Ciliate diversity in rodrigo de freitas lagoon (Rio de Janeiro, Brazil) from an integrative standpoint.

The Rodrigo de Freitas Lagoon is a highly eutrophic lacustrine system and has one of the longest histories of exploration and anthropic alteration in Brazil. Despite its relevance, limited studies explored the diversity of micro-eukaryotes in the lagoon. Ciliates (Alveolata, Ciliophora) are overlooked in environmental microbiology, especially in tropical and subtropical ecosystems, resulting in limited knowledge about their diversity and functional relevance in South American habitats, particularly in coastal lagoons. To fill this gap, here we investigated the diversity of ciliates in a brackish coastal lagoon in an urban area of Rio de Janeiro, Brazil, applying and comparing the performance of morphological and metabarcoding approaches. The metabarcoding analysis, based on high-throughput sequencing of the hipervariable region V4 of the 18S rRNA genes detected 37 molecular operational taxonomic units (MOTUs) assigned to Ciliophora, representing only about a half (56.9%) of the diversity detected by microscopy, which counted 65 ciliate morphotypes. The most representative classes in both approaches were Spirotrichea and Oligohymenophorea. The metabarcoding analysis revealed that 35.3% of the ciliate MOTUs had less than 97% similarity to available sequences in the NCBI database, indicating that more than one-third of these MOTUs potentially represents still not represented or undescribed ciliate species in current databases. Our findings indicate that metabarcoding techniques can significantly enhance the comprehension of ciliate diversity in tropical environments, but the scarcity of reference sequences of brackish ciliates in molecular databases represents a challenge to the taxonomic assignment of the MOTUs. This study provides new insights into the diversity of ciliates in a threatened coastal lagoon, revealing a vast array of still unknown and rare ciliate taxonomic units in tropical environments.

In Vitro Investigation into the Effect of Cryopreservation on the Mechanical Characteristics of Dental Hard Tissues.

Previous research has reported on hidden damage within the dentin introduced by cryopreservation, but the effect on the mechanical properties of the hard tissues at tooth level remains unclear. The main objective of this study is to investigate the effect of cryopreservation on the mechanical properties of teeth. A matched sample of 234 premolars of 117 children (9 ≤ age ≤ 16 years), bilaterally extracted for orthodontic reasons, were included. For each child, one tooth was randomly allocated to the cryopreservation group and the contralateral tooth was assigned to the control group. Static compression tests were performed to determine load to failure, stiffness, and toughness. In a subgroup of 20 teeth, a cyclic preloading or chewing simulation was performed. Additionally, the fracture mode was determined, and the microstructure of the fractured surfaces was examined using a scanning electron microscope (SEM). Linear mixed model analyses could not detect a statistical difference in the mean load to failure (p = 0.549), mean toughness (p = 0.968), or mean stiffness (p = 0.150) between cryopreserved and non-cryopreserved teeth. No significant difference in load to failure after cyclic preloading was detected between groups (p = 0.734). SEM analysis revealed comparable fracture characteristics between groups. It is concluded that cryopreservation does not affect the mean load to failure, stiffness, or toughness of teeth, indicating that hidden damage in the dentin is not critical at tooth level.

Identification of an anti-CRISPR protein that inhibits the CRISPR-Cas type I-B system in Clostridioides difficile.

IMPORTANCE: Clostridioides difficile is the widespread anaerobic spore-forming bacterium that is a major cause of potentially lethal nosocomial infections associated with antibiotic therapy worldwide. Due to the increase in severe forms associated with a strong inflammatory response and higher recurrence rates, a current imperative is to develop synergistic and alternative treatments for C. difficile infections. In particular, phage therapy is regarded as a potential substitute for existing antimicrobial treatments. However, it faces challenges because C. difficile has highly active CRISPR-Cas immunity, which may be a specific adaptation to phage-rich and highly crowded gut environment. To overcome this defense, C. difficile phages must employ anti-CRISPR mechanisms. Here, we present the first anti-CRISPR protein that inhibits the CRISPR-Cas defense system in this pathogen. Our work offers insights into the interactions between C. difficile and its phages, paving the way for future CRISPR-based applications and development of effective phage therapy strategies combined with the engineering of virulent C. difficile infecting phages.

Role of Nitric Oxide in the Altered Calcium Homeostasis of Platelets from Rats with Biliary Cirrhosis.

INTRODUCTION: Previously, we found that intracellular calcium (Ca2+) homeostasis is altered in platelets from an experimental model of liver cirrhosis, namely the bile-duct-ligated (BDL) rat. These alterations are compatible with the existence of a hypercoagulable state. OBJECTIVE: In the present study, we analyzed the role of nitric oxide in the abnormal calcium signaling responses of an experimental cirrhosis model, the bile duct-ligated rat. METHODS: Chronic treatment with L-NAME was used to inhibit NO production in a group of control and BDL animals, and the responses compared to those obtained in a control and BDL untreated group (n = 6 each). The experiments were conducted on isolated platelets loaded with fura-2, using fluorescence spectrometry. RESULTS: Chronic treatment with L-NAME increased thrombin-induced Ca2+ release from internal stores in both control and BDL rats. However, the effect was significantly greater in the BDL rats (p < 0.05). Thrombin-induced calcium entry from the extracellular space was also elevated but at lower doses and, similarly in both control and BDL platelets, treated with the NO synthesis inhibitor. Capacitative calcium entry was also enhanced in the control platelets but not in platelets from BDL rats treated with L-NAME. Total calcium in intracellular stores was elevated in untreated platelets from BDL rats, and L-NAME pretreatment significantly (p < 0.05) elevated these values both in controls and in BDL but significantly more in the BDL rats (p < 0.05). CONCLUSIONS: Our results suggest that nitric oxide plays a role in the abnormal calcium signaling responses observed in platelets from BDL rats by interfering with the mechanism that releases calcium from the internal stores.

Reliability of the novel cleft lip and palate smile esthetic index (CLP-SEI).

BACKGROUND/OBJECTIVES: The aim of this research was to determine the reliability of a novel index for the assessment of smile esthetics in patients with cleft lip and/or palate (CL ± P) at the end of their multidisciplinary treatment, for use in clinical and academic settings. MATERIALS AND METHODS: Five orthodontists, five periodontologists, five general practitioners, five dental students, and five laypeople rated the smiles of 10 patients with CL ± P twice at a 2 weeks interval. The rating scale consisted of four main categories: 1. nasolabial esthetics, 2. gingival esthetics, 3. dental esthetics, and 4. overall esthetics. A total of 15 parameters were rated. Intra- and inter-rater agreements were calculated using SPSS. RESULTS: The inter-rater agreement varied between good and excellent whereby the orthodontists, periodontists, general practitioners, dental students, and laypeople obtained agreement scores of 0.86, 0.92, 0.84, 0.90, and 0.89, respectively. The intra-rater agreement was good with agreement scores of 0.78, 0.84, 0.84, 0.80, and 0.79, respectively. LIMITATIONS: Smile esthetics were rated on static pictures, not in real life or by video recordings in a young adult population. CONCLUSIONS/IMPLICATIONS: The cleft lip and palate smile esthetic index is a reliable tool for the assessment of smile esthetics in patients with CL ± P.

Rapid maxillary expansion vs slow maxillary expansion in patients with cleft lip and/or palate: a systematic review and meta-analysis.

OBJECTIVES: To compare the dentoalveolar outcomes of slow maxillary expansion (SME) and rapid maxillary expansion (RME) used for maxillary expansion before secondary alveolar bone grafting in patients with cleft lip and/or palate (CL/P). Secondarily, the advantages and disadvantages of SME vs RME were reviewed. MATERIALS AND METHODS: A systematic search was conducted up to November 2021, including Medline (via PubMed), Embase (via Ovid), Web of Science, Cochrane Central, and Google Scholar. The Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines were followed. Risk-of-bias assessment was performed using the Risk of Bias (RoB 2.0) and Risk Of Bias In Non-randomized Studies of Interventions (ROBINS I) tool. Overall quality was assessed using the Grading of Recommendations Assessment, Development, and Evaluation tool. RESULTS: Of 4007 records, five studies met the inclusion criteria. The randomized control trial (RCT) had a low risk of bias, the non-RCTs presented with a moderate risk of bias. Arch width and perimeter increased significantly with both SME and RME treatments. No difference in the increase in palatal depth was found. The meta-analysis showed a greater anterior-to-posterior expansion ratio for the Quad Helix (QH) appliance. The results for dental tipping were not conclusive. CONCLUSIONS: SME and RME promote equal posterior expansion in cleft patients. The anterior differential expansion is greater with SME (QH appliance). No clear evidence exists concerning the amount of dental adverse effects of SME and RME in cleft patients.

Robust In Vitro and In Vivo Immunosuppressive and Anti-inflammatory Properties of Inducible Caspase-9-mediated Apoptotic Mesenchymal Stromal/Stem Cell.

Mesenchymal stromal stem/cells (MSC) therapies are clinically used in a wide range of disorders based on their robust HLA-independent immunosuppressive and anti-inflammatory properties. However, the mechanisms underlying MSC therapeutic activity remain elusive as demonstrated by the unpredictable therapeutic efficacy of MSC infusions reported in multiple clinical trials. A seminal recent study showed that infused MSCs are actively induced to undergo apoptosis by recipient cytotoxic T cells, a mechanism that triggers in vivo recipient-induced immunomodulation by such apoptotic MSCs, and the need for such recipient cytotoxic cell activity could be replaced by the administration of ex vivo-generated apoptotic MSCs. Moreover, the use of MSC-derived extracellular vesicles (MSC-EVs) is being actively explored as a cell-free therapeutic alternative over the parental MSCs. We hypothesized that the introduction of a "suicide gene" switch into MSCs may offer on-demand in vivo apoptosis of transplanted MSCs. Here, we prompted to investigate the utility of the iCasp9/AP1903 suicide gene system in inducing apoptosis of MSCs. iCasp9/AP1903-induced apoptotic MSCs (MSCiCasp9+) were tested in vitro and in in vivo models of acute colitis. Our data show a very similar and robust immunosuppressive and anti-inflammatory properties of both "parental" alive MSCGFP+ cells and apoptotic MSCiCasp9+ cells in vitro and in vivo regardless of whether apoptosis was induced in vivo or in vitro before administering MSCiCasp9+ lysates. This development of an efficient iCasp9 switch may potentiate the safety of MSC-based therapies in the case of an adverse event and, will also circumvent current logistic technical limitations and biological uncertainties associated to MSC-EVs.

A non-cephalometric three-dimensional appraisal of soft tissue changes by functional appliances in orthodontics: a systematic review and meta-analysis.

BACKGROUND: Patients with dental and skeletal Class II growth patterns present typical facial characteristics. Functional appliance (FA) therapy is one of the conventional orthodontic treatment modalities used to harmonize these facial imbalances. OBJECTIVES: The aim of this systematic review was to investigate and summarize the effects of FAs on the facial soft tissues captured in three dimensions. SEARCH METHODS: A systematic search was conducted up to December 2020. Databases Medline (via PubMed), Embase (via Ovid), Web of Science, and Cochrane Central were searched without language restrictions. SELECTION CRITERIA: Randomized clinical trial (RCT)'s investigating the influence of FA therapy on the facial soft tissues in a non-cephalometric and three-dimensional (3D) manner, were included. DATA COLLECTION AND ANALYSIS: Data extraction was independently undertaken by two authors. In case of conflict, a third author was consulted. RoB 2.0 tool was used to assess the risk of bias. A meta-analysis was performed for total facial height, upper lip length, lower lip length, commissural width, and facial depth. The single-step activation versus incremental approach was compared using a random effects model. RESULTS: Five studies were included in the qualitative synthesis of the systematic review. Facial height increased after FA therapy. Three studies met the criteria for the meta-analysis. The mean facial depth increased significantly (P < 0.003) with 1.39 mm (0.46 to 2.32). CONCLUSIONS: Current evidence suggests a positive influence of the FA therapy on the soft tissues in Class II patients. However, conclusions were drawn based on a two-dimensional analysis of 3D images, suggesting that more controlled studies using a comprehensive 3D analysis are necessary to confirm these results. REGISTRATION: PROSPERO registration (CRD 42021224016).

Morphological and phylogenetic investigations shed light on evolutionary relationships of the enigmatic genus Copemetopus (Ciliophora, Alveolata), with the proposal of Copemetopus verae sp. nov.

Copemetopus Villeneuve-Brachon, 1940 is a rare, poorly known sapropelic ciliate genus composed of only two valid nominal species. Over time, Copemetopus was taxonomically assigned to Heterotrichea and Armophorea classes, but its phylogenetic affinities remained unknown. Until the present study, there were no molecular data available for Copemetopus representatives. Here, we present the 18S and 28S-rDNA sequences and the phylogenetic position of Copemetopus verae sp. nov., as well as its detailed morphological description based on live observations, protargol impregnation, and scanning electron microscopy. Transmission electron micrographs of the type species C. subsalsus Villeneuve-Brachon, 1940 reveal new morphological traits and a unique somatic ciliature pattern of Copemetopus, composed by short segments of dikinetids with one or two supplementary kinetosomes. The phylogenetic trees recovered Copemetopus as the sister group of the genus Protocruzia, both constituting early-divergent lineages that split first from a common ancestor of Intramacronucleta. Morphological and molecular evidence suggest that Copemetopus is neither a heterotrichean nor an armophorean ciliate, but a distinct clade related to Protocruzia.

A non-cephalometric two-dimensional appraisal of soft tissue changes by functional therapy in Class II patients: a systematic review and meta-analysis.

BACKGROUND: Facial attractiveness has social, psychological, and economic repercussions. Class II malocclusions are associated with a retroposition of the mandible, facial convexity, protruded upper lip, retruded lower lip, and pronounced mentolabial fold. Functional appliances (FA) try to increase mandibular length and normalize lip position, thereby decreasing the facial profile convexity and aesthetics. OBJECTIVES: The aim of this review and meta-analysis is to investigate the effects of FA on the soft tissues in a non-cephalometric two-dimensional manner. SEARCH METHODS: Medline (via PubMed), Embase (via Ovid), Web of Science, and Cochrane Central were used to identify studies describing the soft tissue results of FA in a non-cephalometric way. The search was conducted up to July 2021. SELECTION CRITERIA: Prospective and retrospective studies investigating the effects of FA appliances on the soft tissues, with non-ionizing, two-dimensional imaging, including an attractiveness assessment based on Visual Analogue Scale or Likert scale, were considered eligible. DATA COLLECTION AND ANALYSIS: Data extraction was performed separately by two authors. Risk of bias was assessed using the ROBINS-I tool. RESULTS: From the eight eligible studies, four were included in the meta-analysis to investigate the influence of assessors on facial attractiveness scores. Age range of the included studies varied from 8 to 21 years old including 313 Caucasian patients whereof 64% females. Six out of eight studies used growing patients as treatment group and only two studies reported follow-up after the end of treatment. Overall facial attractiveness increased after treatment with FA, resulting from a decrease in facial convexity and a normalization of the lip relationship. Orthodontists score higher aesthetic improvements after FA treatment as compared to laypeople. CONCLUSIONS: Current evidence suggests the positive effects of FA on the facial attractiveness in growing and non-growing patients. FA normalize the lip relationship. Furthermore, they tend to decrease facial convexity and advance the chin point. More pleasing results can be expected in growing patients. Orthodontists are more generous when assessing treatment changes as compared to laypeople. REGISTRATION: PROSPERO registration (CRD42021254835).

Effects of Flavonoids in Experimental Models of Arterial Hypertension.

Flavonoids are a class of substances of a vegetal origin with many interesting actions from the point of view of human disease. Interest in flavonoids in the diet has increased in recent years due to the publication of basic, clinical and epidemiological studies that have shown a whole array of salutary effects related to intake of flavonols and flavones as well as a lower morbility and mortality of cardiovascular diseases. Since arterial hypertension is the most common modifiable risk factor for cardiovascular diseases, this review will focus mainly on the effects of flavonoids on the cardiovascular system with relation to the elevation of blood pressure. Its antihypertensive effects as well as the many investigations performed in experimental models of arterial hypertension, are reviewed in this mini-review.

Perception of Medical Professionalism among Medical Residents in Spain.

BACKGROUND: Medical professionalism, defined as commitment to the primacy of patient welfare, is the basis for doctor-patient-society relationships, but previous research with medical students has shown that professionalism and social commitment to medicine may be waning. To determine if this trend also appears in recently qualified practicing doctors, we surveyed 90 newly graduated doctors currently working as medical residents in two university hospitals in Murcia, Spain. A previously validated questionnaire that studies the perception of six categories (responsibility, altruism, service, excellence, honesty and integrity, and respect) defining medical professionalism was used. RESULTS: A good perception of professionalism was found among medical residents, with more than 70% positive responses in all these six categories. There is an increasing trend in the number of negative responses as the residency goes on. Altruism was the category with the greatest percentage of negative answers (22.3%) and Respect was the category with the lowest percentage (12.9%). CONCLUSIONS: The results show a good professionalism perception in medical residents, but also a slight decline in positive answers that began during medical school. A significant trend was found when including both students and residents. Although there were some differences between students and residents, these were not statistically significant. Educational interventions are needed both at the level of medical school and postgraduate medical residency.

Digging into the lesser-known aspects of CRISPR biology.

A long time has passed since regularly interspaced DNA repeats were discovered in prokaryotes. Today, those enigmatic repetitive elements termed clustered regularly interspaced short palindromic repeats (CRISPR) are acknowledged as an emblematic part of multicomponent CRISPR-Cas (CRISPR associated) systems. These systems are involved in a variety of roles in bacteria and archaea, notably, that of conferring protection against transmissible genetic elements through an adaptive immune-like response. This review summarises the present knowledge on the diversity, molecular mechanisms and biology of CRISPR-Cas. We pay special attention to the most recent findings related to the determinants and consequences of CRISPR-Cas activity. Research on the basic features of these systems illustrates how instrumental the study of prokaryotes is for understanding biology in general, ultimately providing valuable tools for diverse fields and fuelling research beyond the mainstream.

Macronuclear Plasticity in Two South American Populations of Spirostomum (Ciliophora, Heterotrichea) Warns About its Use for Species Classification: Revision and New Insights.

Spirostomum is a widely distributed heterotrichean genus composed of well-known species with described ecology and phylogenetic affinities. The morphological classification of Spirostomum species is mostly based on the body size/shape, number of cortical granule rows and macronuclear characteristics. These features along with molecular phylogenies based on ribosomal genes divide the genus into two phylogroups, one including species with a compact macronucleus, and another including species with a moniliform macronucleus. Here, we present our observations on atypical Spirostomum specimens with unusually two distinct macronuclei and shortened adoral zone of membranelles. These atypical forms appeared in the cultures of S. minus and S. yagiui, sampled at different sites in South America (Chile and Brazil) and associated with unrelated substrate types. Morphological observations of living and stained cells, 18S rRNA gene analyses, and a thorough investigation of the literature suggest that the atypical phenotype may be a result of uncommon pathways during the conjugative process. Thus, we demonstrate that studies of ciliate natural populations and their morphological variations, especially from undersampled biogeographical regions, can reveal the boundaries of widely used morphological characters for Spirostomum taxonomy and species identification.

Possible role of microbiome in the pathogenesis of endometriosis.

INTRODUCTION: There is an urgent necessity to explore the complex pathophysiological nature of endometriosis, which may enable the rationale for new diagnostic and therapeutic strategies to be discovered. This systematic review aimed to clarify the bidirectional relationship between endometriosis and the microbiome and evaluate if the microbiome may be involved in endometriosis's pathogenesis, establishing a potential connection between the different studies. EVIDENCE ACQUISITION: Studies were identified through a systematic literature search of papers that evaluated the microbiomes of human or other animal species with endometriosis and of those without in the electronic database PubMed/Medline, and Embase without a date restriction. We included all cohort studies focusing on the interaction between endometriosis and the microbiomes of humans or other mammals, evaluating if the microbiome may be involved in endometriosis's pathogenesis. EVIDENCE SYNTHESIS: Endometriosis appears to be associated with elevated levels of different microorganisms across various microbiome sites. An ineffective immune response seems to play a key role in endometriosis pathogenesis, and there is some scientific proof to state that the immune response may be modulated by the microbiome. Interestingly, nine studies of our review detected species belonging to the phyla Proteobacteria, Bacteroidetes, and Negativicutes characterized by Gram-negative staining, that were significantly increased in endometriosis cohorts. CONCLUSIONS: Laboratory and clinical investigations indicate that hosts' microbiome profiles with and without endometriosis can be significantly different. To further our understanding of the relationships between endometriosis and the host microbiome, more studies are necessary.

Platelet function and microvesicle generation in patients with hemophilia A.

Our results do not support any effect of FVIII on platelet function in patients with severe HA treated under the regime of prophylaxis.

Las unidades de educación médica en las facultades de Medicina y de Ciencias de la Salud en España. ¿Son necesarias? / Medical education units in Medical and Health Sciences schools in Spain. Are they needed?

En una mayoría de países, la educación médica es una especialidad médica más y preside la vida académica en el grado, el posgrado y en la formación continuada. Pero la situación en España es muy mejorable. Aunque existe un creciente interés en la educación médica como disciplina o especialidad, la mayor parte de las facultades de Medicina españolas no disponen de una unidad o departamento de educación médica que se encargue del avance de la disciplina. Algunas facultades han dispuesto una unidad, cátedra, departamento o centro de estudios, adscrita o independiente a la dirección del centro, a veces sin relación orgánica alguna con el proceso de formación. En este artículo describiremos por qué creemos que estas estructuras son necesarias, su utilidad, así como sus funciones y el alcance de sus actividades. Analizaremos la situación actual en España con el ánimo de promocionar la creación de estas estructuras en todas las facultades de Medicina. Igualmente, repasaremos los mecanismos de los que se ha dotado a la formación especializada en el posgrado para dar respuestas a sus necesidades de formación

In a majority of countries, medical education is one more medical specialty and presides over academic life in undergraduate, graduate and continuing education. But, the situation in Spain is very improvable. Although there is a growing interest in medical education as a discipline or specialty, most of the Spanish faculties of Medicine do not have a unit or department of medical education, which is in charge of advancing the discipline. Some faculties have arranged a unit, chair, department or study center, attached or independent to the management of the center, sometimes without any organic relationship to the training process. In this article we will describe why we believe these structures are necessary, their usefulness, as well as their functions and the scope of their activities. We will analyze the current situation in Spain with the aim of promoting the creation of these structures in all the faculties of Medicine. We will also review the mechanisms that specialized postgraduate training has been equipped to provide answers to their training needs

Ciliate Diversity From Aquatic Environments in the Brazilian Atlantic Forest as Revealed by High-Throughput DNA Sequencing.

Rainforest aquatic ecosystems include complex habitats with scarce information on their unicellular eukaryote diversity and community structure. We have investigated the diversity of ciliates in freshwater and brackish environments along the Brazilian Atlantic Forest, based on the hypervariable V4 region of the 18S-rDNA obtained by high-throughput DNA sequencing. Our analyses detected 409 ciliate taxonomic units (OTUs), mostly attributed to the classes Oligohymenophorea and Spirotrichea. A total of 11 classes, 12 subclasses, 112 genera, and 144 species were reported. We found the following: (a) the ciliate communities are more diverse in freshwater- than in Atlantic Forest-associated brackish environments; (b) the ciliate communities are composed by a small amount of highly abundant OTUs, but a high number of low-abundant or rare OTUs; (c) nearly one-third of the ciliate OTUs share less than 97% sequence identity to reference sequences and (d) phylogenetic inference supports the hypothesis that the V4 region of the Ciliophora 18S-rDNA is a suitable marker for accurate evolutionary inferences at class level. Our results showed that a considerable fraction of the HTS-detected diversity of ciliates from Brazilian Atlantic Forest is not represented in the currently available molecular databases.

The orthodontic bonding properties of human enamel after cryopreservation.

The aim was to investigate the effect of cryopreservation on the enamel bonding properties of orthodontic brackets. Sixty-six human premolars were randomly allocated to a control group or a cryopreserved group. Conventional stainless-steel orthodontic brackets were bonded with a light cure adhesive on the buccal side of the premolars. The shear bond strength (SBS) was determined at a crosshead speed of 1 mm/min. The SBS and adhesive remnant index (ARI) were evaluated respectively by an independent samples t test and Fisher's exact test (α≤0.05). The mean failure load was lower in the cryopreserved group. However, this difference in SBS was not significant (p=0.443). In both groups, the ARI mostly indicated a failure at the enamel-adhesive interface. The mean ARI scores for both groups were not significantly different (p=0.099). Within the limitations of this macro bond strength testing, it can be concluded that cryopreservation does not significantly affect the bonding properties of enamel.