Bioengineering embryonic stem cell microenvironments for exploring inhibitory effects on metastatic breast cancer cells

The recreation of an in vitro microenvironment to understand and manipulate the proliferation and migration of invasive breast cancer cells may allow one to put a halt to their metastasis capacity. Invasive cancer cells have been linked to embryonic stem (ES) cells as they possess certain similar characteristics and gene signatures. Embryonic microenvironments have the potential to reprogram cancer cells into a less invasive phenotype and help elucidate tumorigenesis and metastasis. In this study, we explored the feasibility of reconstructing embryonic microenvironments using mouse ES cells cultured in alginate hydrogel and investigated the interactions of ES cells and highly invasive breast cancer cells in 2D, 2&1/2D, and 3D cultures. Results showed that mouse ES cells inhibited the growth and tumor spheroid formation of breast cancer cells. The mouse ES cell microenvironment was further constructed and optimized in 3D alginate hydrogel microbeads, and co-cultured with breast cancer cells. Migration analysis displayed a significant reduction in the average velocity and trajectory of breast cancer cell locomotion compared to control, suggesting that bioengineered mouse ES cell microenvironments inhibited the proliferation and migration of breast cancer cells. This study may act as a platform to open up new options to understand and harness tumor cell plasticity and develop therapeutics for metastatic breast cancer.     

Antisolvent crystallisation is a potential technique to prepare engineered lactose with promising aerosolisation properties: Effect of saturation degree

Engineered lactose particles were prepared by anti-solvent crystallisation technique using lactose solutions with different saturation degrees. In comparison to commercial lactose, engineered lactose particles exhibited less elongated and more irregular shape (large aggregates composed of smaller sub-units), rougher surface texture, higher specific surface area, and different anomer form. Engineered lactose powders demonstrated smaller bulk density, smaller tap density, and higher porosity than commercial lactose powder. Dry powder inhaler (DPI) formulations containing engineered lactose and salbutamol sulphate as a model drug demonstrated improved drug content homogeneity and higher amounts of drug delivered to lower airway regions. Higher fine particle fraction of drug was obtained in the case of lactose powders with higher porosity, higher specific surface area and higher fine particle content (<5μm). The results indicated that the higher the saturation degree of lactose solution used during crystallisation the smaller the specific surface area, the higher the amorphous lactose content, and the higher the β-lactose content of engineered lactose particles. Also, lactose powders obtained from lactose solution with higher degree of saturation showed higher bulk and tap densities and smaller porosity. Engineered lactose powders crystallized from lower saturation degree (20% and 30% w/v) deposited higher amounts of drug on lower airway regions. In conclusion, this study demonstrated that it is possible to prepare engineered lactose particles with favourable properties (e.g. higher fine particle fraction and better drug content homogeneity) for DPI formulations by using lactose solutions with lower degree of saturation during crystallisation process.     

Resistance to new chemical insecticides in the house fly, Musca domestica L., from dairies in Punjab, Pakistan

The house fly, Musca domestica L., is one of the major pests in dairy operations that has developed resistance to a number of insecticides with different modes of action. Adult house fly populations from six dairies in Punjab, Pakistan were evaluated for resistance to insecticides with novel modes of action (abamectin, emamectin benzoate, fipronil, imidacloprid, indoxacarb, and spinosad). Significant levels of resistance to most of the insecticides tested were observed in the present study. For avermectins at LC₅₀ level, the resistance ratios were in the range of 38.40 to 94.44-fold for abamectin and 13.16 to 36.30-fold for emamectin benzoate. Fipronil LC₅₀ resistance ratios exceeded 10-fold in three house fly populations, while all the populations had >10-fold resistance ratios for imidacloprid. Indoxacarb and spinosad had the lowest resistance ratios that ranged from 3.02 to 7.12-fold for indoxacarb and 2.91 to 9.0-fold for spinosad. As the resistance to fipronil, indoxacarb, and spinosad are emerging, therefore these chemicals should be used cautiously in management programs to retain the efficacy for longer times.     

<Emphasis Type="Italic">Streptococcus agalactiae</Emphasis> CAMP factor/protein B does not bind to human IgG

CAMP factor is an extracellular cytolytic protein produced by Streptococcus agalactiae. CAMP factor has been reported to bind the Fc fragments of immunoglobulin G (IgG) and has therefore also been called protein B, in analogy to protein A of Staphylococcus aureus. We attempted to characterize the interaction of protein B with IgG in more detail. In contrast to protein A, CAMP factor does not inhibit the activation of complement by hemolysin antibodies bound to sheep red cell surfaces. IgG also failed to inhibit the co-hemolytic activity of CAMP factor, which is in disagreement with previous findings. After co-incubation, CAMP factor and IgG were cleanly separated by gel filtration, indicating that no binding had occurred. Waseem El-Huneidi and Ryan Mui contributed equally to this work.     

Evaluation of Fluoride Release in Chitosan-Modified Glass Ionomer Cements

ObjectiveThis study assessed the influence of chitosan nanoparticles on the fluoride-releasing ability of 4 glass ionomer cement (GIC) through an in vitro analysis.MethodsFour types of GIC (type II light cure universal restorative, type II universal restorative, GC Fuji VII, and type IX) were modified with nanochitosan particles; 10% chitosan was added to the glass ionomer liquid. Six specimens for each of the 4 groups were created, using expendable Teflon moulds. Discs of each type of GIC (n = 6) were immersed in deionised water at various time intervals. Electrodes selective for fluoride ions were employed to analyse the amount of released fluoride at 1, 7, 14, 21, and 28 days.ResultsChitosan-modified GICs showed greater fluoride release than conventional GICs at all time points. All samples showed an initial high release of fluoride that tapered off with time. The total amount of fluoride released increased from the 1st day to the 28th day on adding chitosan to all the 4 types of GIC. Amongst those, type IX high-strength posterior extra with chitosan released a considerably higher quantity of fluoride at all time intervals.ConclusionsIn all the experimental groups, adding chitosan to the glass ionomer liquid had an accelerating effect on its fluoride-releasing property.     

Reactive oxygen species initiate a metabolic collapse in hippocampal slices: potential trigger of cortical spreading depression

Excessive accumulation of reactive oxygen species (ROS) underlies oxidative damage. We find that in hippocampal slices, decreased activity of glucose-based antioxidant system induces a massive, abrupt, and detrimental change in cellular functions. We call this phenomenon metabolic collapse (MC). This collapse manifested in long-lasting silencing of synaptic transmission, abnormal oxidation of NAD(P)H and FADH₂ associated with immense oxygen consumption, and massive neuronal depolarization. MC occurred without any preceding deficiency in neuronal energy supply or disturbances of ionic homeostasis and spread throughout the hippocampus. It was associated with a preceding accumulation of ROS and was largely prevented by application of an efficient antioxidant Tempol (4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl). The consequences of MC resemble cortical spreading depression (CSD), a wave of neuronal depolarization that occurs in migraine, brain trauma, and stroke, the cellular initiation mechanisms of which are poorly understood. We suggest that ROS accumulation might also be the primary trigger of CSD. Indeed, we found that Tempol strongly reduced occurrence of CSD in vivo, suggesting that ROS accumulation may be a key mechanism of CSD initiation.