Shedding light on the polyphyletic behavior of the genus Sterkiella: The importance of ontogenetic and molecular phylogenetic approaches

Present study, investigates a poorly known species of the genus Sterkiella, i.e., S. tricirrata, based on two populations isolated from soil samples collected from the Colfiorito Regional Park, Umbria Region, Italy and from the Silent Valley National Park, India. Both populations showed a highly similar morphology, however different ontogenetic pattern in between. The study confirms the validity of the species S. tricirrata which was considered to be a species within the Sterkiella histriomuscorum complex. The main ontogenetic difference between S. tricirrata and other species of the genus Sterkiella is the different mode of formation of anlagen V and VI of the proter in the former. In the phylogenetic analyses, Sterkiella tricirrata clusters with Sterkiella sinica within the stylonychine oxytrichids, in a clade away from the type species (Sterkiella cavicola) of the genus Sterkiella. The study highlights the importance of ontogenetic as well as molecular data in shedding light on the polyphyletic behavior of the genus Sterkiella. A detailed description of S. tricirrata based on morphology, ontogenesis and molecular phylogenetic methods is presented. Further, the improved diagnosis has been provided for the genus Sterkiella and the poorly known species S. tricirrata

Ciliate diversity and behavioural observations from the chemoautotrophic cave ecosystem of Frasassi, (Marche region, Italy)

Chemoautotrophic cave organisms require specific adaptations to tolerate the stress of living in extreme environmental conditions, such as darkness, nutrient and energy limitations, low temperatures (12-13°C), highly variable sulphide concentrations (from 0 up to 415 μM H2S) and toxic levels of gases (H2S, CO2, CH4). To date, due to the difficulties in sampling in such harsh environment, very few studies were performed in order to describe the ciliate communities from caves; these have remained largely unexplored. Thus, the main aims of our study were to characterize for the first time, the ciliates from the sulfide-rich Frasassi cave complex (Marche region, Italy) and to observe possible behavioural differences with their non-cave-dwelling counterpart ciliate species. Four main sampling sites within Frasassi caves were selected: Pozzo dei Cristalli, Lago Verde, Ramo Solfureo and Grotta Solfurea. The ciliate diversity from the site Pozzo dei Cristalli was studied in greater detail for its spatio-temporal distribution, since it is highly diversified and includes several microhabitats represented by small sulfidic (H2S-rich) ponds, streams and springs as well as, deep and shallow muddy, stagnant lakes. Periodic sampling was realised from 2009 to 2011 in the form of water-sediments, picked up by scraping the surface. Classical culturing, silver staining methods and 18S rRNA gene (for some selected species) for phylogenetic analysis were employed. A total of 31 species belonging to 9 classes, 15 orders and 23 genera were identified. Fluctuation of the ciliate communities were mainly recorded at the Pozzo dei Cristalli sites during various sampling occasions, this could be due to changing environmental conditions (mainly H2S concentrations and water levels). Interestingly, it was observed that some species e.g. Urocentrum turbo, Coleps hirtus hirtus, Oxytricha sp, Euplotes sp, showed adaptation for the cave environment (photo-sensitivity, sulphur tolerance, feeding behaviour, morphological difference). Overall, these results provide a platform for various in-depth studies of ciliates to understand potential role in aquatic microhabitats, nature of chemical compounds secreted, dispersal pattern, and adaptations to cave environment

Morphology, ontogenetic, and molecular phylogenetic studies on species of the genera Sterkiella and Fragmospina

We investigated a poorly known species of the genus Sterkiella based on two populations collected from an agricultural field in Marche Region, Italy and from soil samples collected from the Silent Valley National Park, India. This study represents an example where data on the ontogenesis clarifies the polyphyletic behavior of the genus Sterkiella. Both populations showed a rather similar morphology and ontogenetic pattern and thus supported the polyphyletic behavior of the genus Sterkiella in molecular phylogenetic analyses. In addition, description of a species of the genus Fragmospina has been presented based on the detailed investigations on morphology, ontogenesis, and molecular phylogenetic methods

Effect of angiotensin-converting enzyme inhibitor and angiotensin receptor blocker initiation on organ support-free days in patients hospitalized with COVID-19

IMPORTANCE Overactivation of the renin-angiotensin system (RAS) may contribute to poor clinical outcomes in patients with COVID-19. Objective To determine whether angiotensin-converting enzyme (ACE) inhibitor or angiotensin receptor blocker (ARB) initiation improves outcomes in patients hospitalized for COVID-19. DESIGN, SETTING, AND PARTICIPANTS In an ongoing, adaptive platform randomized clinical trial, 721 critically ill and 58 non–critically ill hospitalized adults were randomized to receive an RAS inhibitor or control between March 16, 2021, and February 25, 2022, at 69 sites in 7 countries (final follow-up on June 1, 2022). INTERVENTIONS Patients were randomized to receive open-label initiation of an ACE inhibitor (n = 257), ARB (n = 248), ARB in combination with DMX-200 (a chemokine receptor-2 inhibitor; n = 10), or no RAS inhibitor (control; n = 264) for up to 10 days. MAIN OUTCOMES AND MEASURES The primary outcome was organ support–free days, a composite of hospital survival and days alive without cardiovascular or respiratory organ support through 21 days. The primary analysis was a bayesian cumulative logistic model. Odds ratios (ORs) greater than 1 represent improved outcomes. RESULTS On February 25, 2022, enrollment was discontinued due to safety concerns. Among 679 critically ill patients with available primary outcome data, the median age was 56 years and 239 participants (35.2%) were women. Median (IQR) organ support–free days among critically ill patients was 10 (–1 to 16) in the ACE inhibitor group (n = 231), 8 (–1 to 17) in the ARB group (n = 217), and 12 (0 to 17) in the control group (n = 231) (median adjusted odds ratios of 0.77 [95% bayesian credible interval, 0.58-1.06] for improvement for ACE inhibitor and 0.76 [95% credible interval, 0.56-1.05] for ARB compared with control). The posterior probabilities that ACE inhibitors and ARBs worsened organ support–free days compared with control were 94.9% and 95.4%, respectively. Hospital survival occurred in 166 of 231 critically ill participants (71.9%) in the ACE inhibitor group, 152 of 217 (70.0%) in the ARB group, and 182 of 231 (78.8%) in the control group (posterior probabilities that ACE inhibitor and ARB worsened hospital survival compared with control were 95.3% and 98.1%, respectively). CONCLUSIONS AND RELEVANCE In this trial, among critically ill adults with COVID-19, initiation of an ACE inhibitor or ARB did not improve, and likely worsened, clinical outcomes. TRIAL REGISTRATION ClinicalTrials.gov Identifier: NCT0273570

Tetrahymena thermophila: A whole cell biosensor for toxicity assessment of Mercury

Ciliated protozoans are ubiquitous, free living, and largely non-pathogenic microorganisms. Being single cell eukaryotic microorganisms they represent an essential component of all ecosystem where they are integral constituents of trophic chains and nutrient cycles (Foissner, 1987,1999, 2004; Darbyshire, 1994; Alpheiet al., 1996; Bonkowski and Schaefer, 1997; Sherr and Sherr, 2002; Cuvelieret al., 2010; Steele et al., 2011). They have been used as model organisms for the discovery of key genomic processes found across the eukaryotic tree of life, e.g., self-splicing RNAs, telomeres, and the role of RNAs in shaping germline and somatic genomes. Unlike bacteria, fungi they lack cell wall and are only separated from external environment via cell membrane, this makes them highly sensitive to any change in the environment. Considering that ciliated protozoa shows high similarity in the conserved genes (more than 800 human genes have orthologs in Tetrahymena and out of these 58 genes are associated with human diseases; Eisenet al., 2006;Fillinghamet al., 2002) between ciliates and several eukaryotes including humans, they represents a better biological tools to detect and diagnose communitylevel impairments in contaminated soil and water ecosystems. Thus, ciliate represents a perfect bioindicators that can be used for assessment of ecotoxicological assays for early warning deterioration of the environment. Furthermore, in response to heavy metal pollution, they express a special protein, i.e., metallothioneins (MT) rich in cysteine (cys) amino acid in which the thiol groups are able to bind heavy metals.Further, the induction of MTs by heavy metals is mainly regulated at transcription level

Complete Genome Analysis of Thermus parvatiensis and Comparative Genomics of Thermus spp. Provide Insights into Genetic Variability and Evolution of Natural Competence as Strategic Survival Attributes

Thermophilic environments represent an interesting niche. Among thermophiles, the genus Thermus is among the most studied genera. In this study, we have sequenced the genome of Thermus parvatiensis strain RL, a thermophile isolated from Himalayan hot water springs (temperature >96°C) using PacBio RSII SMRT technique. The small genome (2.01 Mbp) comprises a chromosome (1.87 Mbp) and a plasmid (143 Kbp), designated in this study as pTP143. Annotation revealed a high number of repair genes, a squeezed genome but containing highly plastic plasmid with transposases, integrases, mobile elements and hypothetical proteins (44%). We performed a comparative genomic study of the group Thermus with an aim of analysing the phylogenetic relatedness as well as niche specific attributes prevalent among the group. We compared the reference genome RL with 16 Thermus genomes to assess their phylogenetic relationships based on 16S rRNA gene sequences, average nucleotide identity (ANI), conserved marker genes (31 and 400), pan genome and tetranucleotide frequency. The core genome of the analyzed genomes contained 1,177 core genes and many singleton genes were detected in individual genomes, reflecting a conserved core but adaptive pan repertoire. We demonstrated the presence of metagenomic islands (chromosome:5, plasmid:5) by recruiting raw metagenomic data (from the same niche) against the genomic replicons of T. parvatiensis. We also dissected the CRISPR loci wide all genomes and found widespread presence of this system across Thermus genomes. Additionally, we performed a comparative analysis of competence loci wide Thermus genomes and found evidence for recent horizontal acquisition of the locus and continued dispersal among members reflecting that natural competence is a beneficial survival trait among Thermus members and its acquisition depicts unending evolution in order to accomplish optimal fitness