Image-based Analysis Research Articles

Rapid and Effective Technology Development for 3D-Model-Based Solar Access Analysis and Comparative Study with Fish-Eye Camera

In this study, we developed a 3D-model-based technology that can evaluate solar access by analyzing solar radiation and shade to find the optimal location for a solar system. We developed an algorithm that can quickly calculate viewshed by applying ray-casting technology, which is useful in the field of computer graphics. To apply the developed technology, an unmanned aerial vehicle (DJI MAVIC 3) was used to create a 3D model by taking 320 photos of the Kangwon National University Samcheok campus. To verify the developed technology, a comparison with image-based analysis using a 360-degree camera was performed for 30 points. As a result of applying the developed technology to the study area, it was possible to calculate the solar access for each point. In general, image-based analysis exaggerates the effects of objects such as trees, whereas the developed technique can produce realistic results if the 3D objects were well built. If the technology is further developed in the future, it can be used to increase the efficiency of solar power generation.

High-content analysis of testicular toxicity of BPA and its selected analogs in mouse spermatogonial, Sertoli cells, and Leydig cells revealed BPAF induced unique multinucleation phenotype associated with the increased DNA synthesis

Bisphenol A is an endocrine disruptor that has been shown to have testicular toxicity in animal models. Its structural analog, including bisphenol S (BPS), bisphenol AF (BPAF), and tetrabromobisphenol A (TBBPA) have been introduced to the market as BPA alternatives. Previously, we developed high-content analysis (HCA) assays and applied machine learning to compare the testicular toxicity of BPA and its analogs in spermatogonial cells and testicular cell co-culture models. There are diverse cell populations in the testis to support spermatogenesis, but their cell type-specific toxicities are still not clear. The purpose of this study is to examine the selective toxicity of BPA, BPS), BPAF, and TBBPA on these testicular cells, including Sertoli cells, Leydig cells, and spermatogonia cells. We developed a high-content image-based single-cell analysis and measured a broad spectrum of adverse endpoints related to the development of reproductive toxicology, including cell number, nuclear morphology, DNA synthesis, cell cycle progression, early DNA damage response, cytoskeleton structure, DNA methylation status, and autophagy. We introduced an HCA index and spectrum to reveal multiple HCA parameters and observed distinct toxicity profiling of BPA and its analogs among three testicular types. The HCA spectrum shows the dynamic, chemical-specific, dose-dependent changes of each HCA parameter. Each chemical displayed a unique dose-dependent profile within each type of cell. All three types of cells showed the highest response to BPAF at 10 μM across all endpoints measured. BPAF targeted spermatogonial cell (C18) more significantly at 5 μM. BPS more likely targeted Sertoli cell (TM4) and Leydig cell (TM3) and less at spermatogonia cells. TBBPA targeted spermatogonia, Sertoli cells, and less at TM3 cells. BPA is mainly targeted at TM4, followed by TM3 cells, and less at spermatogonial cells. Most importantly, we observed that BPAF induced a dose-dependent increase in spermatogonia cells, not in Sertoli and Leydig cells. In summary, our current HCA assays revealed the cell-type-specific toxicities of BPA and its analogs in different testicular cells. Multinucleation induced by BPAF, along with increased DNA damage and synthesis at low doses, could possibly have a profound long-term effect on reproductive systems.

Image-based time series analysis to establish differential disease progression for two Fusarium head blight pathogens in oat spikelets with variable resistance.

Oat-based value-added products have increased their value as healthy foodstuff. Fusarium head blight (FHB) infections and the mycotoxins accumulated to the oat seeds, however, pose a challenge to oat production. The FHB infections are predicted to become more prevalent in the future changing climates and under more limited use of fungicides. Both these factors increase the pressure for breeding new resistant cultivars. Until now, however, genetic links in oats against FHB infection have been difficult to identify. Therefore, there is a great need for more effective breeding efforts, including improved phenotyping methods allowing time series analysis and the identification of molecular markers during disease progression. To these ends, dissected spikelets of several oat genotypes with different resistance profiles were studied by image-based methods during disease progression by Fusarium culmorum or F. langsethiae species. The chlorophyll fluorescence of each pixel in the spikelets was recorded after inoculation by the two Fusarium spp., and the progression of the infections was analyzed by calculating the mean maximum quantum yield of PSII (Fv/Fm) values for each spikelet. The recorded values were (i) the change in the photosynthetically active area of the spikelet as percentage of its initial size, and (ii) the mean of Fv/Fm values of all fluorescent pixels per spikelet post inoculation, both indicative of the progression of the FHB disease. The disease progression was successfully monitored, and different stages of the infection could be defined along the time series. The data also confirmed the differential rate of disease progression by the two FHB causal agents. In addition, oat varieties with variable responses to the infections were indicated.

Particle-tracking-based strategy for the optimization of agitation conditions in a suspension culture of human induced pluripotent stem cells in a shaking vessel

Suspension cultures are widely used for cell expansion in regenerative medicine and production. Shaking culture is one of the useful suspension culture methods that ensures gentle agitation. There are other shaking methods, including orbital shaking, reciprocating, and rocking; however, optimizing the shaking conditions for each method to meet cell culture requirements is time-consuming. In this study, we used a particle-tracking-based strategy for optimizing the agitation conditions. When the average accelerations of aggregates were calculated, high acceleration occurred periodically, and acceleration of the aggregates in orbital shaking was stable. Furthermore, the number of dead cells correlated with the average time of acceleration. We observed that cell growth was ideally maintained by factors such as optimal acceleration, aggregate formation, and cell death. These results indicate that the image-based analyses of aggregates help optimize the agitation conditions for the shaking suspension culture of induced pluripotent stem cells (iPSCs).

Artificial Intelligence for Anterior Segment Diseases: A Review of Potential Developments and Clinical Applications.

Artificial intelligence (AI) technology is promising in the field of healthcare. With the developments of big data and image-based analysis, AI shows potential value in ophthalmology applications. Recently, machine learning and deep learning algorithms have made significant progress. Emerging evidence has demonstrated the capability of AI in the diagnosis and management of anterior segment diseases. In this review, we provide an overview of AI applications and potential future applications in anterior segment diseases, focusing on cornea, refractive surgery, cataract, anterior chamber angle detection, and refractive error prediction.

An image-based shape analysis approach and its application to young women’s waist-hip-leg position

Digital image processing has been widely used for researches in the fashion industry. This study presented a method to classify women’s waist-hip-leg position based on body images. 135 healthy female students were selected as the experimental subjects, and then photo and 3D body measuring methods were used to obtain 40 shape parameters. Through factor analysis, five factors were extracted for the waist-hip position and eight factors for the leg position. The waist-hip and leg were separately classified into four categories after clustering analysis with optimised factors. The distribution of waist-hip-leg shape was analysed by combining the waist-hip and leg classification results. The results showed that 12.31% of the subjects had prominent abdomens, flat buttocks, and round and thin legs. The landmarks and parameters were automatically extracted for waist-hip-leg shape identification. The image-based shape analysis approach (ISA) was finally verified with an accuracy rate of over 90% with 30 new subjects. Practitioner summary: This study will propose an image-based shape analysis approach (ISA) to realise the quick automatic shape identification of the waist-hip-leg position based on body images. In addition, the method can be applied to pants’ pattern alteration for different body types by analysing the relationship between the waist-hip-leg shape and pants.

A multimodal sentiment analysis system for recognizing person aggressiveness in pain based on textual and visual information

This article proposes a multimodal sentiment analysis system for recognizing a person’s aggressiveness in pain. The implementation has been divided into five components. The first three steps are related to a text-based sentiment analysis system to perform classification tasks such as predicting the classes into non-aggressive, covertly aggressive, and overtly aggressive classes. The remaining two components are related to an image-based sentiment analysis system. A deep learning-based approach has been employed to do feature learning and predict the three types of pain classes. An aggression dataset for the text-based system and the UNBC-McMaster database for an image-based system has been employed, respectively. Experimental results have been compared with the state-of-the-art methods, showing the superiority of the proposed approach. Finally, the scores due to text-based and image-based sentiment analysis systems are fused to obtain the performance for the proposed multimodal sentiment analysis system.

Pain in the eye of the beholder: Variations in pain visual representations as a function of face ethnicity and culture.

Pain experienced by Black individuals is systematically underestimated, and recent studies have shown that part of this bias is rooted in perceptual factors. We used Reverse Correlation to estimate visual representations of the pain expression in Black and White faces, in participants originating from both Western and African countries. Groups of raters were then asked to evaluate the presence of pain and other emotions in these representations. A second group of White raters then evaluated those same representations placed over a neutral background face (50% White; 50% Black). Image-based analyses show significant effects of culture and face ethnicity, but no interaction between the two factors. Western representations were more likely to be judged as expressing pain than African representations. For both cultural groups, raters also perceived more pain in White face representations than in Black face representations. However, when changing the background stimulus to the neutral background face, this effect of face ethnic profile disappeared. Overall, these results suggest that individuals have different expectations of how pain is expressed by Black and White individuals, and that cultural factors may explain a part of this phenomenon.

Persistent discrepancies in orbital morphology after surgical treatment of unicoronal craniosynostosis: a critical image-based analysis.

Unicoronal craniosynostosis (UCS) is characterized by complex orbital deformity and is typically treated by asymmetrical fronto-orbital remodeling (FOR) during the 1st year of life. The aim of this study was to elucidate to what extent orbital morphology is corrected by surgical treatment. The extent to which orbital morphology was corrected by surgical treatment was tested by analysis of differences in volume and shape between synostotic, nonsynostotic, and control orbits at two time points. In total, 147 orbits were analyzed from patient CT images obtained preoperatively (mean age 9.3 months), at follow-up (mean age 3.0 years), and in matched controls. Semiautomatic segmentation software was used to determine orbital volume. For analysis of orbital shape and asymmetry, geometrical models, signed distance maps, principal modes of variation, and three objective parameters (mean absolute distance, Hausdorff distance, and dice similarity coefficient) were generated by statistical shape modeling. Orbital volumes on both the synostotic and nonsynostotic sides were significantly smaller at follow-up than volumes in controls and significantly smaller both preoperatively and at follow-up than orbital volumes on the nonsynostotic side. Significant differences in shape were identified globally and locally, both preoperatively and at 3 years of age. Compared with controls, deviations were mostly found on the synostotic side at both time points. Asymmetry between synostotic and nonsynostotic sides was significantly decreased at follow-up, but not compared with the inherent asymmetry of controls. On a group level, the preoperative synostotic orbit was mainly expanded in the anterosuperior and anteroinferior regions and smallest on the temporal side. At follow-up, the mean synostotic orbit was still larger superiorly but also expanded in the anteroinferior temporal region. Overall, the morphology of nonsynostotic orbits was more similar to that of controls than to synostotic orbits. However, the individual variation in orbital shape was greatest for nonsynostotic orbits at follow-up. In this study, the authors presented what is, to their knowledge, the first objective automatic 3D bony evaluation of orbital shape in UCS, defining in greater detail than has been done previously how synostotic orbits differ from nonsynostotic and control orbits, and how orbital shape changes from 9.3 months of age preoperatively to 3 years of age at the postoperative follow-up. Despite surgical treatment, both local and global deviations in shape persist. These findings may have implications for future directions in the development of surgical treatment. Future studies connecting orbital morphology to ophthalmic disorders, aesthetics, and genetics could provide further insight to enable better outcomes in UCS.

Gunshot detection from audio excerpts of urban sounds using transfers learning

In this work, a pre-trained convolutional neural network is employed to detect gunfire from audio excerpts of urban sounds. The pretrained convolutional neural network is fined-tuned with transfer learned features to a new task using a smaller number of training signals. Two CNN methods are applied to the time-frequency representation of the audio signals. The first CNN method is based on classifying specific events in audio signals. The second CNN method is an image-based analysis method. The accuracy of the two CNN results will be compared and analyzed based on gunfire type and retrieved urban multipath conditions. A k-means clustering algorithm is employed to identify gunfire types and parametric modeling to retrieve the urban multipath conditions.

Generation of 3D Tumor Spheroids for Drug Evaluation Studies.

In recent decades, in addition to monolayer-cultured cells, three-dimensional tumor spheroids have been developed as a potentially powerful tool for the evaluation of anticancer drugs. However, the conventional culture methods lack the ability to manipulate the tumor spheroids in a homogeneous manner at the three-dimensional level. To address this limitation, in this paper, we present a convenient and effective method of constructing average-sized tumor spheroids. Additionally, we describe a method of image-based analysis using artificial intelligence-based analysis software that can scan the whole plate and obtain data on three-dimensional spheroids. Several parameters were studied. By using a standard method of tumor spheroid construction and a high-throughput imaging and analysis system, the effectiveness and accuracy of drug tests performed on three-dimensional spheroids can be dramatically increased.

Real-time intelligent classification of COVID-19 and thrombosis via massive image-based analysis of platelet aggregates.

Microvascular thrombosis is a typical symptom of COVID-19 and shows similarities to thrombosis. Using a microfluidic imaging flow cytometer, we measured the blood of 181 COVID-19 samples and 101 non-COVID-19 thrombosis samples, resulting in a total of 6.3 million bright-field images. We trained a convolutional neural network to distinguish single platelets, platelet aggregates, and white blood cells and performed classical image analysis for each subpopulation individually. Based on derived single-cell features for each population, we trained machine learning models for classification between COVID-19 and non-COVID-19 thrombosis, resulting in a patient testing accuracy of 75%. This result indicates that platelet formation differs between COVID-19 and non-COVID-19 thrombosis. All analysis steps were optimized for efficiency and implemented in an easy-to-use plugin for the image viewer napari, allowing the entire analysis to be performed within seconds on mid-range computers, which could be used for real-time diagnosis.

Assessment of Bio-Based Materials as a Sustainable and Scalable Alternative for Detection of Plasmodium spp. (Haemospororida: Plasmodiidae) Sporozoites in Field Deployable Testing.

Malaria is responsible for over 435,000 deaths annually, mostly occurring in sub-Saharan Africa. Detecting Plasmodium spp. sporozoites (spzs) in the salivary glands of Anopheles (Diptera: Culicidae) vectors with circumsporozoite enzyme-linked immunosorbent assay (csELISA) is an important surveillance method. However, current technological advances are intellectual property and often require of distribution and highly trained users. The transition into paper-based rapid plataforms would allow for decentralization of survillance, especially in areas where it was virtually eliminated. The addition of bio-based materials have shown the potential to improve binding of target antigens, while being widely available. Here, we evaluate the use of chitosan and cellulose nanocrystals (CNC) as antibody carriers and substrate coatings on 96-well plates and on wax hydrophobized paper plates for the detection of Plasmodium falciparum (Pf), P. vivax VK210 (Pv210), and P. vivax VK247 (Pv247). To further improve the user-friendliness of the paper plates a quantitative photograph image-based color analysis was done. Interactions between the materials and the assay antibodies were studied by quartz crystal microbalance with dissipation monitoring (QCM-D). Overall, the addition of chitosan increased the interaction with antibodies and enhanced signaling in all tests. This work demonstrated that the adaptation of a PcsELISA shows potential as a cost-effective alternative assay platform easily adaptable in deployable testing sites that also showed reduction in reagent volumes by 80% and assay run time by seventh. While dipstick assays were previously developed, paper-based assays are a cost-effective and field-deployable alternative, reducing volumes of reagents that could be used in malaria control and elimination settings.

Quantitative Characterization of Pore Structure Parameters in Coal Based on Image Processing and SEM Technology

The pore structure parameters of coal have an important influence on the exploration and development of coalbed methane. In this study, a series of pore structure parameters, including porosity, pore radius, pore throat radius, pore coordination number, pore throat ratio, and specific surface area, are identified, extracted, and calculated in the scanning electron microscopy (SEM) images of coal reservoir samples using algorithms and application programs in MATLAB. Constant rate-controlled mercury injection and low-temperature N2 adsorption experiments were carried out to determine the accuracy of the SEM image-based processing analysis results. Characterization results show that the distribution of pore radius in the target coal samples of different organic matters range from 15 nm to 500 μm with porosity of 1.87–8.31% and radius distribution of 12.7 nm to ~100 μm. A noise-reduction system was constructed to eliminate the optical noise of non-porous features and repair the space affected by binarization noise. It is suggested that the characterization processing in this study is suitable for coal or other organic-rich porous materials with porosity > 2% and pore radius > 15 nm.

Remote Sensing Image-Based Analysis of the Urban Heat Island Effect in Relation to the Normalized Difference Vegetation Index (NDVI): A Case Study of Patna Municipal Corporation

Abstract: The consequences of population growth and urbanization on the urban environment, climate, and water supply have caused a wide range of issues. As a result of the rapid urbanization process, vegetation covers are being converted by impervious and dry concrete covers. This has led to the growth of "heat islands," where urban areas experience warmer temperatures than the areas surrounding them. Urban Heat Island (UHI) is a human-caused environmental phenomenon that has a wide range of impacts on city dwellers, including changing the vegetation cover and its usage, which alters the thermal energy flow and raises surface and air temperatures. Such heat islands have far-reaching impacts for cities, the most significant of which is an increase in the expense of maintaining a safe living and working environment. The major objective of this study is to examine multitemporal Land Surface Temperature (LST)/Urban Heat Island (UHI) and Normalized Difference Vegetation Index (NDVI) in the Patna Municipal Corporation (PMC) from 1990 to 2022. The Landsat satellite data sets for the years 1990, 2001, 2011, and 2022 have used to investigate the impact of UHI/LST in relation to NDVI in the study area. The most significant change has been observed in vegetation cover (NDVI), which has declined compared to other types of land use land cover. According to the study, built-up area and barren land have high temperatures, whereas vegetated covers and water bodies have lower temperatures. The LST of some portion of study area is high due to the high population density and high percentage of built-up and concrete cover. The LST over the study area has risen on average by 6.88 °C between 1990 and 2022. The regression line provided a conclusive answer, demonstrating a strong negative relationship between the normalized difference vegetation index (NDVI) and UHI of PMC. According to the findings of this study, a major transition has occurred in PMC in terms of a decline in NDVI due to a rapid growth in urban expansion, and other infrastructure projects. To study urban climate and interactions between people and the environment, land surface temperature (LST) variations within cities are of utmost importance in respect to ascertain the LST/Urban Heat Island (UHI) effect and NDVI variations in PMC. While developing urbanization, the environmental impact must be taken into account.

Scanning Electron-Raman Cryomicroscopy for Characterization of Nanoparticle-Albumin Drug Products.

Nanomaterials have expanded the use of active pharmaceutical ingredients by improving efficacy, decreasing toxicity, and facilitating targeted delivery. To systematically achieve this goal, nanomaterial-containing drugs need to be manufactured with precision in attributes such as size, morphology, surface chemistry, and composition. Their physicochemical characterization is essential as their attributes govern pharmacokinetics yet can be challenging due to the nature of many nanomaterial-based formulations unless advanced sample fixation and in vitro characterization methods are utilized. Here, different cryogenic and other fixation strategies were assessed, and a novel physicochemical characterization method was developed using scanning electron Raman cryo-microscopy (SERCM). A complex nanoparticle albumin bound paclitaxel (nab-paclitaxel) formulation was chosen as a model drug. Plunge freezing (PF), high pressure freezing (HPF), freeze substitution (FS), and membrane filtration were compared for their influence on size and morphology measurements, and formulation-based variations were quantified. SERCM was introduced as a multiattribute physicochemical characterization platform, and the composition of nanoparticles was confirmed as albumin-paclitaxel complexes. By coupling image-based quantitative analysis with chemical analysis, SERCM has the potential to pave the way for the development of comprehensive tools for assessing injectable and ophthalmic nanomaterial-containing drugs in their native-like state.

Concurrent and Angle-Trajectory Validity and Intra-Trial Reliability of a Novel Multi-View Image-Based Motion Analysis System.

Sports-related injuries are the most common in the lower extremities among physical regions. To evaluate impaired functional performance in sports training facilities and sports, a marker-less motion analysis system that can measure joint kinematics in bright indoor and outdoor environments is required. The aim of this study was to establish the concurrent and angle-trajectory validity and intra-trial reliability of a novel multi-view image-based motion analysis system with marker-less pose estimation during lower extremity tasks in healthy young men. Ten healthy young men participated voluntarily in this study. The hip and knee joint angles were collected using a multi-view image-based motion analysis system (marker-less) and a Vicon motion capture system (with markers) during the lower extremity tasks. Intraclass correlation coefficient (ICC) analyses were used to identify the concurrent and angle-trajectory validity and intra-trial reliability of the multi-view image-based motion analysis system. In the concurrent validity, the correlation analysis revealed that the ICC3, k values on the hip and knee flexions during knee bending in sitting, standing, and squat movements were from 0.747 to 0.936 between the two systems. In particular, the angle-trajectory validity was very high (ICC3, 1 = 0.859-0.998), indicating a high agreement between the two systems. The intra-trial reliability of each system was excellent (ICC3, 1 = 0.773-0.974), reflecting high reproducibility. We suggest that this novel marker-less motion analysis system is highly accurate and reliable for measuring joint kinematics of the lower extremities during the rehabilitation process and monitoring sports performance of athletes in training facilities.

Perturbed actin cap as a new personalized biomarker in primary fibroblasts of Huntington's disease patients.

Primary fibroblasts from patient's skin biopsies are directly isolated without any alteration in the genome, retaining in culture conditions their endogenous cellular characteristics and biochemical properties. The aim of this study was to identify a distinctive cell phenotype for potential drug evaluation in fibroblasts from Huntington's Disease (HD) patients, using image-based high content analysis. We show that HD fibroblasts have a distinctive nuclear morphology associated with a nuclear actin cap deficiency. This in turn affects cell motility in a similar manner to fibroblasts from Hutchinson-Gilford progeria syndrome (HGPS) patients used as known actin cap deficient cells. Moreover, treatment of the HD cells with either Latrunculin B, used to disrupt actin cap formation, or the antioxidant agent Mitoquinone, used to improve mitochondrial activity, show expected opposite effects on actin cap associated morphological features and cell motility. Deep data analysis allows strong cluster classification within HD cells according to patients' disease severity score which is distinct from HGPS and matching controls supporting that actin cap is a biomarker in HD patients' cells correlated with HD severity status that could be modulated by pharmacological agents as tool for personalized drug evaluation.

The role of small GTPases in bisphenol AF-induced multinucleation in comparison with dibutyl phthalate in the male germ cells.

The goal of this study is to examine bisphenol AF (BPAF)-induced multinucleation (MNC) in comparison with dibutyl phthalate (DBP), known to induce MNC in mouse gonocytes in vivo. We performed image-based single-cell high content analysis (HCA) in the mouse spermatogonia C18-4 cells treated with various concentrations of BPAF and DBP. BPAF as low as 5 µM was cytotoxic and resulted in 40% cell death of the C18-4 cells after 72 h. HCA revealed that 5 µM of BPAF significantly increased the number of MNC by an average of 3.6-fold. DBP did not induce MNC in the doses we tested. Cytokinesis is tightly regulated by various small GTPase-signaling pathways. We, therefore, tested 5 selective GTPase inhibitors and found that Y27632, a ROCK inhibitor, reduced the BPAF-induced MNC by nearly 30%. Inhibition of Cdc42 by ML141 conversely increased the number of BPAF-induced MNC. We performed a hierarchical cluster analysis of the HCA data and demonstrated that the cytoskeletal disruption by BPAF was reversely modified by Y27632. We found that mRNA expression of genes regulating Rho and Rac GTPase activities, p190RhoGap and MgcRacGap, was altered in BPAF-treated C18-4 cells in a time-dependent manner. Multinucleated gonocytes are often indicators of disease pathologies. Our results provided the first evidence of mechanisms of the dual toxicity by BPAF to male germ cells, which induces chromosome endoreplication without the coordinated cytokinetic cellular components. The unique genotoxic mechanism of forming multinucleated germ cells suggests a novel mode of action in the male repro-toxicity concern over the increasingly ubiquitous presence of BPA analogs.

Morphological characterization of some poorly known populations of the genus Spalerosophis Jan, 1865 (Reptilia: Colubridae): is the geometric morphometrics useful to identify them as taxonomic units?

Despite the use of traditional morphology as a standard method for species identification, additional methods like image-based analysis can be desirable to differentiate species and populations. In this study, we used traditional morphology and geometric morphometrics (GM) to describe three distinct populations of the genus Spalerosophis and test the hypothesis that all populations are morphologically distinct. A total of 132 specimens were analysed and based on morphological traits (morphometric, meristic, and colour pattern) subcaudal scales was the only distinguishable trait between Spalerosophis diadema cliffordii and Spalerosophis diadema schirasianus. In addition, Spalerosophis atriceps showed significant differences in most studied characters compared to the other two populations. In the geometric morphometric analysis, we placed 2D Landmarks on the dorsal view images of the head of 93 specimens. Canonical variate analysis (CVA) showed significant differences among the head shape of all three populations. There was an increase in the head width, snout length, and curvature in the posterior part of parietals from west Africa in S. d. cliffordii to the north of India in S. atriceps. This may be the result of variations in their environmental factors, diet and the amount of time that has passed since their separation.