Adaptation of cyanobacteria and microalgae to extreme environmental changes derived from anthropogenic pollution

Nowadays are living in a global extinction period consequence of human activities that are altering biosphere-level processes. Microalgal populations are being exposed to by-products of human activities for the first time and little is known about their mechanisms of adaptation. Cyanobacteria and microalgae could to adapt to novel environments through selection on spontaneous mutations or through individual responses (i.e. resistant organisms arising by mutations occurring randomly prior to selective agent exposure or by direct and specific adaptation in response to selective agent). We are working on mechanisms of adaptation of these organisms to catastrophic environmental changes coming from residual materials of water pollution. Fluctuation analysis is the procedure to study this adaptive process. Cyanobacteria and microalgal species exposed to several pollutants (herbicides, antibiotics, heavy metals, sulphureous waters and others) showed resistant variants arise from rare spontaneous pre-selective mutations occurring prior to pollutant exposition. Spontaneous mutations seem to be enough to assure the adaptation of large microalgal populations. However, resistant mutants show diminished growth and photosynthesis.Estamos en un periodo de extinción global consecuencia de las actividades humanas y que están alterando los procesos de la biosfera. Las poblaciones de microalgas estan siendo expuestas por primera vez a estos productos fruto de la actividad antropogénica y sus mecanismos de adaptación son muy poco conocidos. Las cianobacterias y microalgas pueden adaptarse a nuevas condiciones ambientales a través de mutaciones espontáneas o a través de respuestas individuales (p.e. los organismos resistentes se pueden producir por mutaciones que ocurren antes de la exposición al agente selectivo, o por una respuesta específica y directa al contaminante). Nosotros hemos trabajado en los mecanismos de adaptación de estos organismos al cambio ambiental brusco debido a contaminantes del agua. El análisis de fluctuación es el procedimiento adecuado para el estudio del proceso adaptativo. Las especies de cianobacterias y microalgas expuestas a distintos contaminantes (herbícidas, antibióticos, metales pesados, aguas sulfurosas y otros…) muestran variantes resistentes que se producen por raras mutaciones espontáneas que ocurren antes de la exposición al contaminante. Estos mutantes resistentes mostraron una disminución del crecimiento y de la fotosíntesis respecto a la población sensible. Las mutaciones espontáneas pueden ser suficientes para asegurar la adaptación en las enormes poblaciones de microalgas

Nutritional status and physical condition in active vs. sedentary elderly people

El objetivo del estudio fue conocer la relación entre el estado nutricional, la adherencia a la dieta mediterránea y el nivel de condición física de personas mayores. Participaron 168 personas mayores de 65 años (grupo control=84 personas activas y grupo experimental=84 personas sedentarias). La adherencia a la dieta mediterránea fue medida con el cuestionario MEDIS-FFQ, el nivel de práctica de actividad física mediante una pregunta creada a tal efecto, y las diferentes pruebas físicas con los instrumentos específicos. Los resultados revelaron que el 63.1% manifestó baja adherencia a la dieta mediterránea y el 34.5% alta, teniendo los sujetos sedentarios mayor adherencia que los activos (p≤0.05; 46.4% vs. 22.6%). Los sujetos activos tienen mejor condición física que los sedentarios (p≤0.001). Por tanto, la mayor parte de las personas mayores deben incrementar su adherencia a la dieta mediterránea y la práctica de actividad física como mecanismo de mejora de su saludThe objective of the study was to know the relationship between nutritional status, adherence to the Mediterranean diet and the level of physical displacement of older people. 168 people older than 65 years participated (control group = 84 active people and experimental group = 84 sedentary people). Adherence to the Mediterranean diet was measured with the MEDIS-FFQ questionnaire, the level of physical activity practice through a question created for that purpose, and the different physical tests with the specific instruments. The results revealed that 63.1% showed low adherence to the Mediterranean diet and 34.5% high, according to the sedentary subject’s greater adherence than the active ones (p≤0.05, 46.4% versus 22.6%). Active subjects have better physical income than sedentary people (p≤0.001). Therefore, most elderly people want to increase their adherence to the Mediterranean diet and the practice of physical activity as a mechanism to reduce their healt

Rapporto tecnico sulla valutazione della biomassa ittioplanctonica mediante l'utilizzo del Multi Plankton Sampler (MPS)

Il piano di campionamento della campagna oceanografica BANSIC'14, condotta a bordo della N/O "URANIA" dal 22 Luglio al 9 Agosto 2014, lungo transetti sotto costa e a largo delle coste meridionali della Sicilia, ha avuto l’obiettivo generale dello studio delle relazioni tra le strutture oceanografiche a mesoscala (vortici verticali ed orizzontali, upwelling, etc.) e le strutture spaziali dei fenomeni biologici relativi ai primi anelli della catena trofica (zooplancton, distribuzione e abbondanza di larve di piccoli pelagici e grandi pelagici) (vedi Rapporto finale BANSIC 2014) per la stima dell’abbondanza dello stock riproduttore. E' una campagna di ricerca nell'ambito del WP3 del progetto SSD-Pesca, finanziato dal MIUR su fondi MISE, a supporto della pesca italiana nelle Regioni Obiettivo 1 e del progetto RITMARE (SP2_WP4_AZ2_UO04). Il campionamento ittioplanctonico è inserito anche nel piano di lavoro del progetto regionale MIPAF-FAO “MedSudMed” (“Assessment and Monitoring of the Fishery Resources and the Ecosystems in the Straits of Sicily”). Il campionamento dell’ittioplancton, durante questa campagna, oltre ai metodi tradizionali quali le reti di tipo Bongo, ha visto l'utilizzo del Multi Plankton Sampler (MPS) MultiNet. I campionatori ittioplanctonici hanno lo scopo di prelevare porzioni di mesozooplancton da un massimo di 100 m fino alla superficie, in quanto le uova di pesci pelagici possiedono una galleggiabilità tale che nonostante le turbolenze superficiali dell’acqua, un campionamento entro i primi metri restituisce un dato affidabile della distribuzione anche se alcune uova possono trovarsi a maggiore profondità (Ahlstrom, 1959). Il campionatore MPS consente, a differenza di altri strumenti, di prelevare la frazione di zooplancton d'interesse con diverse modalità di campionamento: orizzontale, verticale e obliquo e, allo stesso tempo, permette di campionare a differenti quote di profondità. Le informazioni così ottenute sono state utilizzate per valutare la variazione della biomassa ittioplanctonica, l'abbondanza e la composizione delle specie lungo gli strati della colonna d'acqua anche in relazione alle componenti oceanografiche. Grazie all'utilizzo di questo strumento è possibile validare e verificare alcune informazioni e acquisirne delle nuove sull’ecologia delle specie larvali e sul mesozooplancton, sul modo in cui queste si distribuiscono lungo la colonna d'acqua e sulle interazioni intra ed interspecifiche legate anche a fattori oceanografici. Ciò consente di ottenere maggiori informazioni e contribuire al miglioramento della comprensione della biologia e dell'ecologia delle specie rinvenute e, nel contempo, approfondire e migliorare le conoscenze sugli stadi di sviluppo di uova di specie ittiche che allo stadio embrionale sono ancora poco conosciuti

Effect of nitric oxide on mitochondrial activity of human synovial cells

<p>Abstract</p> <p>Background</p> <p>Nitric oxide (NO) is a messenger implicated in the destruction and inflammation of joint tissues. Cartilage and synovial membrane from patients with rheumatoid arthritis (RA) and osteoarthritis (OA) have high levels of NO. NO is known to modulate various cellular pathways and, thus, inhibit the activity of the mitochondrial respiratory chain (MRC) of chondrocytes and induce the generation of reactive oxygen species (ROS) and cell death in multiple cell types. For these reasons, and because of the importance of the synovial membrane in development of OA pathology, we investigated the effects of NO on survival, mitochondrial function, and activity of fibroblastic human OA synovial cells.</p> <p>Methods</p> <p>Human OA synovia were obtained from eight patients undergoing hip joint replacement. Sodium nitroprusside (SNP) was used as a NO donor compound and cell viability was evaluated by MTT assays. Mitochondrial function was evaluated by analyzing the mitochondrial membrane potential (Δψm) with flow cytometry using the fluorofore DePsipher. ATP levels were measured by luminescence assays, and the activities of the respiratory chain complexes (complex I: NADH CoQ₁reductase, complex II: succinate dehydrogenase, complex III: ubiquinol-cytochrome c reductase, complex IV: cytochrome c oxidase) and citrate synthase (CS) were measured by enzymatic assay. Protein expression analyses were performed by western blot.</p> <p>Results</p> <p>SNP at a concentration of 0.5 mM induced cell death, shown by the MTT method at different time points. The percentages of viable cells at 24, 48 and 72 hours were 86.11 ± 4.9%, 74.31 ± 3.35%, and 43.88 ± 1.43%, respectively, compared to the basal level of 100% (*<it>p </it>< 0.05). SNP at 0.5 mM induced depolarization of the mitochondrial membrane at 12 hours with a decrease in the ratio of polarized cells (basal = 2.48 ± 0.28; SNP 0.5 mM = 1.57 ± 0.11; *<it>p </it>< 0.01). The time course analyses of treatment with SNP at 0.5 mM demonstrated that treatment reliably and significantly reduced intracellular ATP production (68.34 ± 14.3% vs. basal = 100% at 6 hours; *<it>p </it>< 0.05). The analysis of the MRC at 48 hours showed that SNP at 0.5 mM increased the activity of complexes I (basal = 36.47 ± 3.92 mol/min/mg protein, SNP 0.5 mM = 58.08 ± 6.46 mol/min/mg protein; *<it>p </it>< 0.05) and III (basal = 63.87 ± 6.93 mol/min/mg protein, SNP 0.5 mM = 109.15 ± 30.37 mol/min/mg protein; *<it>p </it>< 0.05) but reduced CS activity (basal = 105.06 ± 10.72 mol/min/mg protein, SNP at 0.5 mM = 66.88 ± 6.08 mol/min/mg protein.; *<it>p </it>< 0.05), indicating a decrease in mitochondrial mass. Finally, SNP regulated the expression of proteins related to the cellular cycle; the NO donor decreased bcl-2, mcl-1 and procaspase-3 protein expression.</p> <p>Conclusions</p> <p>This study suggests that NO reduces the survival of OA synoviocytes by regulating mitochondrial functionality, as well as the proteins controlling the cell cycle.</p

Effect of a natural extract of chicken combs with a high content of hyaluronic acid (Hyal-Joint®) on pain relief and quality of life in subjects with knee osteoarthritis: a pilot randomized double-blind placebo-controlled trial

<p>Abstract</p> <p>Background</p> <p>Intra-articular hyaluronic acid represents a substantive addition to the therapeutic armamentarium in knee osteoarthritis. We examined the effect of dietary supplementation with a natural extract of chicken combs with a high content of hyaluronic acid (60%) (Hyal-Joint^®) (active test product, AP) on pain and quality of life in subjects with osteoarthritis of the knee.</p> <p>Methods</p> <p>Twenty subjects aged ≥40 years with knee osteoarthritis (pain for at least 15 days in the previous month, symptoms present for ≥6 months, Kellgren/Lawrence score ≥2) participated in a randomized double-blind controlled trial. Ten subjects received AP (80 mg/day) and 10 placebo for 8 weeks. The Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) and quality of life by the Short Form-36 (SF-36v2) were administered at baseline and after 4 and 8 weeks of treatment.</p> <p>Results</p> <p>WOMAC pain (primary efficacy variable) was similar in both study groups (mean [SD]) with 6.6 (4.0) points in the AP group and 6.4 (2.7) in the placebo group (<it>P </it>= 0.943). As compared with baseline, subjects in both groups showed statistically significant improvements in WOMAC pain, stiffness, physical function subscales, and in the aggregate score, but the magnitude of changes was higher in the AP group for WOMAC physical function (-13.1 [12.0] vs. -10.1 [8.6], <it>P </it>= 0.575) and total symptoms (-18.6 [16.8] vs. -15.8 [11.4], <it>P </it>= 0.694). At 4 weeks, statistically significant mean changes compared with baseline were observed in the SF-36v2 scales of role-physical, bodily pain, social functioning and role-emotional among subjects in the AP group, and in physical functioning, bodily pain, and social functioning in the placebo group. At 8 weeks, changes were significant for role-physical, bodily pain, and physical component summary in the AP group, and for physical functioning and role-emotional in the placebo arm. Changes in bodily pain and social functioning were of greater magnitude in subjects given AP.</p> <p>Conclusion</p> <p>This pilot clinical trial showed that daily supplementation with oral hyaluronic acid from a natural extract of chicken combs (Hyal-Joint^®) was useful to enhance several markers of quality of life in adults with osteoarthritis of the knee. The results warrant further study in larger sample sizes.</p

Semen quality in Peruvian pesticide applicators: association between urinary organophosphate metabolites and semen parameters

<p>Abstract</p> <p>Background</p> <p>Organophosphates are broad class of chemicals widely used as pesticides throughout the world. We performed a cross-sectional study of associations between dialkylphosphate metabolites of organophosphates and semen quality among pesticide applicators in Majes (Arequipa), Peru.</p> <p>Methods</p> <p>Thirty-one men exposed to organophosphate (OP) pesticides and 31 non-exposed were recruited (age, 20–60 years). In exposed subjects, semen and a blood sample were obtained one day after the last pesticide application. Subjects were grouped according to levels of OP metabolites in urine. Semen samples were analyzed for sperm concentration, percentage of sperm motility, percentage of normal morphology, semen leucocytes and concentrations of fructose and zinc. Exposure to OP was assessed by measuring six urinary OP metabolites (dimethyl and diethyl phosphates and thiophosphates) by gas chromatography using a single flame photometric detector.</p> <p>Results</p> <p>Diethyldithiophosphate (p = 0.04) and diethylthiophosphate (p = 0.02) better reflected occupational pesticide exposure than other OP metabolites. Semen analysis revealed a significant reduction of semen volume and an increase in semen pH in men with OP metabolites. Multiple regression analysis showed that both occupational exposure to pesticides and the time of exposure to pesticides were more closely related to alterations in semen quality parameters than the single measurement of OP metabolites in urine.</p> <p>Conclusion</p> <p>The study demonstrated that occupational exposure to OP pesticides was more closely related to alterations in semen quality than a single measurement of urine OP metabolites. Current measurement of OP metabolites in urine may not reflect the full risk.</p

Hsp90β inhibition modulates nitric oxide production and nitric oxide-induced apoptosis in human chondrocytes

<p>Abstract</p> <p>Background</p> <p>Hsp90β is a member of the Hsp90 family of protein chaperones. This family plays essential roles in the folding, maturation and activity of many proteins that are involved in signal transduction and transcriptional regulation. The role of this protein in chondrocytes is not well understood, although its increase in osteoarthritic cells has been reported. The present study aimed to explore the role of Hsp90β in key aspects of OA pathogenesis.</p> <p>Methods</p> <p>Human OA chondrocytes were isolated from cartilage obtained from patients undergoing joint replacement surgery, and primary cultured. Cells were stimulated with proinflammatory cytokines (IL-1β or TNF-α) and nitric oxide donors (NOC-12 or SNP). For Hsp90β inhibition, two different chemical inhibitors (Geldanamycin and Novobiocin) were employed, or siRNA transfection procedures were carried out. Gene expression was determined by real-time PCR, apoptosis was quantified by flow cytometry and ELISA, and nitric oxide (NO) production was evaluated by the Griess method. Indirect immunofluorescence assays were performed to evaluate the presence of Hsp90β in stimulated cells.</p> <p>Results</p> <p>Hsp90β was found to be increased by proinflammatory cytokines. Inhibition of Hsp90β by the chemicals Geldanamycin (GA) and Novobiocin (NB) caused a dose-dependent decrease of the NO production induced by IL-1β in chondrocytes, up to basal levels. Immunofluorescence analyses demonstrate that the NO donors NOC-12 and SNP also increased Hsp90β. Chemical inhibition or specific gene silencing of this chaperone reduced the DNA condensation and fragmentation, typical of death by apoptosis, that is induced by NO donors in chondrocytes.</p> <p>Conclusions</p> <p>The present results show how Hsp90β modulates NO production and NO-mediated cellular death in human OA chondrocytes.</p

In vitro analysis of the effects on wound healing of high- and low-molecular weight chains of hyaluronan and their hybrid H-HA/L-HA complexes

Abstract Background: Recent studies have reported the roles of Hyaluronic acid (HA) chains of diverse length in wound repair, especially considering the simultaneous occurrence in vivo of both high- (H-HA) and low-molecular weight (L-HA) hyaluronan at an injury site. It has been shown that HA fragments (5 ≤ MW ≤ 20 kDa) usually trigger an inflammatory response that, on one hand, is the first signal in the activation of a repair mechanism but on the other, when it’s overexpressed, it may promote unwanted side effects. The present experimental research has aimed to investigate H-HA, L-HA and of a newly developed complex of the two (H-HA/L-HA) for stability (e.g. hyaluronidases digestion), for their ability to promote wound healing of human keratinocytes in vitro and for their effect on cellular biomarker expression trends. Results: Time-lapse video microscopy studies proved that the diverse HA was capable of restoring the monolayer integrity of HaCat. The H-HA/L-HA complex (0.1 and 1%w/v) proved faster in regeneration also in co-culture scratch test where wound closure was achieved in half the time of H-HA stimulated cells and 2.5-fold faster than the control. Gene expression was evaluated for transformation growth factor beta 1 (TGF-β1) proving that L-HA alone increased its expression at 4 h followed by restoration of similar trends for all the stimuli. Depending on the diverse stimulation (H-HA, L-HA or the complex), metalloproteinases (MMP-2, -9, -13) were also modulated differently. Furthermore, type I collagen expression and production were evaluated. Compared to the others, persistence of a significant higher expression level at 24 h for the H-HA/L-HA complex was found. Conclusions: The outcomes of this research showed that, both at high and low concentrations, hybrid complexes proved to perform better than HA alone thus suggesting their potential as medical devices in aesthetic and regenerative medicine. Keywords: Wound healing, Hyaluronan, MMPs, Hybrid complexe

Screening mutations in myosin binding protein C3 gene in a cohort of patients with Hypertrophic Cardiomyopathy

<p>Abstract</p> <p>Background</p> <p><it>MyBPC3 </it>mutations are amongst the most frequent causes of hypertrophic cardiomyopathy, however, its prevalence varies between populations. They have been associated with mild and late onset disease expression. Our objectives were to establish the prevalence of <it>MyBPC3 </it>mutations and determine their associated clinical characteristics in our patients.</p> <p>Methods</p> <p>Screening by Single Strand Conformation Polymorphisms (SSCP) and sequencing of the fragments with abnormal motility of the <it>MyBPC3 </it>gene in 130 unrelated consecutive HCM index cases. Genotype-Phenotype correlation studies were done in positive families.</p> <p>Results</p> <p>16 mutations were found in 20 index cases (15%): 5 novel [D75N, V471E, Q327fs, IVS6+5G>A (homozygous), and IVS11-9G>A] and 11 previously described [A216T, R495W, R502Q (2 families), E542Q (3 families), T957S, R1022P (2 families), E1179K, K504del, K600fs, P955fs and IVS29+5G>A]. Maximum wall thickness and age at time of diagnosis were similar to patients with <it>MYH7 </it>mutations [25(7) vs. 27(8), p = 0.16], [46(16) vs. 44(19), p = 0.9].</p> <p>Conclusions</p> <p>Mutations in <it>MyBPC3 </it>are present in 15% of our hypertrophic cardiomyopathy families. Severe hypertrophy and early expression are compatible with the presence of <it>MyBPC3 </it>mutations. The genetic diagnosis not only allows avoiding clinical follow up of non carriers but it opens new possibilities that includes: to take preventive clinical decisions in mutation carriers than have not developed the disease yet, the establishment of genotype-phenotype relationship, and to establish a genetic diagnosis routine in patients with familial HCM.</p