Search

Southeast Asia's human population is expected to rise by 100 million between 2023 and 2050, with an associated rise in animal-product output in the region's low- to middle-income countries. Countries with the largest population are forecast to continue their increasing poultry consumption, with regional pig meat consumption also to rise, but much less than in China to the north, and much less in Muslim-dominant countries. The forecast growth in the regional ruminant population is more modest and the farm-gate greenhouse gas (GHG) cost per unit of human food protein generated is much higher for ruminant meat (203–584 kg carbon dioxide equivalents (CO2e)/kg protein) than for pig meat (18 kg/kg) or poultry (4 kg/kg). Changing human diets away from ruminant or any animal-sourced protein, is being explored to increase the human food supply at a lower GHG cost. However, with small-holder livestock production systems dominant across many regional countries, the social, land-use and broader economic roles of ruminants need consideration. Strategies to expand ruminant production but with a reduced GHG unit cost (emission intensity) are being pursued. Increasing individual animal-product output, largely through simple animal health and nutritional management decisions, can allow future food targets to be met at a lower GHG emission than if this additional food was produced by business-as-usual livestock production systems. Because the Paris Agreement recognises the priority of food provision over emission abatement, it seems reasonable that much of Southeast Asia should pursue emission intensity targets more than absolute emission targets, and reflect this in their nationally determined contributions (NDCs). Emission-intensity intentions are already apparent not just in NDCs but in emerging carbon markets. Southeast Asia's forecast rise in human population and demand for animal-sourced food is likely to increase absolute greenhouse-gas emissions from the region's livestock sector. There are substantial animal and feed management opportunities to suppress the increase in these emissions by improving animal productivity. Targeting a regional reduction in emissions per unit animal product (emission intensity) instead of absolute emissions offers a means to support food security, while controlling emission of livestock derived greenhouse gases. This article belongs to the collection: SAADC 2023. [ABSTRACT FROM AUTHOR]

Objectives To determine women’s perception of the risk of food safety and how it relates to diet, health and decision making as part of formative research for a market-based intervention that aims to improve the safety of animal-source foods sold in informal markets.Design Qualitative study including in-depth personal interviews with 24 caregivers were conducted and complemented with a second follow-up PhotoVoice interview, which allowed the women to photograph their meals and perceptions of food safety and nutrition. Interview data were analysed using thematic analysis in MAXQDA. Participants were purposively sampled from a larger Safe Food, Fair Food for Cambodia study, conducted from May to August 2018.Setting Urban and periurban neighborhoods of Phnom Penh, Cambodia.Participants 24 female caregivers (mothers and grandmothers) of children under age 5, each interviewed twice.Findings A primary food safety concern expressed was that chemicals (pesticides and other agricultural additives) in animal-source foods, fruits and vegetables may impact the health of their families by causing diarrhoea and problems during pregnancy. This fear created a lack of trust in markets, which influenced their food purchasing behaviours and strategies for making the food safer for their families. These mitigation strategies, including food selection and cleaning, vary among the women but are perceived as important to be able to provide their families with what they define as safe meals.Conclusions Interventions that wish to decrease rates of foodborne illness and increase animal source food consumption should also address the belief that the food system has been compromised by the addition of pesticides and agricultural additives.

Animals decrease intake of an indispensable amino acid deficient diet, due in part to decreased dietary limiting amino acid concentrations within the anterior piriform cortex (APC). In addition to studies supporting a primary role for the APC in this phenomenon, recent studies have shown that the lateral hypothalamus (LH), which receives projections from the APC, also mediates the anorectic response to amino acid deficiency. The neurochemical changes within the LH that accompany the anorexia to amino acid deficiency are unclear. We hypothesized that norepinephrine (NE), dopamine (DA) and serotonin, whose levels are altered in response to amino acid deficiency within the APC, also act within the LH to mediate amino acid deficiency-induced anorexia. We determined that ingestion of an amino acid devoid diet increased concentrations of NE and the serotonin metabolite, 5-hydroxyindoleacetic acid in the LH. The 5-hydroxytryptamine metabolite was increased overall, according to analysis by area under the curve. Individual points reached significance at 130 min; NE was elevated at 170 min. These results suggest that the sustained anorectic response following ingestion of an amino acid devoid diet may be associated with increased activity of the NE and 5-hydroxytryptamine systems in the LH. [ABSTRACT FROM AUTHOR]

Social mixing patterns are key determinants of infectious disease transmission. Mathematical models parameterised with empirical data from contact pattern surveys have played an important role in understanding epidemic dynamics and informing control strategies, including for SARS-CoV-2. However, there is a paucity of data on social mixing patterns in many settings. We conducted a community-based survey in Cambodia in 2012 to characterise mixing patterns and generate setting-specific contact matrices according to age and urban/rural populations. Data were collected using a diary-based approach from 2016 participants, selected by stratified random sampling. Contact patterns were highly age-assortative, with clear intergenerational mixing between household members. Both home and school were high-intensity contact settings, with 27.7% of contacts occurring at home with non-household members. Social mixing patterns differed between rural and urban residents; rural participants tended to have more intergenerational mixing, and a higher number of contacts outside of home, work or school. Participants had low spatial mobility, with 88% of contacts occurring within 1 km of the participants' homes. These data broaden the evidence-base on social mixing patterns in low and middle-income countries and Southeast Asia, and highlight within-country heterogeneities which may be important to consider when modelling the dynamics of pathogens transmitted via close contact., (© 2023. The Author(s).)

Background & Aims: Obesity and type 2 diabetes increase hepatocellular carcinoma (HCC) incidence in humans and accelerate diethylnitrosamine (DEN)-induced hepatocarcinogenesis in mice. We investigated whether exercise reduces HCC development in obese/diabetic Alms1 mutant (foz/foz) mice and studied protective mechanisms., Methods: We measured HCC development in DEN-injected male foz/foz and wild-type (WT) littermates housed with or without an exercise wheel from week 4 until 12 or 24 weeks, and in foz/foz mice pair-fed to WT littermates. We also studied HCC development in DEN-injected Jnk1 -/- .foz/foz mice generated by cross breeding, as well as their genetic controls. Dysplastic hepatocytes were identified by glutathione-S-transferase pi form (GST-pi) immunohistochemistry, liver nodules were counted, and HCC was analysed by histopathology., Results: Exercising foz/foz mice maintained similar weight as WT mice up to 10 weeks, but then gained weight and were obese by 24 weeks; a similar body weight profile was obtained by pair-feeding foz/foz mice to WT. At 12 weeks, livers of exercising foz/foz mice exhibited fewer GST-pi positive hepatocytes than sedentary counterparts; by 24 weeks, fewer exercising foz/foz mice developed HCC (15% vs. 64%, p <0.05). Conversely, pair-feeding foz/foz mice failed to reduce HCC incidence. In these insulin-resistant foz/foz mice, exercise failed to activate hepatic AMPK or Akt/mTORC1. Instead, it improved insulin sensitivity, ameliorated steatosis and liver injury, activated p53 to increase p27 expression, and prevented JNK activation. This was associated with suppression of hepatocellular proliferation. DEN-injected Jnk1 -/- .foz/foz mice failed to develop liver tumours or HCC at 24 weeks., Conclusions: Direct effects of exercise dampen proliferation of dysplastic hepatocytes to reduce 3-month dysplastic foci and 6-month incidence of DEN-induced HCC in obese, insulin-resistant mice. The effects of exercise that potentially slow hepatocarcinogenesis include p53-mediated induction of p27 and prevention of JNK activation., Lay Summary: Fatty liver disease commonly occurs alongside obesity and diabetes, contributing to rapidly increasing rates of liver cancer throughout the world. Herein, we show that exercise reduces the incidence and progression of hepatocellular carcinoma in mouse models. The effect of exercise on cancer risk was shown to be independent of changes in weight. Exercise could be a protective mechanism against liver cancer in at-risk individuals., Competing Interests: Conflict of interests The authors have no conflict of interest to disclose. Please refer to the accompanying ICMJE disclosure forms for further details., (Copyright © 2020 European Association for the Study of the Liver. All rights reserved.)

The underlining mechanisms of dietary cholesterol and nonalcoholic steatohepatitis (NASH) in contributing to hepatocellular carcinoma (HCC) remain undefined. Here we demonstrated that high-fat-non-cholesterol-fed mice developed simple steatosis, whilst high-fat-high-cholesterol-fed mice developed NASH. Moreover, dietary cholesterol induced larger and more numerous NASH-HCCs than non-cholesterol-induced steatosis-HCCs in diethylnitrosamine-treated mice. NASH-HCCs displayed significantly more aberrant gene expression-enriched signaling pathways and more non-synonymous somatic mutations than steatosis-HCCs (335 ± 84/sample vs 43 ± 13/sample). Integrated genetic and expressional alterations in NASH-HCCs affected distinct genes pertinent to five pathways: calcium, insulin, cell adhesion, axon guidance and metabolism. Some of the novel aberrant gene expression, mutations and core oncogenic pathways identified in cholesterol-associated NASH-HCCs in mice were confirmed in human NASH-HCCs, which included metabolism-related genes (ALDH18A1, CAD, CHKA, POLD4, PSPH and SQLE) and recurrently mutated genes (RYR1, MTOR, SDK1, CACNA1H and RYR2). These findings add insights into the link of cholesterol to NASH and NASH-HCC and provide potential therapeutic targets.

Dysplastic hepatocytes (DH) represent altered hepatocytes with potential for malignant transformation. To date, most research on pathways to hepatocarcinogenesis has focused on use of "hepatoma" cell lines derived from hepatocellular carcinoma (HCC). We describe a novel technique for deriving/culturing DH and demonstrate their utility for functional studies in vitro, compared to primary hepatocytes (PH) and HCC. PH and DH were prepared by portal vein collagenase perfusion from C57BL/6J mice. DH were subsequently subjected to FACS. HCC from diethylnitrosamine (DEN)-injected mice were mechanically isolated. Cell cycle analyses were performed by flow cytometry and PCNA immunohistochemistry. To establish utility of DH, we studied pathways of p53 turnover, apoptosis and cell proliferation using pfithrin-α (PFT) and nutlin-3. Like PH, DH were minimally proliferative compared to HCC. Only 30±0.03% of DH were in G 2 /M phase versus 51±0.01% of HCC; this difference corroborated with PCNA-immunostaining of dysplastic nodules from DEN-injected mice. In DH and HCC, nutlin-3 suppressed p53 mRNA, induced p53 and mdm2 activation but paradoxically resulted in increased anti-apoptotic and proliferative activity. Primary murine DH display distinctive biological characteristics compared with PH and HCC. As an intermediate cell type to HCC, they offer a new pathobiologically relevant primary cell culture system with which to interrogate the molecular changes in hepatocarcinogenesis., (Copyright © 2017 Elsevier Inc. All rights reserved.)

In a rat model of right free wall replacement, the transplantation of an engineered multilayered myocardial patch fabricated from a polycaprolactone membrane supporting a chitosan/heart matrix hydrogel induces significant muscular and vascular remodeling and results in a significantly higher right ventricular ejection fraction compared to use of a commercially available pericardium patch., (© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

The stiffness of myocardial tissue changes significantly at birth and during neonatal development, concurrent with significant changes in contractile and electrical maturation of cardiomyocytes. Previous studies by our group have shown that cardiomyocytes generate maximum contractile force when cultured on a substrate with a stiffness approximating native cardiac tissue. However, effects of substrate stiffness on the electrophysiology and ion currents in cardiomyocytes have not been fully characterized. In this study, neonatal rat ventricular myocytes were cultured on the surface of flat polyacrylamide hydrogels with elastic moduli ranging from 1 to 25 kPa. Using whole-cell patch clamping, action potentials and L-type calcium currents were recorded. Cardiomyocytes cultured on hydrogels with a 9 kPa elastic modulus, similar to that of native myocardium, had the longest action potential duration. Additionally, the voltage at maximum calcium flux significantly decreased in cardiomyocytes on hydrogels with an elastic modulus higher than 9 kPa, and the mean inactivation voltage decreased with increasing stiffness. Interestingly, the expression of the L-type calcium channel subunit α gene and channel localization did not change with stiffness. Substrate stiffness significantly affects action potential length and calcium flux in cultured neonatal rat cardiomyocytes in a manner that may be unrelated to calcium channel expression. These results may explain functional differences in cardiomyocytes resulting from changes in the elastic modulus of the extracellular matrix, as observed during embryonic development, in ischemic regions of the heart after myocardial infarction, and during dilated cardiomyopathy.

Background and Aim: While gender differences in hepatocellular carcinoma (HCC) are profound, the mechanism is unclear. Using castration and hormone replacement strategies, we tested whether these gender differences are attributable to testosterone or estradiol/progesterone effects on cell cycle regulators and p53., Methods: We studied dysplastic liver and HCCs in intact and castrated diethylnitrosamine-injected C57BL/6J male and female mice, with or without hormonal replacement. Effects of sex steroids on proliferation and survival of primary hepatocytes and primary HCC cells were also characterized., Results: Diethylnitrosamine-injected female mice displayed fewer dysplastic foci and slower onset of HCC than male mice, with smaller/more differentiated tumors and fewer metastases. Castration of diethylnitrosamine-injected male mice reduced cyclin E kinase and augmented hepatocyte apoptosis compared with intact male mice; estradiol/progesterone enhanced these effects. In intact female mice, cyclin E kinase activity was less than in males; testosterone administered to ovariectomized female mice upregulated cyclin E, increased cyclin E kinase, and accelerated hepatocarcinogenesis. In vitro, testosterone increased expression of cell cycle regulators (cyclin D1, cyclin E, and cyclin-dependent kinase 2) and reduced p53 and p21, which enhanced hepatocyte viability. In contrast, estradiol both suppressed hepatocyte cell cycle markers, upregulated p53 and reduced viability of hepatocytes and HCC cells., Conclusions: Testosterone is the positive regulator of hepatocyte cell cycle via cyclin E, while estradiol plays a negative role by effects of p53 and p21. Together, both sex hormones determine the male predominance of gender differences in hepatocarcinogenesis., (© 2015 Journal of Gastroenterology and Hepatology Foundation and John Wiley & Sons Australia, Ltd.)

Introduction: Cochlear implantation is a clinically routine treatment for patients with severe sensorineural hearing loss for over 20 years. Up to now, the general recommendation for speech processor activation was 4 weeks after cochlear implantation. The aim of this study was to determine whether activation is possible at <4 weeks postop and to show the postoperative change in telemetry over time., Material and Methods: The study was retrospectively carried out at the Karl Landsteiner University Hospital St. Pölten, Austria. Patients who routinely received a CI between January and August 2013 were included in this study. Two weeks after the surgery clinical complications, the ability to wear the audio processor and the impedance values were analysed., Results: Forty-five patients were included in this study. Forty patients were examined at the first postoperative visit, scheduled 2 weeks after the surgery. In nine cases, a mild wound-healing complication occurred; just in one patient, the activation could not carry out immediately. There were no statistically significant differences between the intraoperative, the 2- and 6-week postop impedance measurements for each channel., Discussion: We used the minimally invasive approach and soft implantation technique in all of our patients and believe this to have been a major reason that earlier activation was possible. With this technique, you can reduce the wound-healing process, which makes it possible for the wound to heal fully and for the patient to wear the sound processor over the wound without problems. We found out that telemetry is not a predictor for the time of activation., Conclusion: Based on the results in this study, an earlier activation can be recommended. This will shorten the time the patient needs to wait following cochlear implantation, and rehabilitation can begin sooner., (© 2015 John Wiley & Sons Ltd.)

Background: In many low-income countries, the private commercial sector plays an important role in the provision of malaria treatment. However, the quality of care it provides is often poor, with artemisinin combination therapy (ACT) generally being too costly for consumers. Decreasing ACT prices is critical for improving private sector treatment outcomes and reducing the spread of artemisinin resistance. Yet limited evidence exists on the factors influencing retailers' pricing decisions. This study investigates the determinants of price mark-ups on anti-malarial drugs in retail outlets in Cambodia., Methods: Taking an economics perspective, the study tests the hypothesis that the structure of the anti-malarial market determines the way providers set their prices. Providers facing weak competition are hypothesized to apply high mark-ups and set prices above the competitive level. To analyse the relationship between market competition and provider pricing, the study used cross-sectional data from retail outlets selling anti-malarial drugs, including outlet characteristics data (e.g. outlet type, anti-malarial sales volumes), range of anti-malarial drugs stocked (e.g. dosage form, brand status) and purchase and selling prices. Market concentration, a measure of the level of market competition, was estimated using sales volume data. Market accessibility was defined based on travel time to the closest main commercial area. Percent mark-ups were calculated using price data. The relationship between mark-ups and market concentration was explored using regression analysis., Results: The anti-malarial market was on average highly concentrated, suggesting weak competition. Higher concentration was positively associated with higher mark-ups in moderately accessible markets only, with no significant relationship or a negative relationship in other markets. Other determinants of pricing included anti-malarial brand status and generic type, with higher mark-ups on cheaper products., Conclusions: The results indicate that provider pricing as well as other key elements of anti-malarial supply and demand may have played an important role in the limited access to appropriate malaria treatment in Cambodia. The potential for an ACT price subsidy at manufacturer level combined with effective communications directed at consumers and supportive private sector regulation should be explored to improve access to quality malaria treatment in Cambodia.

A major limitation in tissue engineering strategies for congenital birth defects is the inability to provide a significant source of oxygen, nutrient, and waste transport in an avascular scaffold. Successful vascularization requires a reliable method to generate vascular cells and a scaffold capable of supporting vessel formation. The broad potential for differentiation, high proliferation rates, and autologous availability for neonatal surgeries make amniotic fluid-derived stem cells (AFSC) well suited for regenerative medicine strategies. AFSC-derived endothelial cells (AFSC-EC) express key proteins and functional phenotypes associated with endothelial cells. Fibrin-based hydrogels were shown to stimulate AFSC-derived network formation in vitro but were limited by rapid degradation. Incorporation of poly(ethylene glycol) (PEG) provided mechanical stability (65%±9% weight retention vs. 0% for fibrin-only at day 14) while retaining key benefits of fibrin-based scaffolds-quick formation (10±3 s), biocompatibility (88%±5% viability), and vasculogenic stimulation. To determine the feasibility of AFSC-derived microvasculature, we compared AFSC-EC as a vascular cell source and AFSC as a perivascular cell source to established sources of these cell types-human umbilical vein endothelial cells (HUVEC) and mesenchymal stem cells (MSC), respectively. Cocultures were seeded at a 4:1 endothelial-to-perivascular cell ratio, and gels were incubated at 37°C for 2 weeks. Mechanical testing was performed using a stress-controlled rheometer (G'=95±10 Pa), and cell-seeded hydrogels were assessed based on morphology. Network formation was analyzed based on key parameters such as vessel thickness, length, and area, as well as the degree of branching. There was no statistical difference between individual cultures of AFSC-EC and HUVEC in regard to these parameters, suggesting the vasculogenic potential of AFSC-EC; however, the development of robust vessels required the presence of both an endothelial and a perivascular cell source and was seen in AFSC cocultures (70%±20% vessel length, 90%±10% vessel area, and 105%±10% vessel thickness compared to HUVEC/MSC). At a fixed seeding density, the coculture of AFSC with AFSC-EC resulted in a synergistic effect on network parameters similar to MSC (150% vessel length, 147% vessel area, 150% vessel thickness, and 155% branching). These results suggest that AFSC-EC and AFSC have significant vasculogenic and perivasculogenic potential, respectively, and are suited for in vivo evaluation.

Background & Aims: Epithelial-mesenchymal transition (EMT) has been implicated in the processes of embryogenesis, tissue fibrosis and carcinogenesis. Transforming growth factor-β (TGF-β) has been identified as a key driver of EMT and plays a key role in the pathogenesis of cirrhosis and hepatocellular carcinoma (HCC). The aim was to identify microRNA (miR) expression in TGF-β-induced hepatocyte EMT., Methods: We treated a human hepatocyte cell line PH5CH8 with TGF-β to induce an EMT-like change in phenotype and then identified dysregulated miRs using TaqMan Low Density Arrays. MiR expression was altered using miR-181a mimic and inhibitor in the same system and gene changes were identified using TaqMan gene arrays. MiR-181a gene expression was measured in human and mouse cirrhotic or HCC liver tissue samples. Gene changes were identified in rAAV-miR-181a-expressing mouse livers using TaqMan gene arrays., Results: We identified miR-181a as a miR that was significantly up-regulated in response to TGF-β treatment. Over-expression of a miR-181a mimic induced an in vitro EMT-like change with a phenotype similar to that seen with TGF-β treatment alone and was reversed using a miR-181a inhibitor. MiR-181a was shown to be up-regulated in experimental and human cirrhotic and HCC tissue. Mouse livers expressing rAAV-miR-181a showed genetic changes associated with TGF-β signalling and EMT., Conclusions: MiR-181a had a direct effect in inducing hepatocyte EMT and was able to replace TGF-β-induced effects in vitro. MiR-181a was over-expressed in cirrhosis and HCC and is likely to play a role in disease pathogenesis., (© 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

The major limitation of current engineered myocardial patches for the repair of heart defects is that insulating polymeric scaffold walls hinder the transfer of electrical signals between cardiomyocytes. This loss in signal transduction results in arrhythmias when the scaffolds are implanted. We report that small, subtoxic concentrations of single-walled carbon nanotubes, on the order of tens of parts per million, incorporated in a gelatin-chitosan hydrogel act as electrical nanobridges between cardiomyocytes, resulting in enhanced electrical coupling, synchronous beating, and cardiomyocyte function. These engineered tissues achieve excitation conduction velocities similar to native myocardial tissue (22 ± 9 cm/s) and could function as a full-thickness patch for several cardiovascular defect repair procedures, such as right ventricular outflow track repair for Tetralogy of Fallot, atrial and ventricular septal defect repair, and other cardiac defects, without the risk of inducing cardiac arrhythmias.

A novel cardiac scaffold comprised of decellularized porcine heart matrix was investigated for use as a biodegradable patch with a potential for surgical reconstruction of the right ventricular outflow tract. Powdered heart matrix solution was blended with chitosan and lyophilized to form three-dimensional scaffolds. For this investigation, we examined the influence of different blending ratios of heart matrix to chitosan on porosity and mechanical properties, then gene expression and electrophysiological function of invading neonatal rat ventricular myocytes (NRVM) compared to type-A gelatin/chitosan composite scaffolds. Heart matrix/chitosan-blended hydrogels (1.6 mg/mL heart matrix) had similar porosity (109±34 μm), and elastic modulus (13.2±4.0 kPa) as previously published gelatin/chitosan scaffolds. Heart matrix/chitosan hydrogels maintained>80% viability and had higher NRVM retention (∼1000 cells/mm(2)) than gelatin/chitosan scaffolds. There was a significant increase in α-myosin heavy chain and connexin-43 expression in NRVM cultured on heart matrix/chitosan scaffolds after 14 days compared with gelatin/chitosan scaffolds. Further, heart matrix/chitosan scaffolds had significantly higher conduction velocity (12.6±4.9 cm/s) and contractile stress (0.79±0.13 mN/mm(2)) than gelatin/chitosan scaffolds. In summary, NRVM cultured on heart matrix scaffold showed improvements in contractile and electrophysiological function.

Background: There is increased interest in using commercial providers for improving access to quality malaria treatment. Understanding their current role is an essential first step, notably in terms of the volume of diagnostics and anti-malarials they sell. Sales volume data can be used to measure the importance of different provider and product types, frequency of parasitological diagnosis and impact of interventions. Several methods for measuring sales volumes are available, yet all have methodological challenges and evidence is lacking on the comparability of different methods., Methods: Using sales volume data on anti-malarials and rapid diagnostic tests (RDTs) for malaria collected through provider recall (RC) and retail audits (RA), this study measures the degree of agreement between the two methods at wholesale and retail commercial providers in Cambodia following the Bland-Altman approach. Relative strengths and weaknesses of the methods were also investigated through qualitative research with fieldworkers., Results: A total of 67 wholesalers and 107 retailers were sampled. Wholesale sales volumes were estimated through both methods for 62 anti-malarials and 23 RDTs and retail volumes for 113 anti-malarials and 33 RDTs. At wholesale outlets, RA estimates for anti-malarial sales were on average higher than RC estimates (mean difference of four adult equivalent treatment doses (95% CI 0.6-7.2)), equivalent to 30% of mean sales volumes. For RDTs at wholesalers, the between-method mean difference was not statistically significant (one test, 95% CI -6.0-4.0). At retail outlets, between-method differences for both anti-malarials and RDTs increased with larger volumes being measured, so mean differences were not a meaningful measure of agreement between the methods. Qualitative research revealed that in Cambodia where sales volumes are small, RC had key advantages: providers were perceived to remember more easily their sales volumes and find RC less invasive; fieldworkers found it more convenient; and it was cheaper to implement than RA., Discussion/conclusions: Both RA and RC had implementation challenges and were prone to data collection errors. Choice of empirical methods is likely to have important implications for data quality depending on the study context.

Background and Aim: By array-comparative genomic hybridization, we demonstrated cyclin E as one of seven genes associated with hepatocellular carcinoma (HCC) development in Ku70 DNA repair-deficient mice. We therefore explored the hypothesis that during hepatocarcinogenesis, cyclin E kinase can overcome the inhibitory effects of p53 and establish whether abnormal miRNA(mi-R)-34, a co-regulator of cyclin E and p53, can account for their interactions as "drivers" of HCC., Methods: Dysplastic hepatocytes (DNs) and HCCs were generated from diethylnitrosamine (DEN)-injected C57BL/6 male mice at 3-12 months., Results: Cyclin E/cdk2 was barely expressed in normal liver, but was readily detected in dysplastic hepatocytes, localizing to glutathione-S transferase pi-form positive cells dissected by laser-dissection. Cyclin E kinase activity preceded cyclin D1, proliferating cell nuclear antigen expression in DNs and HCCs despite maximal p53 and p21 expression. We confirmed that cyclin E, rather than cyclin D1, is the proliferative driver in hepatocarcinogenesis by immunoprecipitation experiments demonstrating preferential binding of p21 to cyclin D1, allowing cyclin E-mediated "escape" from G1/S checkpoint. We then showed cyclin E was responsible for regulating wild-type p53 by knockdown experiments in primary HCC cells; cyclin E-knockdown increased p53 and p21, diminished anti-apoptotic Bcl-XL and reduced cell viability. Conversely, blocking p53 augmented cyclin E, Bcl-XL expression and increased proliferation. Physiological interactions between cyclin E/p53/p21 were confirmed in primary hepatocytes. miR-34a,c were upregulated in dysplastic murine, human liver and HCCs compared with normal liver, and appeared to be linked to cyclin E/p53., Conclusion: Upregulation of functionally active cyclin E via miR34 with loss of p53 function is associated with cell-cycle checkpoint failure increasing proliferative drive that favors hepatocarcinogenesis., (© 2013 Journal of Gastroenterology and Hepatology Foundation and Wiley Publishing Asia Pty Ltd.)

A three-dimensional scaffold composed of self-assembled polycaprolactone (PCL) sandwiched in a gelatin-chitosan hydrogel was developed for use as a biodegradable patch with a potential for surgical reconstruction of congenital heart defects. The PCL core provides surgical handling, suturability and high initial tensile strength, while the gelatin-chitosan scaffold allows for cell attachment, with pore size and mechanical properties conducive to cardiomyocyte migration and function. The ultimate tensile stress of the PCL core, made from blends of 10, 46 and 80kDa (Mn) PCL, was controllable in the range of 2-4MPa, with lower average molecular weight PCL blends correlating with lower tensile stress. Blends with lower molecular weight PCL also had faster degradation (controllable from 0% to 7% weight loss in saline over 30 days) and larger pores. PCL scaffolds supporting a gelatin-chitosan emulsion gel showed no significant alteration in tensile stress, strain or tensile modulus. However, the compressive modulus of the composite tissue was similar to that of native tissue (∼15kPa for 50% gelatin and 50% chitosan). Electron microscopy revealed that the gelatin-chitosan gel had a three-dimensional porous structure, with a mean pore diameter of ∼80μm, showed migration of neonatal rat ventricular myocytes (NRVM), maintained NRVM viability for over 7 days, and resulted in spontaneously beating scaffolds. This multi-layered scaffold has sufficient tensile strength and surgical handling for use as a cardiac patch, while allowing migration or pre-loading of cardiac cells in a biomimetic environment to allow for eventual degradation of the patch and incorporation into native tissue., (Copyright © 2012 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

The objective of this study was to investigate a bioreactor suitable for human bladder regeneration. Simulations were performed using the computational fluid dynamic tools. The thickness of the bladder scaffold was 3 mm, similar to the human bladder, and overall hold-up volume within the spherical shape scaffold was 755 ml. All simulations were performed using (i) Brinkman equation on porous regions using the properties of 1% chitosan-1% gelatin structures, (ii) Michaelis-Menten type rate law nutrient consumption for smooth muscle cells (SMCs) and (iii) Mackie-Meares relationship for determining effective diffusivities. Steady state simulations were performed using flow rates from 0.5 to 5 ml/min. Two different inlet shapes: (i) straight entry at the centre (Design 1) and (ii) entry with an expansion (Design 2) were simulated to evaluate shear stress distribution. Also, mimicking bladder shape of two inlets (Design 3) was tested. Design 2 provided the uniform shear stress at the inlet and nutrient distribution, which was further investigated for the effect of scaffold locations within the reactor: (i) attached with a 3-mm open channel (Design 2-A), (ii) flow through with no open channel (Design 2-B) and (iii) porous structure suspended in the middle with 1.5-mm open channel on either side (Design 2-C). In Design 2-A and 2-C, fluid flow occurred by diffusion dominant mechanisms. Furthermore, the designed bioreactor is suitable for increased cell density of SMCs. These results showed that increasing the flow rate is necessary due to the decreased permeability at cell densities similar to the human bladder.

This article reviews current applications and novel candidate biomaterials for use as tissue-engineered scaffolds in pediatric cardiac tissue engineering. This overview of different types of biomaterials includes naturally derived and synthetic polymers and their biological, physical, and biomechanical properties for the use as a patch or baffle for surgical reconstruction of congenital heart defects. Applications and characteristics of composite biomaterials are highlighted with their respective feasibilities in use as cardiac scaffolds. Currently, a wide range of biomaterials has been introduced for cardiovascular reconstruction for complex congenital cardiac defects. However, there are still many remaining challenges for engineered tissue implantations.

Background/aims: FGFR4, a member of the fibroblast growth factor receptor family, has been recently associated with progression of melanoma, breast and head and neck carcinoma. Given its uniquely high expression in the liver, we investigated its contributory role to hepatocellular carcinoma (HCC)., Methods: We performed a comprehensive sequencing of full-length FGFR4 transcript in 57 tumor/normal HCC tissue pairs, and quantified their mRNA expressions. Notable mutations and expression patterns were correlated with patient data. Clinically significant trends were examined in in vitro models., Results: We found eight genetic alterations including two highly frequent polymorphisms (V10I and G338R). Secretion of alpha-fetoprotein (AFP), a HCC biomarker, was increased among patients bearing homozygous Arg388 alleles. One-third of these patients exhibited increased FGFR4 mRNA expression in the matched tumor/normal tissue. Subsequent in vitro perturbation of FGFR4 signaling through both FGF19-stimulation and FGFR4 silencing confirmed a mechanistic link between FGFR4 activities and tumor aggressiveness. More importantly, inhibition of FGFR activity with PD173074 exquisitely blocked HuH7 (high FGFR4 expression) proliferation as compared to control cell lines., Conclusions: FGFR4 contributes significantly to HCC progression by modulating AFP secretion, proliferation and anti-apoptosis. Its frequent overexpression in patients renders its inhibition a novel and much needed pharmacological approach against HCC.

Protein tyrosine kinases (PTKs) play a critical role in the manifestation of cancer cell properties, and respective signaling mechanisms have been studied extensively on immortalized tumor cells. To characterize and analyze commonly used cancer cell lines with regard to variations in the primary structure of all expressed PTKs, we conducted a cDNA-based sequence analysis of the entire tyrosine kinase transcriptome of 254 established tumor cell lines. The profiles of cell line intrinsic PTK transcript alterations and the evaluation of 155 identified polymorphisms and 234 somatic mutations are made available in a database designated "Tykiva" (tyrosine kinome variant). Tissue distribution analysis and/or the localization within defined protein domains indicate functional relevance of several genetic alterations. The cysteine replacement of the highly conserved Y367 residue in fibroblast growth factor receptor 4 or the Q26X nonsense mutation in the tumor-suppressor kinase CSK are examples, and may contribute to cell line-specific signaling characteristics and tumor progression. Moreover, known variants, such as epidermal growth factor receptor G719S, that were shown to mediate anticancer drug sensitivity could be detected in other than the previously reported tumor types. Our data therefore provide extensive system information for the design and interpretation of cell line-based cancer research, and may stimulate further investigations into broader clinical applications of current cancer therapeutics.

Objective: To evaluate the possibility of preservation of low-frequency hearing in atraumatic cochlear implant electrode insertion procedures for combined, ipsilateral electric and acoustic stimulation., Material and Methods: A total of 21 patients were implanted with a MED EL C40+ cochlear implant using an atraumatic electrode insertion technique to preserve residual low-frequency hearing. Pure-tone audiometric thresholds were measured pre- and postoperatively to evaluate the degree of preserved hearing. Speech discrimination tests in quiet and with background noise were performed in a patient with successful hearing preservation., Results: Using the atraumatic electrode insertion procedure with an insertion depth of 360 degrees (18-24 mm), hearing preservation could be achieved in 18/21 patients (85.7%). Three patients (14.3%) lost their residual low-frequency hearing after the implantation. Residual hearing was preserved completely in 13 patients (61.9%) and partial hearing preservation was possible in 5 (23.8%). Preliminary speech discrimination tests showed a dramatic benefit for the combined electric and acoustic stimulation mode compared to cochlear implantation alone., Conclusion: Preservation of low-frequency hearing in cochlear implantation is possible in patients implanted because of profound high-frequency deafness. With the development of new, more atraumatic electrode designs, preservation of residual hearing should be further improved.

The aim of this study was to determine the improvement in speech recognition provided by a cochlear implant (CI) in conjunction with a hearing aid (HA) in the opposite ear. The study was a retrospective cohort study in the context of a university teaching hospital CI programme. Seven CI patients who still use their HA in the opposite ear were tested. The scores with the CI alone and the CI in conjunction with an HA were evaluated by using three speech perception tests in quiet (Freiburger Numbers, Freiburger Monosyllables, and Innsbrucker Sentence Test). In the majority of tests and subjects, the CI alone performed better than the HA alone, and the bimodal (CI+HA) condition was superior to the CI alone. On the sentence test, the patients as a group improved from 47-96% (mean: 79%; CI alone) to 50-100% (mean: 88.1%; CI+HA, pv<0.05). With the more difficult monosyllable test, the scores improved from 15-52% (mean: 37.2%; CI alone) to 15-82% (mean: 48.7%; CI+ HA, p<0.05). On the numbers test, scores increased from 65-98% (mean: 83%; CI alone) to 75-98% (mean: 88.7%; CI+ HA, p<0.05). All patients in this study were implanted in the poorer ear. The results of the present study suggest the advantage of CI usage in conjunction with an HA in the opposite ear.

It has been shown in several studies that there is a statistically confirmed connection between cholesterol and Hb levels in the sense that hypocholesterolemia and anemia occur together particular frequently. This was demonstrated in an epidemiological cross-section study of 3958 patient data, and also that in a retrospective case control study of 159 anemias there was a concurrent hypocholesterolemia. It is postulated that a causal connection exists which is explained by the fact that cholesterol deficiency leads to rigidity of the erythrocytes.

Two cases with acute renal failure after prolonged hypothermia are presented. Both patients were found in come, became rapidly uremic and required hemodilaysis treatment. Although the laboratory findings were typical of severe muscle damage, e.g. elevated levels of serum creatinine phosphokinase, serum lactic dehydrogenase and serum aldolase activities, visible "crush-injuries" were not found. Acute renal failure was characterized by extreme catabolism and severe metabolic acidosis. After 4 and 10 hemodialyses respectively, the patients became polyuric and finally were discharges with normal renal and muscle function. Hypotension with diminished renal perfusion and nontraumatic rhabdomyolysis due to prolonged hypothermia are regarded as the dominant pathogenetic factors in the acute renal failure.

It has been shown in several studies that there is a statistically confirmed connection between cholesterol and Hb levels in the sense that hypocholesterolemia and anemia occur together particularly frequently. This was demonstrated in an epidemiological cross-section study of 3958 patient data, and also that in a retrospective case control study of 159 anemias there wasa concurrent hypocholesterolemia. It is explained by the fact that cholesterol deficiency leads to rigidity of the erythrocytes.