BRCA1/BARD1 ubiquitinates PCNA in unperturbed conditions to promote continuous DNA synthesis.

Deficiencies in the BRCA1 tumor suppressor gene are the main cause of hereditary breast and ovarian cancer. BRCA1 is involved in the Homologous Recombination DNA repair pathway and, together with BARD1, forms a heterodimer with ubiquitin E3 activity. The relevance of the BRCA1/BARD1 ubiquitin E3 activity for tumor suppression and DNA repair remains controversial. Here, we observe that the BRCA1/BARD1 ubiquitin E3 activity is not required for Homologous Recombination or resistance to Olaparib. Using TULIP2 methodology, which enables the direct identification of E3-specific ubiquitination substrates, we identify substrates for BRCA1/BARD1. We find that PCNA is ubiquitinated by BRCA1/BARD1 in unperturbed conditions independently of RAD18. PCNA ubiquitination by BRCA1/BARD1 avoids the formation of ssDNA gaps during DNA replication and promotes continuous DNA synthesis. These results provide additional insight about the importance of BRCA1/BARD1 E3 activity in Homologous Recombination.

EXO1 and DNA2-mediated ssDNA gap expansion is essential for ATR activation and to maintain viability in BRCA1-deficient cells.

DNA replication faces challenges from DNA lesions originated from endogenous or exogenous sources of stress, leading to the accumulation of single-stranded DNA (ssDNA) that triggers the activation of the ATR checkpoint response. To complete genome replication in the presence of damaged DNA, cells employ DNA damage tolerance mechanisms that operate not only at stalled replication forks but also at ssDNA gaps originated by repriming of DNA synthesis downstream of lesions. Here, we demonstrate that human cells accumulate post-replicative ssDNA gaps following replicative stress induction. These gaps, initiated by PrimPol repriming and expanded by the long-range resection factors EXO1 and DNA2, constitute the principal origin of the ssDNA signal responsible for ATR activation upon replication stress, in contrast to stalled forks. Strikingly, the loss of EXO1 or DNA2 results in synthetic lethality when combined with BRCA1 deficiency, but not BRCA2. This phenomenon aligns with the observation that BRCA1 alone contributes to the expansion of ssDNA gaps. Remarkably, BRCA1-deficient cells become addicted to the overexpression of EXO1, DNA2 or BLM. This dependence on long-range resection unveils a new vulnerability of BRCA1-mutant tumors, shedding light on potential therapeutic targets for these cancers.

Rasgos funcionales de frutos con importancia particular para los dispersores de semillas en el bosque seco tropical

Introducción: La frugivoría es un proceso ecológico determinante para la estructuración y regeneración de los bosques. En los trópicos, donde la diversidad de plantas y animales frugívoros es alta, las relaciones interespecíficas son complejas y requieren estudio. Objetivo: Identificar las especies de plantas ecológicamente importantes en dos redes de interacción, y el papel de los rasgos funcionales de los frutos en esas interacciones en un bosque seco. Métodos: Recolectamos 10 frutos por planta de 10 plantas de cada especie de interés en un bosque seco colombiano, calculamos el índice de importancia de las plantas a partir de la relevancia de aves y mamíferos frugívoros en la estructura de las redes. Esta relevancia se relaciona directamente con el potencial del animal como dispersor efectivo de semillas. Utilizamos modelos lineales generalizados para estimar el tamaño, color, estrato, y tipo de pulpa, en el índice. Resultados: Las plantas más importantes son especies de los géneros Miconia, Ficus, Cecropia, Bursera, Casearia y Trichilia, también identificadas como recursos importantes para los frugívoros de los trópicos en otros estudios. Las plantas con frutos carnosos, rojos y de menor tamaño son los mejores para dispersores de semillas. El índice de importancia de las plantas tiene alta variación; esto sugiere que un conjunto de especies frugívoras beneficiadas por cada especie de planta tiene una contribución diferenciada en procesos ecológicos derivados de la dispersión de semillas. Conclusiones: Programas de restauración para este tipo de bosque tropical seco debería incluir una variedad de plantas, incluyendo especies con frutos pequeños, rojos y carnosos.

Introduction: Frugivory is a pivotal ecological process for the structure and regeneration of forests. In the tropics, where the diversity of plants and frugivorous animals is high, interspecific relationships in the interaction networks are complex and need study. Objective: To identify ecologically important plant species in two interaction networks, and the role of functional fruit traits in those interactions in a dry forest. Methods: We collected 10 fruits per plant from 10 plants of each species of interest in a Colombian dry forest, we calculated the Plant Importance Index based on the bird and mammal frugivores relevance for network structure. This relevance is directly related to the animal's potential as effective seed dispersers. We used generalized linear models to estimate the effect of fruit size, color, stratum, and type of pulp, on the index. Results: The most important plants are species of the genera Miconia, Ficus, Cecropia, Bursera, Casearia and Trichilia, also identified as important resources for tropical frugivores in other studies. Plants with small, red, and fleshy fruits are the best for seed dispersers. The plant importance index has a high variation; this suggests that the set of frugivore species benefited by each plant species has a differential contribution to the ecological processes derived from seed dispersal. Conclusions: Restoration programs for this kind of tropical dry forest should include a variety of plants, including species with small, red, and fleshy fruits.

Common carp as an ecological indicator of environmental pollution in reservoirs of southern Spain: inferring the environmental risks of anthropogenic activities.

Extraction and mineral processing, as well as the waste generated by old abandoned mining sites, are the main sources of contamination of water bodies and lands by potentially toxic elements (PTEs). The common carp (Cyprinus carpio Linnaeus 1758) has been reported to be a good ecological indicator of environmental pollution in water bodies. Hence, we evaluated the concentration of eleven PTEs (As, Cd, Co, Cr, Cu, Fe, Hg, Mn, Ni, Pb, and Zn) in different tissues of common carp in two reservoirs of the province of Jaén, southern Spain: El Tranco de Beas (S1) and La Fernandina (S2). We also assessed the concentration of PTEs in water and sediment samples. We used inductively coupled plasma mass spectrometry for all the collected samples. We found high concentrations of As and Fe in water in the S2 reservoir, above the maximum limits allowed by the sanitary criteria in Spain; however, the analysis of sediments indicated low ecological risk in S1 and moderate ecological risk for As in S2. The concentration of PTEs in common carp was higher in the S2 reservoir, exceeding the permissible limits in the case of As, Cd, Pb, and Zn. As and Cd showed higher concentrations in the kidney; Cu, Fe, and Zn showed higher concentrations in the liver; and Pb and Mn presented higher concentrations in the gill and gill bone. There was a good correlation between the concentrations found in water/sediment samples and those in common carp, corroborating its usefulness as a good ecological indicator, allowing the detection of environmental pollution and inferring previous or current anthropogenic activities such as mining.

Objective Evaluation of Laparoscopic Experience Based on Muscle Electromyography and Accelerometry Performing Circular Pattern Cutting Tasks: A Pilot Study.

Purpose.The aim of this work is to present a new physical laparoscopy simulator with an electromyography (EMG)/accelerometry-based muscle activity recording system, EvalLap EMG-ACC, and perform objective evaluation of laparoscopic skills based on the quantification of muscle activity of participants with different levels of laparoscopic experience. Methods. EMG and ACC signals were obtained from 14 participants (6 experts, 8 medical students) performing circular pattern cutting tasks using a laparoscopic box trainer with the Trigno (Delsys Inc, Natick, MA) portable wireless system of 16 wireless sensors. Sensors were placed on the proximal and distal muscles of the upper extremities. Seven evaluation metrics were proposed and compared between skilled and novice surgeons. Results. The proximal and distal arm muscles (trapezius, deltoids, biceps, and forearms) were most active while executing laparoscopic tasks. Laparoscopic experience was associated with differences in EMG amplitude (Aavg), muscle activity (iEMG), hand acceleration (iACH), user movement (iAC), and muscle fatigue. For the cutting task, the deltoid, bicep, forearm EMG amplitude, and user movement significantly differed between experience groups. Conclusion. This pilot study demonstrates that different muscle groups are preferentially activated during laparoscopic tasks depending on the level of surgical experience. Expert surgeons showed less muscle activity compared with novices. EvalLap EMG-ACC represents a promising means to distinguish surgeons with basic cutting skills from those who have not yet developed these skills.

Assessment of Functional and Physical Performances of Pre-filled Syringes in Deep Cold Storage Conditions.

The recent emergence of new drug technologies such as messenger ribonucleic acid-based vaccines developed to fight the outbreak of the COVID-19 global pandemic has driven increased demand for delivery solutions capable of withstanding deep cold storage conditions down to -50°C, and even down to -80°C. Although significant data exist for deep cold storage in vials, little evidence is available for pre-filled syringes. Because pre-filled syringes serve as both the storage container and the delivery mechanism, there are additional risks to performance that must be evaluated, such as plunger gliding performance, syringe lubrication, silicone layer stability, and container closure integrity (CCI). In the present study, a comprehensive assessment of functional and physical performances of pre-filled syringes (PFS filled with water) was performed after one or multiple freeze/thaw (F/T) cycles between ambient temperature and various temperature cycles including -40°C, -50°C or -80°C for both 'staked needle' and 'luer lock' configurations. The experiments were guided by historical normative methods such as ISO 11040-4 and USP <1207> and combined with headspace gas analysis for barrel-stopper tightness testing. In addition, they were complemented with a novel approach, namely in situ real-time optical imagery, to track plunger stopper movement during the F/T cycle. The findings indicated that there is no significant impact on the functional performances from F/T down to -80°C, whereas no CCI risk was found after F/T down to -50°C.

High-Efficiency Plasmid DNA Transformation in Yeast.

Transformation of DNA into cells of the budding yeast Saccharomyces cerevisiae and other industrially important yeasts is most commonly performed using chemical-based methods. Current protocols typically involve exposure of the cells to lithium ions in a solution containing the crowding agent polyethylene glycol (PEG), often in conjunction with other reagents such as dimethyl sulfoxide (DMSO) that promote destabilization of the cell wall and/or cell envelope. Recent work has demonstrated that it is possible to achieve high transformation efficiencies with early stationary phase cells, i.e., small overnight liquid cell cultures, using methods that are rapid and readily scalable for high-throughput projects. Herein, we describe carrier DNAs, chemical reagents, and cell growth media that permit transformation of yeast cells with either plasmids or linear DNA fragments with high efficiency.

Manganese is a physiologically relevant TORC1 activator in yeast and mammals.

The essential biometal manganese (Mn) serves as a cofactor for several enzymes that are crucial for the prevention of human diseases. Whether intracellular Mn levels may be sensed and modulate intracellular signaling events has so far remained largely unexplored. The highly conserved target of rapamycin complex 1 (TORC1, mTORC1 in mammals) protein kinase requires divalent metal cofactors such as magnesium (Mg2+) to phosphorylate effectors as part of a homeostatic process that coordinates cell growth and metabolism with nutrient and/or growth factor availability. Here, our genetic approaches reveal that TORC1 activity is stimulated in vivo by elevated cytoplasmic Mn levels, which can be induced by loss of the Golgi-resident Mn2+ transporter Pmr1 and which depend on the natural resistance-associated macrophage protein (NRAMP) metal ion transporters Smf1 and Smf2. Accordingly, genetic interventions that increase cytoplasmic Mn2+ levels antagonize the effects of rapamycin in triggering autophagy, mitophagy, and Rtg1-Rtg3-dependent mitochondrion-to-nucleus retrograde signaling. Surprisingly, our in vitro protein kinase assays uncovered that Mn2+ activates TORC1 substantially better than Mg2+, which is primarily due to its ability to lower the Km for ATP, thereby allowing more efficient ATP coordination in the catalytic cleft of TORC1. These findings, therefore, provide both a mechanism to explain our genetic observations in yeast and a rationale for how fluctuations in trace amounts of Mn can become physiologically relevant. Supporting this notion, TORC1 is also wired to feedback control mechanisms that impinge on Smf1 and Smf2. Finally, we also show that Mn2+-mediated control of TORC1 is evolutionarily conserved in mammals, which may prove relevant for our understanding of the role of Mn in human diseases.

Human TREX1 Repairs 3'-End DNA Lesions in Vitro.

Human three-prime repair exonuclease 1 (TREX1) is the major 3' to 5' exonuclease that functions to deplete the cytosolic DNA to prevent the autoimmune response. TREX1 is upregulated and translocates from cytoplasm to the nucleus in response to genotoxic stress, but the function of nuclear TREX1 is not well understood. Herein, we wish to report our in vitro finding that TREX1 efficiently excises 3'-phospho-α,ß-unsaturated aldehyde and 3'-deoxyribose phosphate that are commonly produced as base excision repair intermediates and also from the nonenzymatic strand incision at abasic sites.

Humoral Response after Three Doses of mRNA-1273 or BNT162b2 SARS-CoV-2 Vaccines in Hemodialysis Patients.

The COVID-19 pandemic continues to be a worldwide health issue. Among hemodialysis (HD) patients, two-dose immunization schemes with mRNA vaccines have contributed to preventing severe COVID-19 cases; however, some have not produced a sufficient humoral response, and most have developed a rapid decline in antibody levels over the months following vaccination. This observational, prospective, multi-center study evaluated the humoral response in terms of presence and levels of IgG antibodies to the receptor-binding domain of the S1 spike antigen of SARS-CoV-2 (anti-S1-RBD IgG) to the third dose of SARS-CoV-2 mRNA vaccines, either the mRNA-1273 (Moderna) or BNT162b2 (Pfizer), in 153 patients from three dialysis units affiliated to Hospital Clínic of Barcelona (Spain). Most hemodialysis patients responded intensely to this third vaccine dose, achieving the seroconversion in three out of four non- or weak responders to two doses. Moreover, 96.1% maintained the upper limit or generated higher titers than after the second. BNT162b2 vaccine, active cancer, and immunosuppressive treatment were related to a worse humoral response. Every hemodialysis patient should be administered a third vaccine dose six months after receiving the second one. Despite the lack of data, immunosuppressed patients and those with active cancer may benefit from more frequent vaccine boosters.

Orthogonal control of gene expression in plants using synthetic promoters and CRISPR-based transcription factors.

BACKGROUND: The construction and application of synthetic genetic circuits is frequently improved if gene expression can be orthogonally controlled, relative to the host. In plants, orthogonality can be achieved via the use of CRISPR-based transcription factors that are programmed to act on natural or synthetic promoters. The construction of complex gene circuits can require multiple, orthogonal regulatory interactions, and this in turn requires that the full programmability of CRISPR elements be adapted to non-natural and non-standard promoters that have few constraints on their design. Therefore, we have developed synthetic promoter elements in which regions upstream of the minimal 35S CaMV promoter are designed from scratch to interact via programmed gRNAs with dCas9 fusions that allow activation of gene expression. RESULTS: A panel of three, mutually orthogonal promoters that can be acted on by artificial gRNAs bound by CRISPR regulators were designed. Guide RNA expression targeting these promoters was in turn controlled by either Pol III (U6) or ethylene-inducible Pol II promoters, implementing for the first time a fully artificial Orthogonal Control System (OCS). Following demonstration of the complete orthogonality of the designs, the OCS was tied to cellular metabolism by putting gRNA expression under the control of an endogenous plant signaling molecule, ethylene. The ability to form complex circuitry was demonstrated via the ethylene-driven, ratiometric expression of fluorescent proteins in single plants. CONCLUSIONS: The design of synthetic promoters is highly generalizable to large tracts of sequence space, allowing Orthogonal Control Systems of increasing complexity to potentially be generated at will. The ability to tie in several different basal features of plant molecular biology (Pol II and Pol III promoters, ethylene regulation) to the OCS demonstrates multiple opportunities for engineering at the system level. Moreover, given the fungibility of the core 35S CaMV promoter elements, the derived synthetic promoters can potentially be utilized across a variety of plant species.

Overview of traditional medicine in the Muisca Reserve of Cota, Colombia: A cross-sectional study. / Panorama de la medicina tradicional en el Resguardo Muisca de Cota, Colombia: estudio transversal.

Introduction: The Muisca Indigenous people in Cota, Colombia, has committed to reviving and strengthening their traditional culture, including the ancestral knowledge associated with their traditional medicine. Objective: To explore the occurrence of traditional medicine and factors associated with its use among the Muisca people in Cota, Colombia. Methods: A participatory cross-sectional study applied a questionnaire to 471 Muisca mothers who had at least one child over 10 in April 2019. The 44 questions inquired demographic, social, and cultural factors of participants and their knowledge, use and practice of traditional medicine. We used the Mantel-Haenszel procedure to measure the associations using Odds Ratio and 95% confidence intervals. Results: 66.2% (312/471) of the mothers knew at least three cases of traditional diseases; 56.8% (267/470) had and used medicinal plants; 15.8% (73/462) had practiced traditional self-care for flu, menstruation and postpartum; and 11.8% (54/458) reported that they had gone to midwives, bonesetters and healers. Four factors had a significant association (p < 0.05) with traditional medicine: consumption of three traditional foods; having traditional gardens and plants; living within the reservation; and parents born in a municipality of indigenous influence. Conclusions: The study showed that the permanence of traditional medicine is strongly associated with traditional food, agricultural vocation and the possession and use of medicinal plants. The results suggest that the strategy for the recovery of traditional medicine could focus on promoting a stronger link between indigenous identity, territory, food and health.

Introducción: La población del Resguardo Indígena Muisca de Cota, Colombia, se encuentra en un proceso de fortalecimiento cultural. Ahora busca la recuperación de sus saberes ancestrales relacionados con la medicina tradicional. Objetivo: Explorar la ocurrencia de la medicina tradicional y factores asociados con su vigencia en el resguardo indígena Muisca de Cota, Colombia. Métodos: Estudio transversal participativo con los miembros del resguardo. Para este estudio se aplicó en abril de 2019 un instrumento de 44 preguntas a 471 madres de familia que tenían por lo menos un hijo mayor de diez años. A través de él se indagaron algunos factores demográficos, sociales y culturales. Además, se plantearon preguntas relacionadas con conocimiento, uso y práctica de medicina tradicional. Usamos el procedimiento de Mantel-Haenszel para establecer las asociaciones mediante el Odds ratio e intervalos de confianza del 95%. Resultados: El 66,2% (312/471) de las madres conocía al menos tres casos de enfermedades tradicionales, el 56,8% (267/470) tenía y usaba plantas medicinales, el 15,8% (73/462) practicaba cuidados tradicionales de la gripa, la menstruación y el puerperio, y el 11,8% (54/458) reportó que han acudido a parteras, sobanderos y curanderos. El consumo de tres alimentos tradicionales, la tenencia de huertos y plantas tradicionales, tener la vivienda dentro del resguardo y padres nacidos en un municipio de influencia indígena, fueron los principales factores asociados con la prevalencia de la medicina tradicional (p < 0,05). Conclusiones: El estudio mostró que la permanencia de la medicina tradicional está fuertemente asociada a la alimentación tradicional, a la vocación agrícola y a la tenencia y uso de plantas medicinales. Los resultados sugieren que la estrategia de recuperación de la medicina tradicional podría enfocarse en promover un mayor vínculo entre identidad indígena, territorio, alimentación y salud.

Panorama de la medicina tradicional en el Resguardo Muisca de Cota, Colombia: estudio transversal / Overview of traditional medicine in the Muisca Reserve of Cota, Colombia: a cross-sectional study

INTRODUCCIÓN: La población del Resguardo Indígena Muisca de Cota, Colombia, se encuentra en un proceso de fortalecimiento cultural. Ahora busca la recuperación de sus saberes ancestrales relacionados con la medicina tradicional. OBJETIVO: Explorar la ocurrencia de la medicina tradicional y factores asociados con su vigencia en el resguardo indígena Muisca de Cota, Colombia. MÉTODOS: Estudio transversal participativo con los miembros del resguardo. Para este estudio se aplicó en abril de 2019 un instrumento de 44 preguntas a 471 madres de familia que tenían por lo menos un hijo mayor de diez años. A través de él se indagaron algunos factores demográficos, sociales y culturales. Además, se plantearon preguntas relacionadas con conocimiento, uso y práctica de medicina tradicional. Usamos el procedimiento de Mantel-Haenszel para establecer las asociaciones mediante el Odds ratio e intervalos de confianza del 95%. RESULTADOS: El 66,2% (312/471) de las madres conocía al menos tres casos de enfermedades tradicionales, el 56,8% (267/470) tenía y usaba plantas medicinales, el 15,8% (73/462) practicaba cuidados tradicionales de la gripa, la menstruación y el puerperio, y el 11,8% (54/458) reportó que han acudido a parteras, sobanderos y curanderos. El consumo de tres alimentos tradicionales, la tenencia de huertos y plantas tradicionales, tener la vivienda dentro del resguardo y padres nacidos en un municipio de influencia indígena, fueron los principales factores asociados con la prevalencia de la medicina tradicional (p < 0,05). CONCLUSIONES: El estudio mostró que la permanencia de la medicina tradicional está fuertemente asociada a la alimentación tradicional, a la vocación agrícola y a la tenencia y uso de plantas medicinales. Los resultados sugieren que la estrategia de recuperación de la medicina tradicional podría enfocarse en promover un mayor vínculo entre identidad indígena, territorio, alimentación y salud.

INTRODUCTION: The Muisca Indigenous people in Cota, Colombia, has committed to reviving and strengthening their traditional culture, including the ancestral knowledge associated with their traditional medicine. OBJECTIVE: To explore the occurrence of traditional medicine and factors associated with its use among the Muisca people in Cota, Colombia. METHODS: A participatory cross-sectional study applied a questionnaire to 471 Muisca mothers who had at least one child over 10 in April 2019. The 44 questions inquired demographic, social, and cultural factors of participants and their knowledge, use and practice of traditional medicine. We used the Mantel-Haenszel procedure to measure the associations using Odds Ratio and 95% confidence intervals. RESULTS: 66.2% (312/471) of the mothers knew at least three cases of traditional diseases; 56.8% (267/470) had and used medicinal plants; 15.8% (73/462) had practiced traditional self-care for flu, menstruation and postpartum; and 11.8% (54/458) reported that they had gone to midwives, bonesetters and healers. Four factors had a significant association (p < 0.05) with traditional medicine: consumption of three traditional foods; having traditional gardens and plants; living within the reservation; and parents born in a municipality of indigenous influence. CONCLUSIONS: The study showed that the permanence of traditional medicine is strongly associated with traditional food, agricultural vocation and the possession and use of medicinal plants. The results suggest that the strategy for the recovery of traditional medicine could focus on promoting a stronger link between indigenous identity, territory, food and health.

Incidence of severe breakthrough SARS-CoV-2 infections in vaccinated kidney transplant and haemodialysis patients.

INTRODUCTION: Given the increased COVID-19 observed in kidney transplant recipients (KTRs) and haemodialysis patients, several studies have tried to establish the efficacy of mRNA vaccines in these populations by evaluating their humoral and cellular responses. However, there is currently no information on clinical protection (deaths and hospitalizations), a gap that this study aims to fill. METHODS: Observational prospective study involving 1,336 KTRs and haemodialysis patients from three dialysis units affiliated to Hospital Clínic of Barcelona, Spain, vaccinated with two doses of mRNA-1273 (Moderna) or BNT162b2 (Pfizer-BioNTech) SARS-CoV-2 mRNA vaccines. The outcomes measured were SARS-CoV-2 infection diagnosed by a positive RT-PCR fourteen days after the second vaccine dose, hospital admissions derived from infection, and a severe COVID-19 composite outcome, defined as either ICU admission, invasive and non-invasive mechanical ventilation, or death. RESULTS: Six per cent (18/302) of patients on haemodialysis were infected, of whom four required hospital admission (1.3%), only one (0.3%) had severe COVID-19, and none of them died. In contrast, 4.3% (44/1034) of KTRs were infected, and presented more hospital admissions (26 patients, 2.5%), severe COVID-19 (11 patients, 1.1%) or death (4 patients, 0.4%). KTRs had a significantly higher risk of hospital admission than HD patients, and this risk increased with age and male sex (HR 3.37 and 4.74, respectively). CONCLUSIONS: The study highlights the need for booster doses in KTRs. In contrast, the haemodialysis population appears to have an adequate clinical response to vaccination, at least up to four months from its administration.

Deforestation by historical lime industry in an arid aeolian sedimentary system: An applied and methodological research.

Traditional land uses have been altering aeolian sedimentary systems for centuries through the removal of plant material for grazing, fuel or farming purposes. However, few studies have been able to quantify the deforestation process associated with these land uses due to the complexity that this entails and the limitations of the historical sources. In this context, the aim of this work is to develop a methodology that allows to reconstruct, evaluate, measure and locate the effects of deforestation processes. The methodology, based on the interpretation of historical documents, oral interviews and publications in the literature; was applied to a case study in Jandía (Fuerteventura, Canary Islands, Spain). On the basis of morphological measurements of the types of plant used to fire lime kilns, the current available biovolume was determined and an estimation made of the surface area affected by plant removal. The data obtained were integrated and analysed through a geographic information system (GIS) in order to quantify the impact of the lime kiln industry on the vegetation in the study area. The main results show that to fire a large-sized lime kiln oven it would be necessary to clear a low-density vegetation area of 21,826.08 m2 (or a a high-density vegetation area of 3075.72 m2) using three main species (Launaea arborescens, Lycium intricatum and Convolvulus caput-madusae). It was also found that distances of up to 38 km had to be travelled to obtain the vegetation required to fire the kilns. It is concluded that a number of impacts resulted from the demands of the limestone industry, particularly on plant communities, the abundance of certain species and flora richness, as well as modifications to geomorphological processes and the eventual collapse of the activity in the 1960s through overexploitation of the plant material. The present research allows us to learn from past experiences in which industries lacked proper planning and thus their activity led to their own collapse and rapid environmental degradation.

Humoral and Cellular Responses to mRNA-1273 and BNT162b2 SARS-CoV-2 Vaccines Administered to Hemodialysis Patients.

RATIONALE & OBJECTIVE: Patients with kidney failure who are receiving maintenance dialysis have a higher risk of infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and worse clinical outcomes after coronavirus disease 2019 (COVID-19) than the general population. Therefore, immunization against SARS-CoV-2 with effective vaccines is an important component of health-maintenance strategies for these patients. This study evaluated the humoral and cellular responses to messenger RNA (mRNA) SARS-CoV-2 vaccines in this population. STUDY DESIGN: Observational prospective multicenter cohort study. SETTING & PARTICIPANTS: 205 patients treated at 3 dialysis units at the Hospital Clínic of Barcelona (Spain) were vaccinated from February 3 to April 4, 2021, and followed until April 23, 2021. EXPOSURE: Immunization with either the mRNA-1273 (Moderna) or BNT162b2 (Pfizer-BioNTech) SARS-CoV-2 mRNA vaccine. OUTCOME: Seroconversion, defined as the detection of IgG antibodies to the receptor-binding domain of the S1 spike antigen of SARS-CoV-2 (anti-S1-RBD IgG), and the identification of activated CD4+T cells 3 weeks after completing vaccination. Anti-S1-RBD IgG levels were also analyzed as a secondary outcome. ANALYTICAL APPROACH: Univariate and multivariable logistic and multiple linear regression models were used to evaluate the associations between vaccination and study outcomes. RESULTS: We found that 97.7% of 175 vaccinated patients who were seronegative at baseline developed a response (humoral, cellular, or both); 95.4% of these patients seroconverted, while 62% of those tested for cellular immunity had a positive response. Greater age and immunosuppressive treatment were associated with lower antibody levels. LIMITATIONS: Mandatory vaccine administration by health authorities. Anti-S1-RBD IgG levels were reported up to 150U/mL and cellular immune responses were characterized qualitatively. Antibody assay and cellular response assessment may not be comparable with previously published laboratory approaches. CONCLUSIONS: Immunization with mRNA vaccines generated a humoral and cellular immune response in a high proportion of patients with kidney failure receiving maintenance dialysis. These findings as well as the high risk of infection and poor clinical outcomes among these patients make their vaccination a health priority.