Oxidative phosphorylation selectively orchestrates tissue macrophage homeostasis.

In vitro studies have associated oxidative phosphorylation (OXPHOS) with anti-inflammatory macrophages, whereas pro-inflammatory macrophages rely on glycolysis. However, the metabolic needs of macrophages in tissues (TMFs) to fulfill their homeostatic activities are incompletely understood. Here, we identified OXPHOS as the highest discriminating process among TMFs from different organs in homeostasis by analysis of RNA-seq data in both humans and mice. Impairing OXPHOS in TMFs via Tfam deletion differentially affected TMF populations. Tfam deletion resulted in reduction of alveolar macrophages (AMs) due to impaired lipid-handling capacity, leading to increased cholesterol content and cellular stress, causing cell-cycle arrest in vivo. In obesity, Tfam depletion selectively ablated pro-inflammatory lipid-handling white adipose tissue macrophages (WAT-MFs), thus preventing insulin resistance and hepatosteatosis. Hence, OXPHOS, rather than glycolysis, distinguishes TMF populations and is critical for the maintenance of TMFs with a high lipid-handling activity, including pro-inflammatory WAT-MFs. This could provide a selective therapeutic targeting tool.

Effectiveness of hallux valgus surgery on improving health-related quality of life: A follow up study.

BACKGROUND: Hallux valgus (HV) negatively impacts health-related quality of life (HRQoL). Patientreported outcome measures (PROMs) are increasingly used in clinical studies of the foot and ankle. We aimed to evaluate the effect of HV surgery on PROMs (i.e., pain scales, general HRQoL, and region-specific scales) and radiological angles. Additionally, we aimed to determine whether the effect on these outcomes depends on the type of surgery (including open and percutaneous techniques) and if it is influenced by potential confounding factors (i.e., age, HVA, 1-2 IMA, body mass index (BMI), and distal metatarsal articular angle (DMAA). METHODS: This was a longitudinal prospective study. We collected the clinical data of all patients who underwent surgery for symptomatic HV deformity in the orthopedic department of the Virgen de la Luz Hospital of Cuenca (Spain).The clinical outcomes were assessed using the American Orthopedic Foot and Ankle Society (AOFAS) Hallux metatarsophalangeal-interphalangeal (HMI) scale, visual analogue scale (VAS), Manchester Oxford foot questionnaire (MOXFQ), short form health survey (SF-12) and European Quality of Life-5 Dimensions (EQ-5D). RESULTS: A total of 72 patients (70 women, 97.2%) were included in the study 72 (72 feet).The AOFAS pre-post-surgery score changed from 42.16 (SD: 10.11) to 83.31 (SD: 6.23). Considering AOFAS domains, the pre-post change was from 14.17 (SD: 9.15) to 33.19 (SD: 4.69) for pain, from 27.22 (SD: 3.90) to 37.94 (SD: 2.78) for function, and from 0.78 (SD: 2.38) to 12.18 (SD: 3.45) for alignment. For other clinical outcomes was VAS score from 5.01(SD: 1.26) to 1.26 (SD: 0.96) and MOXFQ score from 61.44 (SD: 7.09) to 12.35 (SD: 4.85). SF-12 (physical) changed from 36.26 (SD: 5.32) to 47.06 (SD: 4.82), SF-12 (mental) from 38.23 (SD: 8.04) to 46.49 (SD: 4.16), and EQ5-D from 0.64 (SD: 0.008) to 0.90 (SD: 0.10). CONCLUSIONS: Our data confirmed the improvements in the clinical and radiological outcomes after HV surgery, and provided some evidence of these improvements not depending on the type of surgery or on some potential confounding factors such as BMI, HVA, 1-2 IMA, and DMAA.

Genetic Variants of ANGPT1, CD39, FGF2 and MMP9 Linked to Clinical Outcome of Bevacizumab Plus Chemotherapy for Metastatic Colorectal Cancer.

Angiogenesis pathway genes show substantial genetic variability causing inter-individual differences in responses to anti-angiogenic drugs. We examined 20 single nucleotide polymorphisms (SNPs) in 13 of these genes to predict tumour response and clinical outcome measured as progression free survival (PFS) and overall survival (OS) in 57 patients with metastatic colorectal cancer (mCRC) given bevacizumab plus chemotherapy. SNPs were detected (iPLEX® Assay) in genomic DNA extracted from formalin-fixed paraffin-embedded tumour specimens. The variant allele CD39 rs11188513 was associated with a good tumour response (p = 0.024). Patients homozygous for the wild-type allele FGF2 rs1960669 showed a median PFS of 10.95 months versus 5.44 months for those with at least one variant allele-A (HR 3.30; 95% CI: 1.52-7.14; p = 0.001). Patients homozygous for wild-type MMP9 rs2236416 and rs2274755 showed a median PFS of 9.48 months versus 6 and 6.62 months, respectively, for those with at least one variant allele (p = 0.022, p = 0.043, respectively). OS was also lengthened to 30.92 months (p = 0.034) in carriers of wild-type ANGPT1 rs2445365 versus 22.07 months for those carrying at least one variant allele-A. These gene variants were able to predict clinical outcome and tumour response in mCRC patients given bevacizumab-based therapy.

Evaluating the efficacy of an Advanced Care Planning Program for Health Decisions in patients with advanced heart failure: protocol for a Randomized Clinical Trial.

BACKGROUND: An Advanced Care Planning (ACP) program of health decisions is the result of a process of reflection and relationship-building between the patient, their relatives and health professionals. It is based on respect for patients' autonomy, involving them in making decisions about their disease in a way that is shared between the medical team, the patient and their relatives. Up until now, the efficacy of an ACP has not been measured in the existing literature, and therefore it is unknown if these programs reach their goal. The main objective of our study is to evaluate the efficacy of an ACP program for decision-making in patients with advanced heart failure (HF) in comparison to usual follow up and care. This objective will be evaluated by the Patient Activation Measure test, which measures the participation and self-management of the patient in decision-making. Secondary objectives: to evaluate the effect of the program on quality of life, to know if the patients wishes expressed through the ACP program are fulfilled, to measure the impact of the program on patients' caregivers, to determine the satisfaction of patients included in the program and to evaluate the effect on quality of death. METHODS: Randomized multicentre clinical trial at four hospitals in Madrid. Once they are included in the study, patients' allocation to groups (control vs intervention) will be made by alternative sampling. ACP will be applied to the intervention group, whereas in the Control Group usual follow-up will be carried out in HF units. All patients will fulfil questionnaires and tests related to the objectives of the study again after a 12-month follow-up period in order to gauge the effect of ACP in patients with advanced HF. DISCUSSION: The characteristics of patients with advanced HF make them a model for designing ACP programs, given the high prevalence of this disease, the progressive increase in its incidence and it's clinical characteristics. Until now, the efficacy of this type of program has not been measured, so this Clinical Trial can provide relevant data for future ACP projects. Trial registration ClinicalTrials.gov Identifier: NCT04424680. Registered 9 June 2020. Retrospectively registered, https://clinicaltrials.gov/ct2/show/NCT04424680?term=NCT04424680&draw=2&rank=1 .

Effect of bacteria type and sucrose concentration on levan yield and its molecular weight.

BACKGROUND: Levan has been traditionally produced from microorganism. However, there is a continuous effort in looking for new strains that improve levan production yield and uses alternative sugar sources for growth. Despite having a wide range of data about levan yield, there are not papers which allow controlling molecular weight, and that plays an essential role for further applications. RESULTS: The effect of the sucrose concentration on levan yield (and its molecular weight) from Bacillus atrophaeus and Acinetobacter nectaris (Gram positive and Gram negative respectively) was studied in this work. It was found that A. nectaris growth (from 3 to 1.5 g L-1 in 40 h) and its levan production (from 3 to 1.5 g L-1) decreases by increasing sucrose concentration (best results at a concentration of 120 g L-1) whereas B. atrophaeus growth (3.5 g L-1 in 30 h) and its levan production (also 3.5 g L-1) were not affected by modifying that parameter. Levan molecular weight from A. nectaris decreases by increasing sucrose concentration (from 8000 to 2000 kDa) whereas levan molecular weight from B. Atrophaeus remains always around 50 kDa. By performing a kinetic study, it was shown that A. nectaris growth follows a substrate-inhibition model, whereas Monod equation provided a good fit for B. atrophaeus growth. Finally, wastes from orange juice industry were used as a medium culture to cultivate those microorganism, obtaining good results with B. atrophaeus (growth 3 g L-1 in 30 h). CONCLUSIONS: Levan production kinetics was determined and compared between different bacteria types.

Simulations of the gain profile and performance of a diode side-pumped QCW Nd:YAG laser.

A design of a diode side-pumped Nd:YAG laser module and simulations of the gain distribution inside the active medium are presented in this paper. The code is based on a nonsequential ray-tracing Monte Carlo method for the light generated by the laser diodes. The fluorescence image of the active medium was analyzed in order to compare it with the simulations, which were found to be in good agreement with experimental data. The laser was tested in QCW mode and provided a maximum average power of 220 W while maintaining constant energy per pulse in the 100-1000 Hz range.

Dendritic cells as orchestrators of anticancer immunity and immunotherapy.

Dendritic cells (DCs) are a heterogeneous group of antigen-presenting innate immune cells that regulate adaptive immunity, including against cancer. Therefore, understanding the precise activities of DCs in tumours and patients with cancer is important. The classification of DC subsets has historically been based on ontogeny; however, single-cell analyses are now additionally revealing a diversity of functional states of DCs in cancer. DCs can promote the activation of potent antitumour T cells and immune responses via numerous mechanisms, although they can also be hijacked by tumour-mediated factors to contribute to immune tolerance and cancer progression. Consequently, DC activities are often key determinants of the efficacy of immunotherapies, including immune-checkpoint inhibitors. Potentiating the antitumour functions of DCs or using them as tools to orchestrate short-term and long-term anticancer immunity has immense but as-yet underexploited therapeutic potential. In this Review, we outline the nature and emerging complexity of DC states as well as their functions in regulating adaptive immunity across different cancer types. We also describe how DCs are required for the success of current immunotherapies and explore the inherent potential of targeting DCs for cancer therapy. We focus on novel insights on DCs derived from patients with different cancers, single-cell studies of DCs and their relevance to therapeutic strategies.

Characterization of carboxylated cellulose nanofibrils and oligosaccharides from Kraft pulp fibers and their potential elicitor effect on the gene expression of Capsicum annuum.

Biomass-derived oligo- and polysaccharides may act as elicitors, i.e., bioactive molecules that trigger plant immune responses. This is particularly important to increase the resistance of plants to abiotic and biotic stresses. In this study, cellulose nanofibrils (CNF) gels were obtained by TEMPO-mediated oxidation of unbleached and bleached kraft pulps. The molecular structures were characterized with ESI and MALDI MS. Analysis of the fine sequences was achieved by MS and MS/MS of the water-soluble oligosaccharides obtained by acid hydrolysis of the CNF gels. The analysis revealed the presence of two families: one corresponding to homoglucuronic acid sequences and the other composed by alternating glucose and glucuronic acid units. The CNF gels, alone or with the addition of the water-soluble oligosaccharides, were tested on Chili pepper (Capsicum annuum). Based on the characterization of the gene expression with Next Generation Sequencing (NGS) of the C. annuum's total messenger RNA, the differences in growth of the C. annuum seeds correlated well with the downregulation of the pathways regulating photosynthesis. A downregulation of the response to abiotic factors was detected, suggesting that these gels would improve the resistance of the C. annuum plants to abiotic stress due to, e.g., water deprivation and cold temperatures.

Application of Box-Behnken Design in Production of Monoglyceride with Esterification of Glycerol and Oleic Acid.

Monoglyceride MG has a wide function in the food industry, in particular as a natural emulsifier, pharmaceuticals, cosmetics, antioxidant, and antibacterial. Therefore, the production of polyol ester from esterification of acid (OA) and glycerol was investigated. The process optimization was performed using a Box-Behnken design, examining the effects of temperature, molar ratio, and catalyst amount. For predicting the optimal point, a second-order polynomial model was fitted to correlate the relationship between independent variables and response (% MG). The effects of temperature (100, 150, and 200 °C); catalyst amount (4, 10, and 16% w/w); and glycerol/oleic acid ratio (1:1, 1:2, and 1:3) were investigated and found to deeply affect the reaction outcome. At the optimal reaction conditions: 200 °C, 0.2% w/w KSF, and a glycerol/oleic acid ratio (3:1), more than 71.8% monoglycerides with selectivity of 80% were obtained. Confirmation experiments were performed to demonstrate the effectiveness of this approach, and the characterization of monoglycerides was performed using high-performance liquid chromatography (HPLC).

Inactivation of the waterborne marine pathogen Vibrio alginolyticus by photo-chemical processes driven by UV-A, UV-B, or UV-C LED combined with H2O2 or HSO5.

Ultraviolet (UV) radiation is a well-implemented process for water disinfection. The development of emergent UV sources, such as light-emitting diodes (LEDs), has afforded new possibilities for advanced oxidation processes. The emission wavelength is considered to be an important factor for photo-chemical processes in terms of both biological damage and energetic efficiency, as the inactivation mechanisms and mode-of-action may differ according to the wavelength that is applied. In addition, these processes merit exploration for inactivating emerging pathogens, such as marine vibrios, that are important bacteria to control in maritime activities. The main goal of this study was to compare the disinfection efficacy of several UV-LED driven processes with different modes of action. First, the effect of UV-LEDs was assessed at different UV ranges (UV-A, UV-B, or UV-C). Second, the possible enhancement of a combination with hydrogen peroxide (H2O2) or peroxymonosulfate salt (HSO5-) was investigated under two different application strategies, i.e. simultaneous or sequential. The results obtained indicate a high sensitivity of Vibrio alginolyticus to UV radiation, especially under UV-B (kobs = 0.24 cm2/mJ) and UV-C (kobs = 1.47 cm2/mJ) irradiation. The highest inactivation rate constants were obtained for UV/HSO5- (kobs (cm2/mJ)=0.0007 (UV-A); 0.39 (UV-B); 1.79 (UV-C)) with respect to UV/H2O2 (kobs (cm2/mJ)=0.0006 (UV-A); 0.26 (UV-B); and 1.54 (UV-C)) processes, however, regrowth was avoided only with UV/H2O2. Additionally, the disinfection enhancement caused by a chemical addition was more evident in the order UV-A > UV-B > UV-C. By applying H2O2 (10 mg/L) or HSO5- (2.5 mg/L) in a sequential mode before the UV, negligible effects were obtained in comparison with the simultaneous application. Finally, promising electrical energy per order (EEO) values were obtained as follows: UV/HSO5- (EEO (kWh/m3)=1.68 (UV-A); 0.20 (UV-B); 0.04 (UV-C)) and UV/H2O2 (EEO (kWh/m3)=2.15 (UV-A); 0.32 (UV-B); 0.04 (UV-C)), demonstrating the potential of UV-LEDs for disinfection in particular activities such as the aquaculture industry or maritime transport.

West Nile virus emergence in humans in Extremadura, Spain 2020.

In Spain, the largest human West Nile virus (WNV) outbreak among humans was reported in 2020, constituting the second most important outbreak in Europe that season. Extremadura (southwestern Spain) was one of the affected areas, reporting six human cases. The first autochthonous human case in Spain was reported in Extremadura in 2004, and no other human cases were reported until 2020. In this work, we describe the first WNV human outbreak registered in Extremadura, focusing on the most important clinical aspects, diagnostic results, and control actions which followed. In 2020, from September to October, human WNV infections were diagnosed using a combination of molecular and serological methods (an in-house specific qRT-PCR and a commercial ELISA for anti-WNV IgM and IgG antibodies) and by analysing serum, urine, and/or cerebrospinal fluid samples. Serological positive serum samples were further tested using commercial kits against related flaviviruses Usutu and Tick-borne encephalitis in order to analyse serological reactivity and to confirm the results by neutralisation assays. In total, six cases of WNV infection (five with neuroinvasive disease and one with fever) were identified. Clinical presentation and laboratory findings are described. No viral RNA was detected in any of the analysed samples, but serological cross-reactivity was detected against the other tested flaviviruses. Molecular and serological methods for WNV detection in various samples as well as differential diagnosis are recommended. The largest number of human cases of WNV infection ever registered in Extremadura, Spain, occurred in 2020 in areas where circulation of WNV and other flaviviruses has been previously reported in humans and animals. Therefore, it is necessary to enhance surveillance not only for the early detection and implementation of response measures for WNV but also for other emerging flaviviruses that could be endemic in this area.

Persistent Overactive Cytotoxic Immune Response in a Spanish Cohort of Individuals With Long-COVID: Identification of Diagnostic Biomarkers.

Long-COVID is a new emerging syndrome worldwide that is characterized by the persistence of unresolved signs and symptoms of COVID-19 more than 4 weeks after the infection and even after more than 12 weeks. The underlying mechanisms for Long-COVID are still undefined, but a sustained inflammatory response caused by the persistence of SARS-CoV-2 in organ and tissue sanctuaries or resemblance with an autoimmune disease are within the most considered hypotheses. In this study, we analyzed the usefulness of several demographic, clinical, and immunological parameters as diagnostic biomarkers of Long-COVID in one cohort of Spanish individuals who presented signs and symptoms of this syndrome after 49 weeks post-infection, in comparison with individuals who recovered completely in the first 12 weeks after the infection. We determined that individuals with Long-COVID showed significantly increased levels of functional memory cells with high antiviral cytotoxic activity such as CD8+ TEMRA cells, CD8±TCRγδ+ cells, and NK cells with CD56+CD57+NKG2C+ phenotype. The persistence of these long-lasting cytotoxic populations was supported by enhanced levels of CD4+ Tregs and the expression of the exhaustion marker PD-1 on the surface of CD3+ T lymphocytes. With the use of these immune parameters and significant clinical features such as lethargy, pleuritic chest pain, and dermatological injuries, as well as demographic factors such as female gender and O+ blood type, a Random Forest algorithm predicted the assignment of the participants in the Long-COVID group with 100% accuracy. The definition of the most accurate diagnostic biomarkers could be helpful to detect the development of Long-COVID and to improve the clinical management of these patients.

Association between HLA-C alleles and COVID-19 severity in a pilot study with a Spanish Mediterranean Caucasian cohort.

The clinical presentations of COVID-19 may range from an asymptomatic or mild infection to a critical or fatal disease. Several host factors such as elderly age, male gender, and previous comorbidities seem to be involved in the most severe outcomes, but also an impaired immune response that causes a hyperinflammatory state but is unable to clear the infection. In order to get further understanding about this impaired immune response, we aimed to determine the association of specific HLA alleles with different clinical presentations of COVID-19. Therefore, we analyzed HLA Class I and II, as well as KIR gene sequences, in 72 individuals with Spanish Mediterranean Caucasian ethnicity who presented mild, severe, or critical COVID-19, according to their clinical characteristics and management. This cohort was recruited in Madrid (Spain) during the first and second pandemic waves between April and October 2020. There were no significant differences in HLA-A or HLA-B alleles among groups. However, despite the small sample size, we found that HLA-C alleles from group C1 HLA-C*08:02, -C*12:03, or -C*16:01 were more frequently associated in individuals with mild COVID-19 (43.8%) than in individuals with severe (8.3%; p = 0.0030; pc = 0.033) and critical (16.1%; p = 0.0014; pc = 0.0154) disease. C1 alleles are supposed to be highly efficient to present peptides to T cells, and HLA-C*12:03 may present a high number of verified epitopes from abundant SARS-CoV-2 proteins M, N, and S, thereby being allegedly able to trigger an efficient antiviral response. On the contrary, C2 alleles are usually poorly expressed on the cell surface due to low association with ß2-microglobulin (ß2M) and peptides, which may impede the adequate formation of stable HLA-C/ß2M/peptide heterotrimers. Consequently, this pilot study described significant differences in the presence of specific HLA-C1 alleles in individuals with different clinical presentations of COVID-19, thereby suggesting that HLA haplotyping could be valuable to get further understanding in the underlying mechanisms of the impaired immune response during critical COVID-19.

Role of Chromodomain-Helicase-DNA-Binding Protein 4 (CHD4) in Breast Cancer.

Chromodomain-helicase-DNA-binding protein 4 (CHD4) is an epigenetic regulator identified as an oncogenic element that may provide a novel therapeutic target for the treatment of breast cancer (BC). CHD4-the core component of the nucleosome remodeling and deacetylase (NuRD) complex-may be mutated in patients with this disease. However, information on CHD4 mutants that might allow their use as biomarkers of therapeutic success and prognosis is lacking. The present work examines mutations in CHD4 reported in patients with breast cancer and included in public databases and attempts to identify their roles in its development. The databases revealed 81 point mutations across different types of breast cancer (19 of which also appeared in endometrial, intestinal, nervous system, kidney, and lymphoid organ cancers). 71.6% of the detected mutations were missense mutations, 13.6% were silent, and 6.2% nonsense. Over 50% affected conserved residues of the ATPase motor (ATPase and helicase domains), and domains of unknown function in the C-terminal region. Thirty one mutations were classified in the databases as either 'deleterious', 'probably/possibly damaging' or as 'high/medium pathogenic'; another five nonsense and one splice-site variant were predicted to produce potentially harmful truncated proteins. Eight of the 81 mutations were categorized as putative driver mutations and have been found in other cancer types. Some mutations seem to influence ATPase and DNA translocation activities (R1162W), while others may alter protein stability (R877Q/H, R975H) or disrupt DNA binding and protein activity (R572*, X34_splice) suggesting CHD4 function may be affected. In vivo tumorigenecity studies in endometrial cancer have revealed R975H and R1162W as mutations that lead to CHD4 loss-of-function. Our study provides insight into the molecular mechanism whereby CHD4, and some of its mutants could play a role in breast cancer and suggest important implications for the biological comprehension and prognosis of breast cancer, identifying CHD4 as a novel therapeutic target for BC patients.

miR-7, miR-10a and miR-143 Expression May Predict Response to Bevacizumab Plus Chemotherapy in Patients with Metastatic Colorectal Cancer.

PURPOSE: Bevacizumab is a monoclonal antibody that binds to vascular endothelial growth factor A. It is currently used in combination with chemotherapy to treat metastatic colorectal cancer. This therapy is not equally effective in every patient; in some, mechanisms of resistance arise that remain poorly understood. The aim of the present work was to determine whether the expression of 26 miRNAs could be associated with the effectiveness of bevacizumab plus chemotherapy, with progression-free survival (PFS), and with overall survival (OS) in metastatic colorectal cancer. PATIENTS AND METHODS: Paraffin-embedded biopsies from 76 patients with metastatic colorectal cancer were collected to isolate miRNAs. The expression of 26 miRNAs was analyzed by quantitative RT-PCR. For the purpose of analysis, patients were classified as either "responders" (PFS ≥6 months since beginning treatment) or "non-responders" (PFS <6 months). For the analysis of PFS and OS, patients were classified into two groups using the median gene expression value as the cut-off point ("high" [≥50% percentile] or "low" [<50% percentile]). Time-to-event data were analyzed using the Kaplan-Meier method and compared by the log rank test. Cox regression was used to estimate hazard ratios (HR) and their 95% confidence intervals. RESULTS: miR-7-5p and miR-10a-5p were more strongly expressed in non-responders than responders (p=0.049 and p=0.043, respectively), and OS was poorer in patients showing these higher expression levels (HR=2.54, 95% CI 1.42-4.55, p=0. 001, and HR=1.81, 95% CI 1.02-3.20, p=0.039, respectively). The overexpression of miR-143-3p, however, was associated with a better prognosis and significantly better PFS (HR=0.57; 95% CI: 0.33-0.96; p=0.033). CONCLUSION: High expression values for miR-7-5p and miR-10a-5p might be considered markers of a poorer prognosis in patients with metastatic colorectal cancer treated with bevacizumab plus chemotherapy, while the same for miR-143-3p might be a marker of better outcomes.

Efficacy of bevacizumab-containing chemotherapy in metastatic colorectal cancer and CXCL5 expression: Six case reports.

BACKGROUND: In metastatic colorectal cancer (mCRC), the anti-vascular endothelial growth factor drug bevacizumab (BVZ) plus chemotherapy significantly improves progression-free survival compared to chemotherapy (CT) alone. This benefit is not, however, observed in all patients. While increased chemokine CXCL5 gene expression promoting angiogenesis has been proposed as a prognostic mCRC biomarker, few studies have examined its relationship with drug efficacy. This study sought to analyze tumor CXCL5 gene expression in six patients with different efficacy of BVZ-containing CT in terms of the tumor response to treatment. CASE SUMMARY: We report six cases of stage IV KRAS-mutated mCRC. Patients were given first line treatment with BVZ-containing chemotherapy in University Hospital of Fuenlabrada. The six patients differed in terms of primary tumor location (right/left side), tumor burden (mostly hepatic and peritoneal disease) and clinical disease course. Before treatment onset, total RNA was isolated from paraffinated tumor biopsy specimens and CXCL5 gene expression quantified through conventional RT-qPCR procedures. Our main finding was that CXCL5 expression levels were several times higher in three patients with lower progression free survival (under 6 mo) from the start of treatment. CONCLUSION: A higher expression of CXCL5 was observed in the three patients showing worse tumor response to treatment.

Identification of Immunological Parameters as Predictive Biomarkers of Relapse in Patients with Chronic Myeloid Leukemia on Treatment-Free Remission.

BCR-ABL is an aberrant tyrosine kinase responsible for chronic myeloid leukemia (CML). Tyrosine kinase inhibitors (TKIs) induce a potent antileukemic response mostly based on the inhibition of BCR-ABL, but they also increase the activity of Natural Killer (NK) and CD8+ T cells. After several years, patients may interrupt treatment due to sustained, deep molecular response. By unknown reasons, half of the patients relapse during treatment interruption, whereas others maintain a potent control of the residual leukemic cells for several years. In this study, several immunological parameters related to sustained antileukemic control were analyzed. According to our results, the features more related to poor antileukemic control were as follows: low levels of cytotoxic cells such as NK, (Natural Killer T) NKT and CD8±TCRγß+ T cells; low expression of activating receptors on the surface of NK and NKT cells; impaired synthesis of proinflammatory cytokines or proteases from NK cells; and HLA-E*0103 homozygosis and KIR haplotype BX. A Random Forest algorithm predicted 90% of the accuracy for the classification of CML patients in groups of relapse or non-relapse according to these parameters. Consequently, these features may be useful as biomarkers predictive of CML relapse in patients that are candidates to initiate treatment discontinuation.

Immobilization of mesenchymal stem cells and monocytes in biocompatible microcapsules to cell therapy.

The aim of cell therapy is to replace, repair, or enhance the function of damaged tissues or organs. Several factors complicate the development of cellular therapies. Of primary importance is protection of the implanted cells from the host's immune system. Cells are encapsulated in selectively semipermeable and biocompatible membranes that block entry of immune mediators but allow outward diffusion of active molecules produced by the cells. The immobilization of mesenchymal stem cells and monocytes, in micrometric (30-60 microm) alginate-barium microcapsules based on atomization processes, has been achieved successfully. This size is necessary to the administration of microcapsules via injection (Hamilton syringe with a needle size of 100 microm) and aerosol. Microencapsulated cells survive at least 2 weeks after preparation in vitro.

Development of a nanoparticle system based on a fructose polymer: Stability and drug release studies.

New drug delivery systems (DDSs) with levan or its carboxymethylated form, as carriers, and 5-fluorouracil as a drug, are produced in this work. Levan is obtained after cultivating A. nectaris and polymer nanoparticles are created in water by a self-assembled process. The effect of pH and the ionic strength on polymer nanoparticles aggregation is studied. Basic pHs produces a particle size between 300 and 400nm with a Z-potential around -20mV because a basic medium promotes repulsion forces. DDSs of 300-400nm and a Z-potential about -25mV are prepared by taking advantage of the amphiphilic properties of the levan. The drug is bound to either levan or carboxymethyllevan surfaces by electrostatic interactions, obtaining the best results at basic pHs. 45-70% of the drug is released from the levan in 23h depending on the pH preparation, whereas only a low percentage of the drug is released from the carboxymethyllevan.

Understanding and optimizing the addition of phytohormones in the culture of microalgae for lipid production.

Some studies have described the use of phytohormones in microalgal culture for the production of biodiesel or selected fatty acids. However, no study has determined the amount of phytohormones that maximizes lipid yield. We determined the optimal concentration of auxins and gibberellins (which is between 40 and 60 µM) in two strains (Scenedemus abundans and Chlorella ellipsoidea) suitable for biodiesel production. More than 3-fold increment was reached with S. abundans and near 7-fold increment with C. ellipsoidea. Furthermore, this work suggests that the improved growth of the microalgae in the presence of the phytohormones was due to a reduction in the level of reactive oxygen species (ROS) in the cells. An economic analysis showed that, due to its low cost, auxin offers a positive cost-benefit balance and therefore could be used at large scale. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1203-1211, 2016.