Digital Image Analysis Software Research Articles

Tertiary lymphoid structures' pattern and prognostic value in primary adenocarcinoma of jejunum and ileum.

To date, there have been no reports on tertiary lymphoid structures (TLS) in primary adenocarcinoma of jejunum and ileum. In this study, we employed digital pathology image analysis software to classify and quantify TLS, and evaluated the maturity of TLS using immunohistochemistry. Molecular genetics and immunotherapy biomarker detection were performed using next-generation sequencing technology, such as tumor mutational burden (TMB) and microsatellite instability (MSI). The aim of this study was to investigate the presence, location, maturity, association with immunotherapy biomarkers, and prognostic value of TLS in primary adenocarcinoma of jejunum and ileum. Compared to secondary follicle-like TLS (SFL-TLS), intra-tumoral TLS (IT-TLS) were more likely to manifest as early TLS (E-TLS) (P = 0.007). Compared to IT-TLS, SFL-TLS had a higher propensity to occur at the invasive margin(IM) (P = 0.032) and showed a trend towards being more prevalent at the tumor periphery (P = 0.057). In terms of immunotherapy biomarkers, there was a higher trend of IM-TLS density in PD-L1(22C3) score CPS < 1 group compared to PD-L1(22C3) score CPS ≥ 1 group (P = 0.071). TMB-H was significantly associated with MSI-H (P = 0.040). Univariate survival analysis demonstrated a correlation between high SFL-TLS group and prolonged disease free survival (DFS) (P = 0.047). There was also a trend towards prolonged DFS in the E-TLS-high group compared to the E-TLS-low group (P = 0.069). The peri-tumoral TLS (PT-TLS)-high group showed a trend of prolonged overall survival (OS) compared to the PT-TLS-low group (P = 0.090). In conclusion, the majority of TLS were located at the invasive margin and tumor periphery, predominantly consisting of mature TLS, while IT-TLS were mainly immature. Notably, TMB was closely associated with MSI and PD-L1, indicating potential predictive value for immunotherapy in primary adenocarcinoma of jejunum and ileum.

The oculomotor cistern and pituitary adenomas: anatomical and clinical study.

The oculomotor cistern (OMC) is a meningeal cuff filled with CSF that contains the oculomotor nerve (cranial nerve [CN] III) at the level of the lateral wall of the cavernous sinus. Only a few studies have investigated the involvement of the OMC by pituitary adenomas (pituitary neuroendocrine tumors [PitNETs]), mainly with relatively small case series. The aim of this study was to perform a histomorphological description of the OMC and systematically analyze its involvement by PitNETs from radiological, clinical, and surgical perspectives. Ten hemisellae from formalin-fixed specimens were studied with 3-µm sections. Digital image analysis software was used for morphological and quantitative assessments. Clinical, radiological, surgical, and histological data of patients undergoing endoscopic transsphenoidal surgery for PitNETs at the University of Brescia, Italy, between 2014 and 2021 were recorded. OMC involvement was graded as not compressed, compressed, and invaded. The same surgical team operated on all patients. The OMC had an elliptical shape with an average area of 3.1 mm2 and a length of 5.5 mm. No cisternal points of weakness were recognized in the histomorphological study. Of 315 patients, 246 had complete data: apoplexy and CN III palsy were documented in 6.9% and 8.5%, respectively. OMC compression and invasion were recorded in 106 (43.1%) and 23 (9.3%) patients. Significant associations between OMC involvement and PitNET dimensions (p < 0.001), Knosp grade (p < 0.001), preoperative oculomotor palsy (p < 0.001), Ki-67 percentage (p = 0.009), and recurrence/progression of residual tumor (p = 0.008) were found. A new postoperative CN III palsy was evident in 2%: transient in 4 cases, and persistent in 1 patient treated for a recurrent PitNET who experienced a local infection complication. Preoperative CN III palsy improved in 10 cases. Significant OMC involvement by PitNETs might be underrecognized, but it can be treated using the endoscopic transsphenoidal approach, and it affects patient outcomes.

Apatite Fission-Track Dating: A Comparative Study of Ages Obtained by the Automated Counting LA-ICP-MS and External Detector Methodologies

Abstract Laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) in combination with developments in digital microscopy, image analysis, and computer software has allowed the implementation of an automated counting approach for the apatite fission-track (AFT) analysis. We refer to this approach as the “automated counting-LA-ICP-MS” (ACLA) method. Two major components comprise the ACLA method: (i) the digital counting of spontaneous tracks performed on high-resolution images captured from apatite grains and (ii) the measurement of 238U content in apatite by LA-ICP-MS. This study includes ACLA analyses from Fish Canyon Tuff (FCT) and Durango apatite standard crystals. Furthermore, a comparative age study between the ACLA and conventional external detector method (EDM) strategies was performed on a set of thirteen granitoid samples from northwestern Mexico and four granitic samples from the eastern Dharwar craton (EDC), India. ACLA analyses on FCT yielded an AFT age of 28.1 ± 0.6 (1σ) and 28.8 ± 1.1 (1σ) Ma for Durango apatite, whereas reported EDM ages are 27.5 ± 0.5 and 31.4 ± 0.5 Ma, respectively. Calculated AFT ages using the ACLA method from northwestern Mexico samples range from 11.1 ± 1.1 to 42.0 ± 3.6 Ma (EDM ages range from 10.0 ± 0.8 to 54.0 ± 3.0 Ma), whereas AFT ages from the EDC samples range from 147 ± 3.1 to 220.5 ± 12.5 Ma (EDM ages range from 120.9 ± 4.5 to 197.1 ± 19.4 Ma). Based on a statistical comparison with ages previously determined by the conventional EDM on the same samples and considering their 2σ uncertainties, these ages are in good agreement.

Quantitative study of the ossification centers of the body of sphenoid bone in the human fetus

The aim of the present study was to examine the growth dynamics of the two ossification centers of the body of sphenoid bone in the human fetus, based on their linear, planar and volumetric parameters. The examinations were carried out on 37 human fetuses of both sexes aged 18–30 weeks of gestation, which had been preserved in 10% neutral formalin solution. Using CT, digital image analysis software, 3D reconstruction and statistical methods, we evaluated the size of the presphenoid and postsphenoid ossification centers. The presphenoid ossification center grew proportionately in sagittal diameter, projection surface area and volume, and logarithmically in transverse diameter. The postsphenoid ossification center increased logarithmically in sagittal diameter, transverse diameter and projection surface area, while its volumetric growth followed proportionately. The numerical findings of the presphenoid and postsphenoid ossification centers may be considered age-specific reference values of potential relevance in monitoring the normal fetal growth and screening for congenital disorders in the fetus. The obtained results may contribute to a better understanding of the growing fetal skeleton, bringing new numerical information regarding its diagnosis and development.

Relationship between infiltration of tissue-resident memory T cells (TRM) in head and neck squamous cell carcinoma (HNSCC) tissue and localization and age.

6089 Background: Elevated levels of tumor-infiltrating lymphocytes (TILs) have been correlated with improved survival rates in cancer patients. Among TIL subgroups, tissue-resident memory T cells (TRM, CD8+CD103+) are recognized as pivotal contributors to the anti-cancer immune response. In this study, we aimed to evaluate TRM presence in Head and Neck Squamous Cell Carcinoma (HNSCC) tissue utilizing a developed tissue microarray (TMA) and a multiplex immunohistochemistry (MxIHC) approach. Methods: HNSCC cases from Southampton Hospital spanning the years 2000 to 2016 were reviewed, resulting in the identification of approximately 300 cases with sufficient primary tumor material. A TMA was constructed by triplicate coring of marked tumor areas on all slides. Subsequently, MxIHC, incorporating markers such as CD8 and CD103, was performed using a stain-scan-strip methodology. Digital image analysis software was employed to analyze scanned slides, and a quality check (QC) was conducted. Results: Following QC, 193 primary tumors remained (hypopharynx and larynx: n = 28, lip and oral cavity: n = 51, oropharynx: n = 114). Significantly higher counts of CD8 T cells and TRM were observed in the combined group of oropharynx, lip and oral cavity compared to the combined group of larynx and hypopharynx. This significance persisted even after excluding HPV negative oropharynx cases. HNSCC of the lip and oral cavity itself had statistically more TRM than larynx and hypopharynx. No differences were found in the subgroups between tonsil and tongue base. Stratifying the analysis by age revealed a statistically significant increase in TRM infiltration in HNSCC among young patients (≤50 years) compared to the 51-60 years and 61-70 years age groups, a pattern not observed for CD8 alone. Conclusions: This study underscores the importance of HNSCC heterogeneity and highlights the impact of age on immune infiltration, emphasizing the unique contribution of TRM in the anti-cancer immune response.

Abstract 5525: Tissue-resident cytotoxic T cell infiltration predicts metachronous colorectal cancer metastasis

Abstract Background: Identifying patients at high risk for metachronous colorectal cancer metastasis is essential for improving prognosis. The balance between host immune regulation and tumor immune evasion ultimately permits or prevents cancer progression. Lymphocytic infiltration in colorectal cancer can predict long-term clinical outcomes. However, these lymphocytes' origin, subtype and function have yet to be adequately explored. We investigated the association between the development of metastasis and CD8+T cells expressing the integrin protein CD103, a marker of tissue-resident memory T cells. Methods: In this retrospective, case-control matched study, we conducted multiplex immunofluorescence staining of tumor samples from 124 patients with colorectal cancer. Tissue microarrays containing representative cores of the central tumor, invasive margin, and adjacent normal tissue were immunostained for CD3, CD8, CD103, pSMAD3, and cytokeratin, using tyramide signal amplification. Cells were quantified using StrataQuest digital image analysis software, with intratumoral and stromal regions analyzed separately. Kruskal-Wallis one-way analysis of variance with subsequent Dunn’s test for pair-wise comparisons was performed using the Bonferroni method of multiple-comparison correction. Results: Central tumor and invasive margin CD3+ densities were associated with synchronous metastasis. Compared to non-metastatic patients, those who went on to develop metachronous metastasis (n=42) had a low proportion of intra-epithelial CD8+CD103+ T cells at the central tumor (non-metastatic: 32.8%, IQR 15.0-42.7%; metachronous metastasis: 12.9%, IQR 2.08-28.4%; P = 0.013). Similarly, patients with metachronous metastasis had a low proportion of stromal CD8+CD103+ T cells at the invasive margin (non-metastatic: 3.51%, IQR 1.83-6.55%; metachronous metastasis: 1.75%, IQR 0.62-3.72%; P = 0.019). On multivariate Cox regression of histological features, amongst patients with metastasis, significant overall survival prognostic markers included a low proportional CD8+CD103+ T cell infiltration (HR 2.41, 95%CI 1.16-5.01, P = 0.019), mucinous features (HR 4.28, 95%CI 1.98-9.24, P &amp;lt; 0.001), and the presence of perineural invasion (HR 2.75, 95%CI 1.28-5.89, P &amp;lt; 0.001). Conclusion: CD8+CD103+ lymphocytes may protect from the development of colorectal metastasis. Proportional CD8+CD103+ T cell infiltration is a promising prognostic marker in colorectal adenocarcinoma for the development of metachronous metastasis, and, amongst patients with metastasis, it is a prognostic marker of overall survival. Citation Format: Ryan Cohen, Tracey Lee-Pullen, Timothy Miller, Shadi Pirasteh, Cameron Platell, Katie Meehan, Kathy Fuller, Melanie McCoy. Tissue-resident cytotoxic T cell infiltration predicts metachronous colorectal cancer metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5525.

Exploring water-absorbing capacity: a digital image analysis of seeds from 120 wheat varieties

Wheat is a staple food crop that provides a significant portion of the world's daily caloric intake, serving as a vital source of carbohydrates and dietary fiber for billions of people. Seed shape studies of wheat typically involve the use of digital image analysis software to quantify various seed shape parameters such as length, width, area, aspect ratio, roundness, and symmetry. This study presents a comprehensive investigation into the water-absorbing capacity of seeds from 120 distinct wheat lines, leveraging digital image analysis techniques facilitated by SmartGrain software. Water absorption is a pivotal process in the early stages of seed germination, directly influencing plant growth and crop yield. SmartGrain, a powerful image analysis tool, was employed to extract precise quantitative data from digital images of wheat seeds, enabling the assessment of various seed traits in relation to their water-absorbing capacity. The analysis revealed significant transformations in seed characteristics as they absorbed water, including changes in size, weight, shape, and more. Through statistical analysis and correlation assessments, we identified robust relationships between these seed traits, both before and after water treatment. Principal Component Analysis (PCA) and Agglomerative Hierarchical Clustering (AHC) were employed to categorize genotypes with similar trait patterns, providing insights valuable for crop breeding and genetic research. Multiple linear regression analysis further elucidated the influence of specific seed traits, such as weight, width, and distance, on water-absorbing capacity. Our study contributes to a deeper understanding of seed development, imbibition, and the crucial role of water absorption in wheat. These insights have practical implications in agriculture, offering opportunities to optimize breeding programs for improved water absorption in wheat genotypes. The integration of SmartGrain software with advanced statistical methods enhances the reliability and significance of our findings, paving the way for more efficient and resilient wheat crop production. Significant changes in wheat seed shape parameters were observed after imbibition, with notable increases in area, perimeter, length, width, and weight. The length-to-width ratio (LWR) and circularity displayed opposite trends, with higher values before imbibition and lower values after imbibition.

ImageJ in Computational Fractal-Based Neuroscience: Pattern Extraction and Translational Research.

To explore questions asked in neuroscience, neuroscientists rely heavily on the tools available. One such toolset is ImageJ, open-source, free, biological digital image analysis software. Open-source software has matured alongside of fractal analysis in neuroscience, and today ImageJ is not a niche but a foundation relied on by a substantial number of neuroscientists for work in diverse fields including fractal analysis. This is largely owing to two features of open-source software leveraged in ImageJ and vital to vigorous neuroscience: customizability and collaboration. With those notions in mind, this chapter's aim is threefold: (1) it introduces ImageJ, (2) it outlines ways this software tool has influenced fractal analysis in neuroscience and shaped the questions researchers devote time to, and (3) it reviews a few examples of ways investigators have developed and used ImageJ for pattern extraction in fractal analysis. Throughout this chapter, the focus is on fostering a collaborative and creative mindset for translating knowledge of the fractal geometry of the brain into clinical reality.

Photochemical Modification of the Extracellular Matrix to Alter the Vascular Remodeling Process.

Therapeutic interventions for vascular diseases aim at achieving long-term patency by controlling vascular remodeling. The extracellular matrix (ECM) of the vessel wall plays a crucial role in regulating this process. This study introduces a novel photochemical treatment known as Natural Vascular Scaffolding, utilizing a 4-amino substituted 1,8-naphthimide (10-8-10 Dimer) and 450 nm light. This treatment induces structural changes in the ECM by forming covalent bonds between amino acids in ECM fibers without harming vascular cell survival, as evidenced by our results. To further investigate the mechanism of this treatment, porcine carotid artery segments were exposed to 10-8-10 Dimer and light activation. Subsequent experiments subjected these segments to enzymatic degradation through elastase or collagenase treatment and were analyzed using digital image analysis software (MIPAR) after histological processing. The results demonstrated significant preservation of collagen and elastin structures in the photochemically treated vascular wall, compared to controls. This suggests that photochemical treatment can effectively modulate vascular remodeling by enhancing the resistance of the ECM scaffold to degradation. This approach shows promise in scenarios where vascular segments experience significant hemodynamic fluctuations as it reinforces vascular wall integrity and preserves lumen patency. This can be valuable in treating veins prior to fistula creation and grafting or managing arterial aneurysm expansion.

Quantitative anatomy of the primary ossification center of the squamous part of temporal bone in the human fetus

Detailed numerical data about the development of primary ossification centers in human fetuses may influence both better evaluation and early detection of skeletal dysplasias, which are associated with delayed development and mineralization of ossification centers. To the best of our knowledge, this is the first report in the medical literature to morphometrically analyze the primary ossification center of the squamous part of temporal bone in human fetuses based on computed tomography imaging. The present study offers a precise quantitative foundation for ossification of the squamous part of temporal bone that may contribute to enhanced prenatal care and improved outcomes for fetuses with inherited cranial defects and skeletodysplasias. The examinations were carried out on 37 human fetuses of both sexes (16 males and 21 females) aged 18–30 weeks of gestation, which had been preserved in 10% neutral formalin solution. Using CT, digital image analysis software, 3D reconstruction and statistical methods, the size of the primary ossification center of the squamous part of temporal bone was evaluated. With neither sex nor laterality differences, the best-fit growth patterns for the primary ossification center of the squamous part of temporal bone was modelled by the linear function: y = −0.7270 + 0.7682 × age ± 1.256 for its vertical diameter, and the four-degree polynomial functions: y = 5.434 + 0.000019 × (age)4 ± 1.617 for its sagittal diameter, y = −4.086 + 0.00029 × (age)4 ± 2.230 for its projection surface area and y = −25.213 + 0.0004 × (age)4 ± 3.563 for its volume. The CT-based numerical data and growth patterns of the primary ossification center of the squamous part of temporal bone may serve as age-specific normative intervals of relevance for gynecologists, obstetricians, pediatricians and radiologists during screening ultrasound scans of fetuses. Our findings for the growing primary ossification center of the squamous part of temporal bone may be conducive in daily clinical practice, while ultrasonically monitoring normal fetal growth and screening for inherited cranial faults and skeletodysplasias.

Automated Radiographic Measurements of Knee Osteoarthritis.

Herewith, we report the development of Orthopedic Digital Image Analysis (ODIA) software that is developed to obtain quantitative measurements of knee osteoarthritis (OA) radiographs automatically. Manual segmentation and measurement of OA parameters currently hamper large-cohort analyses, and therefore, automated and reproducible methods are a valuable addition in OA research. This study aims to test the automated ODIA measurements and compare them with available manual Knee Imaging Digital Analysis (KIDA) measurements as comparison. This study included data from the CHECK (Cohort Hip and Cohort Knee) initiative, a prospective multicentre cohort study in the Netherlands with 1,002 participants. Knee radiographs obtained at baseline of the CHECK cohort were included and mean medial/lateral joint space width (JSW), minimal JSW, joint line convergence angle (JLCA), eminence heights, and subchondral bone intensities were compared between ODIA and KIDA. Of the potential 2,004 radiographs, 1,743 were included for analyses. Poor intraclass correlation coefficients (ICCs) were reported for the JLCA (0.422) and minimal JSW (0.299). The mean medial and lateral JSW, eminence height, and subchondral bone intensities reported a moderate to good ICC (0.7 or higher). Discrepancies in JLCA and minimal JSW between the 2 methods were mostly a problem in the lateral tibia plateau. The current ODIA tool provides important measurements of OA parameters in an automated manner from standard radiographs of the knee. Given the automated and computerized methodology that has very high reproducibility, ODIA is suitable for large epidemiological cohorts with various follow-up time points to investigate structural progression, such as CHECK or the Osteoarthritis Initiative (OAI).

QuPath Analysis for CD30+ Cutaneous T-Cell Lymphoma.

Mycosis fungoides is the most common subtype of cutaneous T-cell lymphoma, in which the expression of cluster of differentiation 30 (CD30)+ subtype can now be treated with the CD30 antibody conjugate brentuximab vedotin. Diagnostic methods are based on immunohistochemical (IHC) staining followed by manual assessment by pathologists, which is always a subjective calculation. QuPath, an open-source software for digital pathology image analysis, satisfies the requirements of objective approaches. Ten samples from mycosis fungoides patients with CD30 expression at different stages were stained for CD3 and CD30 by IHC staining, scanned, and quantitative analysis was performed using QuPath (version 2.1). Each slide was independently assessed by 3 board-certified dermatopathologists. Individual estimates for CD30+/CD3+ cells varied among the individual histopathologists (mean coefficient of variation, 0.46; range, 0-0.78). QuPath analysis showed excellent separation between the positively stained cells for CD3 and CD30 IHC and other cells and tissue structures, although the results correlated strongly with the respective mean estimates of the 3 histopathologists (Pearson-R 0.93). The results show a high interobserver variability evaluation of IHC markers, although quantitative image analysis offer a significant advantage for comparison. This is not only relevant for clinical routine but also especially critical in therapeutic studies addressing targeted molecules.

Tissue microarrey: a potential cost-effective approach for mismatch repair testing in colorectal cancer

Deficiencies in Mismatch Repair (MMR) proteins are one of the major pathways in the development of colorectal cancer (CRC). MMR status evaluation is recommended in every new CRC patient. However, this is not fully implemented due to high costs. Tissue microarray (TMA) enables allocating tissue cores from few specimens to a single paraffin block. The primary objective of this study was to evaluate the accuracy of TMA MMR immunohistochemistry (IHC) compared to whole slide. The secondary objective was to evaluate and validate automatic digital image analysis software in differentiating pathological and normal TMA cores. Pathological cores were defined if at least one MMR protein was unstained. Tumoral and normal tissue of 11 CRC patients with known MMR status was used to obtain 623 TMA cores. The MMR staining of each core was evaluated by a pathologist and compared to the whole slide result. Digital analysis software by 3DHistech Ltd. was used to identify cell nucleus and quantify nuclear staining in 323 tissue cores. To identifying pathological tissue, cores the cohort was divided into a test (N = 146 cores) and validation sets (N = 177 cores). A staining intensity score (SIS) was developed, and its performance compared to the pathologist review of each core and to the whole slide result. Compared to the whole slide, the pathologist's assessment had 100% sensitivity (n/N = 112/112) and 100% specificity (n/N = 278/278) with 95% lower limit of 97 and 99% respectively. The area under the receiver operating characteristic (ROC) curve of SIS was 77%. A cutoff of 55 was obtained from the ROC curve. By implementing the cutoff in the validation dataset, the SIS had sensitivity and specificity of 98.2% [90.1-100%] and 58.5% [49.3-67.4%] respectively. The MMR status of CRC can be evaluated in TMA tissue cores thus potentially reducing MMR testing costs. The SIS can be used as triage indicator during pathologic review. Institutional ethical approval was granted for the performance of this study (Emek Medical Center Ethics ID: EMC-19-0179).

Orbicularis Oculi Muscle Immunohistochemical, Metabolic, and Morphometric Differences in Affected and Nonaffected Sides in Hemifacial Spasm vs Healthy Subjects.

Subtle morphological alterations have been reported even in the nonaffected side of the orbicularis oculi muscle in patients with hemifacial spasm. However, no previous study assessed immunohistochemical, metabolic, and morphometric alterations in orbicularis oculi muscle fibers in affected and nonaffected sides in patients with this condition, compared with samples obtained from healthy subjects. The purpose of this study is to objectively assess and compare orbicularis oculi muscle (OOM) samples of hemifacial spasm affected and nonaffected sides and healthy subjects. Orbicularis oculi samples from 8 patients with hemifacial spasm who had not been previously treated and 6 healthy subjects were prepared using hematoxylin and eosin, nicotinamide adenine dinucleotide tetrazolium reductase, cytochrome oxidase, succinate dehydrogenase, Gomori staining, and monoclonal antibodies against myosin slow and myosin fast. A digital image analysis software was used for objective analysis. OOM fiber area was significantly greater in both affected ( P = 0.0379) and nonaffected sides ( P = 0.0012) of HFS samples when compared with control subjects' fibers. A significantly greater number of oxidative fibers were observed in both affected and nonaffected sides of patients with HFS when compared with control subjects ( P < 0.001 for both). A significantly greater percentage of slow fibers was observed in the affected side of HFS patients ( P = 0.0012) compared with control subjects. This study's findings suggest that repeated contractions might lead to OOM fiber hypertrophy, increased mitochondrial metabolism, and possible conversion of fast-twitch orbicularis oculi muscle fibers into slow-twitch fibers in patients with HFS. Alterations were observed in affected and nonaffected sides, confirming initial findings that the nonaffected side is not normal in this unique condition.

Comparative analysis of the leaf architectural characters of Saurauia bontocensis Merr. and Saurauia polysperma (Blanco) Merr. with the application of Digital Image Analysis Software

Abstract. Daipan BPO, Buot IE Jr., Daipan HMO. 2022. Comparative analysis of the leaf architectural characters of Saurauia bontocensis Merr. and Saurauia polysperma (Blanco) Merr. with the application of Digital Image Analysis Software. Biodiversitas 23: 3869-3875. Major confusion still exists among several authors concerning the taxonomy of the dagwey plant. This species is economically significant to the Ikalahan tribe of Luzon, Philippines. Several papers claimed that the plant is Saurauia bontocensis, while others stated that it is Saurauia subglabra. Many botanical websites claimed that S. subglabra is synonymous with S. polysperma, while another article distinguished S. subglabra from S. polysperma. Thus, to help resolve the taxonomic ambiguities, this paper performed a comparative analysis of the leaf architectural characters of S. bontocensis and S. polysperma with the aid of digital image analysis software and cluster analysis. Results revealed that the blade characters alone, delineate the two species, particularly, in terms of the laminar size, shape, symmetry, leaf L:W ratio, base and apex angle, and base shape. Additionally, the leaf venation characters delineate the two species, specifically, on the 2º to 5º vein categories, 2º vein angle, 3º vein course, 3º vein angle to 1º, areolation, freely ending ultimate veins, marginal ultimate venation, and leaf rank. Generally, S. bontocensis have larger, longer, and wider leaves compared with the S. polysperma. The leaf architecture analysis performed in this paper produced a very promising result in differentiating the two Saurauia species and could be used to address some of the taxonomic confusion of these species. The study indicated that S. bontocensis and S. polysperma are two distinct species. Moreover, the use of digital image analysis software was very helpful in the computations of the various blade characters, particularly the digital herbarium exsiccate in the absence of actual leaf samples.

A Machine-Learning Method to Assess Growth Patterns in Plants of the Family Lemnaceae.

Numerous new technologies have been implemented in image analysis methods that help researchers draw scientific conclusions from biological phenomena. Plants of the family Lemnaceae (duckweeds) are the smallest flowering plants in the world, and biometric measurements of single plants and their growth rate are highly challenging. Although the use of software for digital image analysis has changed the way scientists extract phenomenological data (also for studies on duckweeds), the procedure is often not wholly automated and sometimes relies on the intervention of a human operator. Such a constraint can limit the objectivity of the measurements and generally slows down the time required to produce scientific data. Herein lies the need to implement image analysis software with artificial intelligence that can substitute the human operator. In this paper, we present a new method to study the growth rates of the plants of the Lemnaceae family based on the application of machine-learning procedures to digital image analysis. The method is compared to existing analogical and computer-operated procedures. The results showed that our method drastically reduces the time consumption of the human operator while retaining a high correlation in the growth rates measured with other procedures. As expected, machine-learning methods applied to digital image analysis can overcome the constraints of measuring growth rates of very small plants and might help duckweeds gain worldwide attention thanks to their strong nutritional qualities and biological plasticity.

Integrating performance indicators in the production rules of shape-shifting grammars for architectural adaptive façade systems

ABSTRACT This paper presents shape-shifting grammar as a computational method that couples the environmental impact of utilizing programable materials in adaptive façades by their design parameters. The proposed grammar relies on physical experiments in a sealed temperature chamber, associated with a digital interface and image analysis software to track motion response for our previously developed composite ‘Hygromorphic Thermo-bimetal (HMTM)’ that responds passively to variation in humidity or temperature. This is linked with Grasshopper to evaluate performance indicators, specifically environmental impact, and percentage of view.The significance of the proposed grammar lies in tracking, analyzing, and predicting the behavior of the programable composite, therefore allowing for an efficient process using programable materials in adaptive façade design without the need for exhaustive physical experimentation and digital simulations. The developed grammar identifies production rules that map embedded and controlled parameters of the composite configuration to a desired deflection and motion response along with its environmental impact.

Is It Worth Using Botulinum Toxin Injections for the Management of Mild to Moderate Blepharoptosis?

Mild ptosis can be aesthetically displeasing for patients, especially in unilateral cases. However, some patients do not desire to undergo a surgical repair. Botulinum toxin injection might be an option in these cases. The authors sought to objectively examine the effects of botulinum neurotoxin A (BoNT-A) injection into the orbicularis oculi muscle in the management of blepharoptosis. In this prospective study, 8 patients with mild to moderate ptosis received application of BoNT-A in the pretarsal orbicularis oculi muscle. Standardized photographs were obtained at baseline and 2, 4, and 24 weeks. Digital image analysis software (Image J) was employed for objective analysis. Primary outcomes were the margin reflex distance-1 and qualitative changes on a 4-point scale. A significant increase in the margin reflex distance-1 on the treated side (baseline: 2.00 ± 1.13 mm; week 2: 2.52 ± 1.13 mm; P = 0.003) and a significant reduction (baseline: 3.23 ± 0.92 mm, week 2: 3.07 ± 0.96; P = 0.0268) on the contralateral eyelid were observed at week 2. After 24 weeks, the effect of BoNT-A diminished, and no significant difference from baseline was observed in either eyelid. Subjectively, 87.5% of patients reported improvement in their eyelid ptosis. Botulinum toxin injection in the pretarsal orbicularis oculi muscle can be an option to temporarily manage ptosis in patients who do not desire to undergo a surgical procedure. This alternative treatment can also be employed to manage temporary ptosis induced by botulinum toxin diffusion to the levator aponeurosis.

Techniques for digital histological morphometry of the pineal gland

IntroductionThe pineal gland is a small photo-neuro-endocrine organ. This study used human post-mortem pineal glands to microscopically assess immunohistochemical marker intensity and percentage of positivity using known and novel digital techniques. Materials and methodsAn experimental non-inferiority study has been performed on 72 pineal glands harvested from post-mortem examinations. The glands have been stained with glial fibrillary acidic protein (GFAP), synaptophysin (SYN), neuron-specific enolase (NSE), and neurofilament (NF). Slides were digitally scanned. Morphometric data were obtained using optical analysis, CaseViewer, ImageJ, and MorphoRGB ResultsStrong and statistically significant correlations were found and plotted using Bland-Altman diagrams between the two image analysis software in the case of mean percentage and intensity of GFAP, NSE, NF, and SYN. DiscussionsSoftware such as SlideViewer and ImageJ, with our novel software MorphoRGB were used to perform histological morphometry of the pineal gland. Digital morphometry of a small organ such as the pineal gland is easy to do by using whole slide imaging (WSI) and digital image analysis software, with potential use in clinical settings. MorphoRGB provides slightly more accurate data than ImageJ and is more user-friendly regarding measurements of parenchyma percentage stained by immunohistochemistry. The results show that MorphoRGB is not inferior in functionality. ConclusionsThe described morphometric techniques have potential value in current practice, experimental small animal models and human pineal glands, or other small endocrine organs that can be fully included in a whole slide image. The software we used has applications in quantifying immunohistochemical stains.

Assessing the prognostic value of tumor-infiltrating CD57+ cells in advanced stage head and neck cancer using QuPath digital image analysis

This study aimed to assess the prognostic value of intratumoral CD57+ cells in head and neck squamous cell carcinoma (HNSCC) and to examine the reproducibility of these analyses using QuPath. Pretreatment biopsies of 159 patients with HPV-negative, stage III/IV HNSCC treated with chemoradiotherapy were immunohistochemically stained for CD57. The number of CD57+ cells per mm2 tumor epithelium was quantified by two independent observers and by QuPath, software for digital pathology image analysis. Concordance between the observers and QuPath was assessed by intraclass correlation coefficients (ICC). The correlation between CD57 and clinicopathological characteristics was assessed; associations with clinical outcome were estimated using Cox proportional hazard analysis and visualized using Kaplan-Meier curves. The patient cohort had a 3-year OS of 65.8% with a median follow-up of 54 months. The number of CD57+ cells/mm2 tumor tissue did not correlate to OS, DFS, or LRC. N stage predicted prognosis (OS: HR 0.43, p = 0.008; DFS: HR 0.41, p = 0.003; LRC: HR 0.24, p = 0.007), as did WHO performance state (OS: HR 0.48, p = 0.028; LRC: 0.33, p = 0.039). Quantification by QuPath showed moderate to good concordance with two human observers (ICCs 0.836, CI 0.805–0.863, and 0.741, CI 0.692–0.783, respectively). In conclusion, the presence of CD57+ TILs did not correlate to prognosis in advanced stage, HPV-negative HNSCC patients treated with chemoradiotherapy. Substantial concordance between human observers and QuPath was found, confirming a promising future role for digital, algorithm driven image analysis.