Deciphering tumour tissue organization by 3D electron microscopy and machine learning

Despite recent progress in the characterization of tumour components, the tri-dimensional (3D) organization of this pathological tissue and the parameters determining its internal architecture remain elusive. Here, we analysed the spatial organization of patient-derived xenograft tissues generated from hepatoblastoma, the most frequent childhood liver tumour, by serial block-face scanning electron microscopy using an integrated workflow combining 3D imaging, manual and machine learning-based semi-automatic segmentations, mathematics and infographics. By digitally reconstituting an entire hepatoblastoma sample with a blood capillary, a bile canaliculus-like structure, hundreds of tumour cells and their main organelles (e.g. cytoplasm, nucleus, mitochondria), we report unique 3D ultrastructural data about the organization of tumour tissue. We found that the size of hepatoblastoma cells correlates with the size of their nucleus, cytoplasm and mitochondrial mass. We also found anatomical connections between the blood capillary and the planar alignment and size of tumour cells in their 3D milieu. Finally, a set of tumour cells polarized in the direction of a hot spot corresponding to a bile canaliculus-like structure. In conclusion, this pilot study allowed the identification of bioarchitectural parameters that shape the internal and spatial organization of tumours, thus paving the way for future investigations in the emerging onconanotomy field.

Nuclear progesterone receptor regulates blood coagulation facilitating ovulation in zebrafish

Ovulation is a remodeling process including blood capillary rupture and coagulation. Until now, there is no regulation and functional studies of coagulation factors in ovulation. Here, we report dramatic increases of coagulation factors (f5, f3a) in zebrafish preovulatory follicles. This upregulation was induced by progestin (DHP: 17α, 20β-dihydroxy-4-pregnen-3-one), a native ligand for nuclear progestin receptor (Pgr) that is essential for ovulation in zebrafish; but was abolished in pgr-/-. In addition, promoter activities of f5 and f3a were significantly enhanced by progestin via zebrafish Pgr. Similarly, we found promoter activities of human F5 were significantly stimulated by progesterone (P4) via human PGRB. Moreover, a dramatic increase of erythrocyte numbers in capillaries on ovarian follicles was associated with ovulation. Importantly, heparin, an anticoagulant, inhibited ovulation. Furthermore, reduced fecundity and impaired ovulation were observed in f5+/- female zebrafish. Together, our results provide plausible evidence for an exceptional function of coagulation factors in ovulation.

A Preliminary Study of Oxygen Transport from Capillary to Tissue using Finite Difference Method

In this study, Finite Difference Method is used to examine oxygen transport from blood capillary to tissue. Tissue is modelled as porous media and oxygen rich blood is transported throughout the tissue from the blood vessel by advection and diffusion. Uniform consumption by tissue is considered for simplicity. Steady state flow patterns and concentration contours are discussed. A simplistic 2-D model was created after reviewing the literature. The model considers interlinkage of capillaries. Stream Function-Vorticity formulation coupled with advection-diffusion equation are solved simultaneously to evaluate velocities and concentration values in the domain. This report focuses on the how various parameters affect flow and mass transfer inside a porous media. Effect of various non dimensional parameters like Darcy number and non-dimensional absorption coefficient are discussed.

Deciphering Tumour Tissue Organization by 3D Electron Microscopy and machine learning

Despite recent progress in the characterization of tumour components, the tri-dimensional (3D) organization of this pathological tissue and the parameters determining its internal architecture remain elusive. Here, we analysed the spatial organization of patient-derived xenograft tissues generated from hepatoblastoma, the most frequent childhood liver tumour, by serial block-face scanning electron microscopy using an integrated workflow combining 3D imaging, manual and machine learning-based semi-automatic segmentations, mathematics and infographics. By digitally reconstituting an entire hepatoblastoma sample with a blood capillary, a bile canaliculus-like structure, hundreds of tumour cells and their main organelles (e.g. cytoplasm, nucleus, mitochondria), we report unique 3D ultrastructural data about the organization of tumoral tissue. We found that the size of hepatoblastoma cells correlates with the size of their nucleus, cytoplasm and mitochondrial mass. We also discovered that the blood capillary controls the planar alignment and size of tumour cells in their 3D milieu. Finally, a set of tumour cells polarized in the direction of a hot spot corresponding to a bile canaliculus-like structure. In conclusion, this pilot study allowed the identification of bioarchitectural parameters that shape the internal and spatial organization of tumours, thus paving the way for new investigations in an emerging field that we call onconanotomy.

Solving the gap between HCV detection and treatment in prison. HCV-RNA testing and treatment in a cohort of new arriving convicts in Southern Italy

Introduction: HCV infection elimination is set to be carried out by 2030. To achieve this goal, the WHO has set minor achievable short-term “mini-goals.” One of these is the treatment of “difficult to reach and treat populations,” such as prisoners. One of the biggest obstacles to reaching this mini goal is the poor knowledge of the real HCV prevalence in such a population and the barriers to its detection, treatment, and follow-up. Even if HCV testing in Italian prisons is feasible and recommended, it is not always carried out. To worsen the picture, the peculiar status of conviction is correlated with the difficulty in carrying out the antiviral therapy due to challenges in follow-up and the refusals by inmates. Aims: A point-of-care test-and-treat program was set up in a penitentiary in Southern Italy to reduce the number of patients lost-to-follow-up (LTFU) between detection and treatment. A secondary aim was to evaluate the prevalence of HCV-infected patients in a cohort of new inmates. Methods: This prospective-observational study was carried out from January 2020 to February 2020. We performed a quick HCV-RNA blood capillary test on all new arriving inmates. As a routine, the new inmates underwent clinical and laboratory assessments. To those who were detected as HCV-RNA positive, the shortest possible antiviral treatment was offered, according to genotype and clinical features. Results: We observed 122 new inmates in the period between January and February of 2020. Overall, 62 (50.8%) subjects accepted HCV-RNA quick testing through blood sampling. Four (6.4%) subjects were found to be HCV-RNA positive; 1 refused antiviral therapy while 3 accepted, obtaining 100% SVR. None of the HCV-active inmates were lost-to-follow-up between detection and treatment proposal. Conclusion: The use of a very fast test-and-treat protocol for HCV infection demonstrated to be effective to avoid LTFU in HCV-positive new inmates in the period between detection and treatment. We observed an apparent prevalence of HCV incident cases in newly arriving inmates of 6.4%. Antiviral therapy was quickly provided, secure, and successful.

Dio2-CreErt2 Knockin Model to Identify T3-Generating Cells That Express Type 2 Deiodinase in Tissues

Background: Thyroid hormone promotes many developmental and homeostatic functions. Apart from adequate circulating levels, the concentration of the active hormone T3 within tissues may be amplified by type 2 deiodinase (Dio2) by conversion from the precursor T4. Dio2 is critical in auditory development, bone maturation, brain function and control of the hypothalamic-pituitary-thyroid axis. Despite its crucial role, an obstacle to studying Dio2 is that the protein has a short half-life, is at low levels and is often transiently expressed, making it difficult to identify Dio2 in tissues at cellular resolution. Methods: We derived a Dio2-CreERt2 knockin mouse that expresses tamoxifen-dependent Cre recombinase from the endogenous Dio2 gene. When crossed onto Ai6 reporter mice, following tamoxifen treatment, Dio2-CreERt2 expression is detected as fluorescent signal in specific cells in brain regions, pituitary, and other tissues. We showed previously that Dio2 is essential for hearing with rising expression levels in the cochlea prior to onset of hearing. The Dio2-CreERt2 model identified positive cell types in the cochlear spiral ligament, septal divisions and modiolus around the sensory epithelium. Dio2-positive fibrocytes were adjacent to and extended projections around blood capillary networks, the source of T4 supply. Transcriptome analysis of isolated positive cells revealed bone lineage-related origins for many of these cells. Conclusion: The Dio2-CreERt2 model detects Dio2 expression sensitively at cellular resolution. In the cochlea, Dio2-positive cell types reside in vascularized support tissues, suggesting combined endocrine and paracrine-like control of the T3 supply. Analysis of cell origins suggests novel interactions between endocrine and skeletal systems in promoting T3 action required for hearing.

The use of whole blood capillary samples to measure 15 analytes for a home-collect biochemistry service during the SARS-CoV-2 pandemic: A proposed model from North West London Pathology

Background The COVID-19 pandemic has drastically changed the delivery of secondary care services. Self-collection of capillary blood at home can facilitate the monitoring of patients with chronic disease to support virtual clinics while mitigating the risk of SARS-CoV-2 infection and transmission. Objective To investigate the comparability of whole blood capillary and plasma venous samples for 15 routinely used biochemical analytes and to develop and pilot a user-friendly home-collection kit to support virtual outpatient clinical services. Methods To investigate the comparability of whole blood capillary and plasma venous samples for 15 routinely requested biochemical analytes, simultaneous samples of venous and capillary blood were collected in EDTA and lithium-heparin plasma separation tubes that were of 4–6 mL and 400–600 µL draw volume, respectively. Venous samples were analysed within 4 h of collection while capillary samples were kept at ambient temperature for three days until centrifugation and analysis. Analyte results that were comparable between the matrices were then piloted in a feasibility study in three outpatient clinical services. Results HbA1c, lipid profile and liver function tests were considered comparable and piloted in the patient feasibility study. The home-collect kit demonstrated good patient usability. Conclusion Home collection of capillary blood could be a clinically-useful tool to deliver virtual care to patients with chronic disease.

Lophiosilurus alexandri, a sedentary bottom fish, adjusts its physiological parameters to survive to hypoxia condition.

We investigated blood gas, hematological and biochemical parameters and gill morphology and morphometry of Lophiosilurus alexandri juveniles submitted to hypoxia for 48 hours followed by recovery for 48 hours. A total of 48 juveniles (360.0 ± 141.6 g) were distributed among eight tanks (120 L) and subjected to hypoxia condition (water with dissolved oxygen at 2.12 ± 0.90 mg L− 1) or normoxia (at 5.60 ± 0.31 mg L− 1). Blood gas values (pH, PvCO2, PvO2, sO2, HCO3−, stHCO3− and base excess) in hypoxia were significantly different from normoxia, while lactate and the electrolytes (K+, Na+, Cl−, Ca2+ and HCO3−) there was no significant change among treatments. The erythrocytes differed significantly between hypoxia and normoxia at 24 h of recovery, while for hemoglobin and hematocrit there were no significant differences. There was a significant difference in glucose, triglycerides, and cholesterol for both normoxia and hypoxia, while plasma protein remained unchanged. All gill components (epithelial cells, erythrocytes, pillar cells, mucus cells, chloride cells, undifferentiated cells, and blood capillary lumen) differed significantly between hypoxia and normoxia. A reduction in the length of the primary lamella was observed in the hypoxia and recovery treatments, when compared to normoxia. The secondary branchial lamella showed no significant difference for both treatments. In general, juveniles of L. alexandri adapted well to hypoxia exposure for 48 h, as they were able to adjust most of their physiological variables to survive this stress condition and return to normoxia within 48 h.

IDENTIFYING THE MENTAL STATUS OF BEDRIDDEN ELDERLY BY THE BODY TEMPERATURE, HEART RATE, AND SKIN CONDUCTIVITY USING AN ARDUINO SENSOR MODULE

Addressing the emotional and mental health of the bedridden elderly is necessary as they are more likely to be depressed being isolated and dependent on a caregiver for a prolonged time. Several studies have been carried out to identify the mental stress of patients through their skin conductivity. The variations in the sympathetic nervous system reflect the emotional state of a person. This is demonstrated by the Galvanic Skin Response and thus can be used as a denotation of psychological or physiological arousal. Such arousal causes the blood capillary dilation, increment of sweat gland activities making the skin further conductive to electricity. In this study we develop a sensor module composed of a Galvanic Skin Response sensor for the bed ridden elderly and identify the relationship between body temperature, heart rate and GSR of them. The experiment is conducted upon 10 bed ridden elderly aged from 60 – 80 years of the Mihinthale region. The observations demonstrate a correlation between the heart rate, body temperature, skin conductivity and the human physiological states.