Methylmercury Effect and Distribution in Two Extremophile Microalgae Strains Dunaliella salina and Coccomyxa onubensis from Andalusia (Spain).

The main entrance point of highly toxic organic Hg forms, including methylmercury (MeHg), into the aquatic food web is phytoplankton, which is greatly represented by various natural microalgal species. Processes associated with MeHg fate in microalgae cells such as uptake, effects on cells and toxicity, Hg biotransformation, and intracellular stability are detrimental to the process of further biomagnification and, as a consequence, have great importance for human health. The study of MeHg uptake and distribution in cultures of marine halophile Dunaliella salina and freshwater acidophilic alga Coccomyxa onubensis demonstrated that most of the MeHg is imported inside the cell, while cell surface adhesion is insignificant. Almost all MeHg is removed from the culture medium after 72 h. Significant processes in rapid MeHg removal from liquid medium are its abiotic photodegradation and volatilization associated with algal enzymatic activity. The maximum intracellular accumulation for both species was in 80 nM MeHg-exposed cultures after 24 h of exposure for D. salina (from 27 to 34 µg/gDW) and at 48 h for C. onubensis (up to 138 µg/gDW). The different Hg intakes in these two strains could be explained by the lack of a rigid cell wall in D. salina and the higher chemical ability of MeHg to pass through complex cell wall structures in C. onubensis. Electron microscopy studies on the ultrastructure of both strains demonstrated obvious microvacuolization in the form of many very small vacuoles and partial cell membrane disruption in 80 nM MeHg-exposed cultures. Results further showed that Coccomyxa onubensis is a good candidate for MeHg-contaminated water reclamation due to its great robustness at nanomolar concentrations of MeHg coupled with its very high intake and almost complete Hg removal from liquid medium at the MeHg levels tested.

Dermatofibrosarcoma Protuberans: A Study of 148 Patients.

INTRODUCTION: Dermatofibrosarcoma protuberans (DFSP) is the most common sarcoma of the skin. Although distant metastases are infrequent, DFSP is highly aggressive locally with frequent local recurrences. It has been reported that the presence within the tumour of areas histopathologically mimicking fibrosarcoma may increase the risk of recurrence. OBJECTIVE: The objective of this study was to review the clinical features of our patients with DFSP and the factors associated with recurrence of the tumour, focussing on the presence of fibrosarcomatous areas. METHODS: Retrospective study of patients with DFSP diagnosed in 1990-2021 in a tertiary university hospital. The medical records were reviewed to obtain the following data: age, sex, tumour location, diameter, evolution time, presence of fibrosarcomatous areas, development of recurrence, and follow-up. Factors possibly associated with disease-free survival were analysed with Kaplan-Meier method and multivariate Cox regression. RESULTS: 148 patients (74 women/74 men, mean age 46.28 years, SD 14.431) were included in the study. Tumours involved the head and neck in 15 cases, thorax in 31, abdomen in 16, upper back in 43, lower back in 10, upper extremities in 10, and lower extremities in 23. Fibrosarcoma-like areas were observed in 16 tumours (10.81%). In 17 patients (11.49%), recurrences were observed (13 local recurrences, 3 lung metastasis, and 1 local recurrence with lung metastasis). Fibrosarcomatous DFSP recurred more frequently than classic DFSP (50% vs. 6.82%, respectively), and its disease-free survival was significantly lower (p < 0.001). In multivariate Cox regression, the presence of fibrosarcomatous areas was the only factor influencing disease-free survival. CONCLUSIONS: It is important to identify the fibrosarcomatous variant since it recurs more frequently and has lower recurrence-free survival. Distant metastases, mainly in the lung, are also more frequent in fibrosarcomatous DFSP.

Dermatomyositis in a donor and receptor of allogenic hematopoietic stem cell transplantation: Auto- or alloimmune disease?

We present the case of a 63-year-old woman who developed dermatomyositis after hematopoietic stem cell transplantation. Anti-melanoma differentiation-associated gene 5 (anti-MDA5) antibodies were positive and pulmonary involvement was severe and progressive. In addition, we also report that the patient's sister and donor also developed dermatomyositis. She had positive anti-PL7 antibodies and negative anti-MDA5 antibodies. The occurrence of autoimmune diseases after allogeneic hematopoietic stem cell transplantation is infrequent and difficult to interpret due to the reconstitution of the immune system and the multifactorial origin of most of these diseases. To our knowledge, this is the first described case of a hematopoietic progenitor transplant donor and recipient developing dermatomyositis. These findings make us wonder whether the dermatomyositis in this case is due to a shared genetic predisposition or to the donor's disease developing in the recipient.

Ultrasound-Based Recovery of Anti-Inflammatory and Antimicrobial Extracts of the Acidophilic Microalga Coccomyxa onubensis.

In the present study, the recovery of valuable molecules of proven anti-inflammatory and antimicrobial activity of the acidophilic microalga Coccomyxa onubensis (C. onubensis) were evaluated using green technologies based on ultrasound-assisted extraction (UAE). Using a factorial design (3 × 2) based on response surface methodology and Pareto charts, two types of ultrasonic equipment (bath and probe) were evaluated to recover valuable compounds, including the major terpenoid of C. onubensis, lutein, and the antimicrobial activity of the microalgal extracts obtained under optimal ultrasound conditions (desirability function) was evaluated versus conventional extraction. Significant differences in lutein recovery were observed between ultrasonic bath and ultrasonic probe and conventional extraction. Furthermore, the antimicrobial activity displayed by C. onubensis UAE-based extracts was greater than that obtained in solvent-based extracts, highlighting the effects of the extracts against pathogens such as Enterococcus hirae and Bacillus subtilis, followed by Staphylococcus aureus and Escherichia coli. In addition, gas chromatography-mass spectrometry was performed to detect valuable anti-inflammatory and antimicrobial biomolecules present in the optimal C. onubensis extracts, which revealed that phytol, sterol-like, terpenoid, and even fatty acid structures could also be responsible for the antibacterial activities of the extracts. Moreover, UAE displayed a positive effect on the recovery of valuable molecules, improving biocidal effects. Our study results facilitate the use of green technology as a good tool in algal bioprocess engineering, improving energy consumption and minimizing environmental impacts and process costs, as well as provide a valuable product for applications in the field of biotechnology.

Interaction of Naturally Occurring Phytoplankton with the Biogeochemical Cycling of Mercury in Aquatic Environments and Its Effects on Global Hg Pollution and Public Health.

The biogeochemical cycling of mercury in aquatic environments is a complex process driven by various factors, such as ambient temperature, seasonal variations, methylating bacteria activity, dissolved oxygen levels, and Hg interaction with dissolved organic matter (DOM). As a consequence, part of the Hg contamination from anthropogenic activity that was buried in sediments is reinserted into water columns mainly in highly toxic organic Hg forms (methylmercury, dimethylmercury, etc.). This is especially prominent in the coastal shallow waters of industrial regions worldwide. The main entrance point of these highly toxic Hg forms in the aquatic food web is the naturally occurring phytoplankton. Hg availability, intake, effect on population size, cell toxicity, eventual biotransformation, and intracellular stability in phytoplankton are of the greatest importance for human health, having in mind that such Hg incorporated inside the phytoplankton cells due to biomagnification effects eventually ends up in aquatic wildlife, fish, seafood, and in the human diet. This review summarizes recent findings on the topic of organic Hg form interaction with natural phytoplankton and offers new insight into the matter with possible directions of future research for the prevention of Hg biomagnification in the scope of climate change and global pollution increase scenarios.

Fe (III)-Mediated Antioxidant Response of the Acidotolerant Microalga Coccomyxa onubensis.

Coccomyxa onubensis (C. onubensis) is an acidotolerant microalga isolated from Tinto River (Huelva), which contains high levels of metal cations in solution, mainly Fe (II) and (III), and Cu (II). Fe is more bioavailable at low pH, mainly because Fe (II) and Fe (III) are far more soluble, especially Fe (III). For this reason, this study aims to evaluate both physiological and biochemical responses of C. onubensis when subjected to Fe (III)-induced stress. Changes in growth, photosynthetic viability and antioxidant responses to the induced oxidative stress were determined. The results obtained suggest that the addition of moderate Fe (III) levels to C. onubensis cultures results in improved growth and photosynthetic viability. Increases in the intracellular levels of the enzyme superoxide dismutase (SOD) and flavonoids, used as antioxidant response biomarkers, a point at Fe (III)-mediated oxidative stress induction. The apparent decrease in the content of other phenolic molecules and polyunsaturated fatty acids might be understood as a sign of antioxidant molecules' involvement in reactive oxygen species (ROS) scavenging. In conclusion, a noticeable antioxidant capacity displayed by C. onubensis allows the use of moderate Fe (III) levels to trigger the accumulation of valuable antioxidant molecules, allowing the production of cell extracts with potential anti-inflammatory activity.

Methodological Optimization of Supercritical Fluid Extraction of Valuable Bioactive Compounds from the Acidophilic Microalga Coccomyxa onubensis.

Microalgae grow in diverse environments and possess a great biotechnological potential as they contain useful bioactive compounds. These bioactive compounds can be obtained by selective and energy-efficient extraction methods. Various industries are using the supercritical fluid extraction (SFE) method to extract these valuable bioactive compounds. Hence, for the first time, we evaluated the effects of SFE on the recovery of bioactive and antioxidant compounds using Coccomyxa onubensis, a eukaryotic acidophilic microalga of potential relevance which can be used in the field of nutraceutical and functional foods. It was isolated from the Tinto River (Pyritic Belt, Huelva, Spain), a mining region in Spain. Variables such as extraction yield, lutein purity (LP) and recovery (LR), total phenols, and antioxidant capacity (Trolox equivalents antioxidant capacity method) were studied using a Box-Behnken design based on a response surface methodology along with the overall extraction curve fitted to a spline linear model. The effects of temperature (30, 50, and 70 °C), pressure (25, 40, and 55 MPa), and the percentage of co-solvent (0, 25%, and 50% v/v ethanol) on SFE were analyzed, resulting in the co-solvent and temperature as the most significant factors followed by the pressure. Under 70 °C, 40 MPa, and 50% v/v ethanol, C. onubensis reached a maximum of 66.98% of LR. The extracts were richest in total phenols and showed the maximum antioxidant activity (36.08 mg GAEs/g extracts and 2.237 mmol TE/g extracts, respectively) under similar pressure and co-solvent percentage values and different temperatures (30 and 70 °C, respectively). The extracts obtained in this study may have potential applications in the food, nutraceutical, and cosmetic industries. SFE is a highly efficient method to valorize microorganisms living in extreme environments, which are so far unexplored using green extraction methods.

Mastitis granulomatosa asociada a eritema nudoso: estudio de 42 casos / Granulomatous mastitis associated with erythema nodosum: A case series of 42 patients

Introducción:La mastitis granulomatosa (MG) se define por la presencia de inflamación granulomatosa en el tejido mamario. El eritema nudoso (EN) es una paniculitis inflamatoria reactiva caracterizada por nódulos subcutáneos dolorosos en las extremidades inferiores. La asociación entre MG y EN ha sido descrita, pero está poco estudiada. Nuestro objetivo fue analizar las características clínicas de las pacientes diagnosticadas de MG en nuestro centro y su asociación con EN.Métodos:Se revisaron retrospectivamente los casos diagnosticados histológicamente de MG entre 1995 y 2020.Resultados:Cuarenta y dos mujeres fueron diagnosticadas de MG. La edad media al diagnóstico fue de 41,619años y el 59,5% tenían ascendencia sudamericana. El EN se asoció con MG en el 11,9% de las pacientes. Las pacientes con EN fueron diagnosticadas precozmente respecto a aquellas con MG aislada (0,4meses frente a 6,81; p<0,05). La ulceración en la MG fue más prevalente en pacientes con EN asociado (60% vs 14,7%; p<0,05).Conclusión:El EN en pacientes con GM puede facilitar el diagnóstico precoz de esta rara condición que imita el cáncer de mama.(AU)

Background:Granulomatous mastitis (GM) is defined by the formation of granulomatous inflammation in breast tissue. Erythema nodosum (EN) is a reactive inflammatory panniculitis characterized by erythematous subcutaneous nodules in the lower limbs. The association of GM with EN has been rarely reported. Our aim was to retrospectively review our series of patients with GM to better characterize their features and their association with EN.Methods:Cases histologically diagnosed as granulomatous inflammation in breast tissue between 1995 and 2020 were retrospectively reviewed.Results:Forty-two women were diagnosed with GM. The average age at diagnosis was 41.619years, and 59.5% were of South-American ethnicity. EN was associated with GM in 11.9% of the patients. Patients with EN were diagnosed earlier than isolated GM (0.4months vs 6.81months; P<.05). Ulceration in the GM was more prevalent in patients with associated EN (60% vs 14.7%; P<.05).Conclusion:EN in patients with GM may reduce the evolution time and may help to diagnose this rare condition that mimics breast carcinoma. (AU)

Cutaneous Leiomyoma: A Clinical Study of 152 Patients.

BACKGROUND: Cutaneous leiomyoma (CL) is a benign smooth muscle tumour included in painful skin tumours. Multiple CLs are cutaneous markers of hereditary leiomyomatosis and renal cell cancer (HLRCC). OBJECTIVES: To retrospectively review our series of patients with CLs to analyse their clinical features and the association with HLRCC. METHODS: Cases coded as CL in the database of the pathology department between 2004 and 2019 were included in the study. Medical records were retrospectively reviewed to obtain the following data: age, sex, location, number of lesions, diameter, evolution time at diagnosis, suspected clinical diagnosis, tenderness, status of resection margins, development of recurrence, follow-up time, and association with HLRCC. RESULTS: 152 patients had CLs, 89 women and 63 men, mean age 56.26, SD 16.030 years. Subtypes were piloleiomyoma in 62 patients, angioleiomyoma in 80, and genital leiomyoma in 10. All of our 11 patients with multiple lesions corresponded to piloleiomyomas, and HLRCC was confirmed in 8 of them (73%). Patients with HLRCC were younger than patients with piloleiomyomas without HLRCC (34.88 vs. 56.17 years, p = 0.009). Vascular and genital leiomyomyomas were solitary and were not associated with HLRCC. CONCLUSION: In patients with multiple piloleiomyomas HLRCC must be ruled out as it is confirmed in a high proportion of cases. The probability of fumarate hydratase mutation is greater in multiple piloleiomyomas involving both the trunk and upper extremities in the same patient.

[Granulomatous mastitis associated with erythema nodosum: A case series of 42 patients]. / Mastitis granulomatosa asociada a eritema nudoso: estudio de 42 casos.

BACKGROUND: Granulomatous mastitis (GM) is defined by the formation of granulomatous inflammation in breast tissue. Erythema nodosum (EN) is a reactive inflammatory panniculitis characterized by erythematous subcutaneous nodules in the lower limbs. The association of GM with EN has been rarely reported. Our aim was to retrospectively review our series of patients with GM to better characterize their features and their association with EN. METHODS: Cases histologically diagnosed as granulomatous inflammation in breast tissue between 1995 and 2020 were retrospectively reviewed. RESULTS: Forty-two women were diagnosed with GM. The average age at diagnosis was 41.619years, and 59.5% were of South-American ethnicity. EN was associated with GM in 11.9% of the patients. Patients with EN were diagnosed earlier than isolated GM (0.4months vs 6.81months; P<.05). Ulceration in the GM was more prevalent in patients with associated EN (60% vs 14.7%; P<.05). CONCLUSION: EN in patients with GM may reduce the evolution time and may help to diagnose this rare condition that mimics breast carcinoma.

Variability of Omega-3/6 Fatty Acid Obtained Through Extraction-Transesterification Processes from Phaeodactylum tricornutum.

The effect of direct transesterification methods on the omega-3/6 composition of extracts from Phaeodactylum tricornutum was studied. The aim of this work was to identify an extraction method which allowed to obtain the most suitable profile of fatty acids in terms of its potential benefits to health, particularly if further used in the food industry. Seven methods using acids, alkalis, and heterogeneous-catalysts, (namely methods from 1 to 7, abbreviated as M1-M7) were performed to determine α-linolenic (ALA), linoleic (LA), docosahexaenoic (DHA) and eicosapentaenoic (EPA) acids. The composition of fatty acids was in all cases characterized by the major abundance of palmitic (23.95-34.08%), palmitoleic (30.94-35.56%), oleic acids (3.00-7.41%), and EPA (0.5-6.45%). Unsaturated fatty acids extraction yield was higher with a two-step transesterification process (M6, 63.65%). The total fatty acid methyl ester content (FAME) obtained with acid-transesterification (M1) reached about 21% wt, and 60% w/w total lipids. ALA higher relative content (ALA/LA ratio) was obtained when a lipid pre-extraction step was performed prior to acid-catalysis (M4). The transesterification method based on alkali-catalyst (M3, KOH catalyst) led to obtain higher DHA relative contents (DHA/EPA ratio up to 0.11), although its FAME content was 3.75-fold lower than that obtained with acid-transesterification (M1). Overall, this study shows that direct transesterification with alkali-catalyst (M3) improves the determination of PUFA content from the diatom through a more efficient transesterification-based extraction process, and thus allow to assess the value of the biomass more accurately for application in the food industry.

Sarcoidosis Induced by Atezolizumab Presenting With Xanthelasma Infiltration.

ABSTRACT: Sarcoidosis induced by anti-PD1 or anti-PDL1 agents such as atezolizumab has recently been reported. It has been suggested that the predilection of sarcoidosis to affect scars is due to the presence of antigens or foreign bodies that can serve as a stimulus for granuloma formation. However, to the best of our knowledge, sarcoidosis-specific skin lesions have not to date been reported involving xanthelasma. We report a patient who developed specific lesions of sarcoidosis infiltrating some xanthelasmata 6 months after starting treatment with atezolizumab. A 69-year-old woman was referred to the dermatology department for infiltration of xanthelasmata. The patient was being treated with atezolizumab for metastatic uterine carcinosarcoma. Cutaneous biopsy from an infiltrated xanthelasma and from still yellow, no infiltrated xanthelasma showed differing proportions of foamy histiocytes and sarcoid granulomas. On physical examination, erythemato-marronaceous papules clustered on both knees, and not previously detected by the patient, were observed. The biopsy showed sarcoid dermal granulomas with foreign bodies. A chest computerized tomography scan was consistent with intrathoracic involvement of sarcoidosis. Endobronchial ultrasound-guided transbronchial needle aspiration of a mediastinal lymphadenopathy showed epithelioid cell granulomas. Histopathologically, the foamy histiocytes of xanthelasma seem to be replaced by or evolve to epithelioid cells to form sarcoid granulomas. The possible pathogenic mechanism is discussed. Dermatologists and dermatopathologists should bear in mind that sarcoidosis can present as infiltration of pre-existing xanthelasmata.

Haloarchaea: A Promising Biosource for Carotenoid Production.

Haloarchaea are halophilic microorganisms belonging to the Archaea domain that inhabit salty environments (mainly soils and water) all around the world. Most of the genera included in this group are able to produce carotenoids at significant concentrations (even wild-type strains). The major carotenoid produced by the cells is bacterioruberin (and its derivatives), which is only produced by this kind of microbes. Nevertheless, the understanding of carotenoid metabolism in haloarchaea, its regulation, and the roles of carotenoid derivatives in this group of extreme microorganisms remains mostly unrevealed. Besides, potential biotechnological uses of haloarchaeal pigments are poorly explored. This work summarizes what it has been described so far about carotenoid production by haloarchaea, haloarchaeal carotenoid production at large scale, as well as the potential uses of haloarchaeal pigments in biotechnology and biomedicine.

Haloferax mediterranei Cells as C50 Carotenoid Factories.

Haloarchaea produce C50 carotenoids such as bacterioruberin, which are of biotechnological in-terest. This study aimed to analyze the effect of different environmental and nutritional conditions on the cellular growth and dynamics of carotenoids accumulation in Haloferax mediterranei. The maximum production of carotenoids (40 µg·mL-1) was obtained during the stationary phase of growth, probably due to nutrient-limiting conditions (one-step culture). By seven days of culture, 1 mL culture produced 22.4 mg of dry weight biomass containing 0.18 % (w/w) of carotenoids. On the other hand, carbon-deficient cultures (low C/N ratio) were observed to be optimum for C50 bacterioruberin production by Hfx. mediterranei, but negatively affected the growth of cells. Thus, a two-steps process was evaluated for optimum carotenoids yield. In the first step, a nutri-ent-repleted culture medium enabled the haloarchaea to produce biomass, while in the second step, the biomass was incubated under osmotic stress and in a carbon-deficient medium. Under the conditions used, the obtained biomass contained 0.27% (w/w) of carotenoids after seven days, which accounts for 58.49 µg·mL-1 of carotenoids for a culture with turbidity 14.0.