Cancer cell sedimentation in 3D cultures reveals active migration regulated by self-generated gradients and adhesion sites.

Cell sedimentation in 3D hydrogel cultures refers to the vertical migration of cells towards the bottom of the space. Understanding this poorly examined phenomenon may allow us to design better protocols to prevent it, as well as provide insights into the mechanobiology of cancer development. We conducted a multiscale experimental and mathematical examination of 3D cancer growth in triple negative breast cancer cells. Migration was examined in the presence and absence of Paclitaxel, in high and low adhesion environments and in the presence of fibroblasts. The observed behaviour was modeled by hypothesizing active migration due to self-generated chemotactic gradients. Our results did not reject this hypothesis, whereby migration was likely to be regulated by the MAPK and TGF-ß pathways. The mathematical model enabled us to describe the experimental data in absence (normalized error<40%) and presence of Paclitaxel (normalized error<10%), suggesting inhibition of random motion and advection in the latter case. Inhibition of sedimentation in low adhesion and co-culture experiments further supported the conclusion that cells actively migrated downwards due to the presence of signals produced by cells already attached to the adhesive glass surface.

Electrically Switchable Polymer Brushes for Protein Capture and Release in Biological Environments.

Interfaces functionalized with polymers are known for providing excellent resistance towards biomolecular adsorption and for their ability to bind high amounts of protein while preserving their structure. However, making an interface that switches between these two states has proven challenging and concepts to date rely on changes in the physiochemical environment, which is static in biological systems. Here we present the first interface that can be electrically switched between a high-capacity (>1 µg cm-2 ) multilayer protein binding state and a completely non-fouling state (no detectable adsorption). Switching is possible over multiple cycles without any regeneration. Importantly, switching works even when the interface is in direct contact with biological fluids and a buffered environment. The technology offers many applications such as zero fouling on demand, patterning or separation of proteins as well as controlled release of biologics in a physiological environment, showing high potential for future drug delivery in vivo.

Structural genome analysis in cultivated potato taxa.

KEY MESSAGE: Twelve potato accessions were selected to represent two principal views on potato taxonomy. The genomes were sequenced and analyzed for structural variation (copy number variation) against three published potato genomes. The common potato (Solanum tuberosum L.) is an important staple crop with a highly heterozygous and complex tetraploid genome. The other taxa of cultivated potato contain varying ploidy levels (2X-5X), and structural variations are common in the genomes of these species, likely contributing to the diversification or agronomic traits during domestication. Increased understanding of the genomes and genomic variation will aid in the exploration of novel agronomic traits. Thus, sequencing data from twelve potato landraces, representing the four ploidy levels, were used to identify structural genomic variation compared to the two currently available reference genomes, a double monoploid potato genome and a diploid inbred clone of S. chacoense. The results of a copy number variation analysis showed that in the majority of the genomes, while the number of deletions is greater than the number of duplications, the number of duplicated genes is greater than the number of deleted ones. Specific regions in the twelve potato genomes have a high density of CNV events. Further, the auxin-induced SAUR genes (involved in abiotic stress), disease resistance genes and the 2-oxoglutarate/Fe(II)-dependent oxygenase superfamily proteins, among others, had increased copy numbers in these sequenced genomes relative to the references.

Neurodevelopmental Outcome in Extremely Low Birth Weight Infants at 2-3 Years of Age.

Background and objectives: The aims of this study were to examine the relationship between neurological outcomes at 3- and 6-months corrected age with the neurodevelopmental outcome at 3 years of age; to identify the perinatal/neonatal risk factors for poor neurodevelopmental outcomes at 3 years of age. Materials and methods: In our single-centre longitudinal cohort study, of the 73 consecutive infants admitted to our Neonatal Intensive Care Unit (NICU), 49 infants (80%) received both Hammersmith Infant Neurological Examination (HINE) at 3- and 6-months corrected age and Bayley-III neurodevelopmental assessment at 2-3 years chronological age. At 3 months follow up, 8.2% had suboptimal scores (below 10th percentile) on the HINE. At 6 months follow up, 4.1% had suboptimal scores (below 10th percentile) on the HINE. The means(±SD) for Bayley-III cognitive, language, and motor subscales were (96.3 ± 9.8), (99.9 ± 11.9), (93.2 ± 9.9). Results: At 3 months corrected age, higher total HINE scores and subscores for function of cranial nerves, posture, tone, were associated with better cognitive scores while poorer scores for function of cranial nerves, posture, movements, tone, and total HINE score were associated with lower motor scores. Infants with a HINE subscore of function of cranial nerves in the suboptimal range have three times higher odds of having a motor delay. Infants with a HINE subscore of function of cranial nerves in the suboptimal range have more than two times higher odds of having a language delay. At 6 months corrected age, poorer scores for function of cranial nerves, movements, tone, reflexes, and total HINE score were associated with worse Bayley-III motor scores whilst infants who have a total HINE score and a subscore of reflexes in the suboptimal range have four and seven times, respectively, higher odds of having a motor delay. Conclusions: Early identification of infants at risk for adverse long-term outcomes is essential in introducing early intervention therapies for optimizing neurodevelopmental outcomes.

Excessive Knee Internal Rotation during Grand Plié in Classical Ballet Female Dancers.

Classical ballet dancers are exposed daily to physically demanding movements. Among these, the Grand Plié stands out for its biomechanical complexity, particularly the stress applied to the knee joint. This study investigates the knee kinematics of healthy professional classical ballet dancers performing the Grand Plié. Twenty dancers were evaluated with a motion analysis system using a marker-based protocol. Before measurements, the self-reported Global Knee Functional Assessment Scale was delivered for the knees' functional ability, and the passive range of knee motion was also assessed. The average score on the Global Knee Functional Assessment Scale was 94.65 ± 5.92. During a complete circle of the Grand Plié movement, executed from the upright position, the average maximum internal rotation of the knee joint was 30.28° ± 6.16°, with a simultaneous knee flexion of 134.98° ± 4.62°. This internal rotation observed during knee flexion exceeds the typical range of motion for the joint, suggesting a potential risk for knee injuries, such as meniscal tears. The findings provide an opportunity for future kinematic analysis research, focusing on the movement of the Grand Plié and other common ballet maneuvers. These data have the potential to yield valuable information about the knee kinematics concerning meniscus damage.

Ηand dexterities assessment in stroke patients based on augmented reality and machine learning through a box and block test.

A popular and widely suggested measure for assessing unilateral hand motor skills in stroke patients is the box and block test (BBT). Our study aimed to create an augmented reality enhanced version of the BBT (AR-BBT) and evaluate its correlation to the original BBT for stroke patients. Following G-power analysis, clinical examination, and inclusion-exclusion criteria, 31 stroke patients were included in this study. AR-BBT was developed using the Open Source Computer Vision Library (OpenCV). The MediaPipe's hand tracking library uses a palm and a hand landmark machine learning model to detect and track hands. A computer and a depth camera were employed in the clinical evaluation of AR-BBT following the principles of traditional BBT. A strong correlation was achieved between the number of blocks moved in the BBT and the AR-BBT on the hemiplegic side (Pearson correlation = 0.918) and a positive statistically significant correlation (p = 0.000008). The conventional BBT is currently the preferred assessment method. However, our approach offers an advantage, as it suggests that an AR-BBT solution could remotely monitor the assessment of a home-based rehabilitation program and provide additional hand kinematic information for hand dexterities in AR environment conditions. Furthermore, it employs minimal hardware equipment.

Relationship between two forms of impulsivity in mice at baseline and under acute and sub-chronic atomoxetine treatment.

RATIONALE: Impulsivity is a symptom of various mental disorders, including attention deficit hyperactivity disorder (ADHD), bipolar disorder, and addiction. Impulsivity is not a unitary construct, but is present in different forms, yet only a few rodent studies have explored the relationship between these forms within individual subjects. OBJECTIVES: In this study, we compared behaviors representing two impulsivity forms, delay discounting (choice impulsivity) and premature responding (waiting impulsivity), within the same mice. METHODS: C57BL/6J male mice were concurrently trained and tested in the delay discounting task and the rodent continuous performance test in a counterbalanced design. The effects of the ADHD medication atomoxetine were tested in both tasks, after both acute (0.3-5.0 mg/kg) and sub-chronic (0.3 mg/kg twice daily for seven days) administration. RESULTS: There was no correlation between the two impulsivity forms at baseline. Acute atomoxetine treatment (1, 3, and 5 mg/kg) significantly reduced premature responding. Furthermore, sub-chronic treatment with 0.3 mg/kg of atomoxetine caused a stable decrease in premature responding. Atomoxetine had no significant effect on delay discounting after acute or sub-chronic administration, although the acute administration of 1 mg/kg showed a trend towards increasing delay discounting. CONCLUSIONS: The present results support that delay discounting and premature responding represent two different forms of impulsivity that show dissimilar responses to atomoxetine treatment. The consistency with findings in humans lends support to the translatability of the results in mice.

Lumbar Kinematics Assessment of Patients with Chronic Low Back Pain in Three Bridge Tests Using Miniaturized Sensors.

Lumbar muscle atrophy, diminished strength, stamina, and increased fatigability have been associated with chronic nonspecific low back pain (LBP). When evaluating patients with LBP, trunk or core stability, provided by the performance and coordination of trunk muscles, appears to be essential. Several clinical tests have been developed to identify deficiencies in trunk performance, demonstrating high levels of validity and reproducibility. The most frequently prescribed tests for assessing the core body muscles are the prone plank bridge test (PBT), the side bridge test (SBT), and the supine bridge test (SUBT). However, quantitative assessments of the kinematics of the lumbar spine during their execution have not yet been conducted. The purpose of our study was to provide objective biomechanical data for the assessment of LBP patients. The lumbar spine ranges of motion of 22 healthy subjects (Group A) and 25 patients diagnosed with chronic LBP (Group B) were measured using two inertial measurement units during the execution of the PBT, SUBT, and SBT. Statistically significant differences between the two groups were found in all three tests' kinematic patterns. This quantitative assessment of lumbar spine motion transforms the three bridge tests into an objective biomechanical diagnostic tool for LPBs that may be used to assess the efficacy of applied rehabilitation programs.

Expert voices in the news reporting of the coronavirus pandemic: A study of UK television news bulletins and their audiences.

The study examines the role of experts in UK television news at the start of the coronavirus pandemic by analysing both how they were used in coverage and perceived by news audiences. Our systematic content analysis of sources (N = 2300) used in the UK's flagship evening news bulletins found a reliance on political sources, principally from the government's perspective. We also discovered health and scientific experts received limited coverage and were only occasionally used to scrutinise public health policy. Yet, our six-week online diary study with 175 participants identified a strong preference for expert views about how the pandemic was being handled. It showed audiences favoured a range of expert sources in routine reporting - balancing government appointed and independent experts - to provide evidence-based scrutiny of the executive's decision-making. Overall, our findings contribute to a greater understanding of audience expectations, opinions, and experiences with broadcast news during a major public health crisis.

Application of Wearable Sensors Technology for Lumbar Spine Kinematic Measurements during Daily Activities following Microdiscectomy Due to Severe Sciatica.

BACKGROUND: The recurrence rate of lumbar spine microdiscectomies (rLSMs) is estimated to be 5-15%. Lumbar spine flexion (LSF) of more than 10° is mentioned as the most harmful load to the intervertebral disc that could lead to recurrence during the first six postoperative weeks. The purpose of this study is to quantify LSFs, following LSM, at the period of six weeks postoperatively. METHODS: LSFs were recorded during the daily activities of 69 subjects for 24 h twice per week, using Inertial Measurement Units (IMU). RESULTS: The mean number of more than 10 degrees of LSFs per hour were: 41.3/h during the 1st postoperative week (P.W.) (29.9% healthy subjects-H.S.), 2nd P.W. 60.1/h (43.5% H.S.), 3rd P.W. 74.2/h (53.7% H.S.), 4th P.W. 82.9/h (60% H.S.), 5th P.W. 97.3/h (70.4% H.S.) and 6th P.W. 105.5/h (76.4% H.S.). CONCLUSIONS: LSFs constitute important risk factors for rLDH. Our study records the lumbar spine kinematic pattern of such patients for the first time during their daily activities. Patients' data report less sagittal plane movements than healthy subjects. In vitro studies should be carried out, replicating our results to identify if such a kinematic pattern could cause rLDH. Furthermore, IMU biofeedback capabilities could protect patients from such harmful movements.

Genome sequencing of adapted diploid potato clones.

Cultivated potato is a vegetatively propagated crop, and most varieties are autotetraploid with high levels of heterozygosity. Reducing the ploidy and breeding potato at the diploid level can increase efficiency for genetic improvement including greater ease of introgression of diploid wild relatives and more efficient use of genomics and markers in selection. More recently, selfing of diploids for generation of inbred lines for F1 hybrid breeding has had a lot of attention in potato. The current study provides genomics resources for nine legacy non-inbred adapted diploid potato clones developed at Agriculture and Agri-Food Canada. De novo genome sequence assembly using 10× Genomics and Illumina sequencing technologies show the genome sizes ranged from 712 to 948 Mbp. Structural variation was identified by comparison to two references, the potato DMv6.1 genome and the phased RHv3 genome, and a k-mer based analysis of sequence reads showed the genome heterozygosity range of 1 to 9.04% between clones. A genome-wide approach was taken to scan 5 Mb bins to visualize patterns of heterozygous deleterious alleles. These were found dispersed throughout the genome including regions overlapping segregation distortions. Novel variants of the StCDF1 gene conferring earliness of tuberization were found among these clones, which all produce tubers under long days. The genomes will be useful tools for genome design for potato breeding.

Phased, chromosome-scale genome assemblies of tetraploid potato reveal a complex genome, transcriptome, and predicted proteome landscape underpinning genetic diversity.

Cultivated potato is a clonally propagated autotetraploid species with a highly heterogeneous genome. Phased assemblies of six cultivars including two chromosome-scale phased genome assemblies revealed extensive allelic diversity, including altered coding and transcript sequences, preferential allele expression, and structural variation that collectively result in a highly complex transcriptome and predicted proteome, which are distributed across the homologous chromosomes. Wild species contribute to the extensive allelic diversity in tetraploid cultivars, demonstrating ancestral introgressions predating modern breeding efforts. As a clonally propagated autotetraploid that undergoes limited meiosis, dysfunctional and deleterious alleles are not purged in tetraploid potato. Nearly a quarter of the loci bore mutations are predicted to have a high negative impact on protein function, complicating breeder's efforts to reduce genetic load. The StCDF1 locus controls maturity, and analysis of six tetraploid genomes revealed that 12 allelic variants of StCDF1 are correlated with maturity in a dosage-dependent manner. Knowledge of the complexity of the tetraploid potato genome with its rampant structural variation and embedded deleterious and dysfunctional alleles will be key not only to implementing precision breeding of tetraploid cultivars but also to the construction of homozygous, diploid potato germplasm containing favorable alleles to capitalize on heterosis in F1 hybrids.

Early versus delayed minimal enteral feeding and risk for necrotizing enterocolitis in preterm growth-restricted infants with abnormal antenatal Doppler results.

We studied the effect of early (< or = 5 days) versus delayed (> or = 6 days) initiation of minimal enteral feeding (MEF) on the incidence of necrotizing enterocolitis (NEC) and feeding intolerance in preterm infants with intrauterine growth restriction (IUGR) and abnormal antenatal Doppler results. We performed a randomized, nonblinded pilot trial of infants receiving early or delayed MEF in addition to parenteral feeding within 48 hours of life. Demographic data, maternal preeclampsia, antenatal steroid exposure, Doppler studies, as well as cases of NEC and feeding intolerance were all recorded. Of the 84 infants enrolled, 81 completed the study: 40 received early (median age: 2 days, range: 1 to 5 days) and 41 delayed (median age: 7 days, range: 6 to 14 days) MEF. The incidence of NEC and feeding intolerance was not significantly different between groups (p = 0.353 and p = 0.533, respectively). Birth weight was an independent risk factor for NEC in both groups. Early MEF of preterm infants with IUGR and abnormal antenatal Doppler results may not have a significant effect on the incidence of NEC or feeding intolerance. Furthermore, birth weight seems to be an independent risk factor for the development of NEC, irrespectively of the timing of MEF introduction.

Genome assembly of six polyploid potato genomes.

Genome assembly of polyploid plant genomes is a laborious task as they contain more than two copies of the genome, are often highly heterozygous with a high level of repetitive DNA. Next Generation genome sequencing data representing one Chilean and five Peruvian polyploid potato (Solanum spp.) landrace genomes was used to construct genome assemblies comprising five taxa. Third Generation sequencing data (Linked and Long-read data) was used to improve the assembly for one of the genomes. Native landraces are valuable genetic resources for traits such as disease and pest resistance, environmental tolerance and other qualities of interest such as nutrition and fiber for breeding programs. The need for conservation and enhanced understanding of genetic diversity of cultivated potato from South America is also crucial to North American and European cultivars. Here, we report draft genomes from six polyploid potato landraces representing five taxa, illustrating how Third Generation Sequencing can aid in assembling polyploid genomes.

Complete plastome assemblies from a panel of 13 diverse potato taxa.

The chloroplasts are a crucial part of photosynthesizing plant cells and are extensively utilized in phylogenetic studies mainly due to their maternal inheritance. Characterization and analysis of complete plastome sequences is necessary to understand their diversity and evolutionary relationships. Here, a panel of thirteen plastomes from various potato taxa are presented. Though they are highly similar with respect to gene order and content, there is also a great extent of SNPs and InDels between them, with one of the Solanum bukasovii plastomes (BUK2) having the highest number of SNPs and InDels. Five different potato plastome types (C, S, A, W, W2) are present in the panel. Interestingly, the S. tuberosum subsp. tuberosum (TBR) accession has a W-type plastome, which is not commonly found in this species. The S-type plastome has a conserved 48 bp deletion not found in other types, which is responsible for the divergence of the S-type from the C-type plastome. Finally, a phylogenetic analysis shows that these plastomes cluster according to their types. Congruence between the nuclear genome and the plastome phylogeny of these accessions was seen, however with considerable differences, supporting the hypothesis of introgression and hybridization between potato species.

Infrared and Raman Spectroscopic Studies of Molecular Disorders in Skin Cancer.

AIM: To investigate the molecular structural disorders of cancerous skin. MATERIALS AND METHODS: Human malignant melanoma and basal cell carcinoma biopsies were used for the investigation. Fourier transform infrared (FT-IR), Raman spectroscopy, and scanning electron microscopy were utilized. Spectral differences between healthy, basal cell carcinoma and melanoma tissues were recorded. RESULTS: The FT-IR bands of vasCH2, vsCH2 and Raman vsCH3 of cell membrane lipids were increased in intensity in melanoma due to an increased lipophilic environment. The FT-IR band at 1,744 cm-1 assigned to malondialdehyde can be used as a band diagnostic of cancer progression. The amide I bands at 1,654 cm-1 and 1,650 cm-1 for Raman and FT-IR, respectively were broader in spectra from melanoma, reflecting changes of protein secondary structure from α-helix to ß-sheet and random coil. The intensity of the FT-IR band at 1,046 cm-1 was increased in melanoma, suggesting glycosylation of the skin upon cancer development. Another band that might be considered as diagnostic was found at about 815 cm-1 in melanoma and was attributed to Z-DNA configuration. As far as we know, this is the first time that scanning electron microscopy revealed that metal components of titanium alloys from tooth implants were transferred to melanoma tissue taken from the back of one patient. CONCLUSION: Vibrational spectroscopy highlighted increased glycosylation in melanoma.

Neurological outcome at 6 and 12 months corrected age in hospitalised late preterm infants -a prospective study.

Late preterm infants (34-0/7 to 36-6/7 weeks" gestation) account for 10-20% of NICU admissions and are at increased risk for morbidity and mortality. Although they are prone to developmental delays, reports on neurological outcome during the first 2 years of life are scarce. The aim of the study was to assess neurological/neuromotor outcome in high risk late preterm infants at 6 and 12 months corrected age and the change in neurological scores over time, and to identify factors associated with the neurological outcome. The Hammersmith Infant Neurological Examination was performed in a cohort of 157 late preterm infants admitted in the NICU. The infants were examined at 6 and 12 months corrected age respectively and scored with the optimality score system including 26 items assessing cranial nerve function, posture, movements, tone and reflexes. Also parents reported neurological milestones in the follow up visit. Infants at 6 months had a global score of 59 (47-76) and optimal scores achieved in 25.4%. At 12 months they had a global score of 70 (58-78) and achieved optimal scores in 63.2%. The subscores of posture, tone and reflexes gradually increased from 6 to 12 months corrected age. Being born small for gestational age was the only factor that adversely influenced HINE score at 6 and 12 months. At 12 months 58.5% achieved independent walking. High risk late preterm infants have suboptimal HINE scores at 6 and 12 months of age, suggesting a need for closer follow up and early intervention programs.

Current Strategies of Polyploid Plant Genome Sequence Assembly.

Polyploidy or duplication of an entire genome occurs in the majority of angiosperms. The understanding of polyploid genomes is important for the improvement of those crops, which humans rely on for sustenance and basic nutrition. As climate change continues to pose a potential threat to agricultural production, there will increasingly be a demand for plant cultivars that can resist biotic and abiotic stresses and also provide needed and improved nutrition. In the past decade, Next Generation Sequencing (NGS) has fundamentally changed the genomics landscape by providing tools for the exploration of polyploid genomes. Here, we review the challenges of the assembly of polyploid plant genomes, and also present recent advances in genomic resources and functional tools in molecular genetics and breeding. As genomes of diploid and less heterozygous progenitor species are increasingly available, we discuss the lack of complexity of these currently available reference genomes as they relate to polyploid crops. Finally, we review recent approaches of haplotyping by phasing and the impact of third generation technologies on polyploid plant genome assembly.

The TOBg Tobacco Treatment Guidelines for Adolescents: A real-world pilot study.

INTRODUCTION: We conducted a pilot study to: 1) obtain feedback from prevention practitioners in terms of their satisfaction, knowledge, and self-efficacy following exposure to the Tobacco Treatment Guidelines for Adolescents (TOBg Guidelines); and 2) examine the effectiveness of a school-based intervention based on the TOBg Guidelines on quit rates among a sample of adolescent tobacco users. METHODS: Two parallel studies were conducted. In Study 1, prevention practitioners were exposed to a 1-day training in the TOBg Guidelines with assessment occurring before, immediately after, and at 6 months following the training. In Study 2, participating adolescent smokers were exposed to a 3-session group-based smoking cessation intervention that drew on the TOBg Guidelines and was delivered by practitioners trained in Study 1. The primary outcome measure was self-reported smoking status assessed at 1 month and at 6 months following baseline. RESULTS: A total of 18 prevention practitioners and 65 adolescent tobacco users participated in the pilot study. The majority of practitioners reported high rates of satisfaction with the TOBg Guidelines and indicated that the guidelines positively influenced the manner in which they addressed tobacco use with adolescents. Prevention practitioners' self-efficacy for intervening with adolescent smokers was also significantly increased following exposure to the TOBg Guidelines and training. Among adolescents exposed to the school-based intervention, 62.5% and 23.1% had reduced smoking by 50% or more at 1 month and at 6 months follow-up, respectively. No significant change in smoking abstinence was documented. CONCLUSIONS: The TOBg Guidelines for adolescent smokers were well received by prevention practitioners and were feasible to implement in a real-world school setting.

Oxidative stress in ageing and disease development studied by FT-IR spectroscopy.

FT-IR spectroscopy was used to investigate the effect of oxidative stress and to approach the mechanism on cancer bone demineralization, aortic valve mineralization and heterotopic ossification on disease development. The FT-IR spectra obtained from paediatric, adult bone and ex vivo irradiated adult healthy bone with a dose of 20Gy were compared with those of healthy bone. The increase of band intensity changes of vasCH2,vsCH2 in the region 3000-2850cm-1 depended on aging, the disease progression and the dose of irradiation. The bands at 3080cm-1 and 1744cm-1, which originate from olefinic terminal bond (v=CH) and ester carbonyl group (vROCO), respectively, indicate the influence of oxidative stress on lipid degradation and peroxidation, respectively. The new bands at about 1690cm-1 and 1516cm-1 denote the presence of ß-sheet conformation of the proteins due to the diseases, confirming the increasing amount of lipophilic environment and fibril formation. Comparison of the FT-IR spectra of calcified aortic valve and hip heterotopic ossification with that of normal bones showed that in the bone-like formation the peroxide anion free radicals play an important role in the disease.