With over 140 vitamin B6 (Pyridoxal 5’-phosphate, PLP) dependent enzymes, serving vital roles in various transamination, decarboxylation, retro-aldol cleavage and synthesis pathways these enzymes constitute the most versatile catalytic systems in nature. Enzymes of this group have an inherent reaction as well as substrate specificity. A single co-factor namely, PLP is used by specific enzymes of this group to serve distinct roles during the catalytic reaction. An ordered evolutionary adaptation in these enzymes has led to specialization achieved by each enzyme for catalyzing specific reactions. L-Threonine aldolase (L-TA) is one such PLP- dependent enzyme that catalyzes the retro-aldol cleavage of several β-hydroxy amino acids, although its natural substrates are L-threonine and L-allo-threonine with the enzyme having significant preference for L-allo-threonine. It also catalyzes racemization and transamination of D-alanine but not of the β-hydroxy amino acids. Thus, the enzyme exhibits both substrate and reaction specificity. Although, L-TA is frequently employed for stereoselective synthesis of pharmaceutically useful compounds, its reaction mechanism and associated specificity is still not clearly understood. L-TA from Escherichia coli (eTA) is being studied in our laboratory. Our objective is to elucidate the catalytic mechanism of eTA and its mode of substrate and reaction specificity using X-ray crystallography. Another objective is to establish evolutionary relationship of L-TA with other B6-dependent enzymes, such as serine hydroxymethyltransferase (SHMT) and Thermatoga maritima L-TA (TTA) that have the same fold and catalyze similar reactions. Our structural studies show that while the crystal structures of the two L-TAs are similar, they are significantly different from that of SHMT, especially at the active site. In the L-TA structures, a loop with proposed important active site residue, His126 is replaced by tetrahydrofolate (THF) in SHMT. The crystal structures of eTA in its native form and in complex with substrate or product have highlighted the importance of His126 in ensuring substrate specificity during retro-aldol cleavage of various β-hydroxy amino acids and His83 or a conserved water molecule to be active site base. Our study emphasizes the molecular level implications of the catalytic mechanism of eTA.

Background: Endothelial dysfunction (ED) is one of the earliest subclinical indicators of impaired cardiovascular health and several non-invasive tools have been developed to evaluate vascular function, including strain gauge plethysmography (SGP), brachial artery flow-mediated dilation (FMD) via ultrasound, and peripheral artery tonometry (PAT). While these tools have extensively been studied during a resting condition, the responses following acute exercise are not as well characterized. Purpose: The purpose of this study was to compare the pre- and post-exercise vascular function values obtained with SGP, FMD, and PAT. Relationships among the primary outcome variables obtained with each assessment tool were also evaluated. Methods: Vascular function was assessed in 17 sedentary, apparently healthy male subjects (24±4 yrs; 24.5±3.2 kg/m2) at rest and following an acute submaximal exercise bout with SGP, FMD, and PAT. Results: During rest, post-occlusion reactive hyperemia resulted in significant (p<0.05) increases in forearm blood flow (FBF; 2.13±1.03 vs 6.35 ± 2.90 mL/min/100 mL tissue) and area under the curve (AUC; 226.77 ± 111.20 vs 588.22 ±283.33 mL/min/100 mL) as determined by SGP. Brachial artery diameter (BAD) as assessed with FMD was increased by 5.3% (p<0.05). Resting reactive hyperemia index (RHI) as assessed by PAT was observed to be 1.73±0.34. Significant exercise-induced increases (p<0.05) were observed in baseline and post-occlusion FBF and baseline AUC values utilizing SGP. Additionally, FMD baseline blood velocity was significantly increased (91.8±11.1 vs 108.0±17.1 cm/sec, p<0.05) and the PAT augmentation index (AI) was significantly more negative (-8.8 ±9.4 vs -18.9±8.4%, p<0.05) after exercise. There were no significant correlations observed among the primary outcome measures obtained from each assessment technique. There was, however, a moderate correlation between pre-exercise vascular reactivity as assessed by SGP and change in blood velocity as assessed by FMD (r= 0.566, p= 0.035). Conclusions: The addition of an exercise stress to vascular function assessment may offer greater insight into the health of the vasculature. This initial study was undertaken to further evaluate the pre- to post-exercise responses obtained using three commonly employed vascular function assessment techniques in healthy individuals. Additional research as to the value of the addition of an exercise stress to vascular function assessment in individuals with traditional cardiovascular disease risk factors or known cardiovascular disease is warranted.

Introduction: Obese women are predisposed to greater risks of insulin resistance and compensatory hyperinsulinemia. Likewise, African-Americans, appear to be inherently insulin resistant and hyperinsulinemic even after controlling for obesity. Hyperinsulinemia has been attributed to insulin resistance and a compensatory insulin hyper-secretion by the pancreas, as well as decreased insulin clearance, notably in obesity. Pharmacological agents that may worsen insulin resistance/hyperinsulinemia in obese women is of clinical relevance. Previous data from our group suggested that combined oral contraceptives (COCs) may worsen insulin sensitivity particularly in obese women, but limited information on insulin clearance is available in obese women or African-American women. Objective: The objective of the study is to evaluate and compare the effect of a COC containing ethinyl estradiol and norgestimate on insulin clearance among lean and obese pre-menopausal women and among African-American obese vs. non African-American obese women. Method: Plasma insulin clearance was calculated from plasma insulin concentrations, following frequently sampled intravenous glucose tolerance test. Changes in insulin clearance, during six months of COC use were analyzed by repeated measures analysis. Result: Six months of COC use showed no significant change in insulin clearance in all women (p=0.3713). Furthermore, there were no divergent effects on insulin clearance among lean (n=13) and obese (n=14) women (p=0.6703) and among African-American obese (n=7) and non African-American obese (n=7) women (p=0.0957). Changes in insulin clearance, following six months of COC administration was found to be positively correlated with changes in insulin sensitivity (r=0.385, p=0.0099) and negatively correlated with changes in acute insulin response to glucose (r=-0.432, p=0.0034). Discussion: In the present study, COC administration did not show any differential effect on insulin clearance in lean vs. obese women. Future studies evaluating the effects of hormonal agents on insulin-glucose dynamics may focus on mechanisms of hormone-mediated insulin resistance and compensatory hyperinsulinemia rather than insulin clearance.

The discovery of heparin in 1916 resulted in a huge impact on the practice of medicine. Heparin has played a major role in alleviating thrombotic disorders and has also exhibited effects on almost every major system in the human body. Over the past few decades, more and more heparin-protein interactions have come to light. It is implicated to modulate several important processes such as cell growth and differentiation, inflammatory response, viral infection mechanism etc. More interesting is the observation that these interactions are considerably specific with regard to oligosaccharide sequences which have specific spatially oriented sulfate groups modulating the responses. However, due to the complex nature of these interactions and lack of effective computational capabilities, predicting these interactions is challenging.An alternative approach to modulating heparin-protein interactions would be to screen a library of molecules having a diverse distribution of the negative charges and screen them against various proteins of interest to obtain valuable information about the binding/selectivity requirements. This approach would not only yield molecules with potential clinical viability, but may also yield molecules that help decipher native mechanisms regulating proteins, which is called chemical biology in today's terms. Since the difficulties associated with carbohydrate synthesis are well known, well characterized highly sulfated oligosaccharide library screening is considered nearly impossible. Thus, the main aim of this project was to develop an effective method for the synthesis of a library of variably sulfated, non-carbohydrate molecules. The library would contain varying in the number of sulfate groups, offer positional variants of the sulfate groups and provide molecules of varying length so as to afford structural diversity necessary to mimic the heparin sequences. Previous attempts in our laboratory to synthesize such a library encountered two major problems: 1) dimerization of polyphenols due to difficult protection / deprotection strategies and 2) ineffective purification of highly water soluble sulfated molecules. To overcome the problem of protection-deprotection, “click” chemistry has been used in this work for dimerization of polyphenols without any protective groups. To overcome the second problem, a non-aqueous method of purification of highly sulfated molecules was developed, which is the first such report.As a proof of concept, a small library of 14 sulfated monomers and dimers and 8 non-sulfated dimers was generated. The protocol for dimerization of free polyphenolic molecules in has been established to use “click” chemistry for coupling the monomers without the need to protect the free hydroxyl groups. Thus by circumventing the inefficient protection-deprotection protocol, there is a tremendous improvement in yields, ease of purification and characterization and greater productivity allowing the synthesis of more number of molecules in a relatively shorter span of time. By masking the charge of the sulfate using an appropriate counter-ion and owing to the inherent lipophilicity of the aromatic scaffold, these highly charged molecules could be purified using normal phase silica gel chromatography. This method reduced the purification time from previous over 48 hours with the aqueous method to approximately 15 minutes. Further, this purification protocol may be possibly automated so as to truly generate a large library of variably sulfated non-carbohydrate molecules for the first time. Screening this library of 22 sulfated and unsulfated molecules against three enzymes of the coagulation cascade – factors IIa, Xa and XIa – has provided a wealth of information with regard to engineering specificity for recognition of these enzymes. The screening led to the identification of CS3 which inhibited factor XIa with an IC 50 of ~ 5 μM and other enzymes with an IC 50 of > 500 μM as a lead candidate with high selectivity. The success of this strategy bodes well for understanding the heparin-protein interactions at a molecular level. Previous attempts in our laboratory to synthesize such a library encountered two major problems: 1) dimerization of polyphenols due to difficult protection / deprotection strategies and 2) ineffective purification of highly water soluble sulfated molecules. To overcome the problem of protection-deprotection, “click” chemistry has been used in this work for dimerization of polyphenols without any protective groups. To overcome the second problem, a non-aqueous method of purification of highly sulfated molecules was developed, which is the first such report.As a proof of concept, a small library of 14 sulfated monomers and dimers and 8 non-sulfated dimers was generated. The protocol for dimerization of free polyphenolic molecules in has been established to use “click” chemistry for coupling the monomers without the need to protect the free hydroxyl groups. Thus by circumventing the inefficient protection-deprotection protocol, there is a tremendous improvement in yields, ease of purification and characterization and greater productivity allowing the synthesis of more number of molecules in a relatively shorter span of time. By masking the charge of the sulfate using an appropriate counter-ion and owing to the inherent lipophilicity of the aromatic scaffold, these highly charged molecules could be purified using normal phase silica gel chromatography. This method reduced the purification time from previous over 48 hours with the aqueous method to approximately 15 minutes. Further, this purification protocol may be possibly automated so as to truly generate a large library of variably sulfated non-carbohydrate molecules for the first time. Screening this library of 22 sulfated and unsulfated molecules against three enzymes of the coagulation cascade – factors IIa, Xa and XIa – has provided a wealth of information with regard to engineering specificity for recognition of these enzymes. The screening led to the identification of CS3 which inhibited factor XIa with an IC 50 of ~ 5 ?M and other enzymes with an IC 50 of > 500 ?M as a lead candidate with high selectivity. The success of this strategy bodes well for understanding the heparin-protein interactions at a molecular level.

Today, more than ever, there exists an overlap between hospitality and healthcare industries; with the impact being felt in the way hospital interiors are designed. This influence impacts the patient’s experience but focuses very little on the experience of the caregiver. Caring for a sick child takes an emotional, mental, and physical toll on a family. It is especially challenging to care for a sick family member when away from the comfort and security of the family’s private home. This project explores that challenge through the interior design of a space that, for both patient and family, is restorative, healing, and encouraging. It is a space that assists a family in caring for their loved one.

Guidelines for the management of asthma in the UK have been published (BMJ, 1990, Thorax, 1993 and 1997) and embraced by many GP practices with improved outcome for patients. The study aims to observe and follow a cohort of adult asthma subjects from differing primary health care settings over a two-year period. Also to assess a newly devised patient focused morbidity index (Q score) by comparison to an established asthma specific quality of life questionnaire (AQLQ, Juniper et al, 1993). One hundred and fourteen subjects from four GP Practices, two inner city and two suburban were studied. Morbidity was assessed by AQLQ and Q score (Rimington et al, 2001), psychological status by the hospital anxiety and depression (HAD) scale (Zigmond and Snaith, 1983). Spirometry values (forced expiratory volume in one second, FEV1), peak expiratory flow (PEF) and details of current treatment as per BTS guidelines treatment step were recorded as markers of asthma severity. Subjects were assessed at baseline, twelve and twenty-four months. A random sub set of patients was asked to repeat certain elements of the study protocol at two weeks in order to assess the reliability of the Q score. The Q score correlated from baseline to two weeks (rs=0.61) as did AQLQ symptom score (rs=0.74) both p<0.01. At baseline AQLQ symptoms correlated with PEF (rs=0.40, p<0.001) and with BTS guidelines treatment step (rs=0.25, p=0.001) as did the Q score. Similar levels of correlation were reported for FEV1 with symptoms. HAD scores also correlated to AQLQ and Q score, but there was little correlation with lung function. At one and two year follow up no significant differences were observed in subjective or objective markers of asthma. There was a significant increase (p<0.001) in the number of subjects in the higher BTS guidelines treatment steps from baseline to twelve and twenty-four months while psychological symptoms remained high for inner city patients. In conclusion the Q score yields similar results to the AQLQ and is quick and easy to use in any busy clinic. The GP practice, at the forefront of asthma care should be offering appropriate therapy and regular review. The Q score used as a patient focused morbidity index can be a useful audit tool. Altering medication can give the impression of treating asthma but with out short-term reassessment the same levels of morbidity can persist.

Research suggests that adoptees are seen in therapeutic settings in a greater proportion than their prevalence in the general population (Kennedy Porch, 2007). Few studies concentrate on adult adoptees, and those that do mainly focus on adjustment and the search for/reunion with the birth family rather than on therapy research. In particular, there seems to be a lack of empirical and theoretical understanding of the complexities of the therapeutic relationship when working with adults who were adopted. Early experiences can affect an adult's ability to form trusting relationships with others, including with a therapist. Counselling psychology, as a discipline, has a particular interest and skill base in understanding the therapeutic relationship and is therefore in the position to offer a valuable contribution to research on the therapeutic relationship with an individual who has been adopted as an infant. Through six semi-structured interviews, adoption counsellors currently working with adult adoptees were asked to share their experiences and understanding of the nature and the role of the therapeutic relationship. A qualitative methodology guided by Interpretative Phenomenological Analysis (IPA) enabled detailed exploration of the adoption counsellors' perspective. Three master themes were identified in the analysis: An awareness of the early rejection in all stages of the therapeutic process; a personal commitment to provide a corrective emotional experience; and the psychological consequences of working with adult adopted clients for therapists. The analysis highlighted the challenges that participants perceived in terms of establishing, maintaining and ending the therapeutic relationship with adoptees. The findings of this study extend the pre-existing research base by emphasising the interconnectedness of the personal and professional experiences of adoption counsellors and the need for flexibility in their practice in each phase of the therapeutic process, predominantly with regard to therapeutic boundaries and engaging in a different ending experience. The themes are discussed in relation to existing literature with an emphasis on attachment theory. The discussion also highlights practical implications, particularly the importance of familiarisation with aspects of adoption due to their likely influence on the therapeutic relationship and the need for reflective practice.

The National Health Service (NHS) in England is changing, not only in terms of the technical aspects of care (such as that afforded by new technology), but also in the way in which care is delivered to patients. For example, due to the increasing pressures being placed upon health services, hospitals need to develop innovative ways to reduce the length of time patients actually spend in hospital. Whilst this improves capacity planning, bed management and patient flow, it also seeks to ensure improved patient safety by delivering care and ongoing treatment in the most appropriate setting. However, discharging patients from hospital can be challenging and fraught with complexities. Therefore, the efficient and effective management of this process can have a considerable impact on subsequent clinical outcomes and readmission rates. As such, effective discharge planning and practice has become an integral part of health service policy and, in recent years, there has been an increased emphasis placed on the contribution nurses make to this process (Department of Health, 2002; 2005a; 2005b). New recommendations from the Kings Fund (Seale, 2016) suggest that a collaborative relationship between patients, carers, third sector parties and communities is central to the future of the NHS. This is because these perspectives are fundamental; indeed, patients are the reason the NHS exists. Since 2010, this has been the aim of my doctoral research ― to explore a shared leadership approach in determining the principles of care. The purpose of this study is to gain an insight into (and a greater understanding of) patient experiences during the discharge process following percutaneous coronary intervention (PCI) after myocardial infarction (MI) ― commonly known as a heart attack. PCI is a relatively new procedure; since its implementation it has resulted in patients staying in hospital for less than three days. Previous management would regularly result in patients remaining in hospital for longer than seven days. However, reducing the length of stay has potentially impacted how information and communication are both provided and understood by patients on discharge from acute hospital care. Previous research such as Hainsworth (2006), Department of Health (DoH) (2004) and Picker (2010) suggests that whilst there has been a great deal of emphasis placed on policy, there still remains scope for improvement in practice. As qualitative research is a form of social enquiry that focuses on the way people make sense of their experiences and the world in which they live, this research focuses on individuals who have had a heart attack and have then undergone the same treatment. The aim is to understand, describe and interpret their experiences, behaviour and feelings by looking at social processes and interactions. This was based on undertaking three separate focus groups involving patients, clinicians and members of the management team from a local NHS acute hospital in West London. Furthermore, in-depth, semi-structured interviews with eight patients were conducted. The idea behind surveying patients', clinical staff and managers narratives/opinions, was to establish a holistic view of discharge planning and practice. Recommendations for change could then be made dependant on these findings, in order to improve the effectiveness and efficiency of this process, in addition to patients’ experiences. Following the completion of the focus groups and semi-structured interviews, analysis was undertaken to identify whether any common themes emerged across the three different groups. Although the narratives identified varying views, the most significant of the emerging themes was that of communication and the attitude of staff. Indeed, further analysis confirmed that communication was a prevalent issue in all three focus groups. The findings suggest patients have specific individual needs; this would not be surprising given the treatment and care received. Communication was also noted to be a major theme for patients; whilst clearly the majority of patients were relatively happy with the care received, they remained keen to make suggestions for improvement. The attitude of staff also featured strongly; patients were keen to suggest the changes they required and their rationale for these suggestions. The clinicians' themes mirrored the patients' themes, whilst also incorporating resource and innovation. The managers' themes commonly focused on service and proposed new ways of working, which included 'joined-up' working across acute and community services. Furthermore, the clinicians' and managers’ themes recognised that the attitudes of all groups of staff remained a challenge. In conclusion, this study has demonstrated the requirement for and the importance of effective communication throughout the entire discharge process. The aims are as follows: to ensure that post-procedure patients are discharged home or into a community setting safely; that they receive the right care, treatment and information in the most appropriate setting; and that they find and use ways to facilitate their recovery. Furthermore, this study has validated the importance of empowering patients to live with their diagnosed condition/disease by enabling the provision of support post-discharge. To this end, there is a need to ensure that positive values (such as empathy and compassion) remain a core part of the discharge planning process. This can be achieved by recognising each patient as an individual who is trying to make sense of a daunting life-changing experience whilst maintaining their own core belief and value systems. As a result, recommendations for improvements and change within this study have been based upon these findings. Further research needs to be defined to ensure that discharging patients from an acute hospital to their home or community setting remains both safe and effective, whilst also being informative and empowering.

Approximately 41,000 young people, under the age of 18, are bereaved of a parent each year within the U.K. (Child Bereavement Network, 2014; Winston’s Wish, 2016), and a significant number of these deaths will have been preceded by a period of terminal illness (TI). Adolescents are considered to be particularly susceptible to negative psychosocial outcomes when a parent is unwell (Grabiak, Bender & Puskar, 2007), yet adolescents as a group are less likely to access mental-health services (Reardon, Harvey, Baranowska, O’Brien, Smith & Creswell, 2017). Thus, whilst adolescents of terminally ill (TI) parents may present a vulnerable group, they may also be difficult to engage and subsequently support (Bremner, 2000). There has also been little consideration of how adolescents experience parental terminal illness (PTI), particularly considering their perceptions and experiences of support during this period. This study qualitatively explores the experiences of six individuals who had a parent diagnosed with a TI during late adolescence. Their experiences are explored in terms of their understanding, processing and adjustment to their parent’s TI, both as an individual, and in the context of their wider family and social setting. Their experiences and perceptions of support are additionally explored; considering the role of supportive individuals, services and interventions. Interpretative Phenomenological Analysis was employed to analyse participant data. Participants were individuals who had a parent diagnosed with a terminal illness aged 16-18. Four superordinate themes emerged from the data: 1) ‘Changing family dynamics’ 2) ‘grappling with adolescence and adjustment to loss’ 3) ‘Barriers to feeling/being supported’ 4) ‘Living with the consequences’. Participants relate the profound impact that PTI during adolescence has had and continues to have on their lives. Whilst there are prominent threads of difficulty and distress, elements of positive change and growth are also highlighted. Implications are discussed in terms of service provision and design, including therapeutic recommendations for counselling psychologists and other professionals working with this group.

7,8-dihydroxyflavone (7,8-DHF), which is a member of the flavonoid family, is a selective tyrosine kinase receptor B (TrkB) agonist that has neurotrophic effects in various neurological diseases such as ischemic stroke and Parkinson’s disease [3]. In this study, we assessed the angiogeneic effect of 7,8-DHF in endothelial cells derived from resistance vessel of the brain. Angiogenesis by 7,8-DHF is an important factor that helps prevent and treat various ischemic diseases. In this study, we found that rat RV cells used in the experiment possess the TrkB receptor. Our data also demonstrates that 7,8-DHF is able to stimulate cell proliferation in RV cells, suggesting that 7,8-DHF is capable of inducing angiogenesis. The 7,8-DHF activates the TrkB receptor which then leads to cell proliferation. In our study we also showed the effects of 7,8-DHF in the presence of the TrkB inhibitor cyclotraxin-B. Addition of cyclotraxin-B blocked the TrkB receptor and counteracted the effects of 7,8-DHF. Cell proliferation occurs in RV cells with the addition of 7,8-DHF, but this proliferation is inhibited by cyclotraxin-B.