Genetic Manipulation of Phytohormone Levels in Sugar beet

The improvement of the commercial sugar beet crop by genetic modification will require the subtle regulation of expression of foreign genes in the transgenic plants. A strategy has been defined for production of high sucrose yield/ high juice purity/ low environmental impact varieties of sugar beet. This requires modification of the phytohormone levels in the outer cambial rings of the storage root. The auxin IAA has been implicated in the development of cambial rings in the sugar beet storage root. The hairy root transformation system was used to introduce the auxin biosynthetic genes (iaaM and iaall) from Agrobacterium rhizogenes strain A4b. Agropine and mannopine were detected in all A4h hairy root clones tested, suggesting transfer (and functional expression) of genes from the Ri plasmid T R T-DNA to the sugar beet genome. It was shown by DNA-DNA hybridisation experiments that part of the TL and TR T-ONA had been integrated into the sugar beet cv. Bella genome. Altered levels of IAA were observed between transformed and non-transformed sugar beet seedling root material and also between different hairy root clones. Preliminary studies on the auxin binding proteins of A4b transformed sugar beet hairy root clones, suggested that there may be a regulatory system to compensate for the excess levels of IAA in transformed tissue. Those clones with the highest level of IAA, in general had lower ABP activity. The hairy root transformation system was a suitable model for introducing the auxin biosynthetic genes of Agrobacterium in order to alter plant growth regulator levels within the sugar beet cv. Bella genome

Evaluating the responsiveness of the Warwick Edinburgh Mental Well-Being Scale (WEMWBS) : group and individual level analysis

Background: Mental well-being now features prominently in UK and international health policy. However, progress has been hampered by lack of valid measures that are responsive to change. The objective of this study was to evaluate the responsiveness of the Warwick Edinburgh Mental Well-being Scale (WEMWBS) at both the individual and group level. Methods: Secondary analysis of twelve different interventional studies undertaken in different populations using WEMWBS as an outcome measure. Standardised response mean (SRM), probability of change statistic (P̂) and standard error of measurement (SEM) were used to evaluate whether WEMWBS detected statistically important changes at the group and individual level, respectively. Results: Mean change in WEMWBS score ranged from −0.6 to 10.6. SRM ranged from −0.10 (95% CI: -0.35, 0.15) to 1.35 (95% CI: 1.06, 1.64). In 9/12 studies the lower limit of the 95% CI for P̂ was greater than 0.5, denoting responsiveness. SEM ranged from 2.4 to 3.1 units, and at the threshold 2.77 SEM, WEMWBS detected important improvement in at least 12.8% to 45.7% of participants (lower limit of 95% CI>5.0%). Conclusions: WEMWBS is responsive to changes occurring in a wide range of mental health interventions undertaken in different populations. It offers a secure base for research and development in this rapidly evolving field. Further research using external criteria of change is warranted

Human sperm ion channel (dys)function:implications for fertilization

BACKGROUND: Intensive research on sperm ion channels has identified members of several ion channel families in both mouse and human sperm. Gene knock-out studies have unequivocally demonstrated the importance of the calcium and potassium conductances in sperm for fertility. In both species, the calcium current is carried by the highly complex cation channel of sperm (CatSper). In mouse sperm, the potassium current has been conclusively shown to be carried by a channel consisting of the pore forming subunit SLO3 and auxiliary subunit leucine-rich repeat-containing 52 (LRRC52). However, in human sperm it is controversial whether the pore forming subunit of the channel is composed of SLO3 and/or SLO1. Deciphering the role of the proton-specific Hv1 channel is more challenging as it is only expressed in human sperm. However, definitive evidence for a role in, and importance for, human fertility can only be determined through studies using clinical samples.OBJECTIVE AND RATIONALE: This review aims to provide insight into the role of sperm ion channels in human fertilization as evidenced from recent studies of sperm from infertile men. We also summarize the key discoveries from mouse ion channel knock-out models and contrast the properties of mouse and human CatSper and potassium currents. We detail the evidence for, and consequences of, defective ion channels in human sperm and discuss hypotheses to explain how defects arise and why affected sperm have impaired fertilization potential.SEARCH METHODS: Relevant studies were identified using PubMed and were limited to ion channels that have been characterized in mouse and human sperm. Additional notable examples from other species are included as appropriate.OUTCOMES: There are now well-documented fundamental differences between the properties of CatSper and potassium channel currents in mouse and human sperm. However, in both species, sperm lacking either channel cannot fertilize in vivo and CatSper-null sperm also fail to fertilize at IVF. Sperm-lacking potassium currents are capable of fertilizing at IVF, albeit at a much lower rate. However, additional complex and heterogeneous ion channel dysfunction has been reported in sperm from infertile men, the causes of which are unknown. Similarly, the nature of the functional impairment of affected patient sperm remains elusive. There are no reports of studies of Hv1 in human sperm from infertile men.WIDER IMPLICATIONS: Recent studies using sperm from infertile men have given new insight and critical evidence supporting the supposition that calcium and potassium conductances are essential for human fertility. However, it should be highlighted that many fundamental questions remain regarding the nature of molecular and functional defects in sperm with dysfunctional ion channels. The development and application of advanced technologies remains a necessity to progress basic and clinical research in this area, with the aim of providing effective screening methodologies to identify and develop treatments for affected men in order to help prevent failed ART cycles. Conversely, development of drugs that block calcium and/or potassium conductances in sperm is a plausible strategy for producing sperm-specific contraceptives.</p

The Impact of EMI on ELT Professional Development

In this paper, it is our intention to discuss the history of CLIL (Content and Language Integrated Learning) and EMI (English Medium Instruction) training at our university and explain how this has led us to realize that there are two possible areas for professional development for the English language teacher (ELT). EMI potentially provides an opportunity for the ELT to become an English language expert (ELE) accompanying EMI instructors or to become an EMI instructor. We consider the steps we have taken in order to foment content classes in English as a means to increasing internationalized education across faculty. The instruction of content courses in English at the higher education level has been the focus of our particular interest for the past few years at our university and the Language Department has been involved in this part of the university’s internationalization project. The university’s interest in raising its international profile has been top-of-mind for some time now. Not only are the authority figures anxious to improve the university rankings, but they are also interested in attracting foreign students to come to Guadalajara to learn Spanish as a foreign language while they continue studying for credits in their undergraduate programs. These credit-bearing classes, which they study in departments such as mathematics, administration, engineering and so on, are taught in English. The content courses in English (EMI classes) are attended by both foreign and Mexican students, thus providing an international experience for both. Some of the EMI lecturers are native English speakers, but the vast majority are Mexican and English is their second language. Over the years, the university has provided several opportunities for CLIL/EMI training. Faculty members who have undergone this training have realized that EMI requires a change in the instructional methods typically used in a higher education setting. Taking a content class in a language that is not the student’s first language implies a greater challenge; therefore, how these classes are given is crucial. Scaffolding learning is of vital importance in order to make input accessible for the students and to help them achieve the required academic production (class related assignments such as, essays, summaries, reports, presentations, discussions in English). This scaffolding process is familiar to the English language teacher but may not be to the EMI instructor; therefore, this is an area of support provided by the ELE. We go on to discuss the importance of this ELE working alongside the faculty instructor to train, advise, observe, and give feedback and support. Becoming an ELE advisor for EMI instructors is one possible area for professional development. The second area of opportunity that we envision is that of the language professional looking at their own background to appreciate if they have a future as a possible EMI lecturer, especially if their first area of academic study is unrelated to a languages background. This paper provides real-life examples of English language teachers in this role at our university.ITESO, A.C

Developing a multi-pollutant conceptual framework for the selection and targeting of interventions in water industry catchment management schemes

In recent years water companies have started to adopt catchment management to reduce diffuse pollution in drinking water supply areas. The heterogeneity of catchments and the range of pollutants that must be removed to meet the EU Drinking Water Directive (98/83/EC) limits make it difficult to prioritise areas of a catchment for intervention. Thus conceptual frameworks are required that can disaggregate the components of pollutant risk and help water companies make decisions about where to target interventions in their catchments to maximum effect. This paper demonstrates the concept of generalising pollutants in the same framework by reviewing key pollutant processes within a source-mobilisation-delivery context. From this, criteria are developed (with input from water industry professionals involved in catchment management) which highlights the need for a new water industry specific conceptual framework. The new CaRPoW (Catchment Risk to Potable Water) framework uses the Source-Mobilisation-Delivery concept as modular components of risk that work at two scales, source and mobilisation at the field scale and delivery at the catchment scale. Disaggregating pollutant processes permits the main components of risk to be ascertained so that appropriate interventions can be selected. The generic structure also allows for the outputs from different pollutants to be compared so that potential multiple benefits can be identified. CaRPow provides a transferable framework that can be used by water companies to cost-effectively target interventions under current conditions or under scenarios of land use or climate change

Complex CatSper-dependent and independent [Ca2⁺]i signalling in human spermatozoa induced by follicular fluid

STUDY QUESTION: Does progesterone in human follicular fluid (hFF) activate CatSper and do other components of hFF modulate this effect and/or contribute separately to hFF-induced Ca2+ signaling?SUMMARY ANSWER: hFF potently stimulates CatSper and increases [Ca2+]i, primarily due to high concentrations of progesterone, however,other components of hFF also contribute to [Ca2+]i signaling, including modulation of CatSper channel activity and inhibition of [Ca2+]i oscillations.WHAT IS KNOWN ALREADY: CatSper, the principal Ca2+ channel in spermatozoa, is progesterone-sensitive and essential for fertility. Both hFF and progesterone, which is present in hFF, influence sperm function and increase their [Ca2+]i.STUDY DESIGN, SIZE, DURATION: This basic medical research study used semen samples from &gt;40 donors and hFF from &gt;50 patients who were undergoing surgical oocyte retrieval for IVF/ICSI.PARTICIPANTS/MATERIALS, SETTING, METHODS: Semen donors and patients were recruited in accordance with local ethics approval (13/ES/0091) from the East of Scotland Research Ethics Service REC1. Activities of CatSper and KSper were assessed by patch clamp electrophysiology. Sperm [Ca2+]i responses were examined in sperm populations and single cells. Computer-assisted sperm analysis (CASA) parameters and penetration into viscous media were used to assess functional effects.MAIN RESULTS AND THE ROLE OF CHANCE: hFF and progesterone significantly potentiated CatSper currents. Under quasiphysiologicalconditions, hFF (up to 50%) failed to alter membrane K+ conductance or current reversal potential. hFF and progesterone (at an equivalent concentration) stimulated similar biphasic [Ca2+]i signals both in sperm populations and single cells. At a high hFF concentration (10%), the sustained (plateau) component of the [Ca2+]i signal was consistently greater than that induced by progesterone alone. In single cell recordings, 1% hFF-induced [Ca2+]i oscillations similarly to progesterone but with 10% hFF generation of [Ca2+]i oscillations was suppressed. After treatment to ‘strip’ lipid-derived mediators, hFF failed to significantly stimulate CatSper currents but induced small [Ca2+]i responsesthat were greater than those induced by the equivalent concentration of progesterone after stripping. Similar [Ca2+]i responses were observed when sperm pretreated with 3 μM progesterone (to desensitize progesterone responses) were stimulated with hFF or stripped hFF. hFF stimulated viscous media penetration and was more effective than the equivalent does of progesterone.LARGE SCALE DATA: N/A.LIMITATIONS, REASONS FOR CAUTION: This was an in vitro study. Caution must be taken when extrapolating these results in vivo.WIDER IMPLICATIONS OF THE FINDINGS: This study directly demonstrates that hFF activates CatSper and establishes that the biologically important effects of hFF reflect, at least in part, action on this channel, primarily via progesterone. However, these experiments also demonstrate that other components of hFF both contribute to the [Ca2+]i signal and modulate the activation of CatSper. Simple in vitro experiments performed out of the context of the complex in vivo environment need to be interpreted with caution