A New Protocol of Computer-Assisted Image Analysis Highlights the Presence of Hemocytes in the Regenerating Cephalic Tentacles of Adult Pomacea canaliculata

In humans, injuries and diseases can result in irreversible tissue or organ loss. This well-known fact has prompted several basic studies on organisms capable of adult regeneration, such as amphibians, bony fish, and invertebrates. These studies have provided important biological information and helped to develop regenerative medicine therapies, but important gaps concerning the regulation of tissue and organ regeneration remain to be elucidated. To this aim, new models for studying regenerative biology could prove helpful. Here, the description of the cephalic tentacle regeneration in the adult of the freshwater snail Pomacea canaliculata is presented. In this invasive mollusk, the whole tentacle is reconstructed within 3 months. Regenerating epithelial, connective, muscular and neural components are already recognizable 72 h post-amputation (hpa). Only in the early phases of regeneration, several hemocytes are retrieved in the forming blastema. In view of quantifying the hemocytes retrieved in regenerating organs, granular hemocytes present in the tentacle blastema at 12 hpa were counted, with a new and specific computer-assisted image analysis protocol. Since it can be applied in absence of specific cell markers and after a common hematoxylin-eosin staining, this protocol could prove helpful to evidence and count the hemocytes interspersed among regenerating tissues, helping to unveil the role of immune-related cells in sensory organ regeneration.

The Second‐to‐Fourth Digit (2D:4D) Ratio of Male Combat Athletes is Associated with the Choice of Sport

The second-to-fourth-digit (2D:4D) ratio has been widely used as a putative marker of prenatal exposure to testosterone in health, behavioral and sport sciences, but it has only been used few times regarding combat athletes. This study involved 200 male elite combat athletes (Olympic wrestling, kickboxing, judo, taekwondo and karate) and 179 males not participating in any sports. The lengths of the index finger (2D) and ring (4D) finger were measured using computer-assisted image analysis (AutoMetric 2.2 software). The 2D:4D ratio of combat athletes was significantly lower than that of the controls. Moreover, a lower 2D:4D ratio was found among wrestlers, judo athletes and kickboxers than in the control group, and a higher 2D:4D ratio was found, but with borderline significance, among karate and taekwondo athletes. Moreover, multivariate analysis adjusted for age showed that judo, Olympic wrestling and kickboxing athletes combined had 2D:4D ratios significantly lower (by 0.035 on average) than those of the rest of the subjects and that karate and taekwondo athletes together had 2D:4D ratios significantly higher (by 0.014 on average) than those of the rest of the subjects. The research results and literature review indicate that knowledge about the functional meaning of the 2D:4D ratio is still too fragmentary and it is too early to use the 2D:4D ratio in the selection of sport talent; however, it may be a useful criteria when screening prospective athletes to be recruited to a team. That is why 2D:4D reference values should be defined for particular sports.

Direct versus image-based digit ratio (2D:4D) measurement compared: Performance indicators and associations with candidate traits

Direct vs computer-assisted image-based measurements of the second-to-fourth digit ratio (2D:4D), a widely studied assumed negative correlate of prenatal testosterone, were compared in a sample of 96 men. Both methods yielded comparable associations of 2D:4D with a range of sexually differentiated personality measures, including the Big Five, aggression, hypermasculinity, and self-esteem (partly additionally assessed with implicit tasks and peer-ratings complementing the self-report measures). Directly ascertained 2D:4D was significantly less precisely measurable and significantly higher than 2D:4D ascertained from images by the same observers. In particular, measurement method impacted fingers differentially: directly measured index vs ring fingers were 1% vs 2% shorter, thereby yielding higher 2D:4D than when measured from images. Finger differences in fingertip size have been hypothesized to account for such discrepancies. However, fingertip size is unrelated to 2D:4D. Alternatively, diminished finger extensibility in supinated hand position (when taking direct measurements), as compared with finger extensibility in pronated hand position (when taking palmar-view hand images), might account for these observations. For anatomical reasons, this effect should be stronger for ring fingers than for index fingers and moreover should increase with age. The present data supported this novel hypothesis for explaining measurement-dependent differences in 2D:4D.

Evaluating Face2Gene as a Tool to Identify Cornelia de Lange Syndrome by Facial Phenotypes

Characteristic or classic phenotype of Cornelia de Lange syndrome (CdLS) is associated with a recognisable facial pattern. However, the heterogeneity in causal genes and the presence of overlapping syndromes have made it increasingly difficult to diagnose only by clinical features. DeepGestalt technology, and its app Face2Gene, is having a growing impact on the diagnosis and management of genetic diseases by analysing the features of affected individuals. Here, we performed a phenotypic study on a cohort of 49 individuals harbouring causative variants in known CdLS genes in order to evaluate Face2Gene utility and sensitivity in the clinical diagnosis of CdLS. Based on the profile images of patients, a diagnosis of CdLS was within the top five predicted syndromes for 97.9% of our cases and even listed as first prediction for 83.7%. The age of patients did not seem to affect the prediction accuracy, whereas our results indicate a correlation between the clinical score and affected genes. Furthermore, each gene presents a different pattern recognition that may be used to develop new neural networks with the goal of separating different genetic subtypes in CdLS. Overall, we conclude that computer-assisted image analysis based on deep learning could support the clinical diagnosis of CdLS.

A New Method of Assessing Sheep Red Blood Cell Types from Their Morphology

Data concerning the morphometric parameters of sheep red blood cells (RBCs) obtained using computer-assisted image analysis have not yet been investigated, and there are no data on any analyses of ovine RBC subpopulations based on their morphometric parameters. The aims of this study are to determine the values of RBC haematological and morphometric size and shape parameters, to form groups according to the obtained values of haematological parameters; to determine the differences in RBC morphometric parameters between the formed groups, and to determine RBC subpopulations and their respective proportions in the formed groups. Thirty-six blood samples were collected from the jugular vein of clinically healthy Lika pramenka sheep, aged between 2 and 5 years. Haematological parameters including haemoglobin (HGB), haematocrit (HTC), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), and RBC distribution width were analysed using a haematology analyser. Haematological parameters were categorized into two groups: those with lower values or values below the physiological range (Groups 1) and groups with higher values or values above the physiological range (Groups 2). Morphometric parameters of RBCs were determined from stained blood smears using SFORM, a computer-assisted program. Significantly higher values of RBC area, outline, convex, minimal and maximal radius, as well as length and breadth were established in Groups 2 compared to Groups 1 of HGB, HCT, MCV, MCH, and MCHC, respectively. Based on the morphometric parameters of RBCs, three RBC subpopulations were obtained using principal component and cluster analysis: ES 1—the smallest and most elongated RBCs, ES 2—the biggest and most rounded RBCs, and ES 3—average size and shape RBCs. Significantly higher proportions of ES 2 and ES 3 subpopulations, as well as a significantly lower proportion of ES 1 subpopulation, were established in Groups 2 compared to Groups 1 of HGB, HTC, MCV, and MCH, respectively. It can be concluded that ovine RBC subpopulations, based on their morphometric parameters, can be obtained by using computer-assisted image analysis of RBC morphometry and multivariate statistical methods, including principal component and cluster analysis. RBC morphometry, including classification into subpopulations, could serve as a basis for future possibilities in the diagnostic interpretation of anaemic syndromes in veterinary medicine, especially in normocytic, macrocytic, and microcytic anaemias in sheep.

A vinca alkaloid effect on microtubules of CHO (China Hamster Ovary)

The study evaluated the effect of selected Vinca alkaloids on the microtubule network of CHO cells line and the recovery of its disruption. Model experiments were used for demonstration of the quantification of microtubule network changes induced by Vinca alkaloids using computer-assisted image analysis cells were treated with Vincristine or Vinblastine at various concentrations from 2 μg/l to 800 μg/l for 60 min, or with Vincristine at a concentration of 4615 μg/l and 9230 μg/l for 60 min. Microtubules were detected by means of indirect immunofluorescence. The damage was examined in a fluorescence microscope. Also, cells were treated for 60 min with Vincristine at concentrations of 20 μg/l or 800 μg/l and the recovery process was studied in time intervals of 6, 7, 8, 9, 10h, or 8 and 12h, respectively. Differences in the arrangement of microtubules were assessed by means of a software for quantification of the cytoskeleton changes in cells treated with Vincristine at a concentration of 20 μg/l and in untreated control cells. The microtubule network recovery process in time intervals of 6, 7, 8, 9, and 10h was quantified after treatment with Vincristine at a concentration of 20 μg/l for 60 min using the software for quantification of the cytoskeleton changes.

Gastrointestinal Stromal Tumors Risk Stratification Utilizing Phospho-Histone H3 Evaluated by Manual Counting and Computer-Assisted Image Analysis

Background. Risk of progressive disease of gastrointestinal stromal tumors (GISTs) relies on mitotic index, size, and location of the tumor. However, manual mitotic counting on hematoxylin and eosin–stained slides (MMC-HE) is inefficient with low reproducibility. Manual count of phospho-histone H3 (MC-PHH3)-positive cells on immunohistochemical stained slides has been shown to have comparable reliability with MMC-HE. This study aims to confirm the reliability of MC-PHH3 in GISTs compared with MMC-HE and then to further compare MC-PHH3 with computer-assisted image analysis of PHH3-positive cells (Comp-PHH3). Methods. The study included 119 patients with GISTs. PHH3 stains were performed. MC-PHH3 was assessed as counts/5 mm2 high-power fields. Whole slide images were captured and the tumor area with greatest mitotic activity was manually identified. The PHH3-positive cells were automatically counted in 0.5 mm2 using Ventana Virtuoso software. Results. MMC-HE ranged from 0 to 157/5 mm2. MC-PHH3 ranged from 0 to 35.6/5 mm2. Comp-PHH3 ranged from 0 to 66/0.5 mm2. Interclass correlation coefficient (ICC) indicates good agreement between the 3 pathologists for MC-PHH3 (ICC = 0.74, P = .42). There is a strong correlation between MMC-HE and MC-PHH3. The Spearman correlation coefficient was 0.63 ( P < .0001). Lin’s concordance further indicated a moderate diagnostic agreement between MC-PHH3 and Comp-PHH3. Conclusion. MC-PHH3 is proposed as a superior alternative to MMC-HE with potential application in GIST reporting and prognostication. Furthermore, Comp-PHH3 may be a valid alternative to MC-PHH3.