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In this randomized phase II study (GEM-KyCyDex, clinicaltrials gov. Identifier: NCT03336073), the combination of weekly carfilzomib 70 mg/m2, cyclophosphamide and dexamethasone (KCd) was compared to carfilzomib and dexamethasone (Kd) in relapsed/refractory multiple myeloma (RRMM) after 1-3 prior lines (PL). One hundred and ninety-seven patients were included and randomized 1:1 to receive KCd (97 patients) or Kd (100 patients) in 28-day cycles until progressive disease or unacceptable toxicity occurred. Patient median age was 70 years, and the median number of PL was one (range, 1-3). More than 90% of patients had previously been exposed to proteasome inhibitors, approximetely 70% to immunomodulators, and approximetely 50% were refractory to their last line (mainly lenalidomide) in both groups. After a median follow-up of 37 months, median progression-free survival (PFS) was 19.1 and 16.6 months in KCd and Kd, respectively (P=0.577). Of note, in the post hoc analysis of the lenalidomide-refractory population, the addition of cyclophosphamide to Kd resulted in a significant benefit in terms of PFS: 18.4 versus 11.3 months (hazard ratio =1.7, 95% confidence interval: 1.1-2.7; P=0.043). The overall response rate and the percentage of patients who achieved complete response was around 70% and 20% in both groups. The addition of cyclophosphamide to Kd did not result in any safety signal, except for severe infections (7% vs. 2%). In conclusion, the combination of cyclophosphamide with Kd 70 mg/m2 weekly does not improve outcomes as compared with Kd alone in RRMM after 1-3 PL, but a significant benefit in PFS was observed with the triplet combination in the lenalidomide-refractory population. The administration of weekly carfilzomib 70 mg/m2 was safe and convenient, and, overall, the toxicity was manageable in both arms.

Multiple myeloma (MM) arises following malignant proliferation of plasma cells in the bone marrow, that secrete high amounts of specific monoclonal immunoglobulins or light chains, resulting in the massive production of unfolded or misfolded proteins. Autophagy can have a dual role in tumorigenesis, by eliminating these abnormal proteins to avoid cancer development, but also ensuring MM cell survival and promoting resistance to treatments. To date no studies have determined the impact of genetic variation in autophagy-related genes on MM risk. We performed meta-analysis of germline genetic data on 234 autophagy-related genes from three independent study populations including 13,387 subjects of European ancestry (6863 MM patients and 6524 controls) and examined correlations of statistically significant single nucleotide polymorphisms (SNPs; p < 1 × 10 -9 ) with immune responses in whole blood, peripheral blood mononuclear cells (PBMCs), and monocyte-derived macrophages (MDM) from a large population of healthy donors from the Human Functional Genomic Project (HFGP). We identified SNPs in six loci, CD46 , IKBKE , PARK2 , ULK4 , ATG5 , and CDKN2A associated with MM risk ( p = 4.47 × 10 -4 -5.79 × 10 -14 ). Mechanistically, we found that the ULK4 rs6599175 SNP correlated with circulating concentrations of vitamin D3 ( p = 4.0 × 10 -4 ), whereas the IKBKE rs17433804 SNP correlated with the number of transitional CD24 + CD38 + B cells ( p = 4.8 × 10 -4 ) and circulating serum concentrations of Monocyte Chemoattractant Protein (MCP)-2 ( p = 3.6 × 10 -4 ). We also found that the CD46 rs1142469 SNP correlated with numbers of CD19 + B cells, CD19 + CD3 - B cells, CD5 + IgD - cells, IgM - cells, IgD - IgM - cells, and CD4 - CD8 - PBMCs ( p = 4.9 × 10 -4 -8.6 × 10 -4 ) and circulating concentrations of interleukin (IL)-20 ( p = 0.00082). Finally, we observed that the CDKN2A rs2811710 SNP correlated with levels of CD4 + EMCD45RO + CD27 - cells ( p = 9.3 × 10 -4 ). These results suggest that genetic variants within these six loci influence MM risk through the modulation of specific subsets of immune cells, as well as vitamin D3 - , MCP-2 - , and IL20-dependent pathways.

Aiming at the development of a greener ethylene removal alternative, the goal of this study was to scale up and ensure the safety of α-cyclodextrin nanosponges (α-CD-NS) for further use as ethylene scavengers. The solvent-free synthesis of α-CD-NS was successfully scaled up using α-cyclodextrin and N,N'-carbonyldiimidazole as cross-linkers (1:4 molar ratio) by means of mechanical alloying using a PM 100 ball mill by focusing on varying the rotation frequency, as determined by FTIR-ATR, X-ray diffraction, and TGA. α-CD-NS washing optimization was performed in water by monitoring the imidazole concentration in the washing solution through the validation of a fast and sensitive HPLC-DAD method. After 6 h at 40 °C, all imidazole was extracted, allowing a faster and less energy-dependent extraction. α-CD-NS absorbent capacity and porosity were also evaluated through BET isotherms and ethylene absorption experiments using α-CD-NS and commercially available absorbents (zeolite and bentonite) were performed by means of gas chromatography (GC) coupled to a flame ionization detector (FID). With a 93 µL h -1 kg adsorbent -1 ethylene removal capacity, α-CD-NS revealed the best ethylene scavenging activity when compared to the other absorbents, opening the doors for a safer, innovative, and eco-friendlier ethylene removal active packaging.

Robust and applicable risk-stratifying genetic factors at diagnosis in pediatric T-cell acute lymphoblastic leukemia (T-ALL) are still lacking, and most protocols rely on measurable residual disease (MRD) assessment. In our study, we aimed to analyze the impact of NOTCH1, FBXW7, PTEN , and RAS mutations, the measurable residual disease (MRD) levels assessed by flow cytometry (FCM-MRD) and other reported risk factors in a Spanish cohort of pediatric T-ALL patients. We included 199 patients treated with SEHOP and PETHEMA consecutive protocols from 1998 to 2019. We observed a better outcome of patients included in the newest SEHOP-PETHEMA-2013 protocol compared to the previous SHOP-2005 cohort. FCM-MRD significantly predicted outcome in both protocols, but the impact at early and late time points differed between protocols. The impact of FCM-MRD at late time points was more evident in SEHOP-PETHEMA 2013, whereas in SHOP-2005 FCM-MRD was predictive of outcome at early time points. Genetics impact was different in SHOP-2005 and SEHOP-PETHEMA-2013 cohorts: NOTCH1 mutations impacted on overall survival only in the SEHOP-PETHEMA-2013 cohort, whereas homozygous deletions of CDKN2A / B had a significantly higher CIR in SHOP-2005 patients. We applied the clinical classification combining oncogenetics, WBC count and MRD levels at the end of induction as previously reported by the FRALLE group. Using this score, we identified different subgroups of patients with statistically different outcome in both Spanish cohorts. In SHOP-2005, the FRALLE classifier identified a subgroup of high-risk patients with poorer survival. In the newest protocol SEHOP-PETHEMA-2013, a very low-risk group of patients with excellent outcome and no relapses was detected, with borderline significance. Overall, FCM-MRD, WBC count and oncogenetics may refine the risk-stratification, helping to design tailored approaches for pediatric T-ALL patients., Competing Interests: MT reports travel and accommodation support from Novartis (outside the submitted work); travel and accommodation support from Jazz Pharma and Shire/Servier (outside the submitted work); and travel and accommodation support from Amgen (outside the submitted work). JLF is a consultant/advisory member for Amgen, Jazz Pharmaceuticals, and Novartis (outside the submitted work), receives honoraria for speaking at symposia from Amgen, Servier, Jazz Pharmaceuticals, and Pfizer (outside the submitted work) and support for attending symposia from Servier and Jazz Pharmaceuticals (outside the submitted work). AM discloses talk fees from Jazz Pharma and Shire/Servier outside the presented work and reports travel and accommodation support from Jazz Pharma, Shire/Servier, and Novartis (outside the submitted work). JD reports advisory board honorarium, speaker fees, and travel and accommodation support from Jazz Pharma and Shire/Servier; personal fees, advisory board honorarium, speaker fees, and travel and accommodation support from Novartis (outside the submitted work); advisory board honorarium, and travel and accommodation support from Amgen (outside the submitted work); advisory board honorarium, speaker fees, and travel and accommodation support from Celgene (outside the submitted work); advisory board honorarium, speaker fees, and travel and accommodation support from Sobi (outside the submitted work). SR reports advisory board honorarium, speaker fees, and travel and accommodation support from Jazz Pharma and Shire/Servier; personal fees, advisory board honorarium, speaker fees, and travel and accommodation support from Novartis (outside the submitted work); advisory board honorarium, and travel and accommodation support from Amgen (outside the submitted work); advisory board honorarium, speaker fees, and travel and accommodation support from Celgene (outside the submitted work). MC discloses talk fees from Shire/Servier outside the presented work. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Vega-García, Perez-Jaume, Esperanza-Cebollada, Vicente-Garcés, Torrebadell, Jiménez-Velasco, Ortega, Llop, Abad, Vagace, Minguela, Pratcorona, Sánchez-Garcia, García-Calderón, Gómez-Casares, Martín-Clavero, Escudero, Riñón Martinez-Gallo, Muñoz, Velasco, García-Morin, Català, Pascual, Velasco, Fernández, Lassaletta, Fuster, Badell, Molinos-Quintana, Molinés, Guerra-García, Pérez-Martínez, García-Abós, Robles Ortiz, Pisa, Adán, Díaz de Heredia, Dapena, Rives, Ramírez-Orellana, Camós and on behalf of the Biological Committee of the Group of Leukaemia of the Spanish Society of Paediatric Haematology Oncology (SEHOP Group).)

Here, we assessed whether 36 single nucleotide polymorphisms (SNPs) within the TNFSF4 and MAPKAPK2 loci influence the risk of developing invasive aspergillosis (IA). We conducted a two-stage case control study including 911 high-risk patients diagnosed with hematological malignancies that were ascertained through the aspBIOmics consortium. The meta-analysis of the discovery and replication populations revealed that carriers of the TNFSF4 rs7526628T/T genotype had a significantly increased risk of developing IA ( p = 0.00022). We also found that carriers of the TNFSF4 rs7526628T allele showed decreased serum levels of TNFSF14 protein ( p = 0.0027), and that their macrophages had a decreased fungicidal activity ( p = 0.048). In addition, we observed that each copy of the MAPKAPK2 rs12137965G allele increased the risk of IA by 60% ( p = 0.0017), whereas each copy of the MAPKAPK2 rs17013271T allele was estimated to decrease the risk of developing the disease ( p = 0.0029). Mechanistically, we found that carriers of the risk MAPKAPK2 rs12137965G allele showed increased numbers of CD38+IgM-IgD- plasmablasts in blood ( p = 0.00086), whereas those harboring two copies of the allele had decreased serum concentrations of thymic stromal lymphopoietin ( p = 0.00097). Finally, we also found that carriers of the protective MAPKAPK2 rs17013271T allele had decreased numbers of CD27-IgM-IgD- B cells ( p = 0.00087) and significantly lower numbers of CD14+ and CD14+CD16- cells ( p = 0.00018 and 0.00023). Altogether, these results suggest a role of the TNFSF4 and MAPKAPK2 genes in determining IA risk.

Background: Stringent complete response (sCR) is used as a deeper response category than complete response (CR) in multiple myeloma (MM) but may be of limited value in the era of minimal residual disease (MRD) testing., Methods: Here, we used 4-colour multiparametric flow cytometry (MFC) or next-generation sequencing (NGS) of immunoglobulin genes to analyse and compare the prognostic impact of sCR and MRD monitoring. We included 193 treated patients in two institutions achieving CR, for which both bone marrow aspirates and biopsies were available., Results: We found that neither the serum free light chain ratio, clonality by immunohistochemistry (IHC) nor plasma cell bone marrow infiltration identified CR patients at distinct risk. Patients with sCR had slightly longer progression-free survival. Nevertheless, persistent clonal bone marrow disease was detectable using MFC or NGS and was associated with significantly inferior outcomes compared with MRD-negative cases., Conclusion: Our results confirm that sCR does not predict a different outcome and indicate that more sensitive techniques are able to identify patients with differing prognoses. We suggest that MRD categories should be implemented over sCR for the future classification of MM responses., Competing Interests: J Martinez-Lopez belongs to the speaker bureau of Adaptive Biotechnologies. This does not alter our adherence to PLOS ONE policies on sharing data and materials. The rest of the authors declare no competing financial interests.

Cardiovascular disease is an important cause of morbidity and mortality in people living with HIV (PLWH), who commonly experience lipid disturbances. The aim of this study was to determine whether the plasma lipidomic profile differs between PLWH receiving a darunavir-based ART and those receiving integrase inhibitor-based ART. This was a cross-sectional study of unselected patients for whom metabolomic analysis was performed using ultra-high-performance liquid chromatography coupled to mass spectrometry. Data for the two subgroups were compared by calculating the log2 of the fold change for each metabolite and then grouping these into the main lipid families. Sixty-two PLWH aged 49.3 ± 8.6 years (82% men) were included: 12 patients (19.4%) had hypertension, 8 (12.9%) had type 2 diabetes, 25 (41.0%) had dyslipidaemia and 9 (14.5%) were taking statins, without significant differences in all these variables between the two groups. Twenty-five (40.3%) received darunavir-based ART and 37 (59.7%) integrase inhibitor-based ART. Although the differences were not statistically significant, patients treated with darunavir-based ART had higher concentrations of total cholesterol (211 mg/dL vs 194 mg/dL), LDL-cholesterol (132 mg/dL vs 117 mg/dL) and triglycerides (155 mg/dL vs 122 mg/dL), and lower HDL-cholesterol concentration (50 mg/dL vs 52 mg/dL). The main lipid families and metabolites differed slightly between groups (log2-fold change; P-value): ceramides (-0.07; 0.49), phosphatidylinositols (-0.05; 0.63), diacylglycerols (0.10; 0.64), phosphatidylethanolamines (0.03; 0.78), triacylglycerols (0.27; 0.18) and lysophosphatidylethanolamines (0.03; 0.83). In the integrase inhibitor-based group, the use of tenofovir alafenamide fumarate significantly increases the majority of lipid fractions, when compared with tenofovir disoproxil fumarate. The lipidomic profile did not differ between PLWH treated with darunavir-based or integrase inhibitor-based ART. This was especially true for ceramides, which are involved in cardiovascular disease. Further studies are needed to study the impact of ART in lipidomic profile.

The marine diatom Thalassiosira pseudonana grown under air (0.04% CO 2 ) and 1 and 5% CO 2 concentrations was evaluated to determine its potential for CO 2 mitigation coupled with biodiesel production. Results indicated that the diatom cultures grown at 1 and 5% CO 2 showed higher growth rates (1.14 and 1.29 div day -1 , respectively) and biomass productivities (44 and 48 mg AFDW L -1  day -1 ) than air grown cultures (with 1.13 div day -1 and 26 mg AFDW L -1  day -1 ). The increase of CO 2 resulted in higher cell volume and pigment content per cell of T. pseudonana. Interestingly, lipid content doubled when air was enriched with 1-5% CO 2 . Moreover, the analysis of the fatty acid composition of T. pseudonana revealed the predominance of monounsaturated acids (palmitoleic-16:1 and oleic-18:1) and a decrease of the saturated myristic acid-14:0 and polyunsaturated fatty acids under high CO 2 levels. These results suggested that T. pseudonana seems to be an ideal candidate for biodiesel production using flue gases.

This proof-of-concept single-arm open-label phase I clinical trial (NCT02481934) studied the safety and efficacy of multiple infusions of activated and expanded natural killer (NKAE) cells in combination with anti-myeloma drugs in multiple myeloma patients. It included five patients with relapsed or refractory MM who had received two to seven prior lines of therapy; NK cells were expanded for 3 weeks with K562-mb15-41BBL cells. Patients received four cycles of pharmacological treatment with two infusions of 7.5 × 10 6 NKAEs/kg per cycle. NKAE generation, expansion, and NK monitoring was assessed using flow cytometry. Eighteen clinical-grade NKAE cell GMP-grade products were generated to obtain 627 × 10 6 NKAEs (range: 315-919 × 10 6 ) for the first infusion and 943 × 10 6 (range: 471-1481 × 10 6 ) for the second infusion with 90% (±7%) purity. Neutropenia grade II occurred in two patients and was related to chemotherapy. Of the five patients, four showed disease stabilization before the end of NKAE treatment, and two showed a 50% reduction in bone marrow infiltration and a long-term (>1 y) response. The NKAE cells had a highly cytotoxic phenotype and high cytotoxicity in vitro . Infused NKAE cells were detected in bone marrow and peripheral blood after infusions. Ex vivo expansion of autologous NK cells is feasible, NKAE cells are clinically active and the multiple infusions are well tolerated in patients with relapsed or refractory myeloma.

Harvesting is one of the key challenges to determine the feasibility of producing biodiesel from algae. This paper presents experimental results for a cost-effective system to harvest Chaetoceros calcitrans, using natural sedimentation, flocculation, and inducing pH. No efficient sedimentation of microalgal cells was observed only by gravity. By alkalinity-induced flocculation, at a pH value of 9.51, 86% recovery of the cells was achieved with a sedimentation rate of 125 cm/h and a concentration factor (CF) of 4 (volume/volume (v/v)) in 10 min. The maximum photochemical quantum yield of photosystem II (Fv/Fm) of concentrated cells was almost the same as fresh culture (0.621). Commercial flocculants, aluminium sulphate and poly-aluminium chloride (PAC), were also successful in harvesting the studied algal cells. Optimum concentration of aluminium sulphate (AS) could be concluded as 10 ppm with 87.6% recovery and 7.10 CF (v/v) in 30 min for cost-efficient harvesting, whereas for PAC it was 20 ppm with 74% recovery and 6.6 CF (v/v). Fv/Fm yields of concentrated cells with AS and PAC showed a 1% reduction compared to fresh culture. Mg+2 was the triggering ion for alkalinity-induced flocculation in the conditions studied. The rheology behaviour of the concentrated cells was Newtonian with values between 2.2×10(-3) and 2.3×10(-3) Pa s at 30°C.

Previous studies have shown the reproducibility of the 2008 World Health Organization (WHO) classification in myelodysplastic syndromes (MDS), especially when multilineage dysplasia or excess of blasts are present. However, there are few data regarding the reproducibility of MDS with unilineage dysplasia. The revised International Prognostic Scoring System R-IPSS described two new morphological categories, distinguishing bone marrow (BM) blast cell count between 0-2 % and >2- < 5 %. This distinction is critical for establishing prognosis, but the reproducibility of this threshold is still not demonstrated. The objectives of our study were to explore the reliability of the 2008 WHO classification, regarding unilineage vs. multilineage dysplasia, by reviewing 110 cases previously diagnosed with MDS, and to study whether the threshold of ≤2 % BM blasts is reproducible among different observers. We used the same methodology as in our previous paper [Font et al. (2013) Ann Hematol 92:19-24], by encouraging investigators to include patients with <5 % BM blasts. Samples were collected from 11 hospitals and were evaluated by 11 morphologists. Each observer evaluated 20 samples, and each sample was analyzed independently by two morphologists. Discordance was observed in 36/108 suitable cases (33 %, kappa test 0.503). Diagnosis of MDS with unilineage dysplasia (refractory cytopenia with unilineage dysplasia (RCUD), refractory anemia with ring sideroblasts (RARS) or unclassifiable MDS) was assessed in 33 patients, by either of the two observers. We combined this series with the cases with RCUD or RARS included in our 2013 paper, thus obtaining 50 cases with unilineage dysplasia by at least one of the observers. The whole series showed very low agreement regarding RCUD (5/23, 21 %) and RARS (5/28, 18 %). Regarding BM blast count, the threshold of ≤2 % was not reproducible (discordance rate 32/108 cases, kappa test 0.277). Our study shows that among MDS WHO 2008 categories, interobserver discordance seems to be high in cases with unilineage dysplasia. We also illustrate that the threshold of ≤2 % BM blasts as settled by the R-IPSS may be not easy to reproduce by morphologists in real practice.

The effect of shear-enhanced filtration by vibratory process in microalgae dewatering is presented in this paper. The aim of this research was to investigate the technical performance and improvement of vibrating membrane filtration compared with conventional tangential cross-flow filtration in microalgae concentration. An industrial-scale available commercial set-up was used. Several membrane materials as polyethersulfone, polyacrylonitrile, etc., and mean pore sizes (from 7000Da to 0.2μm) were tested and compared in both filtration set-ups. Experiments were carried-out with Nannochloropsis gaditana and Phaeodactylum tricornutum microalgae. It has been demonstrated that, even if the choice of the membrane depends on its cut-off, its material and the type of microalgae filtrated, dynamic filtration is always the best technology over a conventional one. If with conventional filtration permeability values were in the vicinity of 10L/h/m(2)/bar in steady state phase, with dynamic filtration these values increased to 30L/h/m(2)/bar or more., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

We compared potential pre-concentration techniques for Nannochloropsis gaditana (Nng) by testing natural sedimentation; flocculation with aluminium sulphate, polyaluminium chloride and chitosan; and induced pH. Promising flocculation efficiencies and concentration factors were obtained in a short time with alkalinity-induced flocculation at an adjusted pH of 9.7 and with chitosan at an adjusted pH of 9.9 using a concentration of 30mgL(-1). The sedimentation rates of alkalinity-induced flocculation were also evaluated. Additionally, viscosity, particle size distribution and Ca/Mg ions were analysed for pre-concentrated samples of N. gaditana (Nng) and the previously studied Phaeodactylum tricornutum (Pht) which were obtained by various different harvesting methods under optimal conditions. The rheological properties of the concentrated algae suspensions of two microalgal species showed Newtonian behaviour. The mean diameters of the flocs were between 39 and 48μm. The Ca/Mg analysis showed that Mg(+2) is the triggering ion for alkalinity-induced flocculation in the conditions studied., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

We extend our previous molecular dynamics experiments [Rodriguez et al., J. Phys. Chem. B 109, 24427 (2005)] to the analysis of the adsorption of catanionic surfactants at water/air interfaces, at a surfactant coverage close to that of the saturated monolayer: 30.3 A(2) per headgroup. The mixture of surfactants investigated corresponds to equal amounts of dodecytrimethylammonium (DTA) and dodecylsulfate (DS). The structure of the interface is analyzed in terms of the local densities and orientational correlations of all relevant interfacial species. In accordance with experimental evidence, the DTA headgroups penetrate deeper into the aqueous substrate than the DS ones, although the average positions of all headgroups, with respect to the interface, lie in positions somewhat more external than the ones observed at lower coverages. Average tail tilts are close to 45 degrees. The characteristics of the headgroup-water substrate correlations are also analyzed using a tessellation procedure of the interface. The density and polarization responses of the interfacial domains closest to the DS headgroups are enhanced, compared to those adjacent to the DTA detergents. Dynamical aspects related to the diffusion and to the orientational correlations of different water layers in close contact with the surfactant are also investigated.

Structural properties pertaining to the solvation of mixtures of dodecytrimethylammonium/dodecylsulfate adsorbed at water/air interfaces were studied using molecular dynamics techniques. Two different surfactant coverages, both in the submonolayer regime, were considered: an infinite-diluted catanionic pair and an equimolar mixture, at a surface concentration of 78.7 A2/headgroup. The most stable solvated structures for the single surfactant pair correspond to contact-head-ion-pairs (CHIP) at a distance close to 5 A. In addition, marginally stable solvent-separated-head-ion-pairs (SSHIP) at distances approximately 7 A were also observed. The mean free energy for the dissociation of CHIP was estimated to be approximately 1 kcal/mol. At finite surfactant concentrations, one observes a considerable degree of clustering between the amphiphiles, due to the strong Coulomb coupling between headgroups. The resulting spatial domains show asymmetric structures with linear dimensions comparable to the simulation box, suggesting the onset of percolative structures. The connectivity pattern of these domains was interpreted in terms of a simplified model consisting of two-dimensional charged Lennard-Jones spheres.

The phylogenetic diversity of oxygenic phototrophic microorganisms in hypersaline microbial mats and their distribution along a salinity gradient were investigated and compared with the halotolerances of closely related cultivated strains. Segments of 16S rRNA genes from cyanobacteria and diatom plastids were retrieved from mat samples by DNA extraction and polymerase chain reaction (PCR), and subsequently analysed by denaturing gradient gel electrophoresis (DGGE). Sequence analyses of DNA from individual DGGE bands suggested that the majority of these organisms was related to cultivated strains at levels that had previously been demonstrated to correlate with characteristic salinity responses. Proportional abundances of amplified 16S rRNA gene segments from phylogenetic groupings of cyanobacteria and diatoms were estimated by image analysis of DGGE gels and were generally found to correspond to abundances of the respective morphotypes determined by microscopic analyses. The results indicated that diatoms accounted for low proportions of cells throughout, that the cyanobacterium Microcoleus chthonoplastes and close relatives dominated the communities up to a salinity of 11% and that, at a salinity of 14%, the most abundant cyanobacteria were related to highly halotolerant cultivated cyanobacteria, such as the recently established phylogenetic clusters of Euhalothece and Halospirulina. Although these organisms in cultures had previously demonstrated their ability to grow with close to optimal rates over a wide range of salinities, their occurrence in the field was restricted to the highest salinities investigated.