The cyclotides: Novel macrocyclic peptides as scaffolds in drug design

Cyclotides are a novel class of circular, disulfide-rich peptides (similar to 30 amino acids) that display a broad range of bioactivities and have exceptionally high stability. Their physical properties, which include resistance to thermal and enzymatic degradation, can be attributed to their unique cyclic backbone and knotted arrangement of disulfide bonds. The applicability of linear peptides as drugs is potentially limited by their susceptibility to proteolytic cleavage and poor bioavailability. Such limitations may be overcome by using the cyclotide framework as a scaffold onto which new activities may be engineered. The potential use of cyclotides for drug design is evaluated here, with reference to rapidly increasing knowledge of natural cyclotides and the emergence of new techniques in peptide engineering.

Structure and function of plant toxins (with emphasis on cystine knot toxins)

Plant toxins are substances produced and secreted by plants to defend themselves against predators. In a broad sense, this includes all substances that have a toxic effect on targeted organisms, whether they are microbes, other plants, insects, or higher animals. Plant toxins have a diverse range of structures, from small organic molecules through to proteins. This review gives an overview of the various classes of plant toxins but focuses on an interesting class of protein-based plant toxins containing a cystine knot motif. This structural motif confers exceptional stability on proteins containing it and is associated with a wide range of biological activities. The biological activities and structural stability offer many potential applications in the pharmaceutical and agricultural fields. One particularly exciting prospect is in the use of protein-based plant toxins as molecular scaffolds for displaying pharmaceutically important bioactivities. Future applications of plant toxins are likely to involve genetic engineering techniques and molecular pharming approaches.

The impact of participation in activities while on vacation on senior's psychological well-being: A path model application

This article extends earlier research regarding the relationships between senior travelers’ participation in activities while on vacation, their overall satisfaction with their travel experiences, and their psychological well-being. A path model was developed to depict the direct and indirect relationships between these variables by using a sample of senior tourists traveling on North American escorted tour itineraries. Whereas the direct effect accounted for 98% of the relationships between the senior tourists’ levels of participation in activities and their psychological well-being, the indirect effect was negligible. The study concluded that senior tourists’activity levels were significantly related to their psychological well-being, but their satisfaction with the tours was not.

Enhanced quantitative resistance to Leptosphaeria maculans conferred by expression of a novel antimicrobial peptide in canola (Brassica napus L.)

The novel antimicrobial peptide MiAMP1, originally isolated from the seeds of Macadamia integrifolia, was constitutively expressed in transgenic tobacco and canola plants to test its effect on disease resistance. Analysis of plants transformed with 35S-MiAMP1 construct by northern and western blot analyses demonstrated the presence of MiAMP1 mRNA and the mature peptide in the transgenic plants. The MiAMP1 purified from the leaves of transgenic plants was biologically active with the same in vitro antifungal activity as native MiAMP1 purified from the seeds of macadamia. The effect of MiAMP1 expression on the economically important canola pathogen Leptosphaeria maculans (causal agent of blackleg disease) was evaluated in comparison with an untransformed control line and an azygous segregant derived from one of the transgenic lines. Lesion development on the cotyledons of the inoculated canola seedlings was significantly reduced in the T-2 progeny of seven independently transformed transgenic lines. These results suggested that, transgenic canola expressing MiAMP1 may be useful for the management of blackleg disease.

The structural and functional diversity of naturally occurring antimicrobial peptides

Antimicrobial peptides occur in a diverse range of organisms from microorganisms to insects, plants and animals. Although they all have the common function of inhibiting or killing invading microorganisms they achieve this function using an extremely diverse range of structural motifs. Their sizes range from approximately 10-90 amino acids. Most carry an overall positive charge, reflecting a preferred mode of electrostatic interaction with negatively charged microbial membranes. This article describes the structural diversity of a representative set of antimicrobial peptides divided into five structural classes: those with agr-helical structure, those with bgr-sheet structure, those with mixed helical / bgr- sheet structure, those with irregular structure, and those incorporating a macrocyclic structure. There is a significant diversity in both the size and charge of molecules within each of these classes and between the classes. The common feature of their three-dimensional structures is, however, that they have a degree of amphipathic character in which there is separate localisation of hydrophobic regions and positively charged regions. An emerging trend amongst antimicrobial proteins is the discovery of more macrocyclic analogues. Cyclisation appears to impart an additional degree of stability on these molecules and minimizes proteolytic cleavage. In conclusion, there appear to be a number of promising opportunities for the development of novel clinically useful antimicrobial peptides based on knowledge of the structures of naturally occurring antimicrobial molecules.

Further evidence for the agreement between patients with stroke and their proxies on the Frenchay Activities Index

Objective: To determine item, subscale and total score agreement on the Frenchay Activities Index (FAI) between stroke patients and proxies six months after discharge from rehabilitation. Design: Prospective study design. Setting/subjects: Fifty patient-proxy pairs, interviewed separately, in the patient's residence. Main outcome measures: Modified FAI using 13 items. Individual FAI items, subscales and total score agreement as measured by weighted kappa and intraclass correlation coefficients (ICC). Results: Excellent agreement was found for the total FAI (ICC 0.87, 95% confidence interval (CI) 0.78-0.93), and domestic (ICC 0.85, 95% CI 0.73-0.91) and outdoor (ICC 0.87, 95% CI 0.78-0.95) subscales, with moderate agreement found for the work/leisure subscale (ICC 0.63, 95% CI 0.34-0.78). For the individual FAI items, good, moderate, fair and poor agreement was found for five, three, four and one item, respectively. The best agreement was for objective items of preparing meals, washing-up, washing clothes, shopping and driving. The poorest agreement was for participation in hobbies, social outings and heavy housework. Scoring biases associated with patient or proxy demographic characteristics were found. Female proxies, and those who were spouses, scored patients lower on domestic activities; male patients, and those who were younger, scored themselves higher on outdoor activities and higher patient FIM scores were positively correlated with higher FAI scores. Conclusions: While total and subscale agreement on the FAI was high, individual item agreement varied. Proxy scores should be used with caution due to bias.

Identifying the communication activities of older people with aphasia: Evidence from naturalistic observation

Background: Increasingly there is a call from clinicians and researchers for measures that document the impact of aphasia on a person's everyday communication. Do existing assessments of communication disability adequately sample communication activities relevant to our clients? Communication skills and networks change with age. A need exists to determine the everyday communication activities of older people and in particular those with aphasia. Aims: The primary aim of this study was to describe and compare the everyday communication activities of older people with aphasia and healthy older people who are living in the community. A secondary aim was to investigate the content validity of the American Speech-Language Hearing Association Functional Assessment of Communication Skills for Adults (ASHA FACS, 1997) for older Australians. Methods & Procedures: Naturalistic observation was the method of choice for detailing the everyday communication of 15 older people with chronic aphasia following stroke and a matched group of 15 healthy older people who were living in the community. Researchers, in the role of participant observer, took field notes for 8 hours, over three occasions within a week. A total of 240 hours of observation have been coded in terms of communication activity, topic, communication partners, and place of communication. A brief 5-day diary served to check the representativeness of the observational data. After each hour of observation, the researcher checked which ASHA FACS items had been observed. Outcomes & Results: Naturalistic observation provided a rich, rigorous, and systematic methodology for detailing the dynamics and complexities of authentic communication. The most common communication activities for both groups were conversations at home and in social groups. Real-life communication was revealed to serve the dual purposes of transaction and interaction. Results indicate that older people with aphasia engage in similar communication activities to healthy older people although differences were evident in the frequency of communication and in specific activities such as story telling, writing, commenting, and acknowledging. ASHA FACS items were generally relevant to older Australians living in the community. Conclusions: This study demonstrated that communication activity is multifaceted in terms of the type of communication and contextual factors. The observational data describe the effects of aphasia on a person's everyday communication activity and reveal the impact of aphasia on the social functions of communication including sharing information, maintaining and establishing relationships, and telling one's story. Functional communication assessment requires a greater focus on the interactional and uniquely interpersonal aspects of social communication.

The developmental activities of elite ice hockey players

The purpose of this study was to analyze the development of four 20 year-old elite hockey players through an in-depth examination of their sporting activities. The theoretical framework of deliberate practice (Ericsson, Krampe, & Tesch-Römer, 1993) and the notion of deliberate play (Côté, 1999) served as the theoretical foundations. Interviews were conducted to provide a longitudinal and detailed account of each participant's involvement in various sporting activities. The interviewer asked questions about the conditions and sporting activities for each year of development. The data obtained were validated through independent interviews conducted with three parents of three different athletes. The results were consistent with Côté's (1999) three stages of development in sport: the sampling (age 6-12), specializing (age 13-15), and investment (age 16+) years.

Decreased coumarin 7-hydroxylase activities and CYP2A6 expression levels in humans caused by genetic polymorphism in CYP2A6 promoter region (CYP2A6*9)

One of seven poor metabolizers of coumarin found in Thai subjects was previously genotyped as heterozygote for the CYP2A6*4 (whole deletion) and CYP2A6*9. Thus, we aimed to investigate the relationship between the genetic polymorphism in the TATA box of the CYP2A6 gene (CYP2A6*9), expression levels of CYP2A6 mRNA and coumarin 7-hydroxylase activities in human livers. Levels of CYP2A6 mRNA were quantified by real-time quantitative reverse transcriptase-polymerase chain reaction. The mean expression levels of CYP2A6 mRNA in individuals with CYP2A6*1/*4, CYP2A6*1/*9 and CYP2A6*4/*9 were 58%, 71% and 21% of the individuals genotyped as CYP2A6*1/*1, respectively. The mean in-vitro coumarin 7-hydroxylase activities in subjects carrying CYP2A6*1/*4, CYP2A6*1/*9 and CYP2A6*4/*9 were 41%, 71% and 12%, respectively, compared to those of the subjects judged as wild-type. Vmax values for coumarin 7-hydroxylation in the liver microsomes from human subjects with genotypes of CYP2A6*1/*1, CYP2A6*1/*4, CYP2A6*1/*9 and CYP2A6*4/*9 were 0.58, 0.26, 0.44 and 0.13 nmol/min/nmol total P450, respectively. CYP2A6 protein levels in human liver microsomes with the CYP2A6*4 and the CYP2A6*9 alleles were markedly decreased. These results suggest that the genetic polymorphism in the promoter region of the CYP2A6 gene (CYP2A6*9) reduced the expression levels of CYP2A6 mRNA and protein in human livers, resulting in the decrease of coumarin 7-hydroxylase activities. Individuals judged as CYP2A6*4/*9 were expected to be poor metabolizers, having extremely low activity of CYP2A6.