Active lupus in Argentina: Results of a multicenter and national registry.

OBJECTIVE: To evaluate the association between patients' characteristics and disease activity in an Argentine lupus registry. METHODS: Cross-sectional study. Disease activity was stratified into: Remission off-treatment: SLEDAI = 0, without prednisone and immunosuppressive drugs. Low disease activity Toronto Cohort (LDA-TC): SLEDAI ≤2, without prednisone or immunosuppressive drugs. Modified lupus low disease activity (mLLDAS): SLEDAI score of ≤4, with no activity in major organ systems and no new features, prednisone of ≤10 mg/day and/or immunosuppressive drugs (maintenance dose) and Active disease: SLEDAI score of >4 and prednisone >10 mg/day and immunosuppressive drugs. A descriptive analysis and logistic regression model were performed. RESULTS: A total of 1346 patients were included. Of them, 1.6% achieved remission off steroids, 0.8% LDA-TC, 12.1% mLLDAS and the remaining 85.4% had active disease. Active disease was associated with younger age (p ≤ 0.001), a shorter time to diagnosis (p ≤ 0.001), higher frequency of hospitalizations (p ≤ 0.001), seizures (p = 0.022), serosal disease (p ≤ 0.001), nephritis (p ≤ 0.001), higher SDI (p ≤ 0.001), greater use of immunosuppressive therapies and higher doses of prednisone compared to those on mLLDAS. In the multivariable analysis, the variables associated with active disease were the presence of pleuritis (OR 2.1, 95% CI 1.2-3.9; p = 0.007), persistent proteinuria (OR 2.5, 95% CI 1.2-5.5; p ≤ 0.011), nephritis (OR 2.5, 95% CI 1.2-5.6; p = .018) and hospitalizations (OR 8.9, 95% CI 5.3-16.0; p ≤ 0.001) whereas age at entry into the registry was negatively associated with it (OR 0.9, 95% CI 0.9-1.0; p = 0.029). CONCLUSION: Active disease was associated with shorter time to diagnosis, worse outcomes (SDI and hospitalizations) and renal, neurological and serosal disease.

Frecuencia de eventos cardiovasculares en pacientes con lupus eritematoso sistémico / Frequency of cardiovascular events in patients with systemic lupus erythematosus

Resumen Introducción: el progreso en los tratamientos para el lupus eritematoso sistémico (LES) resultó en una disminución de la mortalidad; sin embargo, la enfermedad cardiovascular y las complicaciones infecciosas aún son las principales causas de muerte. La evidencia apoya la participación del sistema inmunológico en la generación de la placa aterosclerótica, así como su conexión con las enfermedades autoinmunes. Objetivos: describir la frecuencia de eventos cardiovasculares (ECV) en el Registro de Lupus Eritematoso Sistémico de la Sociedad Argentina de Reumatología (RELESSAR) transversal, así como sus principales factores de riesgo asociados. Materiales y métodos: estudio descriptivo y transversal para el cual se tomaron los pacientes ingresados en el registro RELESSAR transversal. Se describieron las variables sociodemográficas y clínicas, las comorbilidades, score de actividad y daño. ECV se definió como la presencia de al menos una de las siguientes patologías: enfermedad arterial periférica, cardiopatía isquémica o accidente cerebrovascular. El evento clasificado para el análisis fue aquel posterior al diagnóstico del LES. Se conformaron dos grupos macheados por edad y sexo 1:2. Resultados: 1515 pacientes mayores de 18 años participaron del registro. Se describieron 80 pacientes con ECV (5,3%). En este análisis se incluyeron 240 pacientes conformando dos grupos. La edad media fue de 47,8 (14,4) y 47,6 (14,2) en el grupo con y sin ECV respectivamente. Los pacientes con ECV tuvieron mayor duración del LES en meses, mayor índice de Charlson, mayor SLICC (Systemic Lupus International Collaborating Clinics/American College of Rheumatology), mayor frecuencia de manifestaciones neurológicas, síndrome antifosfolípido, hospitalizaciones y uso de ciclofosfamida. Las únicas variables asociadas en el análisis multivariado fueron el índice de Charlson (p=0,004) y el SLICC (p<0,001). Conclusiones: los ECV influyen significativamente en nuestros pacientes, y se asocian a mayor posibilidad de daño irreversible y comorbilidades.

Abstract Introduction: progress in treatments for systemic lupus erythematosus (SLE) has resulted in a decrease in mortality; however, cardiovascular and infectious diseases remain the leading causes of death. Evidence supports the involvement of the immune system in the generation of atherosclerotic plaque, as well as its connection to autoimmune diseases. Objectives: to describe the frequency of cardiovascular disease (CVD) in the cross-sectional RELESSAR registry, as well as its associated variables. Materials and methods: a descriptive and cross-sectional study was performed using patients admitted to the cross-sectional RELESSAR registry. Sociodemographic variables, clinical variables, comorbidities, activity and damage scores were described. CVD was defined as at least one of the following: peripheral arterial disease, ischemic heart disease, or cerebrovascular accident. All patients with at least one CVD were included in our analysis (heart attack, central nervous system vascular disease, and peripheral arteries atherosclerotic disease). The event classified for the analysis was that after the diagnosis of SLE. SLE diagnosis was previous to CVD. Two groups matched by age and sex, 1:2 were formed. Results: a total of 1515 patients older than 18 years participated in the registry. Eighty patients with CVD (5.3%) were described in the registry. Two-hundred and forty patients were included, according to two groups. The mean age was 47.8 (SD 14.4) and 47.6 (SD 14.2) in patients with and without CVD, respectively. Patients with CVD had a longer duration of SLE in months, a higher Charlson index, a higher SLICC, increased frequency of neurological manifestations, antiphospholipid syndrome, hospitalizations, and use of cyclophosphamide. The associated variables in the multivariate were the Charlson Index (p=0.004) and the SLICC (p<0.001). Conclusions: CVDs have a significant influence on our patients, being associated with a greater possibility of damage and comorbidities.

Combination of slurry-bioreactors and actinobacteria consortia as strategy to bioremediate chlordane-contaminated soils.

Soil contamination caused by pesticides poses a significant environmental challenge, and addressing it requires effective solutions. Bioremediation, combining the utilization of slurry-bioreactors and microbial consortia, emerges as an appropiated strategy to tackle this issue. Therefore, this research evaluated the chlordane (CLD) removal efficiency by a Streptomyces consortium through bioaugmentation of polluted soils, and slurry-bioreactors. For that, a Streptomyces defined consortium with CLD removal abilities was inoculated in soil microcosms and soil-slurry bioreactors (SB), with (SB-TSB) and without stimulation (SB-water). In soil, CLD presence has no negative effect on consortium growth. This was supported by comparing its duplication time (7.48 ± 0.14 h) with the obtained in the biotic control (7.45 ± 0.04 h). Furthermore, 17% of pesticide removal by microbial action was detected in the treated microcosms. In SB, the microbial development was not affected by the pesticide presence. In SB-TSB, the microbial growth was higher than in SB-water. This was supported by its lesser duplication time (7.27 ± 0.17 h) with respect to the non-stimulated systems (10.88 ± 0.29 h). However, SB-water showed the highest CLD removal ability (34.8%), with a concomitant increase in the chloride ion release. In the phytotoxicity test, the vigor index showed that the bioremediation in SB-water did not exert adverse effects greater than those generated by the CLD. Indeed, the root length increased after the treatment. These findings demonstrate the versatility of the Streptomyces consortium to remediate solid and semi-solid matrices impacted with pesticides, and the advantage of using bioaugmented SB to enhance the pollutants removal and accelerating the clean-up time required.

Developing a glyphosate-bioremediation strategy using plants and actinobacteria: Potential improvement of a riparian environment.

Glyphosate (Gly) and its principal degradation product, the aminomethylphosphonic acid (AMPA) were found in soils from a riparian environment in Argentina. Sixty-five actinobacteria were isolated from these soils, rhizosphere, and plants (Festuca arundinacea and Salix fragilis). The isolate Streptomyces sp. S5 was selected to be used as bioinoculant in a greenhouse test, in which plants, actinobacteria, and their combinations were assessed to bioremediate the riparian soil. The dissipation of both compounds were estimated. All treatments dissipated similarly the Gly, reaching 87-92 % of dissipation. AMPA, dissipation of 38 % and 42 % were obtained by Salix and Festuca, respectively, while they increased to 57 % and 70 % when the actinobacterium was added to each planted system. Regarding the total dissipation, the higher efficiencies for both compounds were achieved by the non-planted soils bioaugmented with the actinobacterium, with 91 % of Gly dissipated and 56 % for AMPA. According to our study, it could be suggested which strategy could be applied depending on the bioremediation type needed. If in situ bioremediation is necessary, the combination of phytoremediation and actinobacteria bioaugmentation could be convenient. On the other hand, if ex situ bioremediation is needed, the inoculation of the soil with an actinobacterium capable to dissipate Gly and AMPA could be the more efficient and easier alternative.

Seguridad de las drogas biológicas y sintéticas dirigidas utilizadas en pacientes con enfermedades reumáticas inmunomediadas. Datos del registro BIOBADASAR 3.0 / Safety of biological and targeted synthetic drugs in patients with immune-mediated rheumatic diseases. Data from the BIOBADASAR 3.0 registry

Introducción: conocer la seguridad de las drogas actualmente disponibles para el tratamiento de las enfermedades reumáticas es muy importante al momento de tomar decisiones terapéuticas objetivas e individualizadas en la consulta médica diaria. Asimismo, datos de la vida real amplían el conocimiento revelado por los ensayos clínicos. Objetivos: describir los eventos adversos (EA) reportados, estimar su frecuencia e identificar los factores relacionados con su desarrollo. Materiales y métodos: se utilizaron datos BIOBADASAR, un registro voluntario y prospectivo de seguimiento de EA de tratamientos biológicos y sintéticos dirigidos en pacientes con enfermedades reumáticas inmunomediadas. Los pacientes son seguidos hasta la muerte, pérdida de seguimiento o retiro del consentimiento informado. Para este análisis se extrajeron datos recopilados hasta el 31 de enero de 2023. Resultados: se incluyó un total de 6253 pacientes, los cuales aportaron 9533 ciclos de tratamiento, incluyendo 3647 (38,3%) ciclos sin drogas modificadoras de la enfermedad biológicas y sintéticas dirigidas (DME-b/sd) y 5886 (61,7%) con DME-b/sd. Dentro de estos últimos, los más utilizados fueron los inhibidores de TNF y abatacept. Se reportaron 5890 EA en un total de 2701 tratamientos (844 y 1857 sin y con DME-b/sd, respectivamente), con una incidencia de 53,9 eventos cada 1000 pacientes/año (IC 95% 51,9-55,9). La misma fue mayor en los ciclos con DME-b/sd (71,1 eventos cada 1000 pacientes/año, IC 95% 70,7-77,5 versus 33,7, IC 95% 31,5-36,1; p<0,001). Las infecciones, particularmente las de la vía aérea superior, fueron los EA más frecuentes en ambos grupos. El 10,9% fue serio y el 1,1% provocó la muerte del paciente. El 18,7% de los ciclos con DME-b/sd fue discontinuado a causa de un EA significativamente mayor a lo reportado en el otro grupo (11,5%; p<0,001). En el análisis ajustado, las DME-b/sd se asociaron a mayor riesgo de presentar al menos un EA (HR 1,82, IC 95% 1,64-1,96). De igual manera, la mayor edad, el mayor tiempo de evolución, el antecedente de enfermedad pulmonar obstructiva crónica, el diagnóstico de lupus eritematoso sistémico y el uso de corticoides se asociaron a mayor riesgo de EA. Conclusiones: la incidencia de EA fue significativamente superior durante los ciclos de tratamientos que incluían DME-b/sd.

Introduction: knowing the efficacy and safety of the drugs currently available for the treatment of rheumatic diseases is very important when making objective and individualized therapeutic decisions in daily medical consultation. Likewise, real-life data extends the knowledge revealed by clinical trials. Objectives: to describe the reported adverse events (AEs), estimate their frequency and identify factors associated to them. Materials and methods: BIOBADASAR data were used, which is a voluntary, prospective follow-up registry of AEs of biological and synthetic treatments in patients with immune-mediated rheumatic diseases. Patients are followed until death, loss of followup, or withdrawal of informed consent. To carry out this analysis, the data collected up to January 31, 2023 was extracted. Results: a total of 6253 patients were included, who contributed with 9533 treatment periods, including 3647 (38.3%) periods without b/ts-DMARDs and 5886 (61.7%) with b/ts-DMARDs. Among the latter, the most used were TNF inhibitors and abatacept. A total of 5890 AEs were reported in a total of 2701 treatments (844 and 1857 without and with b/ts-DMARDs, respectively), with an incidence of 53.9 events per 1000 patients/ year (95% CI 51.9-55.9). It was higher during the periods with b/ts-DMARDs (71.1 events per 1000 patients/year, 95% CI 70.7-77.5 vs 33.7, 95% CI 31.5-36.1, p<0.001). Infections, particularly those of the upper respiratory tract, were the most frequent AEs in both groups. 10.9% were severe and 1.1% were associated with the death of the patient. 18.7% of the periods with b/ts-DMARDs were discontinued due to an AE, significantly higher than that reported in the other group (11.5%; p<0.001). In the adjusted analysis, b/ts-DMARDs were associated with a higher risk of presenting at least one AE (HR 1.82, 95% CI 1.64-1.96). Similarly, older age, longer evolution time, history of chronic obstructive pulmonary disease, diagnosis of systemic lupus erythematosus, and use of corticosteroids were associated with a higher risk of AE. Conclusions: the incidence of AEs was significantly higher during those treatment periods that included DME-b/sd.

The Bacterial and Fungal Gut Microbiota of the Greater Wax Moth, Galleria mellonella L. Consuming Polyethylene and Polystyrene.

Plastic production has been increasing exponentially in the last 60 years, but plastic disposal is out of control, resulting in the pollution of all ecosystems on Earth. Finding alternative environmentally sustainable choices, such as biodegradation by insects and their associated gut microbiota, is crucial, however we have only begun to characterize these ecosystems. Some bacteria and one fungus have been previously identified in the gut of Greater Wax Moth larvae (Galleria mellonella L., Lepidoptera, Pyralidae) located mainly in the Northern hemisphere. The aim of this study was to describe changes in the gut microbiota associated with the consumption of polyethylene and polystyrene by the Greater Wax Moth in Argentina, considering both bacteria and fungi. Larvae were fed polyethylene, polystyrene and beeswax as control for 7 days. Next generation sequencing revealed changes in the bacterial gut microbiome of the wax moth larvae at the phyla and genus levels, with an increase in two Pseudomonas strains. The fungal communities showed no differences in composition between diets, only changing in relative abundance. This is the first report of both bacterial and fungal communities associated with a plastivore insect. The results are promising and call for more studies concerning a potential multi-kingdom synergy in the plastic biodegradation process.

Multicenter lupus register from Argentina, the RELESSAR database: Influence of ethnicity on disease phenotype.

OBJECTIVE: The objective is to describe the main characteristics of patients with systemic lupus erythematosus (SLE) in Argentina and to examine the influence of ethnicity on the expression of the disease. PATIENTS AND METHODS: RELESSAR is a multicentre register carried out by 106 researchers from 67 rheumatologic Argentine centres. It is a cross-sectional study of SLE (1982/1997 ACR) patients. RELESSAR electronic database includes demographic, cumulative SLE manifestations, SELENA-SLEDAI, SLICC-SDI, Katz's severity and Charlson's comorbidity indexes and treatment patterns. RESULTS: We included 1,610 patients, 91.7% were female with a median age at diagnosis of 28.1 ± 12.8; 96.2% met ≥4 ACR 1982/97 criteria. Frequent manifestations were arthritis (83.5%), malar rash (79.5%), photosensitivity (75.3%), haematological (63.8%) and renal disease (47.4%), antinuclear antibodies (96%), anti-dsDNA (66.5%) and anti-Smith antibodies (29%). The mean Selena-SLEDAI score at last visit was 3.18 (SD 4.3) and mean SDI was 1 (SD 1.3). The accumulated treatments most frequently used were antimalarials (90.4%), corticosteroids (90%), azathioprine (31.8%), intravenous cyclophosphamide (30.2%), mycophenolate mofetil or mycophenolic acid (24.5%), methotrexate (19.3%), belimumab 5.3% and rituximab 5.1%. Refractory lupus was diagnosed in 9.3% of the cases. The main causes of death were lupus activity (25.0%), activity and concomitant infections (25.0%), infections (18.2%), vascular disease (13.6%) and cancer (4.5%). Mortality was associated with higher SLEDAI, Katz, damage indexes and comorbidities. Of the 1610 patients included, 44.6% were Caucasian, 44.5% Mestizo, 8.1% Amerindian and 1.2% Afro-Latin American. Mestizo patients had higher male representation, low socioeconomic status, more inadequate medical coverage, fewer formal years of education and shorter disease duration. Polyadenopathies and Raynaud's phenomenon were more frequent in Caucasians. In the logistic regression analysis higher damage index (OR 1.28, CI 95% 1.02-1.61, p = 0.03) remained associated to mestizo ethnicity. CONCLUSIONS: This study represents the largest number of adult patients with SLE studied in Argentina. Caucasian patients were differentiated by having Raynaud's phenomenon and polyadenopathy more frequently, while patients of Mestizo origin had higher damage indexes.

Enhanced biodegradation of hexachlorocyclohexane (HCH) isomers by Sphingobium sp. strain D4 in the presence of root exudates or in co-culture with HCH-mobilizing strains.

Lindane and other 1,2,3,4,5,6-hexachlorocyclohexane (HCH) isomers are persistent organic pollutants highly hydrophobic, which hampers their availability and biodegradation. This work aimed at (i) investigating genes encoding enzymes involved in HCH degradation in the bacterium Sphingobium sp. D4, (ii) selecting strains, from a collection of environmental isolates, able to mobilize HCHs from contaminated soil, and (iii) analysing the biodegradation of HCHs by strain D4 in co-culture with HCH-mobilizing strains or when cultivated with root exudates. Fragments of the same size and similar sequence to linA and linB genes were successfully amplified. Two isolates, Streptomyces sp. M7 and Rhodococcus erythropolis ET54b able to produce emulsifiers and to mobilize HCH isomers from soil were selected. Biodegradation of HCH isomers by strain D4 was enhanced when co-inoculated with HCH mobilizing strains or when cultivated with root exudates. The degrader strain D4 was able to decompose very efficiently HCHs isomers, reducing their concentration in soil slurries by more than 95% (from an average initial amount of 50 ± 8 mg HCH kg-1 soil) in 9 days. The combination of HCH-degrading and HCH-mobilizing strains can be considered a promising inoculum for future soil bioremediation studies using bioaugmentation techniques or in combination with plants in rhizodegradation assays.

Lupus en Argentina. Pacientes no respondedores al tratamiento estándar y belimumab como posible opción. Datos del registro RELESSAR / Lupus in Argentina. Non-responders to standard treatment and belimumab as a possible option. RELESSAR record data

Introducción: el lupus es una enfermedad compleja y varias veces de difícil abordaje. Alcanzar la remisión es uno de los objetivos, incorporando opciones terapéuticas. Objetivos: describir las características generales de los pacientes según el estado de la enfermedad y el uso de belimumab. Materiales y métodos: estudio de corte transversal, registro RELESSAR. Se definió el estado de la enfermedad como: remisión: SLEDAI=0 y sin corticoides; baja actividad de la enfermedad: SLEDAI >0 y ≤4 y sin corticoides; control no óptimo: SLEDAI >4 y cualquier dosis de corticoides. Resultados: se incluyeron 1.277 pacientes, 23,4% en remisión, 12,6% en baja actividad y 63,8% con control no óptimo. En este último grupo eran más jóvenes y con menor duración de la enfermedad; presentaban mayores índices de actividad y cronicidad, y mayor empleo de inmunosupresores. Solo el 22,3% de los pacientes con criterio potencial de uso de belimumab (lupus eritematoso sistémico activo a pesar del tratamiento estándar) lo recibía en ese momento. Las variables asociadas a hospitalizaciones fueron: terapia con corticoides, ciclofosfamida y mayor SLICC. Conclusiones: se refleja la complejidad del manejo de estos pacientes y se visualizan aspectos estructurales como la desigualdad. El uso del belimumab resultaría beneficioso en los pacientes seleccionados.

Introduction: lupus is a complex disease and often difficult to approach. Achieving remission is one of the objectives, incorporating therapeutic options. Objectives: to describe the characteristics of the patients and the use of belimumab, according to the status of the disease. Materials and methods: cross-sectional study. Patients of the RELESSAR registry. Stratification: Remission: SLEDAI=0 and without corticosteroids. Low disease activity SLEDAI> 0 and ≤4 and without corticosteroids and non-optimal control: SLEDAI> 4 and any dose of corticosteroids. Results: a total of 1,277 patients were included, 23.4% in remission, 12.6% in low disease activity and 63.8% in non-optimal control. The last group was younger and had a shorter duration of the disease. They had higher activity and chronicity indices and greater use of immunosuppressants. Only 22.3% of the patients with potential criteria for the use of belimumab (activity disease despite standard treatment) were receiving it. The variables associated with hospitalizations were: corticosteroids, cyclophosphamide and higher SLICC. Those associated with severe infection: mycophenolate mofetil, azathioprine, corticosteroids, and higher SLICC. Conclusions: the complexity of the management of these patients is reflected, visualizing structural aspects such as inequality. The use of belimumab could be beneficial in selected patients.

Lupus en Argentina. Pacientes no respondedores al tratamiento estándar y belimumab como posible opción. Datos del registro RELESSAR / Lupus in Argentina. Non-responders to standard treatment and belimumab as a possible option. RELESSAR record data

Introducción: el lupus es una enfermedad compleja y varias veces de difícil abordaje. Alcanzar la remisión es uno de los objetivos, incorporando opciones terapéuticas. Objetivos: describir las características generales de los pacientes según el estado de la enfermedad y el uso de belimumab. Materiales y métodos: estudio de corte transversal, registro RELESSAR. Se definió el estado de la enfermedad como: remisión: SLEDAI=0 y sin corticoides; baja actividad de la enfermedad: SLEDAI >0 y ≤4 y sin corticoides; control no óptimo: SLEDAI >4 y cualquier dosis de corticoides. Resultados: se incluyeron 1.277 pacientes, 23,4% en remisión, 12,6% en baja actividad y 63,8% con control no óptimo. En este último grupo eran más jóvenes y con menor duración de la enfermedad; presentaban mayores índices de actividad y cronicidad, y mayor empleo de inmunosupresores. Solo el 22,3% de los pacientes con criterio potencial de uso de belimumab (lupus eritematoso sistémico activo a pesar del tratamiento estándar) lo recibía en ese momento. Las variables asociadas a hospitalizaciones fueron: terapia con corticoides, ciclofosfamida y mayor SLICC. Conclusiones: se refleja la complejidad del manejo de estos pacientes y se visualizan aspectos estructurales como la desigualdad. El uso del belimumab resultaría beneficioso en los pacientes seleccionados.

Introduction: lupus is a complex disease and often difficult to approach. Achieving remission is one of the objectives, incorporating therapeutic options. Objectives: to describe the characteristics of the patients and the use of belimumab, according to the status of the disease. Materials and methods: cross-sectional study. Patients of the RELESSAR registry. Stratification: Remission: SLEDAI=0 and without corticosteroids. Low disease activity SLEDAI> 0 and ≤4 and without corticosteroids and non-optimal control: SLEDAI> 4 and any dose of corticosteroids. Results: a total of 1,277 patients were included, 23.4% in remission, 12.6% in low disease activity and 63.8% in non-optimal control. The last group was younger and had a shorter duration of the disease. They had higher activity and chronicity indices and greater use of immunosuppressants. Only 22.3% of the patients with potential criteria for the use of belimumab (activity disease despite standard treatment) were receiving it. The variables associated with hospitalizations were: corticosteroids, cyclophosphamide and higher SLICC. Those associated with severe infection: mycophenolate mofetil, azathioprine, corticosteroids, and higher SLICC. Conclusions: the complexity of the management of these patients is reflected, visualizing structural aspects such as inequality. The use of belimumab could be beneficial in selected patients.

B-cell depletion with obinutuzumab for the treatment of proliferative lupus nephritis: a randomised, double-blind, placebo-controlled trial.

OBJECTIVE: Randomised trials of type I anti-CD20 antibodies rituximab and ocrelizumab failed to show benefit in proliferative lupus nephritis (LN). We compared obinutuzumab, a humanised type II anti-CD20 monoclonal antibody that induces potent B-cell depletion, with placebo for the treatment of LN in combination with standard therapies. METHODS: Patients with LN receiving mycophenolate and corticosteroids were randomised to obinutuzumab 1000 mg or placebo on day 1 and weeks 2, 24 and 26, and followed through week 104. The primary endpoint was complete renal response (CRR) at week 52. Exploratory analyses through week 104 were conducted. The prespecified alpha level was 0.2. RESULTS: A total of 125 patients were randomised and received blinded infusions. Achievement of CRR was greater with obinutuzumab at week 52 (primary endpoint, 22 (35%) vs 14 (23%) with placebo; percentage difference, 12% (95% CI -3.4% to 28%), p=0.115) and at week 104 (26 (41%) vs 14 (23%); percentage difference, 19% (95% CI 2.7% to 35%), p=0.026). Improvements in other renal response measures, serologies, estimated glomerular filtration rate and proteinuria were greater with obinutuzumab. Obinutuzumab was not associated with increases in serious adverse events, serious infections or deaths. Non-serious infusion-related reactions occurred more frequently with obinutuzumab. CONCLUSIONS: Improved renal responses through week 104 were observed in patients with LN who received obinutuzumab plus standard therapies compared with standard therapies alone. Obinutuzumab was well tolerated and no new safety signals were identified. TRIAL REGISTRATION NUMBER: NCT02550652.

Assessment of the Streptomyces-plant system to mitigate the impact of Cr(VI) and lindane in experimental soils.

Phytoremediation techniques have been proposed as ecological methods to clean up contaminated sites. This study is aimed to evaluate the effect of the Streptomyces sp. Waksman & Henrici and Zea mays L. plant system on the dissipation of Cr(VI) and/or lindane from a co-contaminated soil, being 2 mg kg-1 of lindane and 150 mg kg-1 of chromium used. Lindane dissipation was improved in the presence of plant-microorganism association; however, Cr(VI) removal was higher when plants or the microorganism were separately. In co-contaminated systems, chromium content in plant tissues was lower than metal content in plants grown only with Cr(VI), suggesting that lindane could interfere with metal accumulation in the plant. The high malondialdehyde (MDA) concentration detected in non-inoculated plants grown with chromium could be consequence of high metal concentration in plant tissues. Interestingly, plants inoculated with Streptomyces sp. Z38 growing with Cr(VI) showed decrease in MDA concentration, indicating that the bacterium could activate defense mechanisms in the plant. Also, inoculated plants showed the highest value of superoxide dismutase activity. Lettuce plants used as bioindicators grew better in biologically treated soils compared with lettuce grown on non-treated soil. The results presented in this work provide the basis that will allow the optimization of future trials on a larger scale.

Beneficial traits of root endophytes and rhizobacteria associated with plants growing in phytomanaged soils with mixed trace metal-polycyclic aromatic hydrocarbon contamination.

The diversity of cultivable bacteria associated with plants from phytomanaged soils with mixed trace metal (TM) and polycyclic aromatic hydrocarbon (PAH) contamination in Pierrelaye (France) was evaluated. The emphasis was on the cultivable bacterial community since the overall objective is to obtain inoculants to improve the remediation of this type of contaminated site. Root endophytic and rhizosphere soil bacterial counts were determined, and isolates were pooled by amplified rDNA restriction analysis and identified by 16S rDNA sequencing. Isolates were further characterized for the production of plant growth-promoting (PGP) substances, and resistance to TM. The selected strains were evaluated for their ability to degrade PAHs. The potential of cell-free microbial supernatant to increase the mobilisation of PAHs from the polluted soil of Pierrelaye was also evaluated. Proteobacteria and Actinobacteria dominated the collection of isolates, and differences in taxonomic diversity were observed between plant species (Populus or Zea mays) and depending on the remediation treatment (Populus inoculation with mycorrhizae or Populus intercropping with Alnus). The majority of isolates exhibited at least one of the tested PGP traits, as well as resistance to more than one TM. Several rhizosphere, endophyte and even one bulk soil isolate showed high rates of fluoranthene and pyrene reduction. The endophyte Rhizobium strain MR28 isolated from maize and degrading pyrene produced bioemulsifying molecules capable of improving the availability of PAHs from the soil of Pierrelaye. A selection of the most interesting strains was made for further re-inoculation experiments in order to assess their potential in rhizoremediation processes.

Whole genome sequence of the multi-resistant plant growth-promoting bacteria Streptomyces sp. Z38 with potential application in agroindustry and bio-nanotechnology.

The genus Streptomyces is widely recognized for its biotechnological potential. Due to a need to improve crops, clean up the environment and produce novel antimicrobial molecules exploiting Streptomyces has become a priority. To further explore the biotechnological potential of these organisms we analyzed the genome of the strain Streptomyces sp. Z38 isolated from contaminated roots tissues. Our analysis not only confirmed the ability of the strain to produce plant growth promoting traits but also a range of mechanisms to cope with the toxic effect of heavy metals through genes involved in metal homeostasis and oxidative stress response. The production of silver nanoparticles indicated that Streptomyces sp. Z38 may find utility in Green, Grey and Red biotechnology.

Effectiveness of the Zea mays-Streptomyces association for the phytoremediation of petroleum hydrocarbons impacted soils.

Restoring polluted sites by petroleum hydrocarbons is a challenge because of their complexity and persistence in the environment. The main objective of the present study was to investigate the performance of plant-actinobacteria system for the remediation of crude petroleum and pure-polycyclic aromatic hydrocarbons (PAHs) contaminated soils. The endophytic strain Streptomyces sp. Hlh1 was tested for its ability to degrade model PAHs (phenanthrene, pyrene and anthracene) in liquid minimal medium. Streptomyces sp. Hlh1 demonstrated the ability to grow on PAHs as sole carbon and energy source, reaching hydrocarbons removal of 63%, 93% and 83% for phenanthrene, pyrene and anthracene, respectively. Maize plant was chosen to study the impact of Streptomyces sp. Hlh1 inoculation on the dissipation of contaminants and plant growth. Thus, maize seedlings grown in soils contaminated with crude petroleum and pure-PAHs were inoculated with Streptomyces sp. Hlh1. Results showed that the endophyte inoculation increased contaminants removal. Maximum hydrocarbons removal (70%) was achieved in inoculated and planted soil contaminated with crude oil, while 61%, 59%, and 46% of hydrocarbons dissipation were registered for phenanthrene, pyrene and anthracene, respectively. These degradations rates were significantly higher compared to non-inoculated systems in all the treatments evaluated. Further, it was revealed that hydrocarbons (C8-C30) were efficiently degraded in plant-Streptomyces Hlh1 system. Moreover, the inoculation with the actinobacteria resulted significant plant development and enhanced photosynthetic pigments compared to plants grown in the other experimental conditions. The present study provide evidence that the inoculation of maize plants with Streptomyces sp. Hlh1 play a remarkable role in the removal of petroleum hydrocarbons, enhancing plant development in contaminated soils.

Evaluation of the effectiveness of a bioremediation process in experimental soils polluted with chromium and lindane.

Bioremediation using actinobacterium consortia proved to be a promising alternative for the purification of co-contaminated environments. In this sense, the quadruple consortium composed of Streptomyces sp. M7, MC1, A5, and Amycolatopsis tucumanensis AB0 has been able to remove significant levels of Cr(VI) and lindane from anthropogenically contaminated soils. However, the effectiveness of the bioremediation process could not be evaluated only by analytical monitoring, which is complex mainly due to the characteristics of the matrix, producing non-quantitative analyte recoveries, or interferences in the detection stage and quantification. However, the effectiveness of the bioremediation process cannot be evaluated only through analytical monitoring, which is complex due mainly to the characteristics of the matrix, to the recoveries of non-quantitative analytes or to interferences in the detection and quantification stage. For this reason, it is essential to have tools of ecological relevance to assess the biological impact of pollutants on the environment. In this context, the objective of this work was to establish the appropriate bioassays to evaluate the effectiveness of a bioremediation process of co-contaminated soils. For this, five model species were studied: four plant species (Lactuca sativa, Raphanus sativus, Lycopersicon esculentum, and Zea mays) and one animal species (Eisenia fetida). On plant species, the biomarkers evaluated were inhibition of germination (IG) and the length of hypocotyls/steam and radicles/roots of the seedling. While on E. fetida, mortality (M), weight lost, coelomocyte concentration and cell viability were tested. These bioindicators and the battery of biomarkers quantified in them showed a different level of sensitivity, from maximum to minimum: E. fetida > L. esculentum > L. sativa > R. sativus ≫>Z. mays. Therefore, E. fetida and L. esculentum and their respective biomarkers were selected to evaluate the effectiveness of the bioremediation process due to the capability of assessing the effect on the flora and the fauna of the soil, respectively. The joint application of these bioindicators in a field scale bioremediation process is a feasible tool to demonstrate the recovery of the quality and health of the soil.

Chromium(VI) reduction in Streptomyces sp. M7 mediated by a novel Old Yellow Enzyme.

Old Yellow Enzymes play key roles in several cellular processes and have become an important family of enzymes with biotechnological potential. One of the major challenges of biotechnology consists of the bioremediation of co-polluted soils with organic and inorganic compounds. In co-contaminated areas, chromium normally exists in its more toxic and carcinogenic form Cr(VI). Microorganisms can reduce this metal to the insoluble and less toxic Cr(III). Streptomyces sp. M7 is a strain able to efficiently bioremediate polluted soils with γ-hexachlorocyclohexane and Cr(VI). The complete degradation pathway for γ-hexachlorocyclohexane was recently elucidated in this strain. In the present work, we confirmed the ability of Streptomyces sp. M7 to eliminate a high percentage of Cr(VI) from a synthetic culture medium. After a transcriptional study in the presence of Cr(VI), we also report the molecular cloning of a gene coding for an Old Yellow Enzyme with chromate reductase activity. Our results suggest that the elimination of Cr(VI) by Streptomyces sp. M7 is directly related to the activity of this Old Yellow Enzyme. The importance of our work is in identifying for the first time an Old Yellow Enzyme with chromate reductase activity in Streptomyces and Actinobacteria. Finding this enzyme helps understand chromium homeostasis in Streptomyces sp. M7, in addition to opening a new research window related to Old Yellow Enzymes from Actinobacteria.

Multi-resistant plant growth-promoting actinobacteria and plant root exudates influence Cr(VI) and lindane dissipation.

The aims of this study were (1) to isolate new multi-resistant actinobacteria from soil, rhizosphere and plant samples collected from an ancient illegal pesticide storage and (2) to elucidate the effects of these microorganisms developed with maize root exudates on lindane and Cr(VI) removal. Fifty-seven phenotypically different actinobacteria were isolated and four of them, belonging to the genus Streptomyces exhibit tolerance to a mixture of lindane and Cr(VI). Two rhizospheric strains named as Streptomyces sp. Z38 and Streptomyces sp. Z2 were selected to be grown with root exudates because they showed the highest Cr(VI) and lindane removal in co-contaminated medium. When root exudates were the only carbon source, metal dissipation increased significantly either as single or mixed contaminant, compared to metal dissipation with glucose. No significant differences were found on lindane removal with root exudates or glucose, so a higher lindane concentration was evaluated. Despite of this, lindane removal remained stable while metal dissipation was notoriously lower when lindane concentration was enhanced. In addition to a good performance growing with mixed contaminants, Streptomyces strains showed plant growth promoting traits that could improve plant establishment. The results presented in this study show the importance of the screening programs addressed to find new actinobacteria able to grow in co-contaminated systems. It was also evidenced that root exudates of maize improve the growth of Streptomyces strains when they were used as carbon source, being the dissipation of Cr(VI) considerably improved in presence of lower lindane concentration.

Production of a microbial emulsifier with biotechnological potential for environmental applications.

The present work reports the production of bioemulsifiers (BEs) by an environmental bacterium closely related to Bacillus spp., using agro-industrial wastes and by-products as low-cost carbon sources. Maximum emulsifying activity was detected using crude glycerol (CG) (E24 = 59%), producing 2.8 g/L of BE at 24 h of incubation. The microbial product presented significantly higher biodegradability in comparison to three commercial emulsifying agents (sodium dodecyl sulfate, SDS; Tween 80; and Arabic gum, AG). Interestingly, BE proved to be innocuous for Caco-2 cells and wheat seedlings, used as toxicological indicators. The BE and AG showed (in most of the cases) higher stability to changes in temperature (37 °C-100 °C), pH (3-10), and salt concentration (5% and 10%, w/v) in comparison to the SDS and Tween 80. Finally, the microbial product displayed a large affinity to a wide range of hydrophobic substrates showing emulsifying activities similar to or even better than SDS, Tween 80 and AG. The results presented in this study demonstrate the potential of a bioproduct obtained from CG to be used for environmental purposes.

Cr(VI) and lindane removal by Streptomyces M7 is improved by maize root exudates.

Environmental mixed pollution by both organic and inorganic compounds are detected worldwide. Phytoremediation techniques have been proposed as ecofriendly methods for cleaning up polluted sites. Several studies have demonstrated enhanced dissipation of contaminants at the root-soil interface through an increase in microbial activity caused by the release of plant root exudates (REs). The aim of this study was to evaluate the effectiveness for Cr(VI) and lindane removal by Streptomyces M7 cultured in a co-contaminated system in presence of maize REs. Our results showed when REs were added to the contaminated minimal medium (MM) as the only carbon source, microbial removal of Cr(VI) and lindane increased significantly in comparison to contaminant removal obtained in MM with glucose 1 g L-1 . The maximum removal of 91% of lindane and 49.5% of Cr(VI) were obtained in the co-contaminated system. Moreover, Streptomyces M7 showed plant growth promoting traits which could improve plant performance in contaminated soils. The results presented in this study provide evidence that maize REs improved growth of Streptomyces M7 when REs were used as a carbon source in comparison to glucose. Consequently, lindane and Cr(VI) removal was considerably enhanced making evident the phytoremediation potential of the actinobacteria-plant partnership.