P5 ATPases are eukaryotic pumps important for cellular metal ion, lipid and protein homeostasis; however, their transported substrate, if any, remains to be identified. Ca2+ was proposed to act as a ...ligand of P5 ATPases because it decreases the level of phosphoenzyme of the Spf1p P5A ATPase from Saccharomyces cerevisiae. Repeating previous purification protocols, we obtained a purified preparation of Spf1p that was close to homogeneity and exhibited ATP hydrolytic activity that was stimulated by the addition of CaCl2. Strikingly, a preparation of a catalytically dead mutant Spf1p (D487N) also exhibited Ca2+-dependent ATP hydrolytic activity. These results indicated that the Spf1p preparation contained a co-purifying protein capable of hydrolyzing ATP at a high rate. The activity was likely due to a phosphatase, since the protein i) was highly active when pNPP was used as substrate, ii) required Ca2+ or Zn2+ for activity, and iii) was strongly inhibited by molybdate, beryllium and other phosphatase substrates. Mass spectrometry identified the phosphatase Pho8p as a contaminant of the Spf1p preparation. Modification of the purification procedure led to a contaminant-free Spf1p preparation that was neither stimulated by Ca2+ nor inhibited by EGTA or molybdate. The phosphoenzyme levels of a contaminant-free Spf1p preparation were not affected by Ca2+. These results indicate that the reported effects of Ca2+ on Spf1p do not reflect the intrinsic properties of Spf1p but are mediated by the activity of the accompanying phosphatase.
Although reference genes have previously been used in the expression analysis of genes involved in sugarcane flowering they had not been experimentally validated for stability and consistency of ...expression between different samples over a wide range of experimental conditions. Here we report the analysis of candidate reference genes in different tissue types, at different temporal time-points, in both short and long day photoperiodic treatments. The stability of the candidate reference genes in all conditions was evaluated with NormFinder, BestKeeper, and RefFinder algorithms that complement each other for a more robust analysis. As the Normfinder algorithm was more appropriate for our experimental conditions, greater emphasis was placed on Normfinder when choosing the most stable genes. UBQ1 and TUB were shown to be the most stable reference genes to use for normalizing RT-qPCR gene expression data during floral induction, whilst 25SrRNA1 and GAPDH were the least stable. Their use as a reference gene pair was validated by analyzing the expression of two differentially expressed target genes (PIL5 and LHP1). The UBQ1/TUB reference genes combination was able to reveal small significant differences in gene expression of the two target genes that were not detectable when using the least stable reference gene combination. These results can be used to inform the choice of reference genes to use in the study of the sugarcane floral induction pathway. Our work also demonstrates that both PIL5 and LHP1 are significantly up-regulated in the initial stages of photoperiodic induction of flowering in sugarcane.
The lowest dosage of empagliflozin (10 mg) showed similar benefits on glycated hemoglobin (HbA1c) level, body weight, blood pressure, and total and cardiovascular mortality in comparison with the ...highest available dose (25 mg) in the EMPAREG trial. These findings have not been clearly demonstrated for canagliflozin and dapagliflozin. The objective was to compare the effect of different doses of SGLT2 inhibitors commercially available in Brazil on HbA1c and body weight of patients with type 2 diabetes. MEDLINE, Cochrane and Embase databases were searched from inception until 11
October 2021 for randomized controlled trials of SGLT2 inhibitors in type 2 diabetes patients, lasting at least 12 weeks. HbA
and body weight variations were described using standard mean difference. We performed direct and indirect meta-analysis, as well as a meta-regression with medication doses as covariates. Eighteen studies were included, comprising 16,095 patients. In the direct meta-analysis, SGLT2 inhibitors reduced HbA1c by 0.62% (95% CI -0.66 to -0.59) and body weight by 0.60 kg (95% CI -0.64 to -0.55). In the indirect meta-analysis, canagliflozin 300 mg ranked the highest regarding reductions in HbA
and body weight. The remaining medications and dosages were clinically similar, despite some statistically significant differences among them. Canagliflozin 300 mg seems to be more potent in reducing HbA1c and body weight in patients with type 2 diabetes. The remaining SGLT2 inhibitors at different doses lead to similar effects for both outcomes. Whether these glycemic and weight effects are reflected in lower mortality and cardiovascular events is still uncertain and may be a topic for further studies.
Traditional sugarcane cultivars (
) proved highly susceptible to diseases, and this led breeders to progress to interspecific crosses resulting in disease resistance. A backcrossing program to
was ...then required to boost sucrose content. Clonal selection across generations and incorporation of other germplasm into cultivated backgrounds established the (narrow) genetic base of modern cultivars (
spp.), which have a man-made genome. The genome complexity has inspired several molecular studies that have elucidated aspects of sugarcane genome constitution, architecture, and cytogenetics. However, there is a critical shortage of information on chromosome behavior throughout meiosis in modern cultivars. In this study, we examined the microsporogenesis of a contemporary variety, providing a detailed analysis of the meiotic process and chromosome association at diakinesis, using FISH with centromeric probes. Chromosomal abnormalities were documented by examining high quality preparations of pollen mother cells (700 in total). Approximately 70% of the cells showed abnormalities, such as metaphase chromosomes not lined up at the plate, lagging chromosomes and chromosomal bridges, and tetrad cells with micronuclei. Some dyads with asynchronous behavior were also observed. Due to the hybrid composition of the sugarcane genome, we suggest that bivalent incomplete pairing may occur in the first prophase leading to univalency. The presence of rod bivalents showing the lagging tendency is consistent with a reduction in chiasma frequency. Finally, the presence of chromatin bridges indicates the indirect occurrence of chromosomal inversions, although chromosome fragments were not clearly recognized. Possible reasons for such meiotic abnormalities and the large prevalence of bivalent formation are discussed.
The plasma membrane Ca
2+
pumps (PMCA) are P-ATPases that control Ca
2+
signaling and homeostasis by transporting Ca
2+
out of the eukaryotic cell. Humans have four genes that code for PMCA isoforms ...(PMCA1-4). A large diversity of PMCA isoforms is generated by alternative mRNA splicing at sites A and C. The different PMCA isoforms are expressed in a cell-type and developmental-specific manner and exhibit differential sensitivity to a great number of regulatory mechanisms. PMCA4 has two A splice variants, the forms “x” and “z”. While PMCA4x is ubiquitously expressed and relatively well-studied, PMCA4z is less characterized and its expression is restricted to some tissues such as the brain and heart muscle. PMCA4z lacks a stretch of 12 amino acids in the so-called A-M3 linker, a conformation-sensitive region of the molecule connecting the actuator domain (A) with the third transmembrane segment (M3). We expressed in yeast PMCA4 variants “x” and “z”, maintaining constant the most frequent splice variant “b” at the C-terminal end, and obtained purified preparations of both proteins. In the basal autoinhibited state, PMCA4zb showed a higher ATPase activity and a higher apparent Ca
2+
affinity than PMCA4xb. Both isoforms were stimulated by calmodulin but PMCA4zb was more strongly activated by acidic lipids than PMCA4xb. The results indicate that a PMCA4 intrinsically more active and more responsive to acidic lipids is produced by the variant “z” of the splicing site A.
In this work, the ability of biosurfactant production by Pseudomonas aeruginosa in batch cultivation using cashew apple juice (CAJ) and mineral media was evaluated. P. aeruginosa was cultivated in ...CAJ, which was supplemented with peptone (5.0 g/L) and nutritive broth. All fermentation assays were performed in Erlenmeyer flasks containing 300 mL, incubated at 30 degrees C and 150 rpm. Cell growth (biomass and cell density), pH, and superficial tension were monitored vs time. Surface tension was reduced by 10.58 and 41% when P. aeruginosa was cultivated in nutrient broth and CAJ supplemented with peptone, respectively. These results indicated that CAJ is an adequate medium for growth and biosurfactant production. Best results of biosurfactant production were obtained when CAJ was supplemented with peptone.
New antihyperglycemic medications have been proven to have cardiovascular (CV) and renal benefits in type 2 diabetes mellitus (T2DM); however, an evidence-based decision tree in specific clinical ...scenarios is lacking.
Systematic review and meta-analysis of randomized controlled trials (RCTs), with trial sequential analysis (TSA). Randomized controlled trial inclusion criteria were patients with T2DM from 1 of these subgroups: elderly, obese, previous atherosclerotic CV disease (ASCVD), previous coronary heart disease (CHD), previous heart failure (HF), or previous chronic kidney disease (CKD). Randomized controlled trials describing those subgroups with at least 48 weeks of follow-up were included. Outcomes: 3-point major adverse cardiovascular events (MACE), CV death, hospitalization due to HF, and renal outcomes. We performed direct meta-analysis with the number of events in the intervention and control groups in each subset, and the relative risk of the events was calculated.
Sodium-glucose cotransporter 2 inhibitors (SGLT2i) and glucagon-like peptide 1 receptor agonists (GLP-1 RA) were the only antihyperglycemic agents related to a reduction in CV events in different populations. For obese and elderly populations, GLP-1 RA were associated with benefits in 3-point MACE; for patients with ASCVD, both SGLT2i and GLP-1 RA had benefits in 3-point MACE, while for patients with CHD, only SGLT2i were beneficial.
SGLT2i and GLP-1 RA reduced CV events in selected populations: SGLT2i led to a reduction in events in patients with previous CHD, ASCVD, and HF. GLP-1 RA led to a reduction in CV events in patients with ASCVD, elderly patients, and patients with obesity. Trial sequential analysis shows that these findings are conclusive. This review opens a pathway towards evidence-based, personalized treatment of T2DM.
PROSPERO CRD42019132807.
Aim
To develop and validate a predictive model for falls in hospitalized adult clinical and surgical patients, assessing intrinsic (i.e. patient‐related) and extrinsic factors (i.e. care ...process‐related).
Background
To identify factors predictive of falls and enable appropriate management of fall risk it is necessary to understand patient and environmental factors, along with care delivery processes.
Design
A matched case–control study.
Methods
This study was conducted in the medical and surgical wards of a Brazilian teaching hospital. The sample included 536 patients, with data collected in 2013–2014. Data analysis included descriptive statistics and conditional logistic regression. Cases of patients aged 18 years or older who fell while hospitalized were included. One patient who did not fall during hospitalization, matched by sex, ward and admission date, was selected as a control for each included case.
Results
The SAK Fall Scale (Severo‐Almeida‐Kuchenbecker) was developed and validated. The scale includes seven variables: disorientation/confusion, frequent urination, walking limitations, lack of caregiver, postoperative status, previous falls and number of medications administered within 72 hr prior to the fall. This scale showed acceptable predictive accuracy.
Conclusions
The newly developed SAK Fall Scale includes five intrinsic and two extrinsic variables and differs from other predictive scales for falls. The findings of this study are broad and the scale, which is easy to apply, can be used worldwide by nurses in health services. In advanced practice, the testing of a new model for fall risk contributes to preventive interventions and thus has an impact on patient safety.
目的
建立并验证住院成人临床与外科患者跌倒的预测模型,评估内在(即:与患者相关)和外在因素(即:与护理过程相关)因素。
背景
为确定跌倒的预测因素,对跌倒风险进行妥善管理,有必要了解患者与环境因素,以及护理实施过程。
设计
配对病例对照研究
方法
本研究在一家巴西教学医院的内科及外科病房进行。样本包括536名患者,数据于2013‐2014年收集。数据分析包括描述性统计和条件逻辑回归。包括18岁及以上患者在住院期间跌倒的病例。住院期间未跌倒的患者(按性别、病房和住院日期匹配)作为纳入研究病例的对照。
结果
建立并验证了SAK(Severo‐Almeida‐Kuchenbecker)跌倒量表。量表包含7个变量:迷失方向/困惑、尿频、行走限制、缺乏照料者、术后状态、既往跌倒及跌倒前72小时内服用的药物数量。该量表具有可接受的预测准确性。
结论
最新开发的SAK跌倒量表包含5个内在变量和2个外在变量,与其它跌倒预测量表不同。本研究结果范围广泛,量表易于应用,可供全球医疗卫生服务行业的护士使用。在高级实践中,测试一种新的跌倒风险模型有助于防止干扰,从而影响患者安全。
Fatty acid methyl esters (FAMEs) were obtained from vegetable oils of soybean, corn and sunflower. The current study was focused on evaluating the antifungal activity of FAMEs mainly against ...Paracoccidioides spp., as well as testing the interaction of these compounds with commercial antifungal drugs and also their antioxidant potential. FAMEs presented small IC50 values (1.86-9.42 μg/mL). All three FAMEs tested showed antifungal activity against isolates of Paracoccidioides spp. with MIC values ranging from 15.6-500 µg/mL. Sunflower FAMEs exhibited antifungal activity that extended also to other genera, with an MIC of 15.6 μg/mL against Candida glabrata and C. krusei and 31.2 μg/mL against C. parapsilosis. FAMEs exhibited a synergetic effect with itraconazole. The antifungal activity of the FAMEs against isolates of Paracoccidioides spp. is likely due to the presence of methyl linoleate, the major compound present in all three FAMEs. The results obtained indicate the potential of FAMEs as sources for antifungal and antioxidant activity.
Sugarcane mosaic disease (SMD) caused by sugarcane mosaic virus, is one of the main diseases in sugarcane production areas in Brazil. Thus, the identification of new sources of resistance for use in ...future introgression crosses is key for reliable economic gains. Here, we aimed to screen a diversity panel of 98 sugarcane genotypes for SMD under natural infection conditions, to investigate virus-specific amplicons from SCMV coat protein gene (CP), and identify marker-trait associations via association mapping using Amplified Fragment Length Polymorphism (AFLP) and Simple Sequence Repeats (SSR). The highest SMD incidence (26.53%) was observed eight months after planting, with significant differences (p<0.01) among genotypes and a means-based broad-sense heritability of 62.49% with a noticeable contribution of
Saccharum spontaneum
to SMD resistance. The CP sequence analysis revealed no variation among the four selected plant samples, which phylogenetic analysis revealed clustering with the RIB-1 strain while putative amino acid substitutions indicate a new SCMV isolate. From a subset of 135 SSR and 663 AFLP markers, selected after quality control, 91 markers were associated with response to SCMV (
p
<0.05) by simple linear regression, and 24 were significant at
p
<0.01. Four out these 24 fit in a stepwise regression at
p
<0.05, all contributing for the resistance to SMD, and are present in thirteen genotypes showing no SMD symptoms. These four markers collectively explain 29.95% of trait variation, while individually explain from 5.51 to 14.02%, and may correspond to new genomic regions conferring genetic resistance to SMD which investigation is worthwhile.