SAT-158 Offspring Exposed to Maternal High Fat Diet Exhibits Systemic Inflammation and Pancreatic Islet Dysfunction

Offspring born to overweight mothers are more likely to develop dysregulated immune response1, obesity1 and pancreatic islet dysfunction2. These offspring have increased inflammation at birth3 and at least until childhood4. We hypothesize that heightened inflammation in offspring of overweight mothers increases offspring risks of pancreatic islet dysfunction. We induced maternal overweight by providing 45% high fat diet (HFD) to female mice 2 - 4 weeks before pregnancy until weaning. When compared to controls, P21 weanlings of HFD mothers had impaired glucose tolerance in dose and gender dependent manner [GTT AUC: male 2-week HFD* 30 ± 6% higher; male 4-week HFD* 37± 3% higher: 9-11/group; female 2-week HFD 13 ± 5% higher; female 4-week HFD* 22 ± 3% higher: 3-9/group, *p<0.05 compared to controls]. Glucose intolerance persisted in 8-week-old male from 2-week HFD mothers (p<0.05, n=6-9/group), with decreased pancreatic islets glucose induced calcium response measured using Fura-2AM calcium imaging (F1/F0 Con:2.00 ± 0.06, HFD2W: 1.69±0.12*, HFD4w: 0.71±0.09*, n =3/group). Cytokines production in the serum, macrophage response and metabolic phenotypes of offspring were assessed on postnatal day 21 (P21) and at 8 weeks old. Compared to control pups, weanling of HFD mothers had elevated serum/plasma IL-1b level along with increased polarization of M1 macrophages and decreased M2 macrophages, as well as an increase of IL-1b secretion in LPS-stimulated macrophages. At 8 weeks of age, HFD male offspring had increased activation markers of splenic dendritic cells indicating a development of systemic inflammatory response early in life. Taken together, our findings suggest that mice offspring from HFD mothers have pancreatic dysfunction, and an inflammatory response. This work is funded by the Riley Children’s Foundation. 1. Kelishadi, R., Roufarshbaf, M., Soheili, S., Payghambarzadeh, F. & Masjedi, M. Association of Childhood Obesity and the Immune System: A Systematic Review of Reviews. Child. Obes. Print 13, 332-346 (2017). 2. Graus-Nunes, F. et al. Pregestational maternal obesity impairs endocrine pancreas in male F1 and F2 progeny. Nutrition 31, 380-387 (2015). 3. Dosch, N. C. et al. Maternal Obesity Affects Inflammatory and Iron Indices in Umbilical Cord Blood. J. Pediatr. 172, 20-28 (2016). 4. Leibowitz, K. L. et al. Maternal obesity associated with inflammation in their children. World J. Pediatr. WJP 8, 76-79 (2012)

Convenience or nuisance?: The ‘WhatsApp’ dilemma

WhatsApp sends real-time messages and is one of the world's most popular communication applications in the 21st century.While this study extends the current knowledge on the use of WhatsApp, it also highlights the challenges of WhatsApp use by young people. The purpose of this study is to examine the domestication of WhatsApp among young people, specifically the undergraduates at Universiti Brunei Darussalam. Results showed how young people perceive WhatsApp as a ‘convenient’ communication application in their everyday lives. Some of the critical issues arising from the use of WhatsApp included distractions and exposure to unregulated messages or information

Barriers and facilitators to point-of-care ultrasound use in rural Australia

This study explores the barriers and facilitators to point-of-care ultrasound (POCUS) use and adoption in rural healthcare since POCUS is a useful resource for rural clinicians to overcome the challenges associated with limited on-site clinical support, such as limited diagnostic imaging services and infrastructure. A qualitative descriptive study was employed, interviews with ten rural clinicians were conducted, and the data were analysed using the Walt and Gilson health policy framework to guide interpretation. Barriers include a lack of standardised training requirements, the cost of the devices and challenges recouping the costs of purchase and training, difficulty with the maintenance of skills, and a lack of an effective method to achieve quality assurance. Coupling POCUS with telemedicine could address the issues of the maintenance of skills and quality assurance to facilitate increased POCUS use, leading to positive patient safety and social and economic implications

A case for mandatory ultrasound training for rural general practitioners : a commentary

Context: Point-of-care ultrasound is a rapidly evolving technology that enables rapid diagnostic imaging to be performed at a patient’s bedside, reducing time to diagnosis and minimising the need for patient transfers. This has significant applications for rural emergency and general practice, and could potentially prevent unnecessary transfers of patients from rural communities to more urban centres for the purpose of diagnostic imaging, reducing costs and preventing disruption to patients’ lives. Meta-analyses on point-of-care ultrasound have reported extremely high sensitivity and specificity when detecting lung pathology, and the potential applications of the technology are substantial. A significant application of the technology is in the care of rural paediatric patients, where acute lower respiratory pathology is the most common cause of preventable deaths, hospitalisations, and emergency medical retrievals from remote communities for children under five. Issues: Although widely available, point-of-care ultrasound technology is not widely utilised in Australian emergency departments and general practices. Issues with comprehensive training, maintenance of skills, upskilling and quality assurance programs prevent physicians from feeling confident when utilising the technology. In Canada, point-of-care ultrasound training is part of the core competency training in the Royal College of Physicians of Canada emergency medicine fellowship program. Point-of-care ultrasound is widely used in rural practice, although lack of training, funding, maintenance of skills and quality assurance were still listed as barriers to use. Lessons learned: Point-of-care ultrasound is a highly sensitive and specific technology with wide potential applications. Issues with quality control and maintenance of skills are preventing widespread use. Coupling point-of-care ultrasound with telemedicine could help increase the usability and accessibility of the technology by reducing the issues associated with maintenance of skills and quality assurance

The effect of consequent exposure of stress and dermal application of low doses of chlorpyrifos on the expression of glial fibrillary acidic protein in the hippocampus of adult mice

<p>Abstract</p> <p>Background</p> <p>Chlorpyrifos (CPF), a commonly used pesticide worldwide, has been reported to produce neurobehavioural changes. Dermal exposure to CPF is common in industries and agriculture. This study estimates changes in glial fibrillary acidic protein (GFAP) expression in hippocampal regions and correlates with histomorphometry of neurons and serum cholinesterase levels following dermal exposure to low doses of CPF with or without swim stress.</p> <p>Methods</p> <p>Male albino mice were separated into control, stress control and four treatment groups (n = 6). CPF was applied dermally over the tails under occlusive bandage (6 hours/day) at doses of 1/10th (CPF 0.1) and 1/5th dermal LD₅₀(CPF 0.2) for seven days. Consequent treatment of swim stress followed by CPF was also applied. Serum cholinesterase levels were estimated using spectroflurometric methods. Paraffin sections of the left hippocampal regions were stained with 0.2% thionin followed by the counting of neuronal density. Right hippocampal sections were treated with Dako Envision GFAP antibodies.</p> <p>Results</p> <p>CPF application in 1/10th LD₅₀did not produce significant changes in serum cholinesterase levels and neuronal density, but increased GFAP expression significantly (p < 0.001). Swim stress with CPF 0.1 group did not show increase in astrocytic density compared to CPF 0.1 alone but decreased neuronal density.</p> <p>Conclusions</p> <p>Findings suggest GFAP expression is upregulated with dermal exposure to low dose of CPF. Stress combined with sub-toxic dermal CPF exposure can produce neurotoxicity.</p

BIODIVERSITY AND HABITAT PREFERENCES OF LIVING SHARKS IN THE SOUTHEAST ASIAN REGION

This paper reviews the biodiversity and habitat preferences of living sharks in the Southeast Asian region accumulated from published literatures including journals, books, proceedings, unpublished technical papers, and technical reports as well as authors’ knowledge and experiences working in this field. A total of 196 species of sharks from nine orders and 30 families have been recorded inhabiting from fresh water to deep ocean in this region. Indonesia recorded the highest diversity with 114 species from 27 families followed by the Philippines with 96 species (27 families), Thailand 76 species (21 families), Vietnam 70 species (23 families), Malaysia 68 species (19 families), Myanmar 64 species (19 families), Brunei Darussalam 45 species (15 families), and Cambodia with 26 species from 10 families. Many species still need to be confirmed and are most probably new species. In term of habitat preferences, 83 species of sharks (42.3%) inhabit continental/insular shelves (SHL) followed by continental/insular slopes (SLO) with 48 species (24.5%), shelf to slope (SHS) with 26 species (13.3%), and shelf to oceanic (SHO) with 16 species (8.2%). Only nine species (4.6%) inhabit oceanic and six species (4%) obligate freshwater and euryhaline freshwater/shelves.

Neutralizing human monoclonal antibody against H5N1 influenza HA selected from a Fab-phage display library

Identification of neutralizing antibodies with specificity away from the traditional mutation prone antigenic regions, against the conserved regions of hemagglutinin from H5N1 influenza virus has the potential to provide a therapeutic option which can be developed ahead of time in preparation for a possible pandemic due to H5N1 viruses. In this study, we used a combination of panning strategies against the hemagglutinin (HA) of several antigenic distinct H5N1 isolates to bias selection of Fab-phage from a naïve human library away from the antigenic regions of HA, toward the more conserved portions of the protein. All of the identified Fab clones which showed binding to multiple antigenically distinct HA were converted to fully human IgG, and tested for their ability to neutralize the uptake of H5N1-virus like particles (VLP) into MDCK cells. Five of the antibodies which showed binding to the relatively conserved HA2 subunit of HA, exhibited neutralization of H5N1-VLP uptake in a dose dependant manner. The inhibitory effects of these five antibodies were similar to those observed with a previously described neutralizing antibody specific for the 140s antigenic loop present within HA1 and highlight the exciting possibility that these antibodies may be efficacious against multiple H5N1 strains

Epitope characterization of the protective monoclonal antibody VN04-2 shows broadly neutralizing activity against highly pathogenic H5N1

The monoclonal antibody VN04-2 was previously shown to protect mice against lethal A/Vietnam/1203/04 H5N1 virus challenge when administered pre- and post-infection. In this study, we characterized the binding requirements of this antibody using direct binding to hemagglutinin and neutralization assays with H5N1 virus-like particles (H5N1-VLP) of eight recent H5N1 strains representing the major mutations within the 140s antigenic loop. Binding was clade independent and 3 mutations within this antigenic region are required before escape is possible, suggesting that apart from the H5N1 viruses circulating in Indonesia, VN04-2 may provide protection against H5N1 viruses from all other regions

Optimized Expression of Full-Length IgG1 Antibody in a Common E. coli Strain

Multi-polypeptide proteins such as antibodies are difficult to express in prokaryotic systems such as E. coli due to the complexity of protein folding plus secretion. Thus far, proprietary strains or fermenter cultures have been required for appreciable yields. Previous studies have shown that expression of heterologous proteins in E. coli can be enhanced by the reduction of protein translation rates. In this paper, we demonstrate that useful quantities of full-length IgG can be expressed and purified from the common E. coli laboratory strain HB2151 in standard shaking culture using a simple strategy of reduced inducer concentration combined with delayed induction times to modulate translation rates. Purified IgG had only marginally reduced avidity compared to mammalian derived IgG. This indicates that this technique can be used to derive antibodies of potentially equal utility as those expressed in mammalian cell culture, particularly for applications where effector functions mediated by the glycosylated residues in the Fragment Crystallizable (Fc) portion of the immunoglobulin are not required