused on the discursive constitution of participants' sense of being a nurse educator and on the implications of their ways of 'knowing and doing' for students. The analysis suggests that the dominant subject positions adopted by participants were those of 'academic' and 'victim'. They employed an ideal/real dichotomy to give a sense of coherence to the inconsistencies or contradications that they saw as structuring their daily activities. Being an academic entailed being an educator, specifically a facilitator, as being competent and having some degree of autonomy. However, participants saw the reality of their situation as markedly different from their ideal; as exhibiting traits of the 'outmoded', oppressive, hospital system of nurse training. Within this context most participants positioned themselves as a 'reluctant instructor'. The implications of their practices for the creation of the 'passive student' are suggested in the paper. In this analysis the nurse educator is viewed as implicated in the formation and maintenance of the context in which she or he is located. Such a view challenges the neutrality that is implicated in understandings of the nurse educator that inform nursing texts and that helped constitute participants' sense of self.

The glucose-6 phosphatase gene is expressed in human and rat small intestine: regulation of expression in fasted and diabetic rats.

BACKGROUND & AIMS: Glucose-6 phosphatase (Glc6Pase) is the last enzyme of gluconeogenesis and glycogenolysis, previously assumed to be expressed in the liver and kidney only, conferring on both tissues the capacity to produce endogenous glucose in blood. METHODS: Using Northern blotting and reverse-transcription polymerase chain reaction and a highly specific Glc6Pase assay, we studied expression of the Glc6Pase gene in human and in rat tissues (fasted and diabetic). RESULTS: The Glc6Pase gene is expressed in the duodenum and jejunum in normal fed rats and in the duodenum, jejunum, and ileum in humans. The Glc6Pase messenger RNA (mRNA) abundance was increased eightfold and sixfold in the duodenum and jejunum of streptozotocin diabetic rats. It was normalized in both tissues after 10 hours of insulin treatment. Glc6Pase activity was increased by 300% in the duodenum and jejunum in diabetic rats compared with normal rats. The Glc6Pase mRNA abundances and enzymatic activities were increased in a similar manner in both tissues in 48-hour-fasted rats. Normalization of mRNA abundance was achieved after refeeding for 7 hours. In addition, Glc6Pase mRNA and activity were also expressed in the ileum during fasting in rats. CONCLUSIONS: These data show that the small intestine has the ability to release endogenous glucose and strongly suggest that its contribution to systemic glucose production might be increased in situations of insulinopenia (type 1 diabetes) and insulin resistance (type 2 diabetes and others).

Enzymatic characterization of four new mutations in the glucose-6 phosphatase (G6PC) gene which cause glycogen storage disease type 1a.

Glycogen storage disease type 1a (GSD1a) is caused by mutations in the gene of glucose-6 phosphatase (G6PC), encoding the last enzyme of gluconeogenesis and glycogenolysis. To study the effect of mutations previously identified, but not yet enzymatically characterized, in French GSD1a patients, we used an in vitro expression system of the human glucose-6 phosphatase (hGlc6Pase) cDNA. Wild type hGlc6Pase expressed in COS-7 cells exhibited kinetic features comparable to microsomal Glc6Pase from normal human liver and kidney. Four new mutations inducing aminoacid changes in the coding sequence, e.g. W77R, A124T, G184E and L211P, were inserted into the Glc6Pase cDNA by site-directed mutagenesis, and studied after transient expression in COS-7 cells. All four mutations totally abolished Glc6Pase activity.

Mutation analysis in 24 French patients with glycogen storage disease type 1a.

Both alleles of 24 French glycogen storage disease type 1a patients were sequenced: 14 different mutations allowed the identification of complete genotypes for all the patients. Nine new gene alterations are reported. Five mutations, Q347X, R83C, D38V, G188R, and 158 del C, account for 75% of the mutated alleles. These data show that the molecular pathology of the glucose-6-phosphatase gene is heterogeneous in this population. Complete genotyping of the index case by systematic sequencing is necessary to allow prenatal diagnosis in chorionic villi for at risk couples.