Whole-Cell Bioconversion of Renewable Biomasses-Related Aromatics to cis,cis-Muconic Acid

Lignin and wheat bran represent renewable feedstocks for generation of useful and value-added compounds such as vanillin (a popular flavoring agent) and cis,cis-muconic acid (ccMA, a building block for the synthesis of plastic materials). In the present work, we report on the setup of an efficient and green process for producing such valuable compounds based on (a) the optimization of the extraction procedures for vanillin from lignin and ferulic acid from wheat bran and (b) the genetic engineering of an Escherichia coli strain with up to three plasmids differing in copy numbers to modulate the expression of up to seven recombinant enzymes. In detail, we used two sequential reactions catalyzed by the decarboxylase Fdc and the dioxygenase Ado to convert wheat bran-derived ferulic acid into vanillin: nature-identical vanillin was produced in one pot with a >85% yield in 20 h. Next, the dehydrogenase LigV, the demethylase VanAB, the decarboxylase AroY, and the dioxygenase C12O converted lignin-derived vanillin into ccMA with a >95% conversion yield and a productivity of 4.2 mg of ccMA/g of Kraft lignin in 30 min. Finally, when the optimized E. coli strain expressing all the abovementioned enzymes was used, ccMA was produced with a >95% conversion yield starting from ferulic acid in 10 h following product isolation, corresponding to 0.73 g of ccMA/g of ferulic acid, 1.4 g of ccMA/L, and 2.2 g of ccMA/g of wheat bran biomass. The optimized whole-cell system represents a sustainable and cost-competitive process for producing high value-added products from renewable resources

Residual effect of Fipronil used in rice farming on non-target organisms.

This study evaluated changes in the composition of the benthic community of non-target organisms in rice crop fields due to residual effect one year after planting seeds treated with different Fipronil doses. Macroinvertebrates were collected during crop irrigation months (February, March and April/2012), with the aid of 1 mm hand net placed close to the ground of each plot, for 20 minutes, collected macroinvertebrates were classified according to its dietary habits function. Orizophagus oryzae larvae, the target organisms, were not found in groups treated with different Fipronil doses; they were only observed in the control treatment. Chironominae (Chironomidae: Diptera) larvae had their population reduced in comparison to the control treatment. However, the herein used insecticide doses did not show significant changes in the benthic macroinvertebrate community one year after their application

Apoptosis gene expression profiling in lens development

During lens development, epithelial cells located at the equatorial region of the lens undergo terminal differentiation to form fibre cells resulting in cell elongation and degradation of all intracellular organelles. Failure to complete this process successfully can result in cataract. This process is thought to be an attenuated form of classical apoptosis. This study was completed to give a comprehensive analysis of apoptosis genes in the developing mouse lens. Macroarrays containing 243 immobilised cDNAs with a known role in apoptosis were utilised to examine the gene expression at different developmental stages. The stages examined were postnatal day 7 and postnatal day 14. Over 100 apoptosis genes were shown to be expressed above background with 20 genes demonstrating significantly differential expression (2-fold or greater change in expression, p-value < 0.05), with highest expression at PI4. PCR confirmed expression of all the genes identified from the array results, and differential expression was confirmed for 52% of the genes. Protein expression of two selected genes, axl and mcl-1, was demonstrated using western blotting. Lens morphology was examined in transgenic mice generated to contain an extended CAG repeat in the huntingtin gene (one of the genes identified from the arrays). Morphology was compared between homozygote, heterozygote and wild-type mice. The presence of the mutated gene did not affect denucleation during lens differentiation and no statistically significant difference was seen in the dimensions of the organelle free zone (OFZ) of the wild-type and homozygote mice. A cross-species comparison was completed. Gene expression of the genes shown to be highly expressed and differentially expressed from the array results was examined in embryonic chick lenses. From the results of this part of the study expression was seen for 83% of genes. These results add to the argument that the process of differentiation is similar in both mouse and chick lenses.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Superlattice Growth via MBE and Green’s Function Techniques

A model has been developed to simulate the growth of arrays consisting of a substrate on which alternating layers of quantum dots (QDs) and spacer layers are epitaxially grown. The substrate and spacer layers are modeled as an anisotropic elastic half-space, and the QDs are modeled as point inclusions buried within the half-space. In this model, the strain at the free surface of this half-space due to the buried point QDs is calculated, and a scalar measure of the strain at the surface is subsequently determined. New point QDs are placed on the surface where the previously calculated scalar strain measure is a minimum. Following available DFT results, this scalar strain measure is a weighted average of the in-plane strains. This model is constructed under the assumption that diffusional anisotropy can be neglected, and thus, the results are more in agreement with results from experiments of growth of SiGe QDs than experiments involving QDs of (In,Ga)As

Curcuma longa Extract Exerts a Myorelaxant Effect on the Ileum and Colon in a Mouse Experimental Colitis Model, Independent of the Anti-Inflammatory Effect

Background: Curcuma has long been used as an anti-inflammatory agent in inflammatory bowel disease. Since gastrointestinal motility is impaired in inflammatory states, the aim of this work was to evaluate if Curcuma Longa had any Methods: The biological activity of Curcuma extract was evaluated against Carbachol induced contraction in isolated mice intestine. Acute and chronic colitis were induced in Balb/c mice by Dextran Sulphate Sodium administration (5% and 2.5% respectively) and either Curcuma extract (200 mg/kg/day) or placebo was thereafter administered for 7 and 21 days respectively. Spontaneous contractions and the response to Carbachol and Atropine of ileum and colon were studied after colitis induction and Curcuma administration. Results: Curcuma extract reduced the spontaneous contractions in the ileum and colon; the maximal response to Carbachol was inhibited in a non-competitive and reversible manner. Similar results were obtained in ileum and colon from Curcuma fed mice. DSS administration decreased the motility, mainly in the colon and Curcuma almost restored both the spontaneous contractions and the response to Carbachol after 14 days assumption, compared to standard diet, but a prolonged assumption of Curcuma decreased the spontaneous and Carbachol-induced contractions. Conclusions: Curcuma extract has a direct and indirect myorelaxant effect on mouse ileum and colon, independent of the anti-inflammatory effect. The indirect effect is reversible and non-competitive with the cholinergic agent. These results suggest the use of curcuma extract as a spasmolytic agent

Productivity of male green asparagus genotypes (Asparagus officinalis var. altilis L.) in their seventh year

El espárrago es una hortaliza no tradicional perenne, la que puede ingresar a los sistemas productivos como una alternativa de diversificación productiva, siendo necesario evaluar el comportamiento de distintos genotipos para cada zona productiva en particular. Con el objetivo de determinar la productividad de genotipos masculinos italianos se efectuó un ensayo (22/11/2006) mediante plantines (P) grandes (PG) y chicos (PCH). Se evaluó en el séptimo año de cosecha: productividad fresca total (PFT) y comercial (PFC), turiones totales (NTT) y comerciales (NTC), calibres (J: Jumbo; XL: Extra-Large; L: Large; M: Medium; S: Small y A: Asparagina), defectos (DE: espigados; P: daño plagas; y OD: otros defectos). Se efectuaron 32 cosechas en el período del 01/09/2014-14/11/2014. Se realizó análisis de la varianza ANOVA-LSD test (P≥0.05). No se encontraron diferencias significativas en P.Como resultado de las 32 cosechas realizadas se logró una productividad promedio de 14,5 t.ha-1 y comercial de primera calidad de 5,4 t.ha-1. En de turiones se lograron 553.241 totales y 252.420 comerciales de primera calidad, siendo el peso promedio por turión de 21 g. Respecto del tamaño de plantines, no se encontraron diferencias significativas. Los genotipos italianos superaron al testigo americano destacándose en PFT Eros y en PFC: Ercole, Eros, Italo y H-668. En turiones producidos se lograron diferencias significativas entre híbridos: en NTT se destacaron Zeno, Eros, Ercole y H-668; mientras que en NTC, Ercole. En distribución de calibres se destacaron, en J: Italo, UC-157, H-668, Zeno y Eros; en XL: Eros, Italo, Zeno y H-668; en L: Eros, Italo, Ercole y H 668, UC-157 y Zeno. En M: Ercole; Eros y H-668 y UC-157. En calibres S: UC-157 y Ercole, mientras en A: UC-157 y Ercole. Los híbridos masculinos, productivamente representan una alternativa valiosa para las características productivas de la Provincia de Buenos Aires.Asparagus is a non-traditional perennial vegetable crop for which evaluation of the performance of different genotypes is required. With the aim of determining the productivity of a set of all-male genotypes from Italy, a trial was planted on 22/11/06 using large (PG) and small (PCH) seedlings. The following characters were evaluated: total (PFT) and commercial (PFC) fresh production, total (NTT) and commercial (NTC) spear number, calibre distribution (J: Jumbo; XL: Extra-Large; L: Large; M: Medium; S: Small and A: Asparagina), defects (DE: opened bracts; P: plague damage; and OD: other defects). Thirty-two harvests were carried out between 1/9/14 and 14/11/14. Data were analysed by ANOVA-LSD (P≥0.05). No significant differences were found between the two seedling sizes. From the thirty-two harvests, a mean productivity of 14.5t.ha-1 was obtained, with a commercial prime quality productivity of 5.4 t.ha-1 . A total of 553,241 spears were obtained, of which 252,420 were of commercial prime quality, with an average weight of 21 g per spear. Regarding seedling size, no significant differences were found. The Italian genotypes performed better than the control genotype from the USA, with Eros outstanding for PFT, and Ercole, Eros, Italo and H-668 for PFC. For spears produced, significant differences were observed between hybrids: for NTT, Zeno, Eros, Ercole and H-668 stood out, while for NTC, Ercole. Regarding calibre distribution, Italo, UC-157, H-668, Zeno and Eros stood out for J; Eros, Italo, Zeno and H-668 for XL; Eros, Italo, Ercole and H-668, UC-157 and Zeno for L; Ercole, Eros and H-668, and UC-157 for M; UC157 and Ercole for S; and UC-157 and Ercole for A. In conclusion, all male hybrids represent a valuable production alternative.Fil: Romero, F.. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Agronomía. Centro Regional de Estudios Sistémico de Cadenas Agroalimentarias; ArgentinaFil: Castagnino, Ana Maria. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Agronomía. Centro Regional de Estudios Sistémico de Cadenas Agroalimentarias; Argentina. Pontificia Universidad Católica Argentina; ArgentinaFil: Diaz, K. E.. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Agronomía. Centro Regional de Estudios Sistémico de Cadenas Agroalimentarias; ArgentinaFil: Guisolis, Andrea Paola. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Agronomía. Centro Regional de Estudios Sistémico de Cadenas Agroalimentarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Rosini, M. B.. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Agronomía. Centro Regional de Estudios Sistémico de Cadenas Agroalimentarias; ArgentinaFil: Rogers, William John. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Cientifico Tecnolológico Mar del Plata. Instituto de Investigaciones en Biodiversidad y Biotecnología. Laboratorio de Biología Funcional y Biotecnología; Argentin

Point-of-care diagnosis of endometrial cancer using the surgical intelligent knife (iknife)-a prospective pilot study of diagnostic accuracy

Introduction: Delays in the diagnosis and treatment of endometrial cancer negatively impact patient survival. The aim of this study was to establish whether rapid evaporative ionisation mass spectrometry using the iKnife can accurately distinguish between normal and malignant endometrial biopsy tissue samples in real time, enabling point-of-care (POC) diagnoses. Methods: Pipelle biopsy samples were obtained from consecutive women needing biopsies for clinical reasons. A Waters G2-XS Xevo Q-Tof mass spectrometer was used in conjunction with a modified handheld diathermy (collectively called the ‘iKnife’). Each tissue sample was processed with diathermy, and the resultant surgical aerosol containing ionic lipid species was then analysed, producing spectra. Principal component analyses and linear discriminant analyses were performed to determine variance in spectral signatures. Leave-one-patient-out cross-validation was used to test the diagnostic accuracy. Results: One hundred and fifty patients provided Pipelle biopsy samples (85 normal, 59 malignant, 4 hyperplasia and 2 insufficient), yielding 453 spectra. The iKnife differentiated between normal and malignant endometrial tissues on the basis of differential phospholipid spectra. Cross-validation revealed a diagnostic accuracy of 89% with sensitivity, specificity, positive predictive value and negative predictive value of 85%, 93%, 94% and 85%, respectively. Conclusions: This study is the first to use the iKnife to identify cancer in endometrial Pipelle biopsy samples. These results are highly encouraging and suggest that the iKnife could be used in the clinic to provide a POC diagnosis

Acute diverticulitis management: evolving trends among Italian surgeons. A survey of the Italian Society of Colorectal Surgery (SICCR)

Acute diverticulitis (AD) is associated with relevant morbidity/mortality and is increasing worldwide, thus becoming a major issue for national health systems. AD may be challenging, as clinical relevance varies widely, ranging from asymptomatic picture to life-threatening conditions, with continuously evolving diagnostic tools, classifications, and management. A 33-item-questionnaire was administered to residents and surgeons to analyze the actual clinical practice and to verify the real spread of recent recommendations, also by stratifying surgeons by experience. CT-scan remains the mainstay of AD assessment, including cases presenting with recurrent mild episodes or women of child-bearing age. Outpatient management of mild AD is slowly gaining acceptance. A conservative management is preferred in non-severe cases with extradigestive air or small/non-radiologically drainable abscesses. In severe cases, a laparoscopic approach is preferred, with a non-negligible number of surgeons confident in performing emergency complex procedures. Surgeons are seemingly aware of several options during emergency surgery for AD, since the rate of Hartmann procedures does not exceed 50% in most environments and damage control surgery is spreading in life-threatening cases. Quality of life and history of complicated AD are the main indications for delayed colectomy, which is mostly performed avoiding the proximal vessel ligation, mobilizing the splenic flexure and performing a colorectal anastomosis. ICG is spreading to check anastomotic stumps’ vascularization. Differences between the two experience groups were found about the type of investigation to exclude colon cancer (considering the experience only in terms of number of colectomies performed), the size of the peritoneal abscess to be drained, practice of damage control surgery and the attitude towards colovesical fistula

"Delirium Day": A nationwide point prevalence study of delirium in older hospitalized patients using an easy standardized diagnostic tool

Background: To date, delirium prevalence in adult acute hospital populations has been estimated generally from pooled findings of single-center studies and/or among specific patient populations. Furthermore, the number of participants in these studies has not exceeded a few hundred. To overcome these limitations, we have determined, in a multicenter study, the prevalence of delirium over a single day among a large population of patients admitted to acute and rehabilitation hospital wards in Italy. Methods: This is a point prevalence study (called "Delirium Day") including 1867 older patients (aged 65 years or more) across 108 acute and 12 rehabilitation wards in Italian hospitals. Delirium was assessed on the same day in all patients using the 4AT, a validated and briefly administered tool which does not require training. We also collected data regarding motoric subtypes of delirium, functional and nutritional status, dementia, comorbidity, medications, feeding tubes, peripheral venous and urinary catheters, and physical restraints. Results: The mean sample age was 82.0 \ub1 7.5 years (58 % female). Overall, 429 patients (22.9 %) had delirium. Hypoactive was the commonest subtype (132/344 patients, 38.5 %), followed by mixed, hyperactive, and nonmotoric delirium. The prevalence was highest in Neurology (28.5 %) and Geriatrics (24.7 %), lowest in Rehabilitation (14.0 %), and intermediate in Orthopedic (20.6 %) and Internal Medicine wards (21.4 %). In a multivariable logistic regression, age (odds ratio [OR] 1.03, 95 % confidence interval [CI] 1.01-1.05), Activities of Daily Living dependence (OR 1.19, 95 % CI 1.12-1.27), dementia (OR 3.25, 95 % CI 2.41-4.38), malnutrition (OR 2.01, 95 % CI 1.29-3.14), and use of antipsychotics (OR 2.03, 95 % CI 1.45-2.82), feeding tubes (OR 2.51, 95 % CI 1.11-5.66), peripheral venous catheters (OR 1.41, 95 % CI 1.06-1.87), urinary catheters (OR 1.73, 95 % CI 1.30-2.29), and physical restraints (OR 1.84, 95 % CI 1.40-2.40) were associated with delirium. Admission to Neurology wards was also associated with delirium (OR 2.00, 95 % CI 1.29-3.14), while admission to other settings was not. Conclusions: Delirium occurred in more than one out of five patients in acute and rehabilitation hospital wards. Prevalence was highest in Neurology and lowest in Rehabilitation divisions. The "Delirium Day" project might become a useful method to assess delirium across hospital settings and a benchmarking platform for future surveys