Detection and tracking volumes of interest in 3D printed tissue engineering scaffolds using 4D imaging modalities.

Additive manufacturing (AM) platforms allow the production of patient tissue engineering scaffolds with desirable architectures. Although AM platforms offer exceptional control on architecture, post-processing methods such as sintering and freeze-drying often deform the printed scaffold structure. In-situ 4D imaging can be used to analyze changes that occur during post-processing. Visualization and analysis of changes in selected volumes of interests (VOIs) over time are essential to understand the underlining mechanisms of scaffold deformations. Yet, automated detection and tracking of VOIs in the 3D printed scaffold over time using 4D image data is currently an unsolved image processing task. This paper proposes a new image processing technique to segment, detect and track volumes of interest in 3D printed tissue engineering scaffolds. The method is validated using a 4D synchrotron sourced microCT image data captured during the sintering of bioactive glass scaffolds in-situ. The proposed method will contribute to the development of scaffolds with controllable designs and optimum properties for the development of patient-specific scaffolds

Bone mineral accrual and gain in skeletal width in pre-pubertal school children is independent of the mode of school transportation – one-year data from the prospective observational pediatric osteoporosis prevention (POP) study

Background: Walking and cycling to school could be an important regular source of physical activity in growing children. The aim of this 12 months prospective observational study was thus to evaluate the effect of self-transportation to school on bone mineral accrual and gain in bone width in pre-pubertal children, both traits independently contributing to bone strength. Methods: Ninety-seven girls and 133 boys aged 7-9 years were recruited as a part of the Malmo Pediatric Osteoporosis Prevention (POP) Study in order to evaluate the influence of self-selected school transportation for the accrual of bone mineral and bone width. Children who walked or cycled to school were compared with children who went by bus or car. Bone mineral content (BMC) was measured by dual energy X-ray absorptiometry (DXA) in the lumbar spine (L2-L4), third lumbar vertebra (L3) and hip, and bone width was calculated at L3 and femoral neck (FN). Changes during the first 12 months were compared between the groups. Subjective duration of physical activity was estimated by a questionnaire and objective level of everyday physical activity at follow-up by accelerometers worn for four consecutive days. All children remained in Tanner stage 1 throughout the study. Comparisons were made by independent student's t-tests between means, ANCOVA and Fisher's exact tests. Results: There were no differences in baseline or annual changes in BMC or bone width when the transportation groups were compared. No differences were detected in objectively measured daily level of physical activity by accelerometer. All children reached above 60 minutes of moderate to intense daily physical activity per day, the international recommended level of daily physical activity according to the United Kingdom Expert Consensus Group. Conclusion: The everyday physical activity in these pre-pubertal children seems to be so high that the school transportation contributes little to their total level of physical activity. As a result, the choice of school transportation seems not to influence the accrual of bone mineral or gain in bone size during a I2-month follow-up period

Branding Practices on Four Dairies in Kantale, Sri Lanka.

Hot-iron branding is illegal in Sri Lanka, but is still commonly used to identify dairy herds in extensive farming systems, which are primarily located in the country's Dry Zone. Despite the negative welfare implications of this practice, there is no written documentation of branding in this region. We observed branding on four smallholder farms in Kantale, Eastern Province to understand the welfare implications associated with the procedure and challenges limiting the uptake of more welfare-friendly alternatives, such as ear tagging. Areas of welfare concern included the duration of restraint, the size and location of the brand, and the absence of pain relief. Animals were restrained with rope for an average duration of 12 min (range 8⁻17 min). Farmers used multiple running irons to mark their initials and, in some cases, their address, with the largest brands extending across the ribs and hip. Three farmers applied coconut or neem oil topically to the brand after performing the procedure. No analgesics were given before or after branding. Farmers reported that poor ear tag retention in extensive systems and theft were the main factors impeding the uptake of alternative forms of identification. Branding is also practiced as part of traditional medicine in some cases. Given the clear evidence that hot-iron branding impairs animal welfare and there is no evidence that this can be improved, alternative identification methods are needed, both in Sri Lanka, as well as in other countries engaging in this practice

A Hexylchloride-Based Catch-and-Release System for Chemical Proteomic Applications

Bioorthogonal ligation methods that allow the selective conjugation of fluorophores or biotin to proteins and small molecule probes that contain inert chemical handles are an important component of many chemical proteomic strategies. Here, we present a new catch-and-release enrichment strategy that utilizes a hexylchloride group as a bioorthogonal chemical handle. Proteins and small molecules that contain a hexylchloride tag can be efficiently captured by an immobilized version of the self-labeling protein HaloTag. Furthermore, by using a HaloTag fusion protein that contains a protease cleavage site, captured proteins can be selectively eluted under mild conditions. We demonstrate the utility of the hexylchloride-based catch-and-release strategy by enriching protein kinases that are covalently and noncovalently bound to ATP-binding site-directed probes from mammalian cell lysates. Our catch-and-release system creates new possibilities for profiling enzyme families and for the identification of the cellular targets of bioactive small molecules

A Hexylchloride-Based Catch-and-Release System for Chemical Proteomic Applications

Bioorthogonal ligation methods that allow the selective conjugation of fluorophores or biotin to proteins and small molecule probes that contain inert chemical handles are an important component of many chemical proteomic strategies. Here, we present a new catch-and-release enrichment strategy that utilizes a hexylchloride group as a bioorthogonal chemical handle. Proteins and small molecules that contain a hexylchloride tag can be efficiently captured by an immobilized version of the self-labeling protein HaloTag. Furthermore, by using a HaloTag fusion protein that contains a protease cleavage site, captured proteins can be selectively eluted under mild conditions. We demonstrate the utility of the hexylchloride-based catch-and-release strategy by enriching protein kinases that are covalently and noncovalently bound to ATP-binding site-directed probes from mammalian cell lysates. Our catch-and-release system creates new possibilities for profiling enzyme families and for the identification of the cellular targets of bioactive small molecules

Changes in mental disorder prevalence among conflict-affected populations: a prospective study in Sri Lanka (COMRAID-R).

BACKGROUND: Longitudinal data are lacking on mental health trajectories following conflict resolution and return migration. COMRAID-R is a follow-up study of Muslims displaced by conflict from Northern Sri Lanka 20 years ago who are now beginning to return. METHODS: Of 450 participants in displacement interviewed in 2011, 338 (75.1%) were re-interviewed a year later, and a supplementary random sample (n = 228) was drawn from return migrants with a comparable displacement history. Common mental disorder (CMD; Patient Health Questionnaire) and post-traumatic stress disorder (CIDI-subscale) were measured. RESULTS: A CMD prevalence of 18.8% (95%CI 15.2-22.5) at baseline had reduced to 8.6% (5.6-11.7) at follow-up in those remaining in displacement, and was 10.3% (6.5-14.1) in return migrants. PTSD prevalences were 2.4%, 0.3% and 1.6% respectively. CONCLUSIONS: We observed a substantial decrease in CMD prevalence in this population over a short period, which may reflect the prospect of return migration and associated optimism following conflict resolution

Targeting Diverse Signaling Interaction Sites Allows the Rapid Generation of Bivalent Kinase Inhibitors

The identification of potent and selective modulators of protein kinase function remains a challenge, and new strategies are needed for generating these useful ligands. Here, we describe the generation of bivalent inhibitors of three unrelated protein kinases: the CAMK family kinase Pim1, the mitogen-activated protein kinase (MAPK) p38α, and the receptor tyrosine kinase (RTK) epidermal growth factor receptor (EGFR). These bivalent inhibitors consist of an ATP-competitive inhibitor that is covalently tethered to an engineered form of the self-labeling protein <i>O</i>⁶-alkylguanine-DNA alkyltransferase (SNAP-tag). In each example, SNAP-tag is fused to a peptide ligand that binds to a signaling interaction site of the kinase being targeted. These interactions increase the overall selectivity and potency of the bivalent inhibitors that were generated. The ability to exploit disparate binding sites in diverse kinases points to the generality of the method described. Finally, we demonstrate that ATP-competitive inhibitors that are conjugated to the bio-orthogonal tag <i>O</i>⁴-benzyl-2-chloro-6-aminopyrimidine (CLP) are cell-permeable. The selective labeling of SNAP-tag with CLP conjugates allows the rapid assembly of bivalent inhibitors in living cells

Cell-delivered magnetic nanoparticles caused hyperthermia-mediated increased survival in a murine pancreatic cancer model

Using magnetic nanoparticles to absorb alternating magnetic field energy as a method of generating localized hyperthermia has been shown to be a potential cancer treatment. This report demonstrates a system that uses tumor homing cells to actively carry iron/iron oxide nanoparticles into tumor tissue for alternating magnetic field treatment. Paramagnetic iron/iron oxide nanoparticles were synthesized and loaded into RAW264.7 cells (mouse monocyte/macrophage-like cells), which have been shown to be tumor homing cells. A murine model of disseminated peritoneal pancreatic cancer was then generated by intraperitoneal injection of Pan02 cells. After tumor development, monocyte/macrophage-like cells loaded with iron/iron oxide nanoparticles were injected intraperitoneally and allowed to migrate into the tumor. Three days after injection, mice were exposed to an alternating magnetic field for 20 minutes to cause the cell-delivered nanoparticles to generate heat. This treatment regimen was repeated three times. A survival study demonstrated that this system can significantly increase survival in a murine pancreatic cancer model, with an average post-tumor insertion life expectancy increase of 31%. This system has the potential to become a useful method for specifically and actively delivering nanoparticles for local hyperthermia treatment of cancer