AAV6 mediated Gsx1 expression in neural stem progenitor cells promotes neurogenesis and restores locomotor function after contusion spinal cord injury.

Genomic screened homeobox 1 (Gsx1 or Gsh1) is a neurogenic transcription factor required for the generation of excitatory and inhibitory interneurons during spinal cord development. In the adult, lentivirus (LV) mediated Gsx1 expression promotes neural regeneration and functional locomotor recovery in a mouse model of lateral hemisection spinal cord injury (SCI). The LV delivery method is clinically unsafe due to insertional mutations to the host DNA. In addition, the most common clinical case of SCI is contusion/compression. In this study, we identify that adeno-associated virus serotype 6 (AAV6) preferentially infects neural stem/progenitor cells (NSPCs) in the injured spinal cord. Using a rat model of contusion SCI, we demonstrate that AAV6 mediated Gsx1 expression promotes neurogenesis, increases the number of neuroblasts/immature neurons, restores excitatory/inhibitory neuron balance and serotonergic neuronal activity through the lesion core, and promotes locomotor functional recovery. Our findings support that AAV6 preferentially targets NSPCs for gene delivery and confirmed Gsx1 efficacy in clinically relevant rat model of contusion SCI.

Hexahydroquinolines are antimalarial candidates with potent blood-stage and transmission-blocking activity.

Antimalarial compounds with dual therapeutic and transmission-blocking activity are desired as high-value partners for combination therapies. Here, we report the identification and characterization of hexahydroquinolines (HHQs) that show low nanomolar potency against both pathogenic and transmissible intra-erythrocytic forms of the malaria parasite Plasmodium falciparum. This activity translates into potent transmission-blocking potential, as shown by in vitro male gamete formation assays and reduced oocyst infection and prevalence in Anopheles mosquitoes. In vivo studies illustrated the ability of lead HHQs to suppress Plasmodium berghei blood-stage parasite proliferation. Resistance selection studies, confirmed by CRISPR-Cas9-based gene editing, identified the digestive vacuole membrane-spanning transporter PfMDR1 (P. falciparum multidrug resistance gene-1) as a determinant of parasite resistance to HHQs. Haemoglobin and haem fractionation assays suggest a mode of action that results in reduced haemozoin levels and might involve inhibition of host haemoglobin uptake into intra-erythrocytic parasites. Furthermore, parasites resistant to HHQs displayed increased susceptibility to several first-line antimalarial drugs, including lumefantrine, confirming that HHQs have a different mode of action to other antimalarials drugs for which PfMDR1 is known to confer resistance. This work evokes therapeutic strategies that combine opposing selective pressures on this parasite transporter as an approach to countering the emergence and transmission of multidrug-resistant P. falciparum malaria.

Exploring the use of tablets (iPads) with children and young adults with disabilities in Trinidad.

PURPOSE: This study was conducted to review data gathered during a pilot project which trialed the use of a tablet computer, the iPad. METHODS: Students from a segregated special education school and pre-vocational centre, with a wide range of intellectual and physical disabilities, were previously observed participating in 5-10-min introductory learning sessions with the iPad. This study reviewed quantitative and qualitative data collected during these sessions which included data regarding students' level of engagement and overall ability to learn how to operate the iPad and its applications. RESULTS: Results were positive for level of engagement and ease of use with cause and effect applications. For lower functioning students or students not previously exposed to tablet technology, scores were lower but overall remained high based on the 5-point scaling used in this study. CONCLUSION: Regular use of tablet technology in the classroom with applications appropriate to the level of ability of the student has the potential to enhance engagement in learning as well as maximise independence in the classroom. Implications for Rehabilitation The iPad has the capacity to be used with learners of all different ability levels if applications are selected appropriately and learners are given equal opportunity to access this type of technology. Enjoyment when using the iPad was high overall and this type of technology has the potential to promote more engagement in the learning process. Many applications are easy to use and progress students through step by step increasing the potential for independent learning in the classroom.

Profiling the Essential Nature of Lipid Metabolism in Asexual Blood and Gametocyte Stages of Plasmodium falciparum.

During its life cycle, Plasmodium falciparum undergoes rapid proliferation fueled by de novo synthesis and acquisition of host cell lipids. Consistent with this essential role, Plasmodium lipid synthesis enzymes are emerging as potential drug targets. To explore their broader potential for therapeutic interventions, we assayed the global lipid landscape during P. falciparum sexual and asexual blood stage (ABS) development. Using liquid chromatography-mass spectrometry, we analyzed 304 lipids constituting 24 classes in ABS parasites, infected red blood cell (RBC)-derived microvesicles, gametocytes, and uninfected RBCs. Ten lipid classes were previously uncharacterized in P. falciparum, and 70%-75% of the lipid classes exhibited changes in abundance during ABS and gametocyte development. Utilizing compounds that target lipid metabolism, we affirmed the essentiality of major classes, including triacylglycerols. These studies highlight the interplay between host and parasite lipid metabolism and provide a comprehensive analysis of P. falciparum lipids with candidate pathways for drug discovery efforts.

ATP-binding cassette transporters and sterol O-acyltransferases interact at membrane microdomains to modulate sterol uptake and esterification.

A key component of eukaryotic lipid homeostasis is the esterification of sterols with fatty acids by sterol O-acyltransferases (SOATs). The esterification reactions are allosterically activated by their sterol substrates, the majority of which accumulate at the plasma membrane. We demonstrate that in yeast, sterol transport from the plasma membrane to the site of esterification is associated with the physical interaction of the major SOAT, acyl-coenzyme A:cholesterol acyltransferase (ACAT)-related enzyme (Are)2p, with 2 plasma membrane ATP-binding cassette (ABC) transporters: Aus1p and Pdr11p. Are2p, Aus1p, and Pdr11p, unlike the minor acyltransferase, Are1p, colocalize to sterol and sphingolipid-enriched, detergent-resistant microdomains (DRMs). Deletion of either ABC transporter results in Are2p relocalization to detergent-soluble membrane domains and a significant decrease (53-36%) in esterification of exogenous sterol. Similarly, in murine tissues, the SOAT1/Acat1 enzyme and activity localize to DRMs. This subcellular localization is diminished upon deletion of murine ABC transporters, such as Abcg1, which itself is DRM associated. We propose that the close proximity of sterol esterification and transport proteins to each other combined with their residence in lipid-enriched membrane microdomains facilitates rapid, high-capacity sterol transport and esterification, obviating any requirement for soluble intermediary proteins.

A Prospective Randomized Study to Compare the Effectiveness of Honey Dressing vs. Povidone Iodine Dressing in Chronic Wound Healing.

To compare the healing of chronic wounds with honey dressing vs. Povidone iodine dressing in adult subjects with chronic wounds of ≥6 weeks of duration, attending wound care clinic in Surgical Out Patient Department of All India Institute of Medical Sciences, Surgical Out Patient Department of Jai Prakash Narayan Apex Trauma center, New Delhi. Forty five subjects were randomized into two groups i.e., Honey & Povidone iodine dressing group. Dressing was done on alternate day basis for 6 weeks of followup period. Main outcome measure was complete healing at 6 weeks. Wound healing status was assessed at 2 weekly intervals till 6 weeks. Seven out of 22 subjects in honey treated group achieved complete healing as compared to none out of 20 subjects in Povidone iodine treated group. There was a significant decrease in the wound surface area, pain score & increase in comfort score in Honey dressing group in comparison to the Povidone Iodine group at 0.05 level of significance. Honey dressing is highly effective in achieving healing in chronic wounds as compared to Povidone iodine dressing.

A functional, genome-wide evaluation of liposensitive yeast identifies the "ARE2 required for viability" (ARV1) gene product as a major component of eukaryotic fatty acid resistance.

The toxic subcellular accumulation of lipids predisposes several human metabolic syndromes, including obesity, type 2 diabetes, and some forms of neurodegeneration. To identify pathways that prevent lipid-induced cell death, we performed a genome-wide fatty acid sensitivity screen in Saccharomyces cerevisiae. We identified 167 yeast mutants as sensitive to 0.5 mm palmitoleate, 45% of which define pathways that were conserved in humans. 63 lesions also impacted the status of the lipid droplet; however, this was not correlated to the degree of fatty acid sensitivity. The most liposensitive yeast strain arose due to deletion of the "ARE2 required for viability" (ARV1) gene, encoding an evolutionarily conserved, potential lipid transporter that localizes to the endoplasmic reticulum membrane. Down-regulation of mammalian ARV1 in MIN6 pancreatic ß-cells or HEK293 cells resulted in decreased neutral lipid synthesis, increased fatty acid sensitivity, and lipoapoptosis. Conversely, elevated expression of human ARV1 in HEK293 cells or mouse liver significantly increased triglyceride mass and lipid droplet number. The ARV1-induced hepatic triglyceride accumulation was accompanied by up-regulation of DGAT1, a triglyceride synthesis gene, and the fatty acid transporter, CD36. Furthermore, ARV1 was identified as a transcriptional of the protein peroxisome proliferator-activated receptor α (PPARα), a key regulator of lipid homeostasis whose transcriptional targets include DGAT1 and CD36. These results implicate ARV1 as a protective factor in lipotoxic diseases due to modulation of fatty acid metabolism. In conclusion, a lipotoxicity-based genetic screen in a model microorganism has identified 75 human genes that may play key roles in neutral lipid metabolism and disease.

A key role for lipoic acid synthesis during Plasmodium liver stage development.

The successful navigation of malaria parasites through their life cycle, which alternates between vertebrate hosts and mosquito vectors, requires a complex interplay of metabolite synthesis and salvage pathways. Using the rodent parasite Plasmodium berghei, we have explored the synthesis and scavenging pathways for lipoic acid, a short-chain fatty acid derivative that regulates the activity of α-ketoacid dehydrogenases including pyruvate dehydrogenase. In Plasmodium, lipoic acid is either synthesized de novo in the apicoplast or is scavenged from the host into the mitochondrion. Our data show that sporozoites lacking the apicoplast lipoic acid protein ligase LipB are markedly attenuated in their infectivity for mice, and in vitro studies document a very late liver stage arrest shortly before the final phase of intra-hepaticparasite maturation. LipB-deficient asexual blood stage parasites show unimpaired rates of growth in normal in vitro or in vivo conditions. However, these parasites showed reduced growth in lipid-restricted conditions induced by treatment with the lipoic acid analogue 8-bromo-octanoate or with the lipid-reducing agent clofibrate. This finding has implications for understanding Plasmodium pathogenesis in malnourished children that bear the brunt of malarial disease. This study also highlights the potential of exploiting lipid metabolism pathways for the design of genetically attenuated sporozoite vaccines.

Demands and rewards associated with working in pediatric oncology: a qualitative study of Canadian health care providers.

OBJECTIVE: Despite recent advances in the outcome of children with cancer, the demands on medical professionals caring for these patients can be intense. Our qualitative study explored the work-related demands and rewards experienced by Canadian pediatric oncology staff. STUDY DESIGN: Interviews were conducted with 33 staff members (10 oncologists, 3 subspecialty residents, 9 nurses, 5 social workers, and 6 child life specialists) from 4 hospitals. Participants were asked to describe work-related rewards and demands. Interviews were recorded and transcribed verbatim. Interview transcripts were analyzed to identify all sources of demands and rewards. RESULTS: Pediatric oncology staff described work-related rewards and demands related to the following areas: (1) working with children; (2) working with families; (3) working within a multidisciplinary health care team; (4) working in a pediatric oncology unit; and (5) working within a hospital or academic health center. Overall, health care providers described their job as fulfilling and meaningful. For most health care providers, many work-related issues were described as both rewarding and demanding. CONCLUSIONS: Our study identifies important demands and rewards associated with working in pediatric oncology. Future research could explore the relationship between work-related stress and job satisfaction and how these factors either cause or prevent burnout syndrome.

Health care providers' perspectives about working with parents of children with cancer: a qualitative study.

Research to understand the sources of stress and job satisfaction in pediatric oncology staff is limited. The aim of the authors' qualitative study was to describe the key work-related demands and rewards in relation to working closely with parents from the perspective of health care providers (HCPs). Semistructured interviews were conducted with 13 doctors, 9 nurses, 5 social workers, and 6 child life specialists. Line-by-line, focused, and theoretical coding was used to establish categories and themes. Constant comparisons were used to examine the relationships within and across codes and categories. Interviewing continued until no new themes emerged. HCPs found it rewarding when they established close or long-term relationships with parents and to helped families through the entire cancer journey, including palliative care. HCPs found it challenging to work with complex families; with parents who are demanding, rude, or angry; with parents who have differing views about the treatments and palliative care; and with having to relay bad news to parents. Future research could explore the relationship between the factors that the authors have identified and burnout syndrome.

Understanding the health impact of caregiving: a qualitative study of immigrant parents and single parents of children with cancer.

PURPOSE: Research looking at the health of parents of children with cancer typically uses outcome measures focused on symptoms of anxiety, depression, or post-traumatic stress. Our team builds on this literature to provide a more comprehensive understanding of the health impact of caregiving. METHODS: Interviews were conducted with 79 Canadian parents of children with cancer at least 6 months post-diagnosis. Line-by-line coding was used to establish categories and themes. Constant comparison was used to examine relationships within and across codes and categories. Interviewing continued until no new themes emerged. RESULTS: Parents described health concerns as including sleep disturbance, daytime fatigue, anxiety, depression, social isolation, and changes in social roles. Parents described the positive impacts of caregiving as including gaining a greater appreciation for child and family and developing compassion, empathy, patience, inner strength, and new perspectives on life. CONCLUSION: Parents of children with cancer can experience a range of health problems due to the emotional impact of a cancer diagnosis and the intensive and often prolonged nature of treatment and aftercare. Given the central role parents play as caregivers, it is crucial to understand the health impact of caregiving so that supportive interventions can be implemented as necessary.

Communication and language challenges experienced by Chinese and South Asian immigrant parents of children with cancer in Canada: implications for health services delivery.

BACKGROUND: Language is an important aspect of health literacy and plays a vital role in families' ability to access and use health information and resources. Our study explored the role of communication and language in the healthcare experiences of immigrant parents of children with cancer living in Canada. PROCEDURE: We used a grounded theory approach. Chinese and South Asian parents of children 6 months post-diagnosis were recruited from six Canadian pediatric oncology centers. Semi-structured interviews were conducted in Cantonese, Mandarin, Hindi, Punjabi, Urdu, or English. Questions relevant to communication included: how parents navigated the healthcare system; nature of interpreter services and translated materials; and suggestions about how to improve services. Analysis involved line-by-line, focused and theoretical coding, and constant comparison. RESULTS: Thirty-one (62%) parents reported no difficulty communicating with healthcare providers in English, while 13 (26%) parents struggled with English, and six (12%) parents could not communicate in English. Communication challenges influenced parents' role in caring for their child and made it difficult to learn complex medical terminology. Interpreting services were sometimes inadequate or not accessible. Parents occasionally missed out on services and resources, reported limited availability of linguistically and culturally appropriate information, and experienced a lack of social integration in the healthcare process. CONCLUSION: Language ability played an essential role in parents' healthcare experiences for pragmatic and social purposes. Language challenges can heighten systemic and socio-cultural barriers to accessing health information and resources. The provision of enhanced culturally and linguistically sensitive services may support immigrant families in their caregiving role.

Perceptions about complementary and alternative medicine use among Chinese immigrant parents of children with cancer.

PURPOSE: Research indicates complementary and alternative medicine (CAM) use among children with cancer is common and widespread. CAM use, particularly traditional Chinese medicine (TCM), is prevalent in the country of origin of Chinese immigrant families, yet little is known about its use after immigrating to Canada. This paper describes 25 Chinese immigrant parents' perception about the use of CAM in their child with cancer in Canada. METHODS: This qualitative grounded theory study is part of a larger study of the caregiving experiences of first generation Chinese and South Asian parents of children with cancer. Chinese parents of children at least 6 months post-diagnosis were recruited from four Canadian pediatric oncology centers. Interviews were conducted in English, Cantonese, or Mandarin and transcribed into English. Analysis involved coding and the use of the constant comparison method to identify important themes. RESULTS: In discussing their caregiving experiences, CAM emerged as an important theme. The following sub-themes were identified: (1) trust in conventional medicine to cure cancer; (2) use of dietary modifications and restrictions; (3) extent of communication with healthcare providers about TCM use; and (4) limited availability of culturally relevant information. CONCLUSIONS: While Chinese immigrant parents in this study placed their trust in conventional medicine to treat their child, the use of food as therapy is part of their daily cultural practice. To ensure safe cancer treatment and more culturally sensitive care, it is essential for health providers to offer reliable information and encourage open discussions about CAM use with Chinese immigrant parents.

Immigrant to Canada, newcomer to childhood cancer: a qualitative study of challenges faced by immigrant parents.

BACKGROUND: Given the increasing numbers of immigrant families in Canada, it is imperative that healthcare providers (HCPs) understand the caregiving experiences of immigrant family caregivers. Our study aimed to explore any special challenges faced by immigrant parents of children with cancer and to identify supportive factors. METHODS: A constructivist grounded theory approach was used. Participants included 50 first generation Chinese and South Asian parents of children with cancer who were at least six months post-diagnosis. Recruitment took place at six Canadian pediatric oncology centres. Interviews were conducted in English, Cantonese, Mandarin, Urdu, Punjabi or Hindi. Analysis involved coding and the use of the constant comparison method. Interviewing continued until no new themes emerged. RESULTS: While immigrant parents described many challenges faced by any parent of a child with cancer, the context of being an immigrant made certain experiences particularly challenging. Parents described challenges in the following areas: managing caregiving demand and financial strain, accessing support from others, and interfacing with the healthcare system. Parents described receiving a range of practical, emotional, social and informational support from extended family, their workplace, other cancer families, community organizations and HCPs. CONCLUSIONS: Our study addresses an important gap in the research literature by providing practical insight into the experiences of immigrant family caregivers. Our findings may help to inform the development of pediatric oncology policies and programs in ways that respond to the unique needs and challenges of culturally and linguistically diverse families.

Cultural beliefs and coping strategies related to childhood cancer: the perceptions of South Asian immigrant parents in Canada.

The purpose of this study is to describe cultural beliefs and coping strategies related to dealing with childhood cancer identified through a qualitative study of the caregiving experiences of first-generation South Asian immigrant parents of children with cancer. A constructivist grounded theory approach was employed. Families with a child at least 6 months postdiagnosis were recruited from 5 Canadian pediatric oncology centers. In-depth semistructured interviews were conducted in English, Hindi, Punjabi, or Urdu with a sample of 25 South Asian parents. Analysis of interviews involved line-by-line coding and using the constant comparison method. The following 2 central themes related to culture and coping emerged: (a) cultural beliefs about childhood cancer being incurable, rare, unspeakable, and understood through religion and (b) parental coping strategies included gaining information about the child's cancer, practicing religious rituals and prayers, trusting the health care professionals, and obtaining mutual support from other South Asian parents. These cultural beliefs and coping strategies have important implications for health care providers to understand the variations in the perceptions of childhood cancer and coping in order to implement culturally sensitive health care services.

Adolescent group empowerment: Group-centred occupations to empower adolescents with disabilities in the urban slums of North India.

This study aimed to understand how adolescents with disabilities can assume greater control over their rehabilitation and participation within a community-based rehabilitation programme in the urban slums of North India. A critical ethnographical approach using multiple qualitative and participatory data collection methods was adopted. Fieldwork was conducted from January to May 2005 and October 2006 to March 2007 with 21 adolescents with and 11 adolescents without disabilities (aged 12 to 18 years), and 10 community-based rehabilitation staff members. A conceptual framework called the 'Adolescent Group Empowerment Pyramid' was developed. Four themes informed the framework: group participation, group demonstration, group recognition and the socio-cultural environment's interaction with disability. Group empowerment, achieved through group-centred occupations, encourages adolescents to work together to address their rehabilitation challenges and agendas. Three external support factors and 10 areas for nurturing the group empowerment process also emerged. A limitation of this study is the lack of data on how the familial and local political and economic environment impact adolescents' ability to make decisions about their rehabilitation. Further research might explore group occupations and occupational recognition, and what this means for social change and the personal and collective growth of adolescents in underserviced communities.

Sterols and sphingolipids: dynamic duo or partners in crime?

One manner in which eukaryotic cells respond to their environments is by optimizing the composition and proportions of sterols and sphingolipids in membranes. The physical association of the planar ring of sterols with the acyl chains of phospholipids, particularly sphingolipids, produces membrane micro-heterogeneity that is exploited to coordinate several crucial pathways. We hypothesize that these lipid molecules play an integrated role in human disease; when one of the partners is mis-regulated, pathology frequently ensues. Sterols and sphingolipid levels are not coordinated by the action of a single master regulator, however the cross-talk between their metabolic pathways is considerable. We describe our perspectives on the key components of synthesis, catabolism and transport of these lipid partners with an emphasis on evolutionarily conserved reactions that produce disease states when defective.