To achieve better accuracy in their investigations, biologists have recently been using three-dimensional models as intermediates between two-dimensional cell culture and the in vivo study of tissues. Some of these models are based on spheroids, which are cellular aggregates retaining many characteristics of in vivo behaviors while being still easy to use and implement in labs. To study such objects, multiple observation techniques can be used according to the objective of the study, including those using electromagnetic waves as nondestructive, noninvasive, and label-free analysis. Low-frequency (<1 MHz) ones are currently under investigation as electrochemical impedance spectroscopy. However, unlike microwaves (from 300 MHz to 60 GHz), they cannot penetrate the cell. Furthermore, most of the devices dedicated to microwave dielectric characterization are only focusing on cellular scales lower than spheroids (single cell, cell mats, or suspensions) or on tissues and organs. In this article, a microwave spectroscopy device using a coplanar waveguide adapted to the study of spheroids is presented with the dielectric characterization of fixed spheroids, with capacitance and conductance measurements from 0.5 to 20 GHz, aiming at filling the scale gap in the state of the art.

FLASH radiotherapy is a novel technique that has been shown in numerous preclinical in vivo studies to have the potential to be the next important improvement in cancer treatment. However, the biological mechanisms responsible for the selective FLASH sparing effect of normal tissues are not yet known. An optimal translation of FLASH radiotherapy into the clinic would require a good understanding of the specific beam parameters that induces a FLASH effect, environmental conditions affecting the response, and the radiobiological mechanisms involved. Even though the FLASH effect has generally been considered as an in vivo effect, studies finding these answers would be difficult and ethically challenging to carry out solely in animals. Hence, suitable in vitro studies aimed towards finding these answers are needed. In this review, we describe and summarise several in vitro assays that have been used or could be used to finally elucidate the mechanisms behind the FLASH effect.

Strategies for preventing Fe deficiency include Fe supplementation and Fe fortification of foods. The absorption, metabolism and chemical characteristics of Fe multi-amino acid chelate (IMAAC) are not known. Absorption of IMAAC was compared with FeSO4 in Fe-depleted mice and in vitro chemical studies of the Fe supplement was performed in HuTu 80 cells. Hb repletion study was carried out in Fe-deficient CD1 mice that were fed for 10 d a diet supplemented with ferrous IMAAC or FeSO4. A control group of Fe-replete mice was fed a diet with adequate Fe concentrations throughout the study. Tissues were collected from the mice, and the expression of Fe-related genes was determined by quantitative PCR. Ferric reductase and Fe uptake were evaluated in HuTu 80 cells. Supplementation of the diet with FeSO4 or IMAAC significantly increased Hb levels (P<0·001) in Fe-deficient mice from initial 93·9 (SD 10·8) or 116·2 (SD 9·1) to 191 (SD 0·7) or 200 (SD 0·5) g/l, respectively. Initial and final Hb for the Fe-deficient control group were 87·4 (SD 6·7) and 111 (SD 11·7) g/l, respectively. Furthermore, the liver non-haem Fe of both supplement groups increased significantly (P<0·001). IMAAC was more effective at restoring Fe in the spleen compared with FeSO4 (P<0·005). Gene expression showed the IMAAC supplement absorption is regulated by the body’s Fe status as it significantly up-regulated hepcidin (P<0·001) and down-regulated duodenal cytochrome b mRNA (P<0·005), similar to the effects seen with FeSO4. A significant proportion of Fe in IMAAC is reduced by ascorbic acid. Fe absorption in mice and cells was similar for both IMAAC and FeSO4 and both compounds induce and regulate Fe metabolism genes similarly in the maintenance of homeostasis in mice.

The distribution of Hepatozoon canis mainly encompasses areas where its main tick vector, Rhipicephalus sanguineus sensu lato, is present. However, the detection of this pathogen in dogs, foxes and golden jackals well outside the areas inhabited by this tick species reinforced the hypothesis that additional ixodids are involved in the life cycle and transmission of this protozoon. The present study provides, for the first time, data supporting the sporogonic development of H. canis in specimens of Rhipicephalus turanicus collected from a naturally infected fox from southern Italy. The epidemiological role of R. turanicus as a vector of H. canis is discussed, along with information on the potential use of cell cultures for the experimental infection with H. canis sporozoites. The in vitro infection of canine leucocytes by sporozoites from ticks is proposed as a potential tool for future in-depth studies on the biology of H. canis.

The study of life remote in space has strong parallels with the study of life remote in time. Both are dependent on decoding those historic phenomena called ‘fossils’, here taken to include biogenic traces of activity and waste products. There is the shared problem of data restoration from incomplete data sets; the importance of contextual analysis of potentially viable habitats; the centrality of cell theory; the need to reject the null hypothesis of an abiogenic origin for candidate cells via morphospace analysis; the need to demonstrate biology-like behaviour (e.g., association with biofilm-like structures; tendency to form clusters and ‘mats’; and a preference for certain substrates), and of metabolism-like behaviour (e.g., within the candidate cell wall; within surrounding ‘waste products’; evidence for syntrophy and metabolic cycles; and evidence for metabolic tiers). We combine these ideas into a robust protocol for demonstrating ancient or extra-terrestrial life, drawing examples from Earth's early geological record, in particular from the earliest known freshwater communities of the 1.0 Ga Torridonian of Scotland, from the 1.9 Ga Gunflint Chert of Canada, from the 3.4 Ga Strelley Pool sandstone of Australia, and from the 3.46 Ga Apex Chert also of Australia.

Human clinical trials have shown that fish oils reduce the risk of a variety of disorders including CVD. Despite this, results have been inconsistent. Fish oils are easily oxidised and some fish oils contain higher than recommended levels of oxidised products, but their effects have not been investigated. Recent evidence indicates that dietary oxidised fats can contribute to the development of atherosclerosis and thrombosis. This review summarises findings from cellular, animal and human trials that have examined the effects of oxidised lipids and their potential to affect health outcomes, and proposes that oxidised products in fish oils may attenuate their beneficial effects. More research is required to determine the magnitude of negative effects of fish oil on health outcomes in clinical trials.

Isolation and culture of thymic epithelial cells (TECs) using conventional primary tissue culture techniques under conditions employing supplemented low calcium medium yielded an immortalized cell line derived from the LDA rat (Lewis [Rt1l] cross DA [Rt1a]) that could be manipulated in vitro. Thymi were harvested from 4–5-day-old neonates, enzymically digested using collagenase (1 mg/ml, 37°C, 1 h) and cultured in low calcium WAJC404A medium containing cholera toxin (20 ng/ml), dexamethasone (10 nM), epidermal growth factor (10 ng/ml), insulin (10 μg/ml), transferrin (10 μg/ml), 2% calf serum, 2.5% Dulbecco's Modified Eagle's Medium (DMEM), and 1% antibiotic/antimycotic. TECs cultured in low calcium displayed round to spindle-shaped morphology, distinct intercellular spaces (even at confluence), and dense reticular-like keratin patterns. In high calcium (0.188 mM), TECs formed cobblestone-like confluent monolayers that were resistant to trypsinization (0.05%) and displayed keratin intermediate filaments concentrated at desmosomal junctions between contiguous cells. Changes in cultured TEC morphology were quantified by an analysis of desmosome/membrane relationships in high and low calcium media. Desmosomes were significantly increased in the high calcium medium. These studies may have value when considering the growth conditions of cultured primary cell lines like TECs.

Laser capture microdissection uses a microscope to identify specific cells for microdissection and then a laser-sensitive plastic to capture and remove the cells from their substrate. This efficient capture method was originally developed to capture cells for genetic analysis. However, it has also been used to capture cells for proteonomic analysis. In this article, we extend the uses of laser-capture microdissection by reporting a method for preparing captured cells for ultrastructural analysis by transmission electron microscopy. Cells prepared by our methodology show good fine structure preservation and are easily sectioned by standard ultramicrotomy.

In this work a new concept of designing two degree of freedom (2-DOF) planar parallel manipulators (PPMs) is presented. With this design the manipulator's workspace can be increased by increasing the number of cells in the manipulator. A general dynamic model is formulated for the manipulator with any number of cells. The model is adapted for SCARA and ADEPT configurations, and a new approach for balancing these type of manipulators is proposed.