Glycosyl transferases in family 61 mediate arabinofuranosyl transfer onto xylan in grasses

Xylan, a hemicellulosic component of the plant cell wall, is one of the most abundant polysaccharides in nature. In contrast to dicots, xylan in grasses is extensively modified by alpha-(1,2)- and alpha-(1,3)-linked arabinofuranose. Despite the importance of grass arabinoxylan in human and animal nutrition and for bioenergy, the enzymes adding the arabinosyl substitutions are unknown. Here we demonstrate that knocking-down glycosyltransferase (GT) 61 expression in wheat endosperm strongly decreases alpha-(1,3)-linked arabinosyl substitution of xylan. Moreover, heterologous expression of wheat and rice GT61s in Arabidopsis leads to arabinosylation of the xylan, and therefore provides gain-of-function evidence for alpha-(1,3)-arabinosyltransferase activity. Thus, GT61 proteins play a key role in arabinoxylan biosynthesis and therefore in the evolutionary divergence of grass cell walls.

Affinity purification of 61- and 65-kDa rat brain corticotropin-releasing factor receptors and receptor-associated G proteins.

Corticotropin-releasing factor (CRF) has been shown to have a central role in physiological adaptation to stress. It is recognized for stimulating the release of adrenocorticotropin from the anterior pituitary gland, and has more recently been implicated as a regulator of autonomic and immunological responses to stress. Much confusion has surrounded the characterization of CRF receptors, with proteins of varying molecular weights having been identified but never purified and characterized. Recently, two CRF receptors have been cloned from brain and pituitary gland, but evidence from in-situ hybridization studies suggests that further CRF receptor types exist. We therefore developed two techniques which enable the isolation of CRF receptors from whole rat brain. The use of a solid-phase CRF analogue affinity column and elution using a competing ligand resulted in the purification of a single protein of 61 kDa. A second technique was devised which allowed the co-isolation of associated signalling proteins and the identification of CRF bound species following purification. CRF was covalently cross-linked to receptors and the complex purified using antibodies specific for the ligand. This enabled the purification of a CRF receptor of approximately 65 kDa and associated alpha and beta gamma G protein subunits. This study demonstrates the successful isolation of CRF receptors which are of different molecular weights to those previously observed from affinity cross-linking studies or predicted from cloned genes. In addition, we confirm the involvement of G proteins in CRF stimulated cell signalling by demonstrating their association with purified CRF receptor.

Valorisation of side streams from wheat milling and confectionery industries for consolidated production and extraction of microbial lipids

Crude enzymes produced via solid state fermentation (SSF) using wheat milling by-products have been employed for both fermentation media production using flour-rich waste (FRW) streams and lysis of Rhodosporidium toruloides yeast cells. Filter sterilization of crude hydrolysates was more beneficial than heat sterilization regarding yeast growth and microbial oil production. The initial carbon to free amino nitrogen ratio of crude hydrolysates was optimized (80.2 g/g) in fed-batch cultures of R. toruloides leading to a total dry weight of 61.2 g/L with microbial oil content of 61.8 % (w/w). Employing a feeding strategy where the glucose concentration was maintained in the range of 12.2 – 17.6 g/L led to the highest productivity (0.32 g/L∙h). The crude enzymes produced by SSF were utilised for yeast cell treatment leading to simultaneous release of around 80% of total lipids in the broth and production of a hydrolysate suitable as yeast extract replacement.

Additional Saturday rehabilitation improves functional independence and quality of life and reduces length of stay: a randomised controlled trial

Background

Many inpatients receive little or no rehabilitation on weekends. Our aim was to determine what effect providing additional Saturday rehabilitation during inpatient rehabilitation had on functional independence, quality of life and length of stay compared to 5 days per week of rehabilitation.

This was a multicenter, single-blind (assessors) randomized controlled trial with concealed allocation and 12-month follow-up conducted in two publically funded metropolitan inpatient rehabilitation facilities in Melbourne, Australia. Patients were eligible if they were adults (aged ≥18 years) admitted for rehabilitation for any orthopedic, neurological or other disabling conditions excluding those admitted for slow stream rehabilitation/geriatric evaluation and management. Participants were randomly allocated to usual care Monday to Friday rehabilitation (control) or to Monday to Saturday rehabilitation (intervention). The additional Saturday rehabilitation comprised physiotherapy and occupational therapy. The primary outcomes were functional independence (functional independence measure (FIM); measured on an 18 to 126 point scale), health-related quality of life (EQ-5D utility index; measured on a 0 to 1 scale, and EQ-5D visual analog scale; measured on a 0 to 100 scale), and patient length of stay. Outcome measures were assessed on admission, discharge (primary endpoint), and at 6 and 12 months post discharge.

We randomly assigned 996 adults (mean (SD) age 74 (13) years) to Monday to Saturday rehabilitation (n = 496) or usual care Monday to Friday rehabilitation (n = 500). Relative to admission scores, intervention group participants had higher functional independence (mean difference (MD) 2.3, 95% confidence interval (CI) 0.5 to 4.1, P = 0.01) and health-related quality of life (MD 0.04, 95% CI 0.01 to 0.07, P = 0.009) on discharge and may have had a shorter length of stay by 2 days (95% CI 0 to 4, P = 0.1) when compared to control group participants. Intervention group participants were 17% more likely to have achieved a clinically significant change in functional independence of 22 FIM points or more (risk ratio (RR) 1.17, 95% CI 1.03 to 1.34) and 18% more likely to have achieved a clinically significant change in health-related quality of life (RR 1.18, 95% CI 1.04 to 1.34) on discharge compared to the control group. There was some maintenance of effect for functional independence and health-related quality of life at 6-month follow-up but not at 12-month follow-up. There was no difference in the number of adverse events between the groups (incidence rate ratio = 0.81, 95% CI 0.61 to 1.08).

Providing an additional day of rehabilitation improved functional independence and health-related quality of life at discharge and may have reduced length of stay for patients receiving inpatient rehabilitation. 

Systematic approach to the quantitative voltammetric analysis of the Fe III /Fe II component of the [α 2 -Fe(OH 2 )P 2 W 17 O 61 ] 7-/8- reduction process in buffered and unbuffered aqueous media

The one-electron reduction of [α2-FeIII(OH2)P2W17O61]7- at a glassy carbon electrode was investigated using cyclic and rotating-disk-electrode voltammetry in buffered and unbuffered aqueous solutions over the pH range 3.45−7.50 with an ionic strength of approximately 0.6 M maintained. The behavior is well-described by a square-scheme mechanism P + e- ↔ Q (E10/ = −0.275 V, k10/ = 0.008 cm s-1, and α1 = 1/2), PH+ + e- ↔ QH+ (E20/ = −0.036 V, k20/ = 0.014 cm s-1, and α2 = 1/2), PH+ ↔ P + H+ (KP = 3.02 × 10-6 M), and QH+ ↔ Q + H+ (KQ = 2.35 × 10-10 M), where P, Q, PH+, and QH+ correspond to [α2-FeIII(OH)P2W17O61]8-, [α2-FeII(OH)P2W17O61]9-, [α2-FeIII(OH2)P2W17O61]7-, and [α2-FeII(OH2)P2W17O61]8-, respectively; E10‘ and E20‘ are the formal potentials, k10‘ and k20‘ are the formal (standard) rate constants, and KP and KQ are the acid dissociation constants for the relevant reactions. The analysis for the buffered media is based on the approach of Laviron who demonstrated that a square scheme with fully reversible protonations, reversible or quasi reversible electron transfers with the assumption that α1 = α2, can be well-described by the behavior of a simple redox couple, ox + e- ↔ red, whose formal potential, Eapp0‘, and standard rate constant, kapp0‘, are straightforwardly derived functions of pH, as are the values of E10‘, k10‘, E20‘, k20‘, and KP (only three of the four thermodynamic parameters in a square scheme can be specified). It was assumed that αapp = 1/2, and the simulation program DigiSim was used to determine the values of Eapp0‘ and kapp0‘, which are required to describe the cyclic voltammograms obtained in buffered media in the pH range from 3.45 to 7.52 (buffer-related reactions which effect general acid−base catalysis are included in the simulations). DigiSim simulations of cyclic voltammograms obtained in unbuffered media yielded the values of E10‘ and k10‘; KQ was then directly computed from thermodynamic constraints. These simulations included additional reactions between the redox species and H2O. The value of the diffusion coefficient of the [α2-FeIII(OH2)P2W17O61]7-, 2.92 × 10-6 cm2 s-1, was determined using DigiSim simulations of voltammograms at a rotating disk electrode in buffered and unbuffered media at pH 3.45. The diffusion coefficients of all redox species were assumed to be identical. When the pH is greater than 6, instability of P (i.e., [α2-FeIII(OH)P2W17O61]8-) led to the loss of the reactant and precluded lengthy experimentation.

Densidades e manejo químico da tiririca na produtividade de cana-de-açúcar

Em dois experimentos instalados e conduzidos a campo, foram avaliados os efeitos de densidades da tiririca e do seu controle químico na produtividade da cana-de-açúcar (var. SP 86042). No primeiro, três densidades foram estudadas: baixa (58 a 246), média (318 a 773) e alta (675 a 1.198 manifestações epígeas m-2). As reduções na produção foram de 13,5, 29,3 e 45,2% em relação à testemunha sem tiririca, respectivamente. No segundo experimento, foram testados os herbicidas sulfentrazone (0,8 kg ha-1), imazapic (0,105 kg ha-1) e flazasulfuron (0,150 kg ha-1), em pré-emergência, além da mistura de trifloxysulfuron-sodium + ametryne e Agral® (1,5 kg ha-1 + 0,20%), halosulfuron + Aterbane® (0,105 kg ha-1 + 0,25%) e flazasulfuron + Aterbane® (0,150 kg ha-1 + 0,25%), em pós emergência. Nesta área, a variação de densidade foi de 66 a 154 plantas m-2. Aos 90 dias após o plantio (DAP), as porcentagens de controle foram de 79,6; 70,6; 30,3; 84,8; 89,1; e 61,1%, respectivamente, para os herbicidas testados. A redução na produtividade foi de 16,4% entre as testemunhas infestada e capinada e de 11,9 e 6,0% onde se aplicou o flazasulfuron em pós e pré-emergência, respectivamente.

Carga de trabalho de enfermagem e infecções relacionadas à assistência à saúde na unidade de terapia intensiva

In recent decades, the search for quality care has been widely discussed by the institutions and health professionals. In this context, it is the nurse coordinator of the process of providing nursing staff, reflecting the commitment to quality of care. In this process, it is the appearance of Infections Related to health care and its potential association with the workload in nursing as a valuable indicator of quality of care. Thus, this research contributes to studies to characterize the demand of nursing work to promote a safe healthcare practice. This study aimed to identify the association of nursing workload with the number of cases of Ventilator-Associated Pneumonia, urinary tract infection and central venous catheter infection in the intensive care unit. This is a quantitative research approach, descriptive, cross-sectional and prospective, held at Unimed Hospital in Natal-RN. The study population consisted of all patients treated in the Intensive Care Unit, Hospital for a period of 90 consecutive days in 2011. The convenience sample was compostapelos patients admitted to the ICU during the period of data collection, a total sample of 286 patients. To perform the data analysis software were used: Statistica 6.0, SPPS (Statistical Package for Social Sciences) version 17.0 (2004) and Excel 2007. In the descriptive analysis, we used Measures of Central Tendency and Measures of Dispersion or Variability and the use of nonparametric tests. Of the 286 patients, 88 were from the ICU and 198 ICU II II. Males predominated in the ICU I (51.1%) and female ICU II (57.6%) patients in the ICU I were aged 61-80 years (39.8%) followed by greater than 80 years (39.8%). In the ICU II, most of the patients were aged 61-80 years (38.9%) and then from 41 to 60 years (24.2%). In relation to the class of TISS inlet predominant class II in the two ICUs (59.1%), followed by Class III also in the two units (34.6%). Most patients (70.6%) out of the ICUs belonging to class II TISS. In the ICU I, the average number of forms of the TISS 28 was 6, has in ICU II this value drops to 3.2 forms. The overall mean was 19.9 TISS points in ICU patients I and ICU II.the 17 points in the average hours required to provide adequate nursing care to patients in the ICU I found that is 10 , 7 hours, and the ICU II 9.2 hours. It was found that the time provided by the nursing staff were higher in ICU II, with an average of 19 hours available for nurses in this sector. In the ICU I, which showed higher need of available hours, it was found that the mean value of 12.7 available hours. It was found that only 2.4% of patients had these units Ventilator-Associated Pneumonia, 1.0% were infected central venous catheter and 1.4% of patients had urinary tract infection. Infection associated with health care occurs, on average, on the tenth day of hospitalization. In the ICU II, this average value extends to the twelfth day with an excess of 2.7 hours of nursing care while in ICU I value decays to the ninth day of hospitalization with a deficiency of 12-hour assistance. It is concluded that patients generally showed a need for classification of semi-intensive care and has been assisted in their need to load. As for his association with the Related Infections Health will assist this analysis could not be performed due to the small number of notifications in this period. It is suggested further study how other factors related to infections me a longer period of analysis

Sorologia Anti PGL-1 e risco de ocorrência de hanseníase em área de alta endemicidade do Estado de São Paulo: quatro anos de seguimento

Os testes sorológicos para diagnóstico de hanseníase, usando o glicolipídeo-fenólico-1 (PGL-1), considerado antígeno específico do M. leprae, têm aberto algumas possibilidades de estudo do comportamento epidemiológico desta doença. Algumas questões, como tempo de latência da doença, infecção subclínica e importância do contato intra-domiciliar (contatos) no controle da endemia, puderam ser melhor analisadas usando este instrumental. Este estudo teve por objetivo verificar a existência de associação entre a situação sorológica e a ocorrência de hanseníase. Foram seguidas, durante 4 anos, 6.520 pessoas com idade igual ou superior a 5 anos, submetidas no início do seguimento ao teste sorológico Anti PGL-1, pertencentes ao universo de 7.416 habitantes da área urbana de um município paulista caracterizado por elevada endemicidade de hanseníase. Foi identificado um grupo de 590 indivíduos soropositivos (9,0 %). Foram diagnosticados, no período, 82 casos novos de hanseníase, 26 no grupo de soropositivos (441 casos novos/10.000 indivíduos) e 48 no de soronegativos (81/10.000). Entre os que não fizeram sorologia, surgiram 8 casos novos (89/10.000). Procurou-se controlar, na análise, a condição de contato, dado que a taxa de soropositividade padronizada por idade e sexo era de 9,61% no grupo de contatos e 7,65% no de não-contatos. Tomando-se os não-contatos soronegativos como o grupo de não expostos, foram calculados os riscos relativos de adoecimento no período, a partir das taxas de detecção padronizadas por idade, resultando no seguinte: os contatos ID soropositivos apresentaram a taxa de 1.704/10.000, 27 vezes maior que a dos não-expostos, igual a 63/10.000; os não-contatos soropositivos e os contatos soronegativos apresentaram taxas, respectivamente, de 274 e 198/10.000, ambas maiores que as dos não-expostos e iguais entre si. A soropositividade associou-se à elevação de 8,6 vezes do risco de hanseníase entre os contatos e de 4,4 entre os não-contatos. Na situação epidemiológica estudada, caracterizada por elevada endemicidade de hanseníase, 50% dos casos novos surgiram entre os não-contatos soronegativos, ou seja, sem fonte de infecção conhecida. Portanto, o teste anti-PGL-1 usado revela-se, na prática, de pouca aplicabilidade. Resta estudar ainda o comportamento da sorologia anti-PGL-1 em áreas de média e baixa endemicidade para que se possa tirar conclusões mais consubstanciadas sobre sua utilidade no controle da endemia. Recomenda-se o aprofundamento das pesquisas sorológicas e de outras que aprimorem o diagnóstico precoce da infecção subclínica, inclusive para detecção de formas paucibacilares, para se ampliar as possibilidades de influir no controle endêmico.

Ophiuroidea das regiões Antártica e Subantártica: 2. Variação em Gorgonocephalus chilensis ( Philippi) (Echinodermata, Ophiuroidea, Gorgonocephalidae)

Foram examinados 198 exemplares de Gorgonocephalus chilensis das regiões antártica e subantartica, tendo-se em vista a variação de caracteres morfológicos externos. Foi constatada uma grande variação nas características morfológicas externas, que parece independer de localização geográfica.