Exposure of children to heavy metals from artisanal gold mining in Nigeria:evidences from bio-monitoring of hairs and nails

In recent times,there had been reported cases of Pb poisoning in Anka gold mining area,Northwest Nigeria.Therefore,this study was carried out to determine the extent of bioaccumulation of heavy metals in the hairs and nails of children in the area.Forty samples(twenty nails and twenty hairs)samples were collected from ten boys and ten girls of ages 5–9 residing in the area.To ascertain the sources of heavy metals in children,15 soils samples,15 groundwater samples,5 samples of mine tailings,and 5 plants samples were collected.Hair and nails of the subjects were collected using internationally acceptable techniques.All samples were kept in uncontaminated ziplock bags prior to laboratory preparation and analysis.The samples were cleaned using nonionic detergent(triton X-100)and deionized water.The hairs and nails were digested with 10 mL of 6:1 mixture of nitric acid and perchloric acid.The soils,mine tailings,and plants were air-dried at room temperature,sieved,and chemically digested using the aqua regia method.The concentrations of metals in all the samples were determined using highperformance liquid chromatography–inductively coupled plasma–mass spectrometry.Statistical analysis was employed to unravel potential sources of metals in the media.Results showed that heavy metals in children of the area are above results from similar studies and pathological ranges for heavy metals in hairs and nails.Also,heavy metals in environmental media are above the recommended standards.Multivariate analysis showed that the metals are mainly from mining and other anthropogenic sources.Results of correlation between heavy metals in hairs and nails with those in geological samples revealed that heavy metal that bioaccumulates in the children of this area are mostly from contaminated environmental media.It is recommended that complete remediation and effective health education be carried out in the area.

Mineral Particles Found on the Hairs of Holy Maria-Magdalena Studied by Scanning Electron Microscopy and Elemental Analysis

The presumed hairs of Holy Maria-Magdalena are studied here for mineral particles found on their surfaces, by the techniques of scanning electron microscopy coupled with energy dispersive X-ray. Exploration by these techniques of the hair surfaces allows us to observe and study some marble fragments that can originate from the walls of the sarcophagus in which Maria-Magdalena’s remains were kept. These new findings support the authenticity of the hair as traditionally reported in the French “tradition des Saints de Provence”.

Cell-type-specific transcriptomics reveals that root hairs and endodermal barriers play important roles in beneficial plant-rhizobacterium interactions

Growth-and health-promoting bacteria can boost crop productivity in a sustainable way.Pseudomonas simiae WCS417is sucha bacterium that efficiently colonizes roots,modifiesthe architecture of the root systemto increase its size,and induces systemic resistance to make plants more resistant to pests and pathogens.Our previous work suggested that WCS417-induced phenotypes are controlled by root cell-type-specific mechanisms.However,it remains unclear how WCS417 affects these mechanisms.In this study,we transcriptionally profiled five Arabidopsis thaliana root cell types following WCS417 colonization.We found that the cortex and endodermis have the most differentially expressed genes,even though they are not in direct contact with this epiphytic bacterium.Many of these genes are associated with reduced cell wall biogenesis,and mutant analysis suggests that this downregulation facilitates WCS417-driven root architectural changes.Furthermore,we observed elevated expression of suberin biosynthesis genes and increased deposition of suberin in the endodermis of WCS417-colonized roots.Using an endodermal barrier mutant,we showed the importance of endodermal barrier integrity for optimal plant-beneficial bacterium association.Comparison of the transcriptome profiles in the two epidermal cell types that are in direct contact with WcS417-trichoblasts that form root hairs and atrichoblasts that do not-implies a difference in potential for defense gene activation.While both cell types respond to WCS417,trichoblasts displayed both higher basal and WCS417-dependent activation of defense-related genes compared with atrichoblasts.This suggests that root hairs may activate root immunity,a hypothesis that is supported by differential immune responses in root hair mutants.Taken together,these results highlight the strength of cell-type-specific transcriptional profiling to uncover"masked"biological mechanisms underlying beneficial plant-microbe associations.

Plasticity of wheat seedling responses to K^(+) deficiency highlighted by integrated phenotyping of roots and root hairs over the whole root system

The availability in the soil of potassium(K^(+)),a poorly mobile macronutrient required in large quantities for plant growth,is generally suboptimal for crop production in the absence of fertilization,making improvement of the ability of crops to adapt to K^(+)deficiency stress a major issue.Increasing the uptake capacity of the root system is among the main strategies to achieve this goal.Here,we report an integrative approach to examine the effect of K^(+)deficiency on the development of young plant entire root system,including root hairs which are known to provide a significant contribution to the uptake of poorly mobile nutrients such as K^(+),in two genetically distant wheat varieties.A rhizobox-type methodology was developed to obtain highly-resolved images of root and root hairs,allowing to describe global root and root hair traits over the whole root system via image analysis procedures.The two wheat varieties responded differently to the K^(+)shortage:Escandia,a wheat ancestor,reduced shoot biomass in condition of K^(+)shortage and substantially increased the surface area of its root system,specifically by increasing the total root hair area.Oued Zenati,a landrace,conversely appeared unresponsive to the K^(+)shortage but was shown to constitutively express,independently of the external K^(+)availability,favorable traits to cope with reduced K^(+)availability,among which a high total root hair area.Thus,valuable information on root system adaptation to K^(+)deficiency was provided by global analyses including root hairs,which should also be relevant for other nutrient stresses.

Mechanical upper bowel obstruction caused by a large trichobezoar in a young woman: A very unusual case report

BACKGROUND Bezoars usually compile human fibers and debris.A special form of bezoar in case of psychologically altered individuals is the trichobezoar.It consists of voluntarily swallowed hair bulks and is normally removed via gastroscopy.Trichobezoars leading to ileus have rarely been reported.CASE SUMMARY A 24-year-old female patient presented to the emergency room with abdominal pain,nausea,and vomiting for 3 d.Her previous medical and psychiatric history was unremarkable.Laboratory analysis showed iron deficiency anemia,leukocytosis,and elevated liver enzymes.An abdominal CT scan revealed a dense structure in the patients’stomach which turned out to be a huge trichobezoar completely obstructing the pylorus.The trichobezoar had to be removed surgi-cally.During her postoperative course,a subcutaneous seroma formed.After a single puncture,the rest of the recovery process was unremarkable,and the patient recovered fully.CONCLUSION A mechanical bowel obstruction is a potentially life-threatening event for every patient.In our case a young female was suffering from severe symptoms of an obstruction which might have resulted in serious harm without successful surgical management.

Recent progress in hair follicle stem cell markers and their regulatory roles

Hair follicle stem cells(HFSCs)in the bulge are a multipotent adult stem cell population.They can periodically give rise to new HFs and even regenerate the epidermis and sebaceous glands during wound healing.An increasing number of biomarkers have been used to isolate,label,and trace HFSCs in recent years.Considering more detailed data from single-cell transcriptomics technology,we mainly focus on the important HFSC molecular markers and their regulatory roles in this review.

Bhlhe40 protects cochlear hair cell-like HEI-OC1 cells against H_(2)O_(2) ‑triggered oxidative injury

Background:Cochlear hair cell injury is a common pathological feature of hearing loss.The basic helix-loop-helix family,member e40(Bhlhe40),a gene belonging to the basic helix-loop-helix(bHLH)family,exhibits strong transcriptional repression activity.Methods:Oxidative damage,in House Ear Institute-Organ of Corti 1(HEI-OC1)cells,was caused using hydrogen peroxide(H2O2).The Ad-Bhlhe40 particles were constructed to overexpress Bhlhe40 in HEI-OC1 cells.Various assays including cell counting kit-8(CCK-8),terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay(TUNEL),flow cytometry,immunofluorescence,and corresponding commercial kits were employed to investigate the impacts of Bhlhe40 on cell viability,apoptosis,oxidative stress levels,mitochondrial membrane potential and cellular senescence.Additionally,a dual-luciferase reporter assay was performed to confirm the targeting of the histone deacetylases 2(Hdac2)by Bhlhe40.Results:The results revealed that Bhlhe40 was downregulated in H_(2)O_(2)-treated HEI-OC1 cells,but its overexpression improved cell viability and mitigated H_(2)O_(2)-induced oxidative injury in HEI-OC1 cells with increase of superoxide dismutase(SOD),catalase(CAT)and glutathione peroxidase(GPx)activities and decrease of reactive oxygen species(ROS)levels.Besides,overexpression of Bhlhe40 suppressed H_(2)O_(2)-triggered cell senescence,as evidenced by the fact that the upregulation of P53,P21,and P16 in HEI-OC1 cells treated with H2O2 were all alleviated by Bhlhe40 overexpression.And we further verified that overexpression of Bhlhe40 could inhibit the expression of Hdac2,which may be related to the repression of Hdac2 transcription.Conclusion:This study suggests that Bhlhe40 plays a protective role against senescence and oxidative damage in cochlear hair cells exposed to H2O2.

Mercury Levels Assessment in the Population of Aby and Frambo Villages in the Vinicity of Aby Lagoon in Côte d’Ivoire (West Africa)

The Minamata Convention in the aim to protect human life and the environment, seeks to reduce mercury (Hg) by monitoring it concentrations in the environment. Artisanal and Small-scale Gold Mining (ASGM) has been identified as the most important anthropogenic source of human exposure to Hg. In this context, the main goal of this study was to assess the level of mercury in hair of two (2) populations living along two lagoons respectively Aby and Tendo, in Ivory Coast. To reach this goal, hair samples of 138 residents were collected and analyzed by using Cold Vapor Atomic Fluorescence Spectrophotometry (CV-AFS) technique for mercury concentration. Results showed that for the entire population the Hg mean was 2.34 μg/g. Also, they were ranged between 0.15 to 8.53 μg/g and presented substantial differences amongst the villages. In Aby village, we observed the highest Hg concentrations (Mean = 2.62 μg/g). Our findings showed that almost the entire sample group (82%) exceed the USEPA recommended limit, furthermore 56% of them exceed the normal level of WHO and 2% of the respondent has the unhealthy levels of mercury (≥6 μg Hg/g) of hair by WHO standards. Gender differences in hair mercury varies greatly among reports. Lower levels in women’s hair compared to men were reported in the both village. Considering age, the lowest concentrations were observed with children. However, when we take in account the age groups, data suggested that the most exposed sub-population of [18-29] years old is from Aby village in opposite at Frambo village, the same case those who were ≥ 40 years old. It’s convenient to note that, the proportion of Mercury levels would not to be neglected among studied population especially with the resident from Aby village. So, some measures need to be taken at the political level to control mercury contamination.

Determinants of Occipital Involvement in Males and Females with Pattern Hair Loss: A Retrospective Study

Background: Pattern hair loss (PHL) is the most common form of hair loss affecting both men and women. The occipital region, located at the back of the scalp, has been considered resistant to hair loss in PHL. It is often used as a donor site for hair transplantation procedures. However, evidence suggests that the occipital region may also be affected by PHL, although to a lesser extent compared to the frontal and vertex regions. Objective: To assess the levels and determinants of occipital involvement in PHL, focusing on gender and disease severity. Such data would enhance the comprehensive understanding of PHL progression and inform development of improved treatment strategies. Methods: This retrospective study involved 1993 adults (1000 males, 993 females) following for PHL at a specialized hair clinic in Jeddah, Saudi Arabia. Any patient who had hair transplantation in the past and any male who had balding areas (Hamilton-Norwood scales 4 to 7) at the initial consultation was excluded. The primary outcome was occipital involvement, defined as >10% thin hairs in the trichometry assessments of the occipital region. Additional data included patients age, treatment status, disease severity, and other trichometry parameters. Results: The prevalence of occipital involvement was found to be 23.8% overall, significantly more frequent in females (35.6%) than males with early PHL (Hamilton-Norwood scales 2 and 3) (12.1%) (p Conclusion: This study underscored the relevance of gender and disease severity in occipital involvement in PHL. Females had significantly higher rates of occipital involvement, suggesting that gender differences are critical in understanding PHL progression. Future research should aim to validate these findings, with an aim to refine therapeutic and surgical approaches to improve patient outcomes.

Black Hair is Beautiful An On-the-spot Report of 1994 XVⅢ Asian Hairstyling & Make-up Competition

From June 7-10 hair and beauty specialists from 20 countries in Asia gathered in Beijing to take part in the 1994 ⅩⅧ Asian Hairstyling and Make-up Competition. This was the first time the event was held in China. At the same time an international cosmetic, hair and skin care products oriental beauty exposition also was taking place. Throughout history men and women have paid attention to physical beauty, both in the West and the East,

Three CNGC Family Members, CNGC5, CNGC6, and CNGC9, Are Required for Constitutive Growth of Arabidopsis Root Hairs as Ca2+-Permeable Channels

The genetic identities of Ca2+ channels in root hair (RH) tips essential for constitutive RH growth have remained elusive for decades. Here, we report the identification and characterization of three cyclicnucleotide-gated channel (CNGC) family members, CNGC5, CNGC6, and CNGC9, as Ca2+ channels essential for constitutive RH growth in Arabidopsis. We found that the cngc5-1cngc6-2cngc9-1 triple mutant(designated shrh1) showed significantly shorter and branching RH phenotypes as compared with thewild type. The defective RH growth phenotype of shrh1 could be rescued by either the expression ofCNGC5, CNGC6, or CNGC9 single gene or by the supply of high external Ca2+, but could not be rescuedby external K+ supply. Cytosolic Ca2+ imaging and patch-clamp data in HEK293T cells showed that thesethree CNGCs all function as Ca2+-permeable channels. Cytosolic Ca2+ imaging in growing RHs furthershowed that the Ca2+ gradients and their oscillation in RH tips were dramatically attenuated in shrh1compared with those in the wild type. Phenotypic analysis revealed that these three CNGCs are Ca2+ channels essential for constitutive RH growth, with different roles in RHs from the conditional player CNGC14.Moreover, we found that these three CNGCs are involved in auxin signaling in RHs. Taken together, ourstudy identified CNGC5, CNGC6, and CNGC9 as three key Ca2+ channels essential for constitutive RHgrowth and auxin signaling in Arabidopsis.

STATIC AND STATIONARY BLACK HOLES WITH QCD HAIRS

The coupled SU(5) Einstein-Yang-Mills-Higgs system of fields is investigated. A familyof static spherically symmetric and stationary axisymmetric black hole solutions is obtained.Our result shows that the black hole can carry not only the abelian electric and magneticcharges but also the non-Abelian SU(3)_c color charge.

A Comparative Study of Human and Animal Hairs:Microscopic Hair Comparison and Cytochrome c Oxidase I Species Identification

Human and animal hairs have been used in forensic investigations for over a century.Hair is stable under adverse natural conditions;hence,it is often recovered at the crime scene,and it is necessary to determine whether the hair is of human or animal origin.Morphological and genetic characteristics are useful to differentiate human hair from animal hair.In the present study,we analyzed the distinguishing characteristics of hair of various species.In addition,we explore species identification by cytochrome c oxidase I mitochondrial gene analysis.We confirm that both the microscopic and molecular analyses of hairs are useful in forensic investigations.

Melatonin promotes the development of the secondary hair follicles by regulating circMPP5

Background The quality and yield of cashmere fibre are closely related to the differentiation and development of secondary hair follicles in the skin of cashmere goats.The higher the density of secondary hair follicles,the higher the quality and yield of cashmere from the fleece.Development of secondary hair follicles commences in the embryonic stage of life and is completed 6 months after birth.Preliminary experimental results from our laboratory showed that melatonin(MT)treatment of goat kids after their birth could increase the density of secondary hair follicles and,thus,improve the subsequent yield and quality of cashmere.These changes in the secondary hair follicles resulted from increases in levels of antioxidant and expression of anti-apoptotic protein,and from a reduction in apoptosis.The present study was conducted to explore the molecular mechanism of MT-induced secondary hair follicle differentiation and development by using whole-genome analysis.Results MT had no adverse effect on the growth performance of cashmere kids but significantly improved the character of the secondary hair follicles and the quality of cashmere,and this dominant effect continued to the second year.Melatonin promotes the proliferation of secondary hair follicle cells at an early age.The formation of secondary hair follicles in the MT group was earlier than that in the control group in the second year.The genome-wide data results involved KEGG analysis of 1044 DEmRNAs,91 DElncRNAs,1054 DEcircRNAs,and 61 DEmiRNAs which revealed that the mitogen-activated protein kinase(MAPK)signaling pathway is involved in the development of secondary hair follicles,with key genes(FGF2,FGF21,FGFR3,MAPK3(ERK1))being up-regulated and expressed.We also found that the circMPP5 could sponged miR-211 and regulate the expression of MAPK3.Conclusions We conclude that MT achieves its effects by regulating the MAPK pathway through the circMPP5 sponged the miR-211,regulating the expression of MAPK3,to induce the differentiation and proliferation of secondary hair follicle cells.In addition there is up-regulation of expression of the anti-apoptotic protein causing reduced apoptosis of hair follicle cells.Collectively,these events increase the numbers of secondary hair follicles,thus improving the production of cashmere from these goats.

Artificial nerve graft constructed by coculture of activated Schwann cells and human hair keratin for repair of peripheral nerve defects

Studies have shown that human hair keratin(HHK) has no antigenicity and excellent mechanical properties. Schwann cells, as unique glial cells in the peripheral nervous system, can be induced by interleukin-1β to secrete nerve growth factor, which promotes neural regeneration. Therefore, HHK with Schwann cells may be a more effective approach to repair nerve defects than HHK without Schwann cells. In this study, we established an artificial nerve graft by loading an HHK skeleton with activated Schwann cells. We found that the longitudinal HHK microfilament structure provided adhesion medium, space and direction for Schwann cells, and promoted Schwann cell growth and nerve fiber regeneration. In addition, interleukin-1β not only activates Schwann cells, but also strengthens their activity and increases the expression of nerve growth factors. Activated Schwann cells activate macrophages, and activated macrophages secrete interleukin-1β, which maintains the activity of Schwann cells. Thus, a beneficial cycle forms and promotes nerve repair. Furthermore, our studies have found that the newly constructed artificial nerve graft promotes the improvements in nerve conduction function and motor function in rats with sciatic nerve injury, and increases the expression of nerve injury repair factors fibroblast growth factor 2 and human transforming growth factor B receptor 2. These findings suggest that this artificial nerve graft effectively repairs peripheral nerve injury.

Hair growth-promotion effects and antioxidant activity of the banana flower extract HappyAngel^(■):double-blind, placebo-controlled trial

Banana flowers contain various bioactive components, including several antioxidants with anti-inflammatory effects. However, it is unclear whether they can reduce and prevent hair loss. This study examines the effect of banana flower extracts on preventing hair loss and strengthening hair roots. The banana flower extract(HappyAngel^(■))was used to treat human hair follicle dermal papilla cells(HFDPCs)and the expression of reactive oxygen species(ROS), dihydrotestosterone(DHT), and hair-related genes(SRD5A1, SRD5A2, AR, and KROX20)were monitored. Fifty subjects were divided into a placebo group and a banana flower group. The experimental group consumed banana flower extract daily for twelve weeks and then underwent hair testing, hair-related genes analysis, collection of hair loss, and questionnaires. The results showed that the banana flower extract significantly increased hair cell growth and decreased the expression of ROS, DHT, and hair follicle growth inhibition-related SRD5A1, SRD5A2, and AR genes, and significantly increased the expression of hair growth-related KROX20 gene in HFDPCs. Consuming banana flower extract for twelve weeks increased the hair root diameter and reduced hair loss and scalp redness compared to the placebo group. Thus, banana flower extract(HappyAngel^(■))can stimulate hair growth and inhibit the activation of hair loss genes.

PTEN inhibitor bisperoxovanadium protects against noise-induced hearing loss

Studies have shown that phosphatase and tensin homolog deleted on chromosome ten(PTEN)participates in the regulation of cochlear hair cell survival.Bisperoxovanadium protects against neurodegeneration by inhibiting PTEN expression.However,whether bisperoxovanadium can protect against noise-induced hearing loss and the underlying mechanism remains unclear.In this study,we established a mouse model of noise-induced hearing loss by exposure to 105 dB sound for 2 hours.We found that PTEN expression was increased in the organ of Corti,including outer hair cells,inner hair cells,and lateral wall tissues.Intraperitoneal administration of bisperoxovanadium decreased the auditory threshold and the loss of cochlear hair cells and inner hair cell ribbons.In addition,noise exposure decreased p-PI3K and p-Akt levels.Bisperoxovanadium preconditioning or PTEN knockdown upregulated the activity of PI3K-Akt.Bisperoxovanadium also prevented H_(2)O_(2)-induced hair cell death by reducing mitochondrial reactive oxygen species generation in cochlear explants.These findings suggest that bisperoxovanadium reduces noise-induced hearing injury and reduces cochlear hair cell loss.

Flavonoids in safflower extract reduce cisplatin-induced damage to human follicle dermal papilla cells by inhibiting DNA damage and Rad17/Chk1/Cdc25C signaling

Background:Cisplatin is a chemotherapeutic agent commonly used clinically for the treatment of various human cancers.Patients often reduce the use of cisplatin due to its side effects,which in turn affects its treatment.This study explored the mechanism of action of safflower extract as an adjuvant traditional Chinese medicine for chemotherapy.Methods:Primary human follicle dermal papilla cells(HFDPCs)were used as target cells for cisplatininduced damage to hair cells.Western blotting was used to investigate the molecular targets of cisplatin and safflower extract in causing HFDPCs damage.Cell survival and cell cycle were analyzed by mitochondrial staining reagent WST-1 and propidium iodide.Results:Cisplatin could reduce the viability of HFDPCs without causing cell death.Cisplatin increased the level of phospho-Rad17 in HFDPCs and activated the Chk1/Cdc25C signaling to reduce the expression of Cdc2 protein,thereby arresting the cells in the G2/M phase.The combination of safflower extract and the flavonoids could effectively inhibit the signal transduction of Rad17/Chk1/Cdc25 in cisplatin-treated cells and reduce the cell population in the G2/M phase.Finally,we also confirmed that safflower extract could effectively inhibit the damage to HFDPCs caused by cisplatin,mainly at the level of reducing the DNA damage caused by cisplatin.Conclusions:Safflower extract can be used as an adjuvant Chinese medicine for chemotherapy to reduce the damage caused by chemotherapy to normal hair follicle cells.

The role of Rho GTPase family in cochlear hair cells and hearing

Rho GTPases are essential regulators of the actin cytoskeleton.They are involved in various physiological and biochemical processes such as the regulation of cytoskeleton dynamics,development,proliferation,survival,and regeneration.During the development of cochlear hair cells,Rho GTPases are activated by various extracellular signals through membrane receptors to further stimulate multiple downstream effectors.Specifically,RhoA,Cdc42,and Rac1,members of the classical subfamily of the Rho GTPase family,regulate the development and maintenance of cilia by inducing the polymerization of actin monomers and stabilizing actin filaments.In addition,they also regulate the normal morphology orientation of ciliary bundles in auditory hair cells,which is an important element of cell polarity regulation.Moreover,the actin-related pathways mediated by RhoA,Cdc42,and Rac1 also play a role in the motility of outer hair cells,indicating that the function of Rho GTPases is crucial in the highly polar auditory sensory system.In this review,we focus on the expression of RhoA,Cdc42,and Rac1 in cochlear hair cells and how these small molecules participate in ciliary bundle morphogenesis and cochlear hair cell movement.We also discuss the progress of current research investigating the use of these small molecules as drug targets for deafness treatment.

A New Hybrid Model for Segmentation of the Skin Lesion Based on Residual Attention U-Net

Skin segmentation participates significantly in various biomedical applications,such as skin cancer identification and skin lesion detection.This paper presents a novel framework for segmenting the skin.The framework contains two main stages:The first stage is for removing different types of noises from the dermoscopic images,such as hair,speckle,and impulse noise,and the second stage is for segmentation of the dermoscopic images using an attention residual U-shaped Network(U-Net).The framework uses variational Autoencoders(VAEs)for removing the hair noises,the Generative Adversarial Denoising Network(DGAN-Net),the Denoising U-shaped U-Net(D-U-NET),and Batch Renormalization U-Net(Br-U-NET)for remov-ing the speckle noise,and the Laplacian Vector Median Filter(MLVMF)for removing the impulse noise.In the second main stage,the residual attention u-net was used for segmentation.The framework achieves(35.11,31.26,27.01,and 26.16),(36.34,33.23,31.32,and 28.65),and(36.33,32.21,28.54,and 27.11)for removing hair,speckle,and impulse noise,respectively,based on Peak Signal Noise Ratio(PSNR)at the level of(0.1,0.25,0.5,and 0.75)of noise.The framework also achieves an accuracy of nearly 94.26 in the dice score in the process of segmentation before removing noise and 95.22 after removing different types of noise.The experiments have shown the efficiency of the used model in removing noise according to the structural similarity index measure(SSIM)and PSNR and in the segmentation process as well.