Pigmented Tissues Research Articles

A comparative study of diode laser and CO2 laser in the treatment of gingival hyperpigmentation

Background: Pigmented gingival tissue, many a times forces the patients to seek treatment for esthetic periodontal therapy. Gingival hyperpigmenta- tion is caused by excessive deposition of melanin in the basal and suprabasal layers of epithelium. Materials and methods: This double blinded prospective study was undertaken to evaluate the effectiveness of diode laser and CO2 laser in gingival depigmentation in ten patients using split mouth technique by evaluating the parameters like colour of the gingiva, bleeding, difficulties of the operator, pain, esthetic perception by the patient and post-operative wound healing. Out of 10 patients in the study group 6 were males and 4 were females. Gingival depigmentation was done from canine to canine in the upper anterior region. A total of 20 an- terior segments were treated: 10 randomly to Carbon dioxide & Diode Laser group using Flip coin method. The data collected were tabulated and statistical analysis was performed. Results & conclusion: The results of this study showed that the degree of pigmentation at the end of 4th week was less in diode group with 20% of the patient showing no pigmentation compared to CO2 group (10%). From our study, the first of its kind to compare the two lasers, we come to the conclusion that on comparing both the group, diode laser study group had appeared to be ef- fective in the management of gingival hyperpigmentation in terms of pain, bleeding intra operatively, difficulty of the operator, wound healing and esthetics.

Novel loss-of-function variant in DENND5A impedes melanosomal cargo transport and predisposes to familial cutaneous melanoma

PurposeMore than half of the familial cutaneous melanomas have unknown genetic predisposition. This study aims at characterizing a novel melanoma susceptibility gene. MethodsWe performed exome and targeted sequencing in melanoma-prone families without any known melanoma susceptibility genes. We analyzed the expression of candidate gene DENND5A in melanoma samples in relation to pigmentation and UV signature. Functional studies were carried out using microscopic approaches and zebrafish model. ResultsWe identified a novel DENND5A truncating variant that segregated with melanoma in a Swedish family and 2 additional rare DENND5A variants, 1 of which segregated with the disease in an American family. We found that DENND5A is significantly enriched in pigmented melanoma tissue. Our functional studies show that loss of DENND5A function leads to decrease in melanin content in vitro and pigmentation defects in vivo. Mechanistically, harboring the truncating variant or being suppressed leads to DENND5A losing its interaction with SNX1 and its ability to transport the SNX1-associated vesicles from melanosomes. Consequently, untethered SNX1-premelanosome protein and redundant tyrosinase are redirected to lysosomal degradation by default, causing decrease in melanin content. ConclusionOur findings provide evidence of a physiological role of DENND5A in the skin context and link its variants to melanoma susceptibility.

Rhodopsins at a glance.

Rhodopsins are photoreceptive membrane proteins consisting of a common heptahelical transmembrane architecture that contains a retinal chromophore. Rhodopsin was first discovered in the animal retina in 1876, but a different type of rhodopsin, bacteriorhodopsin, was reported to be present in the cell membrane of an extreme halophilic archaeon, Halobacterium salinarum, 95 years later. Although these findings were made by physiological observation of pigmented tissue and cell bodies, recent progress in genomic and metagenomic analyses has revealed that there are more than 10,000 microbial rhodopsins and 9000 animal rhodopsins with large diversity and tremendous new functionality. In this Cell Science at a Glance article and accompanying poster, we provide an overview of the diversity of functions, structures, color discrimination mechanisms and optogenetic applications of these two rhodopsin families, and will also highlight the third distinctive rhodopsin family, heliorhodopsin.

Ultraviolet photoacoustic microscopy with tissue clearing for high-contrast histological imaging

Ultraviolet photoacoustic microscopy (UV-PAM) has been investigated to provide label-free and registration-free volumetric histological images for whole organs, offering new insights into complex biological organs. However, because of the high UV absorption of lipids and pigments in tissue, UV-PAM suffers from low image contrast and shallow image depth, hindering its capability for revealing various microstructures in organs. To improve the UV-PAM imaging contrast and imaging depth, here we propose to implement a state-of-the-art optical clearing technique, CUBIC (clear, unobstructed brain/body imaging cocktails and computational analysis), to wash out the lipids and pigments from tissues. Our results show that the UV-PAM imaging contrast and quality can be significantly improved after tissue clearing. With the cleared tissue, multilayers of cell nuclei can also be extracted from time-resolved PA signals. Tissue clearing-enhanced UV-PAM can provide fine details for organ imaging.

Genotyping Analysis by RAD-Seq Reads Is Useful to Assess the Genetic Identity and Relationships of Breeding Lines in Lavender Species Aimed at Managing Plant Variety Protection.

Lavender species are widely distributed in their wild forms around the Mediterranean Basin and they are also cultivated worldwide as improved and registered clonal varieties. The economic interest of the species belonging to the Lavandula genus is determined by their use as ornamental plants and important source of essential oils that are destinated to the production of cosmetics, pharmaceuticals and foodstuffs. Because of the increasing number of cases of illegal commercialization of selected varieties, the protection of plant breeders’ rights has become of main relevance for the recognition of breeding companies’ royalties. With this aim, genomic tools based on molecular markers have been demonstrated to be very reliable and transferable among laboratories, and also much more informative than morphological descriptors. With the rising of the next-generation sequencing (NGS) technologies, several genotyping-by-sequencing approaches are now available. This study deals with a deep characterization of 15 varietal clones, belonging to two distinct Lavandula species, by means of restriction-site associated DNA sequencing (RAD-Seq). We demonstrated that this technology screens single nucleotide variants that enable to assess the genetic identity of individual accessions, to reconstruct genetic relationships among related breeding lines, to group them into genetically distinguishable main subclusters, and to assign their molecular lineages to distinct ancestors. Moreover, a number of polymorphic sites were identified within genes putatively involved in biosynthetic pathways related to both tissue pigmentation and terpene production, useful for breeding and/or protecting newly registered varieties. Overall, the results highlighted the presence of pure ancestries and interspecific hybrids for the analyzed Lavandula species, and demonstrated that RAD-Seq analysis is very informative and highly reliable for characterizing Lavandula clones and managing plant variety protection.

Abnormal Arm Tone, Cigarette Smoking and Use of Blood Pressure Medication in a Sight Enhancement Clinic Population

There are three pigmented tissues originating from neural crest which frequently degenerate during aging in industrialized societies: the pigmented epithelium of the eye, the substancia nigra of the thalmus and the pigmented cells of the organ of Corti. Each of these tissues once developed does not regenerate following loss. They must last the insults of a lifetime for the elderly to see clearly, hear well, and have smooth coordination. Is degeneration of these tissues interrelated? Are they influenced by common risk factors?

Uveal Malignant Melanoma in a Hybrid Macaw Parrot (Ara chloropterus × Ara militaris).

An adult male green-winged (Ara chloropterus) × military (Ara militaris) hybrid macaw of unknown age was presented to the Texas A&M Veterinary Medical Teaching Hospital for evaluation of left periocular swelling, blepharospasm, and ocular discharge. Complete ophthalmic examination and B mode ocular ultrasonography were performed to evaluate the affected eye. Ocular examination revealed a blind left globe with exophthalmos, periocular swelling, diffuse corneal edema, and severe anterior fibrinous uveitis obscuring visualization of the intraocular structures. An ultrasound examination revealed a hyperechoic mass that filled the posterior segment of the globe. Focal discontinuity of the posterior sclera suggested scleral rupture. Enucleation with histopathology was recommended and performed 1.5 months later. Intraoperatively, a heavily pigmented and friable soft tissue mass extruded through the posterior sclera and extended into the orbit. Histologically, the mass was consistent with a malignant melanoma. The patient died 18 hours after the surgical procedure because of unknown complications. Postmortem examination did not identify evidence of metastasis. This study described the clinical appearance and histopathologic findings of a rare ocular neoplasm with extrascleral extension in a hybrid macaw. Although uncommon in psittacine birds, primary ocular melanocytic neoplasms may display features of malignancy with scleral or orbital invasion, or both.

Disruption of Photomorphogenesis Leads to Abnormal Chloroplast Development and Leaf Variegation in Camellia sinensis.

Camellia sinensis cv. ‘Yanlingyinbiancha’ is a leaf-variegated mutant with stable genetic traits. The current study aimed to reveal the differences between its albino and green tissues, and the molecular mechanism underlying the variegation. Anatomic analysis showed the chloroplasts of albino tissues to have no intact lamellar structure. Photosynthetic pigment in albino tissues was significantly lower than that in green tissues, whereas all catechin components were more abundant in the former. Transcriptome analysis revealed most differentially expressed genes involved in the biosynthesis of photosynthetic pigment, photosynthesis, and energy metabolism to be downregulated in albino tissues while most of those participating in flavonoid metabolism were upregulated. In addition, it was found cryptochrome 1 (CRY1) and phytochrome B (PHYB) genes that encode blue and red light photoreceptors to be downregulated. These photoreceptors mediate chloroplast protein gene expression, chloroplast protein import and photosynthetic pigment biosynthesis. Simultaneously, SUS gene, which was upregulated in albino tissues, encodes sucrose synthase considered a biochemical marker for sink strength. Collectively, we arrived to the following conclusions: (1) repression of the biosynthesis of photosynthetic pigment causes albinism; (2) destruction of photoreceptors in albino tissues suppresses photomorphogenesis, leading to abnormal chloroplast development; (3) albino tissues receive sucrose from the green tissues and decompose their own storage substances to obtain the energy needed for survival; and (4) UV-B signal and brassinosteroids promote flavonoid biosynthesis.

Tissue Spectroscopy and Optical Clearing of Colorectal Mucosa in the Pursuit of New Cancer Diagnostic Approaches

In this paper we present three studies that demonstrate the applicability of spectroscopy methods and optical clearing treatments in pathology identification and monitoring. In the first study, by obtaining the absorption spectra of human healthy and pathological (adenocarcinoma) colorectal mucosa tissues, it was possible to identify a higher content of a pigment in the diseased tissues. This study also shows that machine learning methods can be used to reach the same differentiated results in vivo through diffuse reflectance spectroscopy. In the second study, the combination of collimated transmittance spectroscopy with optical clearing treatments allowed to obtain the diffusion coefficients of glucose in healthy and pathological colorectal mucosa as: D glucose =5.8x 10 – 7 cm 2 /s and D glucose =4.4x 10 – 7 cm 2 /s, respectively. This study also demonstrated that the diseased tissues contains about 5% more mobile water than the healthy tissues. The third study was performed to evaluate the protein dissociation mechanism of optical clearing. By treating both healthy and pathological colorectal mucosa tissues with 93%-glycerol, a protein dissociation rate of about 3 times higher was obtained for the pathological mucosa. All the discriminating parameters that result from these studies can be obtained in the in vivo situation through diffuse reflectance spectroscopy and further studies to evaluate their values in different stages of cancer progression are of great importance to develop disease monitoring protocols.

Voltammetric Electrochemical Sensor for Phylogenetic Study in Acer Linn.

Acer Linn. is a highly divergent species morphology in the maple family (Aceraceae). It is one of the genera facing a very difficult taxonomic situation. The phylogeny of the genus and the taxonomic system under the genus remain unclear. The use of electrochemical fingerprints for plant phylogenetic study is an emerging application in biosensors. In this work, leaves of 18 species of Acer Linn. with an exo-taxa were selected for electrochemical fingerprint recording. Two different conditions were used for improving the data abundance. The fingerprint of all species showed a series of oxidation peaks. These peaks can be ascribed to the oxidation of flavonols, phenolic acids, procyanidins, alkaloids, and pigments in plant tissue. These electrochemical fingerprints can be used for the identification of plant species. We also performed a phylogenetic study with data from electrochemical fingerprinting. The phylogenetic tree of Acer is divided into three main clades. The result is in full agreement with A. shangszeense var. anfuense, A. pictum subsp. mono, A. amplum, A. truncatum, and A. miaotaiense, belonging to the subsection Platanoidea. A. nikoense and A. griseum were clustered together in the dendrogram. Another group that fits the traditional classification results is in the subsection Integrifolia.

An Ayurvedic approach in the management of Koshta-Shakharshita Kamala with special reference to Hepatocellular Jaundice: A Case study

Modern lifestyle and advanced technology have given life easier, but this has led to many diseases. In advanced lifestyle irregular eating habits, eating unhealthy foods and eating spicy fast food has become a fashion and alcohol consumption is increasing day by day. All of these factors lead to different disorders. Kamala is one among the diseases which are caused due to excessive intake of sour food, alcohol, unhealthy food and when a person with Panduroga continues intake of Pittakara Aahara then he may develop Kamala. According to modern science, Kamala can be correlated with Jaundice. Clinically the Jaundice is a sign of an ongoing disease process with common signs and symptoms like yellowish discolourations of the skin, mucous membranes, the eyes, urine etc. It is characterized by increase deposition of bile pigments in body fluids and tissues. It is perceptible only when the bilirubin level and its conjugates exceeds 1.5 mg/ 100ml plasma. Here is the case study of a patient who appeared to Parul Ayurved Hospital with the history of oedema over the bilateral lower limbs, heaviness in abdomen and chest region in the last 1 year, fever since 4-5 months and yellowish discolourations of eyes, nails and urine are present. In the present study, the patient was treated with Ayurvedic treatments i.e. Virechana Karma and Shamana Chikitsa.

Development of an Effective Therapy for Alkaptonuria - Lessons for Osteoarthritis.

Osteoarthritis (OA) is one of the major causes of disability and pain worldwide, yet despite a massive international research effort, no effective disease-modifying drugs have been identified to date. In this review, we put forward the proposition that greater focus on rarer forms of OA could lead to a better understanding of the pathogenesis of more common OA. We have investigated the severe osteoarthropathy of the ultra-rare disease alkaptonuria (AKU). In addition to the progress made in finding a treatment for AKU, our research has revealed important lessons for more common OA, including the identification of high-density mineralized protrusions (HDMPs), new pathoanatomical structures which may play an important role in joint destruction and pain in AKU and in OA. AKU is an inherited disorder of tyrosine metabolism, caused by genetic lack of the enzyme homogentisate 1,2 dioxygenase (HGD), which leads to failure to breakdown homogentisic acid (HGA). While most HGA is excreted over time, some of it is deposited as a pigment in connective tissues, a process described as ochronosis. Ochronotic pigment alters the mechanical properties of tissues, leading to inevitable joint destruction and frequently to cardiac valve disease. Until recently, there was no effective therapy for AKU, but preclinical studies demonstrated that upstream inhibition of tyrosine metabolism by nitisinone, a drug previously used in hereditary tyrosinaemia 1 (HT1), completely prevented ochronosis in AKU mice. This was followed by successful clinical trials which have resulted in nitisinone being approved for therapy of AKU by the European Medicines Agency, making AKU the only cause of OA for which there is an effective therapy to date. Study of other rare causes of OA should be a higher priority for researchers and funders to ensure further advances in understanding and eventual therapy of OA.

POS1393 QUANTITATIVE AUTOFLUORESCENCE FINDINGS IN PATIENTS UNDERGOING HYDROXYCHLOROQUINE TREATMENT

Background:Hydroxychloroquine (HCQ) is a relatively safe and effective drug widely used as primary or adjunctive treatment for several rheumatological and dermatological disorders1. HCQ modulates immune response through several mechanisms and has a tropism for pigmented ocular tissues, particularly retinal pigment epithelium (RPE)2. Its accumulation within RPE cells can lead to sight threatening retinal toxicity, with bull’s eye maculopathy (BEM) representing its advanced phenotype. 3 Quantitative Auto-Fluorescence (qAF) is an imaging modality that allows the measurement of retinal auto-fluorescence following short-wavelength light (488nm) excitation of retinal fluorophores (lipofuscin). 4 Two recent studies have focused on qAF values in patients treated with HCQ 5,6. In both cases qAF was increased in eyes with BEM. Furthermore, Reichel et al.6 were able to detect increased values of qAF in patients without BEM as early as 6 months after the start of HCQ treatment using an experimental imaging analysis procedure.Objectives:To measure quantitative autofluorescence (qAF) in patients under treatment with hydroxychloroquine (HCQ) with no apparent signs of retinal toxicity and to compare it with that of untreated subjects.Methods:Consecutive patients at risk for the development of HCQ retinal toxicity (duration of treatment >5 years or daily HCQ dose >5 mg/kg of actual body weight (ABW) and/or renal insufficiency)7 but no alterations on Spectral Domain - Optical Coherence Tomography, Short-Wavelength Autofluorescence and 10-2 Visual Field examination were recruited. Healthy subject matched by age and sex were also enrolled in the study. All subjects underwent qAF measurements in one eye. Images were analyzed using the conventional qAF grid by Delori calculating the qAF of 8 sectors of the intermediate ring and the mean of those values (qAF8).Results:Thirty-nine patients treated with HCQ (38 females, mean age 52,1 ± 8,6 years) and 39 untreated subjects (38 females, mean age 51,2 ± 8,6 years). In both HCQ patients and untreated subjects, qAF8 was positively correlated with age (p=0.004) (Figure 1). Although HCQ patients showed a higher mean qAF8 compared to untreated subjects (294,7 ±65,3 vs 268,9 ± 57,5), the difference was not significant (p=0.068). HCQ patients showed significantly higher mean qAF values in the inferior-temporal, inferior and inferior-nasal sectors of the intermediate ring of qAF grid compared to untreated subjects (all p<0.05).Figure 1.Visual representation of a model predicting the standardized qAF values as influenced by age and HCQ daily dose/ABW, calculated for a treatment duration of 15 years.Conclusion:These results suggest a possible preclinical increase of qAF values in inferior parafoveal sectors probably induced by HCQ exposure. Further studies are required to improve our understanding of preclinical stages of HCQ retinopathy and the possible role of qAF in the HCQ toxicity screening.

Violet-blue light exposure of the skin: is there need for protection?

Advocates of skin protection against blue light express concern about exposure to indoor lighting and electronic screens as well as natural outdoor exposure. However, the nature of adverse effects in skin is unclear and the doses to induce effects are unknown. We aimed to reveal whether there is a scientific basis for promoting skin protection against violet-blue light (400–500 nm, VBL). Based on published literature, we determined the time to reach a threshold dose that induced a biological response in human skin. In the absence of an action spectrum for effects on skin, we used a hand held probe with a defined spectral response and measurements of the unweighted exposure between 400 and 500 nm to estimate the exposure by a selection of artificial light sources and solar light. For comparison, an outdoor threshold erythemally weighted UV dose was set to 1 SED (standard erythema dose). Outdoor, weighted irradiances were obtained using a radiative transfer model. Induction of pigmentation in human skin tissue was the only consistently reported endpoint after VBL exposure of about 65 Jcm−2. This threshold dose was reached in 0.5 to 20 months of exposure to indoor lighting sources. In comparison, specialised medical sources reached this dose in 0.5 min to 45 h. The time outdoors to reach 1 SED was shorter than the time to reach a VBL threshold dose throughout all seasons. Skin protection against VBL is superfluous for exposures to domestic lighting sources or screens and for solar radiation; however, it may be advantageous for patients suffering from photosensitive diseases or taking photosensitising medication.

Effects of heat stress on photosystem II activity and antioxidant enzymes in two maize cultivars.

The main reason for the maize genotype "DKC7221" to be heat tolerant is to have higher photosynthetic activity under heat stress conditions. The genotype "P3167" is sensitive to high temperature because of the heat-induced inhibition in photosynthetic electron transport reactions. In the present study, the effect of heat stress (45ºC for 20min) on some physiological changes was investigated through a chlorophyll afluorescence technique, and some endogenous resistance mechanisms (activities of some antioxidant enzymes, free proline, and reduced ascorbate contents) in two maize cultivars (Zea mays L. cvs. P3167 and DKC7221). Chlorophyll fluorescence measurements demonstrated that heat stress led to the reduction in the efficiency of the Hill reaction, accumulation of inactive reaction centers, inhibition of electron flow from reaction centers to the plastoquinone pool, and induction of non-photochemical dissipation of absorbed light energy. Changes in Φo/(1 - Φo), SFIABS and PIABS indicated that electron transport reactions in P3167 were almost completely inhibited by heat stress. In DKC7221, however, photosynthetic electron transport reactions were maintained under heat stress conditions. As a result of impairment in the photosynthetic efficiency in P3167 under heat stress, oxidative stress appeared as shown by lower antioxidant activity, accumulation of H2O2, malondialdehyde, and formazon and photooxidative injuries in chlorophyll pigments in the leaf tissue. DKC7221, on the other hand, had a higher antioxidant efficiency and lower oxidative damage under heat stress. FeSOD activity was found to be responsible for the dismutation of superoxide radicals in both maize genotypes under heat stress. As a result, it may be concluded that the genotype DKC7221 is more tolerant to heat stress than P3167.

Escleras azules e hiperlaxitud ligamentaria ¿Enfermedad del tejido conectivo?

Introduction: the bluish coloration of sclera, frequently, can be associated with multiple systemic syndromes like the ligamentous hyperlaxity, which is a connective tissue disease currentely affecting population, joints, tendons and also some organs, all due to tegumentary weaknesses. Objective: to describe all epidemiologic and clinical characteristics of joint hypermobility syndrome associated with blue sclera in pediatric patients. Method: a review of the literature was conducted on published articles from May 2002 up to August 2020. Databases checked were Scopus, Wiley Online Library, SciELO, Dialnet, EBSCO, PubMed/Medline. Keywords used were: “blue scleras”, “joints”, “joint hypermobility syndrome”, “connective tissue”, all in spanish and english language. A total of 21 bibliographic references were selected. Results: the uvea is a highly pigmented tissue, and when this pigmented layer increase or when the sclera decrease, allows the uvea to show through the sclera, giving rise to blue sclera. If extraocular manifestations are obvious, so blue sclera becomes an additional data supporting the diagnosis of joint hypermobility syndrome, may even be the only guide to an accurate diagnosis. Conclusions: it becomes necessary, from an ophthalmologic point of view, consider blue sclera a possible sign of onset acute diseases, cause of severe visual loss and systemic syndromes like joint hypermobility syndrome.

Microsurgical Dissection and Tissue Clearing for High Resolution Intact Whole Retina and Vitreous Imaging.

Neuronal and vascular structures of the retina in physiologic and pathologic conditions can be better visualized and characterized by using intact whole retina imaging techniques compared to conventional retinal flat mount preparations and sections. However, immunofluorescent imaging of intact whole retina is hindered by the opaque coatings of the eyeball, i.e., sclera, choroid, and retinal pigment epithelium (RPE) and the light scattering properties of retinal layers that prevent full thickness high resolution optical imaging. Chemical bleaching of the pigmented layers and tissue clearing protocols have been described to address these obstacles; however, currently described methods are not suitable for imaging endogenous fluorescent molecules such as green fluorescent protein (GFP) in intact whole retina. Other approaches bypassed this limitation by surgical removal of pigmented layers and the anterior segment of the eyeball allowing intact eye imaging, though the peripheral retina and hyaloid structures were disrupted. Presented here is an intact whole retina and vitreous immunofluorescent imaging protocol that combines surgical dissection of the sclera/choroid/retina pigment epithelium (RPE) layers with a modified tissue clearing method and light sheet fluorescent microscopy (LSFM). The new approach offers an unprecedented view ofunperturbed vascular and neuronal elements of the retina as well as the vitreous and hyaloid vascular system in pathologic conditions.

Morphea: progress to date and the road ahead.

Morphea is a rare autoimmune condition causing inflammation and sclerosis of the skin and underlying soft tissue. It is characterized by periods of activity (inflammation admixed with fibrosis), ultimately resulting in permanent damage (pigment change and tissue loss). Damage resulting from unchecked activity can lead to devastating, permanent cosmetic and functional sequelae including hair loss; cutaneous, soft tissue and bony atrophy; joint contractures; and growth restriction of the affected body site in children. This makes the early identification of activity and initiation of appropriate treatment crucial to limiting damage in morphea. To this end, recent investigative work has focused on validation of clinical, biomarker, imaging, and histologic outcomes aimed at accurately quantifying activity and damage. Despite promising results, further work is needed to better validate these measures before they can be used in the clinic and research settings. Although there has been recent approval of less toxic, targeted therapies for many inflammatory skin conditions, none have been systematically investigated in morphea. The mainstays of treatment for active morphea are corticosteroids and methotrexate. These are often limited by substantial toxicity. The paucity of new treatments for morphea is the result of a lack of studies examining its pathogenesis, with many reviews extrapolating from research in systemic sclerosis. Recent studies have demonstrated the role of dysregulated immune and fibrotic pathways in the pathogenesis of morphea, particularly interferon (IFN) gamma related pathways. Active morphea lesions have been found to display an inflammatory morphea signature with CXCR3 receptor ligands, as well as a distinct fibrotic signature reflecting fibroblast activation and collagen production. CXCL9 and 10 have been associated with increased measures of disease activity. While immune dysfunction is thought to play the primary role in morphea pathogenesis, there are other factors that may also contribute, including genetic predisposition, environmental factors, and vascular dysregulation. There remains an essential need for further research to elucidate the pathogenesis of morphea and the mode of action of dysregulated upstream and downstream immune and fibrotic pathways. These studies will allow for the discovery of novel biomarkers and targets for therapeutic development.

Drought increases chlorophyll content in stems of Vitis interspecific hybrids

Plants respond to drought stress through different physiological mechanisms. To highlight a specific adaptation strategy related to stem photosynthetic activity, woody tissue pigmentation was studied. In this work, 25 Vitis hybrids were studied in two experimental vineyards: one of them with sufficient water availability and the other showing drought stress. 1500 reflectance spectra of woody tissues were collected. Beside the spectral elaboration and interpretation, indexes for the pigment quantification were calculated. The content and the proportion in chlorophylls (a and b) in stems were significantly modified in response to water stress. Genotypes were classified based on their plasticity/elasticity and adaptive/susceptible behavior. The cluster analysis produced 4 main groups based on their average spectra and on the spectral variations between watered and stress conditions. The most interesting genotypes concerning this trait were pointed out. Nevertheless, this adaptation mechanism could also be due to an increased detoxification necessity related to other physiological disfunctions caused by water stress. A multi-parameter approach is encouraged for phenotyping of abiotic stress tolerant plant selection. In the next future, this work will support the release of new drought tolerant rootstocks for viticulture.

Culturing of Melanocytes from the Equine Hair Follicle Outer Root Sheath

Hair follicles harbor a heterogeneous regenerative cell pool and represent a putative low-to-non-invasively available source of stem cells. We previously reported a technology for culturing human melanocytes from the hair follicle outer root sheath (ORS) for autologous pigmentation of tissue engineered skin equivalents. This study translated the ORS technology to horses. We de-veloped a culture of equine melanocytes from the ORS (eMORS) from equine forelock hair follicles cultured by means of an analogue human hair follicle-based in vitro methodology. The procedure was adjusted to equine physiology by addition of equine serum to the culture medium. The hair follicles were isolated by macerating forelock skin rests, enzymatically digested and subjected to air-medium-interface cultivation method. The procedure resulted in differentiated equine melanocytes, which exhibited typical morphology, presence of melanosomes, expression of cytoskeleton proteins vimentin, α-SMA, Sox2, S100ß and tyrosinase as well as tyrosinase activity followed by production of melanin. According to all assessed parameters, eMORS could be ranked as partially melanotic melanocytes. The results of the study offer an experimental base for further insight into hair follicle biology in equine and for comparative studies of hair follicles across different species.