Shotguns vs Lasers: Identifying barriers and facilitators to scaling-up plant molecular farming for high-value health products.

Plant molecular farming (PMF) is a convenient and cost-effective way to produce high-value recombinant proteins that can be used in the production of a range of health products, from pharmaceutical therapeutics to cosmetic products. New plant breeding techniques (NPBTs) provide a means to enhance PMF systems more quickly and with greater precision than ever before. However, the feasibility, regulatory standing and social acceptability of both PMF and NPBTs are in question. This paper explores the perceptions of key stakeholders on two European Union (EU) Horizon 2020 programmes-Pharma-Factory and Newcotiana-towards the barriers and facilitators of PMF and NPBTs in Europe. One-on-one qualitative interviews were undertaken with N = 20 individuals involved in one or both of the two projects at 16 institutions in seven countries (Belgium, France, Germany, Italy, Israel, Spain and the UK). The findings indicate that the current EU regulatory environment and the perception of the public towards biotechnology are seen as the main barriers to scaling-up PMF and NPBTs. Competition from existing systems and the lack of plant-specific regulations likewise present challenges for PMF developing beyond its current niche. However, respondents felt that the communication of the benefits and purpose of NPBT PMF could provide a platform for improving the social acceptance of genetic modification. The importance of the media in this process was highlighted. This article also uses the multi-level perspective to explore the ways in which NPBTs are being legitimated by interested parties and the systemic factors that have shaped and are continuing to shape the development of PMF in Europe

Clinicopathological correlation and prognostic significance of VEGF-A, VEGF-C, VEGFR-2 and VEGFR-3 expression in colorectal cancer.

BACKGROUND: Colorectal cancer (CRC) is the third most common type of cancer and the fourth most frequent cause of cancer death. Literature indicates that vascular endothelial growth factor is a predominant angiogenic factor and that angiogenesis plays an important role in the progression of CRC. PATIENTS AND METHODS: The present series consisted of tissue samples obtained from 672 patients who had undergone large bowel resection between 2005 and 2010 at the Braga Hospital, Portugal. Archival paraffin-embedded CRC tissue and normal adjacent samples were used to build up tissue microarray blocks and VEGF-A, VEGF-C, VEGFR-2 and VEGFR-3 expression was immunohistochemically assessed. RESULTS: We observed an overexpression of VEGF-C in CRC when tumour cells and normal-adjacent tissue were compared (p=0.004). In tumour samples, VEGF-C-positive cases were associated with VEGFR-3 expression (p=0.047). When assessing the correlation between VEGF-A, VEGF-C, VEGFR-2 and VEGFR-3 expressions and the clinicopathological data, it was revealed that VEGF-A positive cases were associated with male gender (p=0.016) and well-differentiated tumours (p=0.001); VEGF-C with colon cancers (p=0.037), exophytic (p=0.048), moderately-differentiated (p=0.007) and T3/T4 (p=0.010) tumours; VEGFR-2 with invasive adenocarcinoma (p=0.007) and VEGFR-3 with the presence of hepatic metastasis (p=0.032). Overall survival curves for CRC were statistically significant for rectal cancer, VEGF-C expression and stage III (p=0.019) and VEGFR-3 expression and stage IV (p=0.047). CONCLUSION: Quantification of VEGF-A, VEGF-C, VEGFR-2 and VEGFR-3 expression seems to provide valuable prognostic information in CRC and the correlation with clinicopathological data revealed an association with characteristics that contribute to progression, invasion and metastasis leading to poorer survival rates and prognosis

Total and segmental colonic transit time in constipated patients with Chagas' disease without megaesophagus or megacolon

Manometric and pharmacological tests have shown that motor abnormalities may occur in the non-dilated colons of chagasic patients. In order to investigate the presence of abnormalities of colonic function in constipated patients with Chagas' disease (ChC) without megaesophagus or megacolon, studies of total and segmental colonic transit time with radiopaque markers were performed on 15 ChC patients, 27 healthy volunteers and 17 patients with idiopathic constipation (IC). The values obtained for the control group were similar to those reported in the literature (total colonic time: 34.1 +/- 15.6 h; right colon: 9.9 +/- 7.3 h; left colon: 10.8 +/- 10 h, and rectosigmoid: 12.6 +/- 9.9 h). Colonic transit time data permitted us to divide both IC and ChC patients into groups with normal transit and those with slow colonic transit. Colonic inertia was detected in 41% of IC patients and in 13% of ChC patients; left colon isolated stasis (hindgut dysfunction) was detected in 12% of IC patients and 7% of ChC patients, and outlet obstruction was detected in 6% of IC patients and 7% of ChC patients. There were no significant differences in total or segmental colonic transit times between slow transit IC and slow transit ChC patients. In conclusion, an impairment of colonic motility was detected in about 30% of constipated patients with Chagas' disease without megaesophagus or megacolon. This subgroup of patients presented no distinctive clinical feature or pattern of colonic dysmotility when compared to patients with slow transit idiopathic constipation.331434

Effect of cinnamon powder addition to a Portuguese custard tart (Pastel de Nata) on healthy adults' postprandial glycemia

Background and Objective: Cinnamon is a spice used over the years in cooking to impart aromatic, flavor and taste properties to food and beverages. Moreover, cinnamon has been used for its medicinal properties due to its potential phenolic content, which can protect against cardio-metabolic diseases. Previous studies reported an improvement of postprandial glycemia after addition of cinnamon powder to a high-sugar meal. The study aims at investigating the effect of adding cinnamon powder to a Portuguese custard tart (Pastel de Nata) on the postprandial glycemic response in healthy subjects. Subjects and Methods: After review board and Ethic Committee of the State approval, thirty-two healthy human subjects were assigned in a controlled study and randomly allocated into 2 groups: 16 subjects ingested a custard tart with cinnamon powder (cinnamon group) and 16 subjects ingested a custard tart alone (control group). Blood glucose concentrations were measured before interventions and after 30, 60, 90 and 120 minutes. Chemical analysis was performed to quantify the total phenolic content and antioxidant activity. Results: The postprandial blood glucose (PBG) area under the curve (AUC) was significantly lower (p = 0.0005) in the cinnamon group (599.2 ± 9.1) compared to the AUC of the control group (645.7 ± 7.7). The administration of cinnamon powder to the custard tarts slightly decreased PBG mean values compared to custard tart without cinnamon powder, although it did not reach statistical significance (p = 0.273). Cinnamon addition to custard tart improved the total phenolic content (1278.7 ± 0.7 compared to 253.7 ± 22.8 mg/L gallic acid) and antioxidant properties, increasing 4.4 times the capacity of free-radical scavenger compared with custard tart without cinnamon (IC50). Conclusion: The addition of cinnamon powder to custard tart could be beneficial to glycemic control

An investigation into the effect of MLSS on the effluent quality and sludge settleability in an aerobic-anoxic sequencing batch reactor (AASBR)

The aim of this study is to enhance the effluent quality and improve the sludge settleability by determining the effects of the mixed liquor suspended solids (MLSS) on the solid’s settling behaviour and the treatment efficiency in an aerobic-anoxic sequencing batch reactor (AASBR). The results obtained from this study revealed that raising the MLSS concentration from 2 to 3 g/l improved the chemical oxygen demand (COD), ammonia-nitrogen (NH3-N) and nitrate-nitrogen NO3-N removal efficiency, and led to an increase in the sludge volume index (SVI) value. Moreover, increasing the MLSS concentration from 3 to 4 g/l did not significantly affect the COD, NH3-N and NO3-N removal rates or the solid’s settling behaviour. However, increasing the MLSS concentration from 4 to 6 g/l significantly reduced the COD and nitrate removal efficiency and the sludge settling rate slowed down. The results proved that the optimal MLSS concentration for COD, NH3-N and NO3-N removal is between 3 and 4 g/l. In this range the removal rates for COD, NH3-N and NO3-N were 93%, 95% and 96% respectively, and the effluent quality was 35 mg/l, 0.43 mg/l and 0.75 mg/l for COD, NH3-N and NO3-N respectively. In addition, a good solid separation occurred during that range with SVI value of 81 ml/g; this finding was supported by a morphological study along with scanning electron microscopy (SEM)

Quantitative image analysis of polyhydroxyalkanoates inclusions from microbial mixed cultures under different SBR operation strategies

Polyhydroxyalkanoates (PHAs) produced from mixed microbial cultures (MMC), regarded as potential substitutes of petrochemical plastics, can be found as intracellular granules in various microorganisms under limited nutrient conditions and excess of carbon source. PHA is traditionally quantified by laborious and time-consuming chromatography analysis, and a simpler and faster method to assess PHA contents from MMC, such as quantitative image analysis (QIA), is of great interest. The main purpose of the present work was to upgrade a previously developed QIA methodology (Mesquita et al., 2013a, 2015) for MMC intracellular PHA contents quantification, increase the studied intracellular PHA concentration range and extend to different sequencing batch reactor (SBR) operation strategies. Therefore, the operation of a new aerobic dynamic feeding (ADF) SBR allowed further extending the studied operating conditions, dataset, and range of the MMC intracellular PHA contents from the previously reported anaerobic/aerobic cycle SBR. Nile Blue A (NBA) staining was employed for epifluorescence microscope visualization and image acquisition, further fed to a custom developed QIA. Data from each of the feast and famine cycles of both SBR were individually processed using chemometrics analysis, obtaining the correspondent partial least squares (PLS) models. The PHA concentrations determined from PLS models were further plotted against the results obtained in the standard chromatographic method. For both SBR the predicted ability was higher at the end of the feast stage than for the famine stage. Indeed, an independent feast and famine QIA data treatment was found to be fundamental to obtain the best prediction abilities. Furthermore, a promising overall correlation (R2 of 0.83) could be found combining the overall QIA data regarding the PHA prediction up to a concentration of 1785.1 mgL-1 (37.3 wt%). Thus, the results confirm that the presented QIA methodology can be seen as promising for estimating higher intracellular PHA concentrations for a larger reactors operation systems and further extending the prediction range of previous studies.This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684) and BioTecNorte operation (NORTE01-0145-FEDER-000004) funded by European Regional Development Fundunder the scope ofNorte2020 - ProgramaOperacional Regional do Norte.The authors also acknowledge the financial support to Cristiano S. Leal (PTDC/EBB-EBI/103147/2008, FCOMP-01-0124-FEDER009704) and Daniela P. Mesquita through the FCT postdoctoral grant (SFRH/BPD/82558/2011).info:eu-repo/semantics/publishedVersio

Quantitative image analysis for the characterization of microbial aggregates in biological wastewater treatment : a review

Quantitative image analysis techniques have gained an undeniable role in several fields of research during the last decade. In the field of biological wastewater treatment (WWT) processes, several computer applications have been developed for monitoring microbial entities, either as individual cells or in different types of aggregates. New descriptors have been defined that are more reliable, objective, and useful than the subjective and time-consuming parameters classically used to monitor biological WWT processes. Examples of this application include the objective prediction of filamentous bulking, known to be one of the most problematic phenomena occurring in activated sludge technology. It also demonstrated its usefulness in classifying protozoa and metazoa populations. In high-rate anaerobic processes, based on granular sludge, aggregation times and fragmentation phenomena could be detected during critical events, e.g., toxic and organic overloads. Currently, the major efforts and needs are in the development of quantitative image analysis techniques focusing on its application coupled with stained samples, either by classical or fluorescent-based techniques. The use of quantitative morphological parameters in process control and online applications is also being investigated. This work reviews the major advances of quantitative image analysis applied to biological WWT processes.The authors acknowledge the financial support to the project PTDC/EBB-EBI/103147/2008 and the grant SFRH/BPD/48962/2008 provided by Fundacao para a Ciencia e Tecnologia (Portugal)

Predicting response to holistic breathlessness services: a pooled analysis of trial data

Background: Holistic breathlessness services have been developed for people with advanced disease and chronic breathlessness, leading to improved psychological aspects of breathlessness and health. The extent to which patient characteristics influence outcomes is unclear. Aim: To identify patient characteristics predicting outcomes of mastery and distress around breathlessness following holistic breathlessness services. Design: Secondary analysis of pooled individual patient data from three clinical trials. Our primary analysis assessed predictors of clinically important improvements in Chronic Respiratory Questionnaire mastery scores (+0.5 point), and our secondary analysis predictors of improvements in Numerical Rating Scale distress due to breathlessness (-1 point). Variables significantly related to improvement in univariate models were considered in separate backwards stepwise logistic regression models. Participants: The dataset comprised 259 participants (118 female; mean (standard deviation) age 69.2(10.6) years) with primary diagnoses of chronic obstructive pulmonary disease (49.8%), cancer (34.7%) and interstitial lung disease (10.4%).Results: Controlling for age, sex, and trial, baseline mastery remained the only significant independent predictor of improvement in mastery (odds ratio 0.57, 95% confidence intervals 0.43 to 0.74; p <0001), and baseline distress remained the only significant predictor of improvement in distress (odds ratio 1.64; 95% confidence intervals 1.35 to 2.03; p<001). Baseline lung function, breathlessness severity, health status, mild anxiety and depression, and diagnosis did not predict outcomes. Conclusions: Outcomes of mastery and distress following holistic breathlessness services are influenced by baseline scores for these variables, and not by diagnosis, lung function, or health status. Stratifying patients by levels of mastery and/or distress appears appropriate for clinical trials and services

Short-term effects of repetitive transcranial magnetic stimulation on sleep bruxism:a pilot study

The purpose of this study was to investigate the effects of repetitive transcranial magnetic stimulation (rTMS) on patients with sleep bruxism (SB). Twelve patients with SB were included in an open, single-intervention pilot study. rTMS at 1 Hz and an intensity of 80% of the active motor threshold was applied to the ‘hot spot' of the masseter muscle representation at the primary motor cortex bilaterally for 20 min per side each day for 5 consecutive days. The jaw-closing muscle electromyographic (EMG) activity during sleep was recorded with a portable EMG recorder at baseline, during rTMS treatment and at follow-up for 5 days. In addition, patients scored their jaw-closing muscle soreness on a 0–10 numerical rating scale (NRS). Data were analysed with analysis of variance. The intensity of the EMG activity was suppressed during and after rTMS compared to the baseline (P = 0.04; P = 0.02, respectively). The NRS score of soreness decreased significantly during and after rTMS compared with baseline (P < 0.01). These findings indicated a significant inhibition of jaw-closing muscle activity during sleep along with a decrease of muscle soreness. This pilot study raises the possibility of therapeutic benefits from rTMS in patients with bruxism and calls for further and more controlled studies