Molecular Analysis of the Bacterial Communities in Crude Oil Samples from Two Brazilian Offshore Petroleum Platforms

Crude oil samples with high- and low-water content from two offshore platforms (PA and PB) in Campos Basin, Brazil, were assessed for bacterial communities by 16S rRNA gene-based clone libraries. RDP Classifier was used to analyze a total of 156 clones within four libraries obtained from two platforms. The clone sequences were mainly affiliated with Gammaproteobacteria (78.2% of the total clones); however, clones associated with Betaproteobacteria (10.9%), Alphaproteobacteria (9%), and Firmicutes (1.9%) were also identified. Pseudomonadaceae was the most common family affiliated with these clone sequences. The sequences were further analyzed by MOTHUR, yielding 81 operational taxonomic units (OTUs) grouped at 97% stringency. Richness estimators also calculated by MOTHUR indicated that oil samples with high-water content were the most diverse. Comparison of bacterial communities present in these four samples using LIBSHUFF and Principal Component Analysis (PCA) indicated that the water content significantly influenced the community structure only of crude oil obtained from PA. Differences between PA and PB libraries were observed, suggesting the importance of the oil field as a driver of community composition in this habitat

miR-214 coordinates melanoma progression by upregulating ALCAM through TFAP2 and miR-148b downmodulation

Malignant melanoma is one of the most aggressive human cancers, but the mechanisms governing its metastatic dissemination are not fully understood. Upregulation of miR-214 and ALCAM and the loss of TFAP2 expression have been implicated in this process, with TFAP2 a direct target of miR-214. Here, we link miR-214 and ALCAM as well as identify a core role for miR-214 in organizing melanoma metastasis. miR- 214 upregulated ALCAM, acting transcriptionally through TFAP2 and also posttranscriptionally through miR-148b (itself controlled by TFAP2), both negative regulators of ALCAM. We also identified several miR-214–mediated prometastatic functions directly promoted by ALCAM. Silencing ALCAM in miR-214–overexpressing melanoma cells reduced cell migration and invasion without affecting growth or anoikisin vitro, and it also impaired extravasation and metastasis formation in vivo. Conversely, cell migration and extravasation was reduced in miR-214–overexpressing cells by upregulation of either miR 148b or TFAP2. These findings were consistent with patterns of expression of miR-214, ALCAM, and miR-148b in human melanoma specimens. Overall, our results define a pathway involving miR-214, miR-148b, TFAP2, and ALCAM that is critical for establishing distant metastases in melanoma

Palytoxin and an Ostreopsis Toxin Extract Increase the Levels of mRNAs Encoding Inflammation-Related Proteins in Human Macrophages via p38 MAPK and NF-κB

BACKGROUND: Palytoxin and, likely, its analogues produced by the dinoflagellate genus Ostreopsis, represent a class of non-proteinaceous compounds displaying high toxicity in animals. Owing to the wide distribution and the poisonous effects of these toxins in humans, their chemistry and mechanism of action have generated a growing scientific interest. Depending on the exposure route, palytoxin and its Ostreopsis analogues may cause several adverse effects on human health, including acute inflammatory reactions which seem more typical of cutaneous and inhalation contact. These observations have led us to hypothesize that these toxins may activate pro-inflammatory signalling cascades. METHODOLOGY AND PRINCIPAL FINDINGS: Here we demonstrate that palytoxin and a semi-purified Ostreopsis cf. ovata toxin extract obtained from a cultured strain isolated in the NW Adriatic Sea and containing a putative palytoxin and all the ovatoxins so far known--including the recently identified ovatoxin-f--significantly increase the levels of mRNAs encoding inflammation-related proteins in immune cells, i.e. monocyte-derived human macrophages, as assessed by Real-Time PCR analysis. Western immunoblot and electrophoretic mobility shift assays revealed that nuclear transcription factor -κB (NF-κB) is activated in cells exposed to toxins in coincidence with reduced levels of the inhibitory protein IκB-α. Moreover, Mitogen-Activated Protein Kinases (MAPK) were phosphorylated in response to palytoxin, as also reported by others, and to the Ostreopsis toxin extract, as shown here for the first time. By using specific chemical inhibitors, the involvement of NF-κB and p38 MAPK in the toxin-induced transcription and accumulation of Cycloxigenase-2, Tumor Necrosis Factor-α, and Interleukin-8 transcripts has been demonstrated. CONCLUSIONS AND SIGNIFICANCE: The identification of specific molecular targets of palytoxin and its Ostreopsis analogues, besides contributing to expand the still limited knowledge of the intracellular signalling cascades affected by these toxins, may have important implications in setting up focused pharmacological interventions, replacing currently used symptomatic treatments

Growth hormone-releasing hormone attenuates cardiac hypertrophy and improves heart function in pressure overload-induced heart failure

It has been shown that growth hormone-releasing hormone (GHRH) reduces cardiomyocyte (CM) apoptosis, prevents ischemia/reperfusion injury, and improves cardiac function in ischemic rat hearts. However, it is still not known whether GHRH would be beneficial for life-threatening pathological conditions, like cardiac hypertrophy and heart failure (HF). Thus, we tested the myocardial therapeutic potential of GHRH stimulation in vitro and in vivo, using GHRH or its agonistic analog MR-409. We show that in vitro, GHRH(1-44)NH2attenuates phenylephrine-induced hypertrophy in H9c2 cardiac cells, adult rat ventricular myocytes, and human induced pluripotent stem cell-derived CMs, decreasing expression of hypertrophic genes and regulating hypertrophic pathways. Underlying mechanisms included blockade of Gq signaling and its downstream components phospholipase Cβ, protein kinase Ce, calcineurin, and phospholamban. The receptor-dependent effects of GHRH also involved activation of Gαsand cAMP/PKA, and inhibition of increase in exchange protein directly activated by cAMP1 (Epac1). In vivo, MR-409 mitigated cardiac hypertrophy in mice subjected to transverse aortic constriction and improved cardiac function. Moreover, CMs isolated from transverse aortic constriction mice treated with MR-409 showed improved contractility and reversal of sarcolemmal structure. Overall, these results identify GHRH as an antihypertrophic regulator, underlying its therapeutic potential for HF, and suggest possible beneficial use of its analogs for treatment of pathological cardiac hypertrophy

Assessment of spatio-temporal variability of faecal pollution along coastal waters during and after rainfall events

© The Author(s), 2022. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Manini, E., Baldrighi, E., Ricci, F., Grilli, F., Giovannelli, D., Intoccia, M., Casabianca, S., Capellacci, S., Marinchel, N., Penna, P., Moro, F., Campanelli, A., Cordone, A., Correggia, M., Bastoni, D., Bolognini, L., Marini, M., & Penna, A. Assessment of spatio-temporal variability of faecal pollution along coastal waters during and after rainfall events. Water, 14(3), (2022): 502, https://doi.org/10.3390/w14030502.More than 80% of wastewaters are discharged into rivers or seas, with a negative impact on water quality along the coast due to the presence of potential pathogens of faecal origin. Escherichia coli and enterococci are important indicators to assess, monitor, and predict microbial water quality in natural ecosystems. During rainfall events, the amount of wastewater delivered to rivers and coastal systems is increased dramatically. This study implements measures capable of monitoring the pathways of wastewater discharge to rivers and the transport of faecal bacteria to the coastal area during and following extreme rainfall events. Spatio-temporal variability of faecal microorganisms and their relationship with environmental variables and sewage outflow in an area located in the western Adriatic coast (Fano, Italy) was monitored. The daily monitoring during the rainy events was carried out for two summer seasons, for a total of five sampling periods. These results highlight that faecal microbial contaminations were related to rainy events with a high flow of wastewater, with recovery times for the microbiological indicators varying between 24 and 72 h and influenced by a dynamic dispersion. The positive correlation between ammonium and faecal bacteria at the Arzilla River and the consequences in seawater can provide a theoretical basis for controlling ammonium levels in rivers as a proxy to monitor the potential risk of bathing waters pathogen pollution.This research was funded by WATERCARE project (Water management solutions for reducing microbial environment impact in coastal areas, project ID 10044130, https://www.italy-croatia.eu/web/watercare, accessed on 17 October 2021) funded by the European Union under the Interreg Italy–Croatia CBC Programme

Inflammatory Indices during and after a Randomized Controlled Trial on Exercise in Old Adults: Could Moderate-intensity Exercise be Safe Enough? (Active Elderly and Health – Clinicaltrials.Gov, NCT03858114)

Background: Aging is characterized by a progressive increase in chronic diseases. According to WHO’s guidance, there is a need to develop strategies to monitor, maintain and rehabilitate the health of older people. The present research aims to elucidate whether or not a moderate physical activity intervention in older adults can cause an increase in inflammatory biomarker indices such as C-reactive Protein (CRP) and erythrocyte sedimentation rate (ESR). It is a valuable tool for promoting health that can be considered tolerable in the over-65 population. Methods: 120 males and females over 65 years of age participated in a randomized controlled trial (RCT NCT03858114) in two groups of 60 individuals of similar size and underwent moderate physical activity or cultural and recreational activities. The exclusion criteria were being younger than 65 years old, being unable to participate in physical activity for medical reasons, and a Body Mass Index of greater than 35. Blood samples from participants were collected for the pre-treatment period (t0), at the end of the study of 12 weeks (t1), and 20 weeks after the end of the study (t2) for measurement of inflammatory indices CRP and ESR. Results: The commonly routine tests for inflammatory reactions (ERS and CPR) showed no change at the end of an RCT on mild-to-moderate exercise. Conclusion: The results on inflammatory indices confirm the safety of this type of intervention and encourage its long-term use and testin

NFĸB signaling drives myocardial injury via CCR2+ macrophages in a preclinical model of arrhythmogenic cardiomyopathy

Nuclear factor κ-B (NFκB) is activated in iPSC-cardiac myocytes from patients with arrhythmogenic cardiomyopathy (ACM) under basal conditions, and inhibition of NFκB signaling prevents disease in Dsg2mut/mut mice, a robust mouse model of ACM. Here, we used genetic approaches and single-cell RNA-Seq to define the contributions of immune signaling in cardiac myocytes and macrophages in the natural progression of ACM using Dsg2mut/mut mice. We found that NFκB signaling in cardiac myocytes drives myocardial injury, contractile dysfunction, and arrhythmias in Dsg2mut/mut mice. NFκB signaling in cardiac myocytes mobilizes macrophages expressing C-C motif chemokine receptor-2 (CCR2+ cells) to affected areas within the heart, where they mediate myocardial injury and arrhythmias. Contractile dysfunction in Dsg2mut/mut mice is caused both by loss of heart muscle and negative inotropic effects of inflammation in viable muscle. Single nucleus RNA-Seq and cellular indexing of transcriptomes and epitomes (CITE-Seq) studies revealed marked proinflammatory changes in gene expression and the cellular landscape in hearts of Dsg2mut/mut mice involving cardiac myocytes, fibroblasts, and CCR2+ macrophages. Changes in gene expression in cardiac myocytes and fibroblasts in Dsg2mut/mut mice were dependent on CCR2+ macrophage recruitment to the heart. These results highlight complex mechanisms of immune injury and regulatory crosstalk between cardiac myocytes, inflammatory cells, and fibroblasts in the pathogenesis of ACM

Principal Component Analysis of the Social and Behavioral Rhythms Scale in elderly

Background: Changes in social and behavioral rhythms (SBR) in elderly are related to health status. Nevertheless, there is no data on factor analysis of the Brief Social Rhythm Scale (BSRS) an internationally well-known tool in this field. The aim was to analyze, in elderly, the factorial structure of the Italian version of BSRS.Design and Methods: Principal Component Analysis of the BSRS carried out in elderly living at home.Results: Sample of 141 participants (83 Females, 58,9%), aged 72.3±4.8. All the items of the questionnaire were related and could compose a single factor, explaining 56% of variance. A solution adopting two factors, the first (including items 1,2,3,4,9,10), the second (including items 5,6,7,8), covered cumulatively 78.8% of the variance.Conclusion: The study confirms that the BSRS is consistent with the idea for which it was built and can be useful for the study of regularity of SBR in old adults

Exercise Improves the Impact of Chronic Pain in Older Adults: Results of an RCT

Background: Chronic Pain (CP) is a crucial determinant for disability in older adults. CP amplifies the impact of other common age-related diseases and increases cardiovascular risk. Physical exercise can improve CP. Randomized Controlled Trials (RCTs) with high-intensity exercise in older adults excluded people with Moderate Chronic Illness (MCI) and CP. Objective: This study aimed at evaluating in an RCT whether moderate exercise training can improve chronic pain in a sample of older adults, including people with MCI, and if any modification persists over time. Methods: A sample of 120 older adults was randomly selected for a moderate-intensity exercise program or cultural activities (control group). Chronic pain was assessed at t0, at t12 (end of the trial), and t48 weeks, by means of the Italian version of the SIP-Roland Scale. Results: Seventy-nine participants completed the follow-up (age 72.3±4.7, women 55.3%). At the end of RCT, an improvement in the SIP scale score was found in the exercise group (p=0.035), showing a lower score than the control group; this difference was not maintained at 48 weeks (p=0.235). Conclusion: Our study highlighted that a moderate-intensity exercise intervention reduced chronic pain in older adults, but this effect disappeared at follow-up after 36 weeks from the end of the training program. These findings suggested that such kinds of programs, easily accessible to old people even with MCI, should be implemented and supported over time, thus promoting active aging and preventing CP of age-related diseases. Clinical Trial Registration: Clinical.Trials.gov.NCT03858114