Axonal damage is a prominent cause of disability and yet its pathogenesis is incompletely understood. Using a xenogeneic system, here we define the bioenergetic changes induced in rat neurons by ...exposure to cerebrospinal fluid samples from patients with multiple sclerosis compared to control subjects. A first discovery cohort of cerebrospinal fluid from 13 patients with multiple sclerosis and 10 control subjects showed that acute exposure to cerebrospinal fluid from patients with multiple sclerosis induced oxidative stress and decreased expression of neuroprotective genes, while increasing expression of genes involved in lipid signalling and in the response to oxidative stress. Protracted exposure of neurons to stress led to neurotoxicity and bioenergetics failure after cerebrospinal fluid exposure and positively correlated with the levels of neurofilament light chain. These findings were validated using a second independent cohort of cerebrospinal fluid samples (eight patients with multiple sclerosis and eight control subjects), collected at a different centre. The toxic effect of cerebrospinal fluid on neurons was not attributable to differences in IgG content, glucose, lactate or glutamate levels or differences in cytokine levels. A lipidomic profiling approach led to the identification of increased levels of ceramide C16:0 and C24:0 in the cerebrospinal fluid from patients with multiple sclerosis. Exposure of cultured neurons to micelles composed of these ceramide species was sufficient to recapitulate the bioenergetic dysfunction and oxidative damage induced by exposure to cerebrospinal fluid from patients with multiple sclerosis. Therefore, our data suggest that C16:0 and C24:0 ceramides are enriched in the cerebrospinal fluid of patients with multiple sclerosis and are sufficient to induce neuronal mitochondrial dysfunction and axonal damage.
•Coupling of the degradation of acetylated and deacetilated units in chitosan with varying deacetylation degree.•Model fitting of degradation in overlapped degradation mechanisms.•Previous activation ...energy determination by applying “free model” methods allows subsequent model fitting of TGA traces.•Sestack-Berggren model yields the best fit to the experimental data in chitosan.
Thermal degradation of chitosan with varying deacetylation degree (DD) ranging between 50 and 85% was analyzed by dynamic thermogravimetric analysis at different heating rates. The present study focused on the temperature range between 500 and 800K, above water evaporation. Thermal degradation showed a main degradation stage in this temperature interval with a second stage that appeared in the weight derivative curves as a shoulder in the high temperature side of the main peak with increasing intensity as the DD decreased. The Kissinger and isoconversional Ozawa-Flynn-Wall models were employed to evaluate the Ea of both thermal degradation processes. Different kinetic models were tested to computer simulate the thermogravimetric traces calculating the model parameters with a non-linear least squares fitting routine. The Sestack-Berggren model allowed reproducing accurately the overlapping of the two degradation mechanisms and calculating the mass fraction lost in each of them revealing the coupling between the two degradation mechanisms.
The metabolic intimacy of symbiosis often demands the work of specialists. Natural products and defensive secondary metabolites can drive specificity by ensuring infection and propagation across host ...generations. But in contrast to bacteria, little is known about the diversity and distribution of natural product biosynthetic pathways among fungi and how they evolve to facilitate symbiosis and adaptation to their host environment. In this study, we define the secondary metabolism of Escovopsis and closely related genera, symbionts in the gardens of fungus-farming ants. We ask how the gain and loss of various biosynthetic pathways correspond to divergent lifestyles. Long-read sequencing allowed us to define the chromosomal features of representative Escovopsis strains, revealing highly reduced genomes composed of seven to eight chromosomes. The genomes are highly syntenic with macrosynteny decreasing with increasing phylogenetic distance, while maintaining a high degree of mesosynteny. An ancestral state reconstruction analysis of biosynthetic pathways revealed that, while many secondary metabolites are shared with non-ant-associated Sordariomycetes, 56 pathways are unique to the symbiotic genera. Reflecting adaptation to diverging ant agricultural systems, we observe that the stepwise acquisition of these pathways mirrors the ecological radiations of attine ants and the dynamic recruitment and replacement of their fungal cultivars. As different clades encode characteristic combinations of biosynthetic gene clusters, these delineating profiles provide important insights into the possible mechanisms underlying specificity between these symbionts and their fungal hosts. Collectively, our findings shed light on the evolutionary dynamic nature of secondary metabolism in Escovopsis and its allies, reflecting adaptation of the symbionts to an ancient agricultural system.IMPORTANCEMicrobial symbionts interact with their hosts and competitors through a remarkable array of secondary metabolites and natural products. Here, we highlight the highly streamlined genomic features of attine-associated fungal symbionts. The genomes of Escovopsis species, as well as species from other symbiont genera, many of which are common with the gardens of fungus-growing ants, are defined by seven chromosomes. Despite a high degree of metabolic conservation, we observe some variation in the symbionts’ potential to produce secondary metabolites. As the phylogenetic distribution of the encoding biosynthetic gene clusters coincides with attine transitions in agricultural systems, we highlight the likely role of these metabolites in mediating adaptation by a group of highly specialized symbionts.
We examined in a prospective, randomized, international clinical trial the performance of a previously defined 30-gene predictor (DLDA-30) of pathologic complete response (pCR) to preoperative weekly ...paclitaxel and fluorouracil, doxorubicin, and cyclophosphamide (T/FAC) chemotherapy, and assessed if DLDA-30 also predicts increased sensitivity to FAC-only chemotherapy. We compared the pCR rates after T/FAC versus FACx6 preoperative chemotherapy. We also did an exploratory analysis to identify novel candidate genes that differentially predict response in the two treatment arms.
Two hundred and seventy-three patients were randomly assigned to receive either weekly paclitaxel × 12 followed by FAC × 4 (T/FAC, n = 138), or FAC × 6 (n = 135) neoadjuvant chemotherapy. All patients underwent a pretreatment fine-needle aspiration biopsy of the tumor for gene expression profiling and treatment response prediction.
The pCR rates were 19% and 9% in the T/FAC and FAC arms, respectively (P < 0.05). In the T/FAC arm, the positive predictive value (PPV) of the genomic predictor was 38% 95% confidence interval (95% CI), 21-56%, the negative predictive value was 88% (95% CI, 77-95%), and the area under the receiver operating characteristic curve (AUC) was 0.711. In the FAC arm, the PPV was 9% (95% CI, 1-29%) and the AUC was 0.584. This suggests that the genomic predictor may have regimen specificity. Its performance was similar to a clinical variable-based predictor nomogram.
Gene expression profiling for prospective response prediction was feasible in this international trial. The 30-gene predictor can identify patients with greater than average sensitivity to T/FAC chemotherapy. However, it captured molecular equivalents of clinical phenotype. Next-generation predictive markers will need to be developed separately for different molecular subsets of breast cancers.
Background:
Cerebrospinal fluid (CSF) is in contact with brain parenchyma and ventricles, and its composition might influence the cellular physiology of oligodendrocyte progenitor cells (OPCs) ...thereby contributing to multiple sclerosis (MS) disease pathogenesis.
Objective:
To identify the transcriptional changes that distinguish the transcriptional response induced in proliferating rat OPCs upon exposure to CSF from primary progressive multiple sclerosis (PPMS) or relapsing remitting multiple sclerosis (RRMS) patients and other neurological controls.
Methods:
We performed gene microarray analysis of OPCs exposed to CSF from neurological controls, or definitive RRMS or PPMS disease course. Results were confirmed by quantitative reverse transcriptase polymerase chain reaction, immunocytochemistry and western blot of cultured cells, and validated in human brain specimens.
Results:
We identified common and unique oligodendrocyte genes for each treatment group. Exposure to CSF from PPMS uniquely induced branching of cultured progenitors and related transcriptional changes, including upregulation (P<0.05) of the adhesion molecule GALECTIN-3/Lgals3, which was also detected at the protein level in brain specimens from PPMS patients. This pattern of gene expression was distinct from the transcriptional programme of oligodendrocyte differentiation during development.
Conclusions:
Despite evidence of morphological differentiation induced by exposure to CSF of PPMS patients, the overall transcriptional response elicited in cultured OPCs was consistent with the activation of an aberrant transcriptional programme.
The objective of this research was to evaluate the
Anadenanthera peregrina
wood cultivated in five spacing and pyrolyzed in two different heating rates. Three trees were collected from each spacing ...(3 × 2, 3 × 3, 4 × 3, 4 × 4 and 5 × 5 m), and samples were taken for the determination of the physical and chemical properties of the wood, the pyrolysis under the two heating rates (1.67 and 0.83 °C min
−1
) and the subsequent analysis of the charcoal produced. Planting spacing did not influence the properties of
A. peregrina
wood. The heating rate influenced the charcoal properties, interacting with the spacing for the ash content and apparent density, yields of pyroligneous liquid and non-condensable gases.
Dysregulated inflammation and coagulation are underlying mechanisms driving organ injury after trauma and hemorrhagic shock. Heparan sulfates, cell surface glycosaminoglycans abundantly expressed on ...the endothelial surface, regulate a variety of cellular processes. Endothelial heparan sulfate containing a rare 3-
-sulfate modification on a glucosamine residue is anticoagulant and anti-inflammatory through high-affinity antithrombin binding and sequestering of circulating damage-associated molecular pattern molecules. Our goal was to evaluate therapeutic potential of a synthetic 3-
-sulfated heparan sulfate dodecasaccharide (12-mer, or dekaparin) to attenuate thromboinflammation and prevent organ injury.
Male Sprague-Dawley rats were pre-treated subcutaneously with vehicle (saline) or dekaparin (2 mg/kg) and subjected to a trauma/hemorrhagic shock model through laparotomy, gut distention, and fixed-pressure hemorrhage. Vehicle and dekaparin-treated rats were resuscitated with Lactated Ringer's solution (LR) and compared to vehicle-treated fresh-frozen-plasma-(FFP)-resuscitated rats. Serial blood samples were collected at baseline, after induction of shock, and 3 hours after fluid resuscitation to measure hemodynamic and metabolic shock indicators, inflammatory mediators, and thrombin-antithrombin complex formation. Lungs and kidneys were processed for organ injury scoring and immunohistochemical analysis to quantify presence of neutrophils.
Induction of trauma and hemorrhagic shock resulted in significant increases in thrombin-antithrombin complex, inflammatory markers, and lung and kidney injury scores. Compared to vehicle, dekaparin treatment did not affect induction, severity, or recovery of shock as indicated by hemodynamics, metabolic indicators of shock (lactate and base excess), or metrics of bleeding, including overall blood loss, resuscitation volume, or hematocrit. While LR-vehicle-resuscitated rodents exhibited increased lung and kidney injury, administration of dekaparin significantly reduced organ injury scores and was similar to organ protection conferred by FFP resuscitation. This was associated with a significant reduction in neutrophil infiltration in lungs and kidneys and reduced lung fibrin deposition among dekaparin-treated rats compared to vehicle. No differences in organ injury, neutrophil infiltrates, or fibrin staining between dekaparin and FFP groups were observed. Finally, dekaparin treatment attenuated induction of thrombin-antithrombin complex and inflammatory mediators in plasma following trauma and hemorrhagic shock.
Anti-thromboinflammatory properties of a synthetic 3-
-sulfated heparan sulfate 12-mer, dekaparin, could provide therapeutic benefit for mitigating organ injury following major trauma and hemorrhagic shock.
The present investigation was carried out to evaluate the antibacterial effect of Aloe vera and Croton lechleri resin in water for human consumption. The study method was quasi-experimental with ...pre-test and post-test. Aloe vera resin and Croton lechleri at concentrations of 25%, 50% and 75% volume were used as biological samples, each concentration corresponding to a treatment, i.e., T1 as control with calcium hypochlorite at 0.5 ppm, T2, T3 and T4. It was determined that there is water consumption with high levels of total coliforms (925 CFU/100mL), thermotolerant coliforms (845 CFU/100mL) and Escherichia coli (1110 CFU/100mL), whose values exceed the maximum permissible limits according to Peruvian Supreme Decree regulations. The research concludes that the treatment with a higher concentration of aloe resin is able to inhibit the concentration of microbial load to a greater extent. Thus, Treatment (T4) achieved a greater bactericidal effect of 100% on total Coliforms, thermotolerant Coliforms and Escherichia coli, followed by Sangre de grado (Croton lechleri) resin, decreasing by 99%. It is important to mention that plants and their derivatives are an efficient and effective alternative for the elimination of bacteria present in water samples, being an ecological proposal for the care of natural resources and human health.
The objective of the research was to apply compost based on municipal organic waste to recover degraded soils in the Peruvian Amazon. An area of 2500 m2 was considered, where five points were sampled ...and two calicata were obtained for each one. The depth of the calicata was 20 cm. and, subsequently, 1 kg of degraded soil was taken for physical, chemical and biological analysis. Four treatments were used (T1: Control, T2: MOW, 250 mg., T3: MOW, 500 mg and T4: MOW, 750 mg.) with 5 replications for each treatment with a total of 20 experimental units. The organic matter was composed of egg shell, orange, banana, stems, leaves and roots to be segregated and crushed until 12,000 kg. were obtained. Mountain microorganisms (MM) were added to obtain 3,000 kg., after which the chemical analysis was carried out. Maize (Zea mays. L) was used as an indicator plant, which germinated for 30 days to demonstrate the recovery of degraded soils and variability in agronomic parameters. Significant differences between treatments were measured with Duncan. It is concluded that there is a significant improvement in the recovery of degraded soils using MOW. Treatments 3 and 4 show significant relevance and differences with respect to macronutrients. Finally, in terms of morpho-physiological analysis, treatment 4 shows greater growth in stem, number, length and width of leaves. Therefore, the use of municipal organic waste could be an efficient and effective alternative for the recovery of contaminated soils.
In relapsing-remitting multiple sclerosis (RRMS) subtype, the patient's brain itself is capable of repairing the damage, remyelinating the axon and recovering the neurological function. Cerebrospinal ...fluid (CSF) is in close proximity with brain parenchyma and contains a host of proteins and other molecules, which influence the cellular physiology, that may balance damage and repair of neurons and glial cells. The purpose of this study was to determine the pathophysiological mechanisms underpinning myelin repair in distinct clinical forms of MS and neuromyelitis optica (NMO) patients by studying the effect of diseased CSF on glucose metabolism and ATP synthesis. A cellular model with primary cultures of oligodendrocyte progenitor cells (OPCs) from rat cerebrum was employed, and cells were treated with CSF from distinct clinical forms of MS, NMO patients and neurological controls. Prior to comprehending mechanisms underlying myelin repair, we determine the best stably expressed reference genes in our experimental condition to accurately normalize our target mRNA transcripts. The
and
algorithms showed that mitochondrial ribosomal protein (
, hypoxanthine guanine phosphoribosyl transferase (
), microglobulin β2 (
), and transferrin receptor (
) were identified as the best reference genes in OPCs treated with MS subjects and were used for normalizing gene transcripts. The main findings on microarray gene expression profiling analysis on CSF treated OPCs cells revealed a disturbed carbohydrate metabolism and ATP synthesis in MS and NMO derived CSF treated OPCs. In addition, using STRING program, we investigate whether gene-gene interaction affected the whole network in our experimental conditions. Our findings revealed downregulated expression of genes involved in carbohydrate metabolism, and that glucose metabolism impairment and reduced ATP availability for cellular damage repair clearly differentiate more benign forms from the most aggressive forms and worst prognosis in MS patients.
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