Anticancer activity of amphipathic barbiturates

Cancer is one of the leading causes of death worldwide. Challenges related to drug resistance, side effects, and poor response rates makes cancer treatment complicated. However, different forms of immunotherapy have been effective for some cancer patients. Oncolytic therapies which can be administered directly intratumorally represent a promising form of immunotherapy. The aim of oncolytic therapies is to kill cancer cells by inducing immunogenic cell death, which can activate a natural anti-tumor immune response. The current project focused on exploring the potential of amphipathic barbiturates, herein referred to as marine product mimics (MPMs), as novel oncolytic compounds for cancer treatment. The MPMs were developed with inspiration from antimicrobial peptides and the eusynstyelamides, which are a group of natural compounds that have previously been isolated from marine animals. It was demonstrated that the compound MPM-1 could kill a range of different cell types and induced a necrosis like death in the head and neck squamous cell carcinoma cell line HSC-3. Nine additional MPMs were synthesized and included in further studies. In vitro experimentation indicated that the MPMs have an intracellular target and that they accumulate in lysosomes, which could be part of their mechanism of inducing cell death. It was demonstrated that the MPMs could cause the release and exposure of damage-associated molecular patterns which are associated with immunogenic cell death, indicating that they could have the potential to be used in oncolytic cancer therapy. An in vivo study was performed where intratumoral injections of MPM-1 were administered to mice bearing B16F1 melanoma tumors. Complete tumor remission was achieved. However, significant protective effects against a rechallenge with the same cancer cells was not observed. In summary, the MPMs demonstrate potent anticancer activity but whether they have the potential to be used in treatment of human cancer remains to be seen

Exploring the in vitro expansion of CD4 T cells. For improved culturing of CD4 T cells linked to FNAIT

Fetal and neonatal alloimmune thrombocytopenia (FNAIT) is a rare disease that may cause serious bleedings in the fetus or neonate of a woman who has developed antibodies against the fetus’ platelets. Development of FNAIT has been linked to the presence of platelet reactive CD4 T cells that help B cells to develop into antibody producing plasma cells. To be able to conduct research on such T cells, the Immunology research group must be able to expand and keep them in long term cultures. Recent work revealed that several established T cell clones had started proliferating poorly. In an attempt to understand why and to possibly improve the culturing of future T cell clones, this study looked into some of the conditions that may influence the growth of these cells when expanded in vitro. The established anti- CD3 expansion culture protocol was compared to one that used PHA, but no advantage of using the latter was detected. It was demonstrated that different B-LCLs used as growth promoting feeder cells expressed varying levels of the surface molecules B7-1 and B7-2. This did however not seem to influence their feeder capacity despite the fact that expanding CD4 T cells were shown to express high levels of CD28, which costimulates growth when bound by B7. Expanding CD4 T cells also expressed the inhibitory molecule PD-1, and it was revealed that expression of its ligand, PD-L1, was induced in B-LCLs when used as feeder cells along with PBMCs. Whether this influences the efficiency of an expansion culture is yet to be determined

Reaching strategies of very preterm infants at 8 months corrected age

Reaching strategies and kinematics for a group of very preterm infants were investigated and compared with a group of full-term infants when reaching for a moving object. Eight-month-old (corrected-age) infants were presented with small toys moving on a semicircular path in the vertical plane. The trajectories of the target and the hands of the infants were measured using a 3D motion analysis system. No differences were found in how often the infants encountered the target. The very preterm group, however, used bimanual strategies more often and had more curved reaching paths than the full-term group. These results suggest that very preterm infants are equally successful as healthy full-term infants in catching a moving object but their reaching strategies are less efficient compared with full-term infants at 8 months (corrected age)

Occlusion Is Hard: Comparing Predictive Reaching for Visible and Hidden Objects in Infants and Adults

Infants can anticipate the future location of a moving object and execute a predictive reach to intercept the object. When a moving object is temporarily hidden by darkness or occlusion, 6-month-old infants’ reaching is perturbed, but performance on darkness trials is significantly better than occlusion trials. How does this reaching behavior change over development? Experiment 1 tested predictive reaching of 6- and 9-month-old infants. While there was an increase in the overall number of reaches with increasing age, there were significantly fewer predictive reaches during the occlusion compared to visible trials and no age-related changes in this pattern. The decrease in performance found in Experiment 1 is likely to apply not only to the object representations formed by infants but also those formed by adults. In Experiment 2 we tested adults with a similar reaching task. Like infants, the adults were most accurate when the target was continuously visible and performance in darkness trials was significantly better than occlusion trials, providing evidence that there is something specific about occlusion that makes it more difficult than merely lack of visibility. Together, these findings suggest that infants’ and adults’ capacities to represent objects have similar signatures throughout development.Psycholog

Quantitative Microanalysis of Nitrogen Distribution in Carpospores of Gracilaria secundata with Scanning Auger Microscopy

Quantitative microanalysis has been performed with a JAMP 10 Scanning Auger Microscope at 0.5 μm spot size, on carpospores of Gracilaria secundata Harv.. Samples were fixed in glutaraldehyde and osmium tetroxide and embedded in plastic material. Thin sections were placed on solid copper discs. the surfaces of which had been smoothed by electropolishing. Quantification of the nitrogen content was performed using a standard with known content of nitrogen. Nitrogen was localized in the nucleus, chloroplasts and osmiophilic inclusions of the carpospore. Trace amounts of nitrogen were also found in cell walls, starch grains and in the embedding material. The use of Scanning Auger Microscopy on biological material is discussed

Absence of spontaneous action anticipation by false belief attribution in children with autism spectrum disorder

Recently, a series of studies demonstrated false belief understanding in young children through completely nonverbal measures. These studies have revealed that children younger than 3 years of age, who consistently fail the standard verbal false belief test, can anticipate others' actions based on their attributed false beliefs. The current study examined whether children with autism spectrum disorder (ASD), who are known to have difficulties in the verbal false belief test, may also show such action anticipation in a nonverbal false belief test. We presented video stimuli of an actor watching an object being hidden in a box. The object was then displaced while the actor was looking away. We recorded children's eye movements and coded whether they spontaneously anticipated the actor's subsequent behavior, which could only have been predicted if they had attributed a false belief to her. Although typically developing children correctly anticipated the action, children with ASD failed to show such action anticipation. The results suggest that children with ASD have an impairment in false belief attribution, which is independent of their verbal ability

Molecular characterization and expression pattern of zona pellucida proteins in gilthead seabream (Sparus aurata)

The developing oocyte is surrounded by an acellular envelope that is composed of 2–4 isoforms of zona pellucida (ZP) proteins. The ZP proteins comprise the ZP1, ZP2, ZP3, and ZPX isoforms. While ZP1 (ZPB) and ZP3 (ZPC) are present in all species, ZP2 (ZPA) is not found in teleost fish and ZPX is not found in mammals. In the present study, we identify and characterize the ZP1, ZP3 and ZPX isoforms of gilthead seabream. Furthermore, by analyzing the conserved domains, which include the external hydrophobic patch and the internal hydrophobic patch, we show that ZP2 and ZPX are closely related isoforms. ZP proteins are synthesized in either the liver or ovary of most teleosts. Only in rainbow trout has it been shown that zp3 has dual transcription sites. In gilthead seabream, all four mRNA isoforms are transcribed in both the liver and ovary, with zp1a, zp1b, and zp3 being highly expressed in the liver, and zpx being primarily expressed in the ovary. However, determination of the ZP proteins in plasma showed high levels of ZP1b, ZP3, and ZPX, with low or non-detectable levels of ZP1a. In similarity to other teleost ZPs, the hepatic transcription of all four ZP isoforms is under estrogenic control. Previously, we have shown that cortisol can potentiate estrogen-induced ZP synthesis in salmonids, and now we show that this is not the case in the gilthead seabream. The present study shows for the first time the endocrine regulation of a teleost ZPX isoform, and demonstrates the dual-organ transcriptional activities of all the ZP proteins in one species

Imaging of Vascular Smooth Muscle Cells with Soft X-Ray Spectromicroscopy

Using X-ray microscopy and spectromicroscopy, vascular smooth muscle cells (VSMCs) were imaged, prepared without using additional embedding material or staining, but by applying simple, noncryo fixation techniques. The cells were imaged with a compact source transmission X-ray microscope and a scanning transmission X-ray microscope (STXM). With the STXM, spectromicroscopy was performed at the C K-edge and the Ca LIII,II-edges. VSMCs were chosen because of their high amount of actin stress fibers, so that the actin cytoskeleton should be visible. Other parts of the cell, such as the nucleus and organelles, were also identified from the micrographs. Both in the spectra and the images, the effects of the different preparation procedures were observable. Furthermore, Ca hotspots were detected and their density is determine

Using a head-mounted camera to infer attention direction

A head-mounted camera was used to measure head direction. The camera was mounted to the forehead of 20 6- and 20 12-month-old infants while they watched an object held at 11 horizontal (-80 degrees to + 80 degrees) and 9 vertical (-48 degrees to + 50 degrees) positions. The results showed that the head always moved less than required to be on target. Below 30 degrees in the horizontal dimension, the head undershoot of object direction was less than 5 degrees. At 80 degrees, however, the undershoot was substantial or between 10 degrees and 15 degrees. In the vertical dimension, the undershoot was larger than in the horizontal dimension. At 30 degrees, the undershoot was around 25% in the downward direction and around 40% in the upward direction. The size of the undershoot was quite consistent between conditions. It was concluded that the head-mounted camera is a useful indicator of horizontal looking direction in a free looking situation where the head is only turned moderately from a straight ahead position