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Abstract This systematic review and network meta-analysis aimed to assess the impact of combining professional mechanical plaque removal (PMPR) with probiotics compared to PMPR + placebo on probing pocket depth (PPD) and clinical attachment level (CAL). Randomized controlled trials published until November 2023 were searched across electronic databases, peer-reviewed journals, and grey literature. Two authors independently selected, extracted data, and assessed bias risk. Primary outcomes were mean changes in PPD and CAL. Secondary outcomes included mean changes in bleeding on probing (BOP), plaque index, and colony-forming units. Network meta-analysis with the frequentist weighted least squares approach evaluated the data quantitatively, and CINeMA framework evaluated the quality of evidence. In 33 articles involving 1290 patients, results were stratified by follow-up period (short and long-time studies) and sensitivity analyses conducted based on probiotic therapy duration (1 month reference). Network meta-analysis revealed significant mean differences in PPD for nine probiotic interventions, CAL for eighteen interventions, and BOP for eight interventions, with Lactobacillus demonstrating the most substantial effects. Combining PMPR with probiotics as adjuvants to subgingival instrumentation may be more effective in improving PPD and CAL. Lactobacillus emerged as the most comprehensive and effective among the studied probiotic.

Peri-implantitis continues to be one of the major reasons for implant failure. We propose a new approach to the incorporation of MTA into zirconia implant surfaces with Nd:YAG laser and investigate the biological and the microbiological responses of peri-implant cells. Discs of zirconia stabilized with yttria and titanium were produced according to the following four study groups: Nd:YAG laser-textured zirconia coated with MTA (Zr MTA), Nd:YAG laser-textured zirconia (Zr textured), polished zirconia discs, and polished titanium discs (Zr and Ti). Surface roughness was evaluated by contact profilometry. Human osteoblasts (hFOB), gingival fibroblasts (HGF hTERT) and S. oralis were cultured on discs. Cell adhesion and morphology, cell differentiation markers and bacterial growth were evaluated. Zr textured roughness was significantly higher than all other groups. SEM images reveal cellular adhesion at 1 day in all samples in both cell lines. Osteoblasts viability was lower in the Zr MTA group, unlike fibroblasts viability, which was shown to be higher in the Zr MTA group compared with the Zr textured group at 3 and 7 days. Osteocalcin and IL-8 secretion by osteoblasts were higher in Zr MTA. The Zr textured group showed higher IL-8 values released by fibroblasts. No differences in S. oralis CFUs were observed between groups. The present study suggests that zirconia implant surfaces coated with MTA induced fibroblast proliferation and osteoblast differentiation; however, they did not present antibacterial properties.

Barium titanate (BaTiO3) piezoelectric ceramic may be a potential alternative for promoting osseointegration due to its piezoelectric properties similar to bone electric potentials generated in loading function. In this sense, the aim of this in vitro study was to evaluate the cellular response of human osteoblasts and gingival fibroblasts as well as the impact on S. oralis when in contact with BaTiO3 functionalized zirconia implant surfaces with piezoelectric properties. Zirconia discs with BaTiO3 were produced and contact poling (piezo activation) was performed. Osteoblasts (hFOB 1.19), fibroblasts (HGF hTERT) and S. oralis were culture on discs. Cell viability and morphology, cell differentiation markers, bacterial adhesion and growth were evaluated. The present study suggests that zirconia composite surfaces with the addition of piezoelectric BaTiO3 are not cytotoxic to peri-implant cells. Also, they seem to promote a faster initial osteoblast differentiation. Moreover, these surfaces may inhibit the growth of S. oralis by acting as a bacteriostatic agent over time. Although the piezoelectric properties do not affect the cellular inflammatory profile, they appear to enable the initial adhesion of bacteria, however this is not significant over the entire testing period. Furthermore, the addition of non-poled BaTiO3 to zirconia may have a potential reduction effect on IL-6 mediated-inflammatory activity in fibroblasts.

Monoallelic expression of variant surface glycoprotein genes (VSGs) is essential for immune evasion by Trypanosoma brucei. Here, Faria et al. show that the VEX protein complex controls VSG allelic exclusion, and that CAF‐1 sustains inheritance of the VEX‐complex in association with the active VSG.

Background: Takotsubo cardiomyopathy is characterized by transient left systolic dysfunction that can mimic acute myocardial infarction. Atrioventricular (AV) block associated with Takotsubo is rare, but a few cases have been reported in recent years. Methods: We present the case of a 77-year-old woman presenting with second-degree AV and Takotsubo syndrome. Results: The diagnosis of Takotsubo syndrome was based on echocardiogram changes and the absence of coronary artery obstruction on coronary angiography. Conclusions: We describe a patient with a diagnosis of Takotsubo syndrome and AV conduction defect. These conditions rarely occur simultaneous, but when they do, a dilemma arises regarding pacemaker implantation.

Abstract Adding a biological apatite layer to the implant surface enhances bone healing around the implant. Objective This study aimed to characterize the mechanical properties and test human gingival fibroblasts behavior in contact with Zirconia and Titanium bioactive-modified implant materials. Methodology 6 groups were considered: Titanium (Ti6Al4V), Ti6Al4V with 5% HA and 5% ßTCP, Zirconia (YTZP), YTZP with 5% HA and 5% ßTCP. For each group, we produced discs using a novel fabrication method for functionally graded materials, under adequate conditions for etching and grit-blasting to achieve equivalent surface microroughness among the samples. Surface roughness (Ra, Rz), water contact angle, shear bond strength, and Vickers hardness were performed. Human gingival fibroblasts immortalized by hTERT gene from the fourth passage, were seeded on discs for 14 days. Cell viability and proliferation were assessed using a resazurin-based method, and cellular adhesion and morphology using field emission gun scanning electron microscopy (FEG-SEM). After 3 days of culture, images of fluorescent nucleic acid stain were collected by confocal laser scanning microscopy (CLSM). Results Results were presented as mean ± standard deviation (SD). We compared groups using one-way ANOVA with Tukey’s post-hoc test, and significance level was set at p

Introduction: Minimally invasive gynecological surgery such as hysteroscopy has a small risk of complications. These include uterine perforation (with or without adjacent pelvic organ lesion), bleeding and infection, and are more common in the presence of risk factors such as smoking, history of pelvic inflammatory disease (PID) and endometriosis. Case Presentation: A patient submitted to a diagnostic hysteroscopy with no immediate complications was admitted five days later to the emergency department in septic shock. The diagnosis of ruptured tubal abscess was made, requiring emergency laparotomy with sub-total hysterectomy and bilateral adnexectomy. Despite multiple organ failure requiring admission to the intensive care unit, the patient made a full recovery. Conclusion: Ascending infection can be a life-threatening complication of hysteroscopy, even in the absence of previously known risk factors. Keywords: Hysteroscopy, Complications, Risk factors, Multiple organ failure, Sepsis

Background: The increased use of dental implants in oral rehabilitation has been followed by the development of new biomaterials as well as improvements in the performance of biomaterials already in use. This triggers the need for appropriate analytical approaches to assess the biological and, ultimately, clinical benefits of these approaches. Aims: To address the role of physical, chemical, mechanical, and biological characteristics in order to determine the critical parameters to improve biological responses and the long-term effectiveness of dental implant surfaces. Data sources and methods: Web of Science, MEDLINE and Lilacs databases were searched for the last 30 years in English, Spanish and Portuguese idioms. Results: Chemical composition, wettability, roughness, and topography of dental implant surfaces have all been linked to biological regulation in cell interactions, osseointegration, bone tissue and peri-implant mucosa preservation. Conclusion: Techniques involving subtractive and additive methods, especially those involving laser treatment or embedding of bioactive nanoparticles, have demonstrated promising results. However, the literature is heterogeneous regarding study design and methodology, which limits comparisons between studies and the definition of the critical determinants of optimal cell response.

The aim of this study was to evaluate gingival fibroblasts and human osteoblasts’ response to textured Nd:YAG laser microgrooves, with different dimensions, on zirconia implant surfaces. A total of 60 zirconia disks (8 mm in diameter and 2 mm in thickness) were produced and divided between four study groups (N = 15): three laser-textured (widths between 125.07 ± 5.29 μm and 45.36 ± 2.37 μm and depth values from 50.54 ± 2.48 μm to 23.01 ± 3.79 μm) and a control group without laser treatment. Human osteoblasts and gingival fibroblasts were cultured on these surfaces for 14 days. FEG-SEM (Field Emission Gun–Scanning Electron Microscope) images showed cellular adhesion at 24 h, with comparable morphology in all samples for both cell types. A similar cell spreading within the grooves and in the space between them was observed. Cell viability increased over time in all study groups; however, no differences were found between them. Additionally, proliferation, ALP (Alkaline phosphatase) activity, collagen type I, osteopontin and interleukin levels were not significantly different between any of the study groups for any of the cell types. Analysis of variance to compare parameters effect did not reveal statistically significant differences when comparing all groups in the different tests performed. The results obtained revealed similar cell behavior based on cell viability and differentiation on different microtopographic laser grooves, compared to a microtopography only established by sandblasting and acid-etching protocol, the reference surface treatment on zirconia dental implants.

ObjectiveThe objective of this paper was to evaluate the structural validity and convergent validity of the first Portuguese version of the Connor-Davidson Resilience Scale (CD-RISC).SettingsThe data sets come from two studies conducted in Portugal, respectively, from the Resilience Effect in Coping with Trauma (RECT) project and from the Health Impact Assessment of Employment Strategies (HIAES) project.ParticipantsThe sample is composed of 476 participants from the RECT project and 405 participants from the HIAES project. In both projects, convenience samples were used.MeasuresThe original CD-RISC items were translated to Portuguese and were used in a survey along with additional psychosocial and biomedical measures.ResultsIndependent exploratory factorial analysis (EFA) with each of the two samples revealed that the best solution in both samples had three factors consistent with the self-efficacy, spirituality and social support factors from the original scale. A Confirmatory factor analysis using the two samples together and specifying the three factors from the EFA revealed a good overall fit and, comparatively, better fit than a model specified with the five factors from the original scale. The study of the convergent validity revealed that bivariate correlations between the three factors and validated measures of stress, life satisfaction, mental health and physical health are globally consistent with previous research.ConclusionsThis study makes available to the broad Community of Portuguese Language Countries a validated measure of resilience extensively used for research and intervention. The results encourage future studies using this translated version of CD-RISC to explore further the three-factor structure found here and to test the convergent validity with new samples.

Maintaining a salivary metabolic profile upon sample collection and preparation is determinant in metabolomics. Nuclear magnetic resonance (NMR) spectroscopy was used to identify metabolite changes during short-term storage, at room temperature (RT)/4 °C/−20 °C, and after sample preparation, at RT/4 °C (mimicking typical clinical/laboratory settings). Interestingly, significant metabolic inter-individual and inter-day variability were noted, probably determining sample stability to some extent. After collection, no changes were noted at −20 °C (at least for 4 weeks). RT storage induced decreases in methylated macromolecules (6 h); lactate (8 h); alanine (12 h); galactose, hypoxanthine, pyruvate (24 h); sarcosine, betaine, choline, N-acetyl-glycoproteins (48 h), while acetate increased (48 h). Less, but different, changes were observed at 4 °C, suggesting different oral and microbial status at different temperatures (with a possible contribution from inter-individual and inter-day variability), and identifying galactose, hypoxanthine, and possibly, choline esters, as potential general stability indicators. After preparation, addition of NaN3 did not impact significantly on saliva stabilization, neither at RT nor at 4 °C, although its absence was accompanied by slight increases in fucose (6.5 h) and proline (8 h) at RT, and in xylose (24 h) at 4 °C. The putative metabolic origins of the above variations are discussed, with basis on the salivary microbiome. In summary, after collection, saliva can be stored at RT/4 °C for up to 6 h and at −20 °C for at least 4 weeks. Upon preparation for NMR analysis, samples are highly stable at 25 °C up to 8 h and at 4 °C up to 48 h, with NaN3 addition preventing possible early changes in fucose, proline (6–8 h), and xylose (24 h) levels.

Bloodstream form African trypanosomes are thought to rely exclusively upon glycolysis, using glucose as a substrate, for ATP production. Indeed, the pathway has long been considered a potential therapeutic target to tackle the devastating and neglected tropical diseases caused by these parasites. However, plasma membrane glucose and glycerol transporters are both expressed by trypanosomes and these parasites can infiltrate tissues that contain glycerol. Here, we show that bloodstream form trypanosomes can use glycerol for gluconeogenesis and for ATP production, particularly when deprived of glucose following hexose transporter depletion. We demonstrate that Trypanosoma brucei hexose transporters 1 and 2 (THT1 and THT2) are localized to the plasma membrane and that knockdown of THT1 expression leads to a growth defect that is more severe when THT2 is also knocked down. These data are consistent with THT1 and THT2 being the primary routes of glucose supply for the production of ATP by glycolysis. However, supplementation of the growth medium with glycerol substantially rescued the growth defect caused by THT1 and THT2 knockdown. Metabolomic analyses with heavy-isotope labelled glycerol demonstrated that trypanosomes take up glycerol and use it to synthesize intermediates of gluconeogenesis, including fructose 1,6-bisphosphate and hexose 6-phosphates, which feed the pentose phosphate pathway and variant surface glycoprotein biosynthesis. We used Cas9-mediated gene knockout to demonstrate a gluconeogenesis-specific, but fructose-1,6-bisphosphatase (Tb927.9.8720)-independent activity, converting fructose 1,6-bisphosphate into fructose 6-phosphate. In addition, we observed increased flux through the tricarboxylic acid cycle and the succinate shunt. Thus, contrary to prior thinking, gluconeogenesis can operate in bloodstream form T. brucei. This pathway, using glycerol as a physiological substrate, may be required in mammalian host tissues.

Protein abundance differs from a few to millions of copies per cell. Trypanosoma brucei presents an excellent model for studies on codon bias and differential gene expression because transcription is broadly unregulated and uniform across the genome. T. brucei is also a major human and animal protozoal pathogen. Here, an experimental assessment, using synthetic reporter genes, revealed that GC3 codons have a major positive impact on both mRNA and protein abundance. Our estimates of relative expression, based on coding sequences alone (codon usage and sequence length), are within 2-fold of the observed values for the majority of measured cellular mRNAs (n > 7000) and proteins (n > 2000). Our estimates also correspond with expression measures from published transcriptome and proteome datasets from other trypanosomatids. We conclude that codon usage is a key factor affecting global relative mRNA and protein expression in trypanosomatids and that relative abundance can be effectively estimated using only protein coding sequences.

The de novo crystal structure of the Leishmania infantum Silent Information Regulator 2 related protein 1 (LiSir2rp1) has been solved at 1.99Å in complex with an acetyl-lysine peptide substrate. The structure is broadly commensurate with Hst2/SIRT2 proteins of yeast and human origin, reproducing many of the structural features common to these sirtuin deacetylases, including the characteristic small zinc-binding domain, and the larger Rossmann-fold domain involved in NAD+-binding interactions. The two domains are linked via a cofactor binding loop ordered in open conformation. The peptide substrate binds to the LiSir2rp1 protein via a cleft formed between the small and large domains, with the acetyl-lysine side chain inserting further into the resultant hydrophobic tunnel. Crystals were obtained only with recombinant LiSir2rp1 possessing an extensive internal deletion of a proteolytically-sensitive region unique to the sirtuins of kinetoplastid origin. Deletion of 51 internal amino acids (P253-E303) from LiSir2rp1 did not appear to alter peptide substrate interactions in deacetylation assays, but was indispensable to obtain crystals. Removal of this potentially flexible region, that otherwise extends from the classical structural elements of the Rossmann-fold, specifically the β8-β9 connector, appears to result in lower accumulation of the protein when expressed from episomal vectors in L. infantum SIR2rp1 single knockout promastigotes. The biological function of the large serine-rich insertion in kinetoplastid/trypanosomatid sirtuins, highlighted as a disordered region with strong potential for post-translational modification, remains unknown but may confer additional cellular functions that are distinct from their human counterparts. These unique molecular features, along with the resolution of the first kinetoplastid sirtuin deacetylase structure, present novel opportunities for drug design against a protein target previously established as essential to parasite survival and proliferation.

A growing interest in asparagine (Asn) metabolism has currently been observed in cancer and infection fields. Asparagine synthetase (AS) is responsible for the conversion of aspartate into Asn in an ATP-dependent manner, using ammonia or glutamine as a nitrogen source. There are two structurally distinct AS: the strictly ammonia dependent, type A, and the type B, which preferably uses glutamine. Absent in humans and present in trypanosomatids, AS-A was worthy of exploring as a potential drug target candidate. Appealingly, it was reported that AS-A was essential in Leishmania donovani, making it a promising drug target. In the work herein we demonstrate that Leishmania infantum AS-A, similarly to Trypanosoma spp. and L. donovani, is able to use both ammonia and glutamine as nitrogen donors. Moreover, we have successfully generated LiASA null mutants by targeted gene replacement in L. infantum, and these parasites do not display any significant growth or infectivity defect. Indeed, a severe impairment of in vitro growth was only observed when null mutants were cultured in asparagine limiting conditions. Altogether our results demonstrate that despite being important under asparagine limitation, LiAS-A is not essential for parasite survival, growth or infectivity in normal in vitro and in vivo conditions. Therefore we exclude AS-A as a suitable drug target against L. infantum parasites.

Ribose 5-phosphate isomerase is an enzyme involved in the non-oxidative branch of the pentose phosphate pathway, and catalyzes the inter-conversion of D-ribose 5-phosphate and D-ribulose 5-phosphate. Trypanosomatids, including the agent of African sleeping sickness namely Trypanosoma brucei, have a type B ribose-5-phosphate isomerase. This enzyme is absent from humans, which have a structurally unrelated ribose 5-phosphate isomerase type A, and therefore has been proposed as an attractive drug target waiting further characterization. In this study, Trypanosoma brucei ribose 5-phosphate isomerase B showed in vitro isomerase activity. RNAi against this enzyme reduced parasites' in vitro growth, and more importantly, bloodstream forms infectivity. Mice infected with induced RNAi clones exhibited lower parasitaemia and a prolonged survival compared to control mice. Phenotypic reversion was achieved by complementing induced RNAi clones with an ectopic copy of Trypanosoma cruzi gene. Our results present the first functional characterization of Trypanosoma brucei ribose 5-phosphate isomerase B, and show the relevance of an enzyme belonging to the non-oxidative branch of the pentose phosphate pathway in the context of Trypanosoma brucei infection.

Asparagine synthetase (AS) catalyzes the ATP-dependent conversion of aspartate into asparagine using ammonia or glutamine as nitrogen source. There are two distinct types of AS, asparagine synthetase A (AS-A), known as strictly ammonia-dependent, and asparagine synthetase B (AS-B), which can use either ammonia or glutamine. The absence of AS-A in humans, and its presence in trypanosomes, suggested AS-A as a potential drug target that deserved further investigation. We report the presence of functional AS-A in Trypanosoma cruzi (TcAS-A) and Trypanosoma brucei (TbAS-A): the purified enzymes convert L-aspartate into L-asparagine in the presence of ATP, ammonia and Mg(2+). TcAS-A and TbAS-A use preferentially ammonia as a nitrogen donor, but surprisingly, can also use glutamine, a characteristic so far never described for any AS-A. TbAS-A knockdown by RNAi didn't affect in vitro growth of bloodstream forms of the parasite. However, growth was significantly impaired when TbAS-A knockdown parasites were cultured in medium with reduced levels of asparagine. As expected, mice infections with induced and non-induced T. brucei RNAi clones were similar to those from wild-type parasites. However, when induced T. brucei RNAi clones were injected in mice undergoing asparaginase treatment, which depletes blood asparagine, the mice exhibited lower parasitemia and a prolonged survival in comparison to similarly-treated mice infected with control parasites. Our results show that TbAS-A can be important under in vivo conditions when asparagine is limiting, but is unlikely to be suitable as a drug target.

This systematic review and network meta-analysis aimed to assess the impact of combining professional mechanical plaque removal (PMPR) with probiotics compared to PMPR + placebo on probing pocket depth (PPD) and clinical attachment level (CAL). Randomized controlled trials published until November 2023 were searched across electronic databases, peer-reviewed journals, and grey literature. Two authors independently selected, extracted data, and assessed bias risk. Primary outcomes were mean changes in PPD and CAL. Secondary outcomes included mean changes in bleeding on probing (BOP), plaque index, and colony-forming units. Network meta-analysis with the frequentist weighted least squares approach evaluated the data quantitatively, and CINeMA framework evaluated the quality of evidence. In 33 articles involving 1290 patients, results were stratified by follow-up period (short and long-time studies) and sensitivity analyses conducted based on probiotic therapy duration (1 month reference). Network meta-analysis revealed significant mean differences in PPD for nine probiotic interventions, CAL for eighteen interventions, and BOP for eight interventions, with Lactobacillus demonstrating the most substantial effects. Combining PMPR with probiotics as adjuvants to subgingival instrumentation may be more effective in improving PPD and CAL. Lactobacillus emerged as the most comprehensive and effective among the studied probiotic. [ABSTRACT FROM AUTHOR]

Purpose: The aim of this study was to characterize and compare the behavior of human osteoblasts and human gingival fibroblasts in contact with polyetheretherketone (PEEK), zirconia, and titanium implant surface materials. Materials and Methods: PEEK, yttria-stabilized zirconia (YTZP), and titanium disks were produced under appropriate and similar conditions to achieve controlled surface features. Human osteoblasts and human gingival fibroblasts were cultured on disks for 14 days. Cell viability and proliferation were evaluated using a resazurin-based method. Morphology and cellular adhesion were observed using field emission gun-scanning electron microscopy (FEG-SEM). Alkaline phosphatase (ALP) activity and bone cell mineralization was evaluated on osteoblasts. Confocal laser scanning microscopy (CLSM) images of fluorescent-stained fibroblasts were obtained at 7 and 14 days of the culture. Results were presented as mean and standard deviation (SD). Group comparisons were tested using analysis of variance (ANOVA) (Tukey's post hoc) with appropriate statistical software, and significance was set at P < .05. Results: Cell viability and proliferation were higher in PEEK and YTZP groups compared with titanium on osteoblast cells (P < .05, all time points) and on fibroblasts (P < .05, 7 and 14 days). All groups showed an increase in ALP activity over time, which was not significant. Mineralization patterns demonstrated an increase in mineral content over time, which was more apparent in the YTZP group. Cell spreading was more evident on PEEK and YTZP specimens. Conclusion: The results suggest increased adhesion, viability, and proliferation of osteoblasts and gingival fibroblasts on zirconia and PEEK surfaces compared with titanium. These results are correlated with the increased wettability of these materials. [ABSTRACT FROM AUTHOR]