Targeting Biomolecular Condensation and Protein Aggregation against Cancer.

Biomolecular condensates, membrane-less entities arising from liquid-liquid phase separation, hold dichotomous roles in health and disease. Alongside their physiological functions, these condensates can transition to a solid phase, producing amyloid-like structures implicated in degenerative diseases and cancer. This review thoroughly examines the dual nature of biomolecular condensates, spotlighting their role in cancer, particularly concerning the p53 tumor suppressor. Given that over half of the malignant tumors possess mutations in the TP53 gene, this topic carries profound implications for future cancer treatment strategies. Notably, p53 not only misfolds but also forms biomolecular condensates and aggregates analogous to other protein-based amyloids, thus significantly influencing cancer progression through loss-of-function, negative dominance, and gain-of-function pathways. The exact molecular mechanisms underpinning the gain-of-function in mutant p53 remain elusive. However, cofactors like nucleic acids and glycosaminoglycans are known to be critical players in this intersection between diseases. Importantly, we reveal that molecules capable of inhibiting mutant p53 aggregation can curtail tumor proliferation and migration. Hence, targeting phase transitions to solid-like amorphous and amyloid-like states of mutant p53 offers a promising direction for innovative cancer diagnostics and therapeutics.

Morphological, cellular, and molecular basis of brain infection in COVID-19 patients.

Although increasing evidence confirms neuropsychiatric manifestations associated mainly with severe COVID-19 infection, long-term neuropsychiatric dysfunction (recently characterized as part of "long COVID-19" syndrome) has been frequently observed after mild infection. We show the spectrum of cerebral impact of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, ranging from long-term alterations in mildly infected individuals (orbitofrontal cortical atrophy, neurocognitive impairment, excessive fatigue and anxiety symptoms) to severe acute damage confirmed in brain tissue samples extracted from the orbitofrontal region (via endonasal transethmoidal access) from individuals who died of COVID-19. In an independent cohort of 26 individuals who died of COVID-19, we used histopathological signs of brain damage as a guide for possible SARS-CoV-2 brain infection and found that among the 5 individuals who exhibited those signs, all of them had genetic material of the virus in the brain. Brain tissue samples from these five patients also exhibited foci of SARS-CoV-2 infection and replication, particularly in astrocytes. Supporting the hypothesis of astrocyte infection, neural stem cell-derived human astrocytes in vitro are susceptible to SARS-CoV-2 infection through a noncanonical mechanism that involves spike-NRP1 interaction. SARS-CoV-2-infected astrocytes manifested changes in energy metabolism and in key proteins and metabolites used to fuel neurons, as well as in the biogenesis of neurotransmitters. Moreover, human astrocyte infection elicits a secretory phenotype that reduces neuronal viability. Our data support the model in which SARS-CoV-2 reaches the brain, infects astrocytes, and consequently, leads to neuronal death or dysfunction. These deregulated processes could contribute to the structural and functional alterations seen in the brains of COVID-19 patients.

NMDA glutamate receptor antagonist MK-801 induces proteome changes in adult human brain slices which are partially counteracted by haloperidol and clozapine.

Deciphering the molecular pathways associated with N-methyl-D-aspartate receptor (NMDAr) hypofunction and its interaction with antipsychotics is necessary to advance our understanding of the basis of schizophrenia, as well as our capacity to treat this disease. In this regard, the development of human brain-derived models that are amenable to studying the neurobiology of schizophrenia may contribute to filling the gaps left by the widely employed animal models. Here, we assessed the proteomic changes induced by the NMDA glutamate receptor antagonist MK-801 on human brain slice cultures obtained from adult donors submitted to respective neurosurgery. Initially, we demonstrated that MK-801 diminishes NMDA glutamate receptor signaling in human brain slices in culture. Next, using mass-spectrometry-based proteomics and systems biology in silico analyses, we found that MK-801 led to alterations in proteins related to several pathways previously associated with schizophrenia pathophysiology, including ephrin, opioid, melatonin, sirtuin signaling, interleukin 8, endocannabinoid, and synaptic vesicle cycle. We also evaluated the impact of both typical and atypical antipsychotics on MK-801-induced proteome changes. Interestingly, the atypical antipsychotic clozapine showed a more significant capacity to counteract the protein alterations induced by NMDAr hypofunction than haloperidol. Finally, using our dataset, we identified potential modulators of the MK-801-induced proteome changes, which may be considered promising targets to treat NMDAr hypofunction in schizophrenia. This dataset is publicly available and may be helpful in further studies aimed at evaluating the effects of MK-801 and antipsychotics in the human brain.

Computed tomography versus ultrasound for the diagnosis of acute cholecystitis: a systematic review and meta-analysis.

OBJECTIVES: Some patients undergo both computed tomography (CT) and ultrasound (US) sequentially as part of the same evaluation for acute cholecystitis (AC). Our goal was to perform a systematic review and meta-analysis comparing the diagnostic performance of US and CT in the diagnosis of AC. MATERIALS AND METHODS: Databases were searched for relevant published studies through November 2023. The primary objective was to compare the head-to-head performance of US and CT using surgical intervention or clinical follow-up as the reference standard. For the secondary analysis, all individual US and CT studies were analyzed. The pooled sensitivities, specificities, and areas under the curve (AUCs) were determined along with 95% confidence intervals (CIs). The prevalence of imaging findings was also evaluated. RESULTS: Sixty-four studies met the inclusion criteria. In the primary analysis of head-to-head studies (n = 5), CT had a pooled sensitivity of 83.9% (95% CI, 78.4-88.2%) versus 79.0% (95% CI, 68.8-86.6%) of US (p = 0.44). The pooled specificity of CT was 94% (95% CI, 82.0-98.0%) versus 93.6% (95% CI, 79.4-98.2%) of US (p = 0.85). The concordance of positive or negative test between both modalities was 82.3% (95% CI, 72.1-89.4%). US and CT led to a positive change in management in only 4 to 8% of cases, respectively, when ordered sequentially after the other test. CONCLUSION: The diagnostic performance of CT is comparable to US for the diagnosis of acute cholecystitis, with a high rate of concordance between the two modalities. CLINICAL RELEVANCE STATEMENT: A subsequent US after a positive or negative CT for suspected acute cholecystitis may be unnecessary in most cases. KEY POINTS: When there is clinical suspicion of acute cholecystitis, patients will often undergo both CT and US. CT has similar sensitivity and specificity compared to US for the diagnosis of acute cholecystitis. The concordance rate between CT and US for the diagnosis of acute cholecystitis is 82.3%.

Mechanical pump complication after HeartMate 3 implantation.

BACKGROUND: With the improvement in postoperative complications and long-term survival post LVAD, continuing to improve clinical outcomes will require efforts to decrease long-term complications. The purpose of this study is to describe the incidence of mechanical pump failure requiring surgery, which we define as pump failure secondary to either outflow graft compression, outflow graft obstruction, or pump thrombosis requiring surgical intervention. METHODS: 141 consecutive adult patients who underwent HeartMate3 Implantation using the "cut-then-sew" implantation technique between September 2015 and September 2021 were included in our study. The primary outcome measure was mechanical pump complication (outflow graft obstruction and or pump thrombosis) requiring surgical intervention. Secondary outcome measures included incidence of bleeding, stroke, renal failure, length of stay, and overall survival. Median follow up was 27.3 months. RESULTS: Eleven (7.8%) of patients developed mechanical pump complications. Six patients developed outflow graft obstruction. Five patients developed acute pump thrombosis. Median time to a mechanical complication was 828 days. Of the 11 patients who underwent surgery, 10 patients (90%) survived to discharge. Overall survival at 1, 3, and 5 years was 82.9%, 69.1% and 55.2% respectively for the entire cohort. CONCLUSION: The mechanical pump complication rate of 7.8% which is quite high may be related to duration of follow up, as the median time to mechanical complication was 828 days. This study highlights an important late complication that occurs post LVAD implantation.

Wound-healing agents for palatal donor area: A network meta-analysis.

BACKGROUND: The aim of this systematic review and network meta-analysis (NMA) was to assess the efficacy of different wound-healing agents for palatal donor area management after soft tissue graft harvesting. METHODS: Electronic searches in six databases were conducted for publications up to October 2023. Studies with data from patients undergoing therapeutic approaches using agents for palatal healing after gingival graft harvesting were included. Data about postoperative pain, wound-healing and postoperative complications reported for each agent were extracted. Three different tools were used for the risk of bias within studies evaluation (Murad tool for case series and case report, RoB 2.0 tool for randomized studies and ROBINS-I tool for non-randomized studies). A Bayesian random effects NMA was conducted for postoperative pain levels and wound healing. RESULTS: Eighty-four publications were included in the systematic review (qualitative analysis), with 14 of these subjected to NMA (quantitative analysis). The summarized results from the qualitative and quantitative analysis showed that all wound-healing agents evaluated promoted better pain control and wound healing compared to spontaneous healing and hemostatic sponges alone. The NMA outcomes reveal that leucocyte- and platelet-rich fibrin (L-PRF) was the most effective agent in reducing postoperative pain in all analyzed periods. Moreover, the L-PRF seems to accelerate wound healing and reduce postoperative complications compared to other agents. CONCLUSION: In conclusion, the L-PRF was the most effective agent in reducing postoperative pain, accelerating wound healing and reducing postoperative complications after harvesting soft tissue grafts from the palatal area.

Ontogeny of daily rhythms in the expression of metabolic factors in Nile tilapia (Oreochromis niloticus) kept at two different temperature regimes: Thermocycle and constant temperature.

The daily variations of temperature are one of the main synchronizers of the circadian rhythms. In addition, water temperature influences the embryonic and larval development of fish and directly affects their metabolic processes. The application of thermocycles to fish larvae has been reported to improve growth and the maturation of the digestive system, but their effects on metabolism are poorly understood. The aim of the present study was to evaluate the effect of two different temperature regimes, cycling versus constant, on the daily rhythms of metabolic factors of Nile tilapia (Oreochromis niloticus) larvae. For this purpose, fertilized eggs were divided into two groups: one reared in a 31 °C:25 °C day:night thermocycle (TCY) and another group maintained in a constant 28 °C temperature (CTE). The photoperiod was set to a 12:12 h light/dark cycle. Samples were collected every 4 h during a 24-h cycle on days 4, 8 and 13 post fertilization (dpf). The expression levels of alanine aminotransferase (alt), aspartate aminotransferase (ast), malic enzyme, glucose-6-phosphate dehydrogenase (g6pd), phosphofructokinase (pfk) and pyruvate kinase (pk) were analyzed by qPCR. Results showed that, in 13 dpf animals, most of the genes analyzed (alt, ast, malic, g6pd and pfk) showed daily rhythms in TCY, but not in the group kept at constant temperature, with most acrophases detected during the feeding period. An increase in nutrient metabolism around feeding time can improve food utilization and thus increase larval performance. Therefore, the use of thermocycles is recommended for tilapia larviculture.

Assessing bone formation on hydrophilic and hydrophobic implant surfaces in a murine model treated with bisphosphonates.

OBJECTIVE: To evaluate the osseointegration of implants with hydrophobic (HFB) and hydrophilic (HFL) surfaces in a murine model of high-dose bisphosphonates (BPs). MATERIALS AND METHODS: Sixty-four rats were randomly allocated into four groups: control group with HFB implants (CG-HFB), control group with HFL implants (CG-HFL), BP group with HFB implants (BP-HFB), and BP group with HFL implants (BP-HFL). Animals were euthanized after 15 and 45 days (n=8). The dependent variables assessed were the removal torque (biomechanical analysis), the bone volume around the implants (%BV/TV) (microtomographic analysis), the bone-implant contact (%BIC), the bone between the threads (%BBT) (histomorphometric analysis), and the expression of bone metabolism markers (immunohistochemistry analysis). RESULTS: The CG-HFL and BP-HFL groups presented higher removal torque than the CG-HFB and BP-HFB implants. The %BIC of the CG-HFL surfaces was slightly higher than that of the CG-HFB implants. The BP-HFB and BP-HFL groups presented a higher %BIC than that of the CG-HFB and CG-HFL groups (p<0.001). BP therapy also increased the %BBT at both implant surfaces. Higher levels of ALP were observed in the matrix region of bone tissue on the HFL surfaces than on the HFB surfaces. CONCLUSION: Both surfaces enable osseointegration in rats under BP therapy. CLINICAL RELEVANCE: The study demonstrates that hydrophobic (HFB) and hydrophilic (HFL) implant surfaces can promote osseointegration in rats undergoing bisphosphonate therapy. The HFL surfaces exhibited improved biomechanical performance, higher bone-implant contact, and increased bone volume, suggesting their potential clinical relevance for implant success in individuals on bisphosphonate treatment.

Water Leakage Pathway Leads to Internal Hydration of the p53 Core Domain.

The gene encoding the p53 tumor suppressor protein is the most frequently mutated oncogene in cancer patients; yet, generalized strategies for rescuing the function of different p53 mutants remain elusive. This work investigates factors that may contribute to the low inherent stability of the wild-type p53 core domain (p53C) and structurally compromised Y220C mutant. Pressure-induced unfolding of p53C was compared to p63C, the p53 family member with the highest stability, the engineered superstable p53C hexamutant (p53C HM), and lower stability p53C Y220C cancer-associated mutant. The following pressure unfolding values (P50% bar) were obtained: p53C 3346, p53C Y220C 2217, p53C HM 3943, and p63C 4326. Molecular dynamics (MD) simulations revealed that p53C Y220C was most prone to water infiltration, followed by p53C, whereas the interiors of p53C HM and p63C remained comparably dry. A strong correlation (r2 = 0.92) between P50% and extent of interior hydration was observed. The pathways of individual water molecule entry and exit were mapped and analyzed, revealing a common route preserved across the p53 family involving a previously reported pocket, along with a novel surface cleft, both of which appear to be targetable by small molecules. Potential determinants of propensity to water incursion were assessed, including backbone hydrogen bond protection and combined sequence and structure similarity. Collectively, our results indicate that p53C has an intrinsic susceptibility to water leakage, which is exacerbated in a structural class mutant, suggesting that there may be a common avenue for rescuing p53 function.

Mortality in children under 5 years of age with congenital syphilis in Brazil: A nationwide cohort study.

BACKGROUND: Congenital syphilis (CS) is a major and avoidable cause of neonatal death worldwide. In this study, we aimed to estimate excess all-cause mortality in children under 5 years with CS compared to those without CS. METHODS AND FINDINGS: In this population-based cohort study, we used linked, routinely collected data from Brazil from January 2011 to December 2017. Cox survival models were adjusted for maternal region of residence, maternal age, education, material status, self-declared race and newborn sex, and year of birth and stratified according to maternal treatment status, non-treponemal titers and presence of signs and symptoms at birth. Over 7 years, a total of 20 057 013 live-born children followed up (through linkage) to 5 years of age, 93 525 were registered with CS, and 2 476 died. The all-cause mortality rate in the CS group was 7·84/1 000 person-years compared with 2·92/1 000 person-years in children without CS, crude hazard ratio (HR) = 2·41 (95% CI 2·31 to 2·50). In the fully adjusted model, the highest under-five mortality risk was observed among children with CS from untreated mothers HR = 2·82 (95% CI 2·63 to 3·02), infants with non-treponemal titer higher than 1:64 HR = 8·87 (95% CI 7·70 to 10·22), and children with signs and symptoms at birth HR = 7·10 (95% CI 6·60 to 7·63). Among children registered with CS, CS was recorded as the underlying cause of death in 33% (495/1 496) of neonatal, 11% (85/770) of postneonatal, and 2·9% (6/210) of children 1 year of age. The main limitations of this study were the use of a secondary database without additional clinical information and the potential misclassification of exposure status. CONCLUSIONS: This study showed an increased mortality risk among children with CS that goes beyond the first year of life. It also reinforces the importance of maternal treatment that infant non-treponemal titers and the presence of signs and symptoms of CS at birth are strongly associated with subsequent mortality. TRIAL REGISTRATION: Observational study.

Osteoprotective effect by interleukin-4 (IL-4) on lipoprotein-induced periodontitis.

Lipoproteins are immunostimulatory bacterial components suggested to participate in inflammation-induced bone loss in periodontal disease through stimulation of osteoclast differentiation. Toll-like receptor 2 activation by Pam2CSK4 (PAM2), known to mimic bacterial lipoproteins, was previously shown to enhance periodontal bone resorption in mice. The anti-inflammatory cytokine interleukin-4 (IL-4) is a known inhibitor of RANKL-induced bone resorption in vitro. Here, we have investigated whether IL-4 could decrease PAM2-induced periodontal bone loss and osteoclastogenesis in vivo. In a model of periodontitis induced by gingival injections of PAM2 in mice, concomitant injections of IL-4 reduced bone loss. Histologically, IL-4 reduced the recruitment of inflammatory cells and the formation of TRAP+ osteoclasts stimulated by PAM2. Mouse bone marrow macrophages (BMMs) and neonatal calvarial osteoblasts were used to assess the effect of IL-4 on PAM2-induced osteoclastogenesis in vitro. In RANKL-primed BMMs stimulated by PAM2 Nfatc1, Ctsk, and Acp5 gene expression was up-regulated and resulted in robust formation of TRAP+ multinucleated osteoclasts, effects which were impaired by IL-4. These effects were mediated by impairment in PAM2-induced c-fos expression. In primary calvarial osteoblast cultures, IL-4 decreased PAM2-induced Tnfsf11 (encoding RANKL) mRNA and enhanced Tnfrsf11b (encoding OPG) expression. Our data demonstrate that the osteoprotective effect by IL-4 on lipoprotein-induced periodontal disease occurs through the inhibition of osteoclastogenesis by three mechanisms, one by acting directly on osteoclast progenitors, another by acting indirectly through decreasing the expression of osteoclast-regulating cytokines in osteoblasts and a third by decreasing inflammation.

Correction: Ten simple rules for organizing a bioinformatics training course in low- and middle-income countries.

[This corrects the article DOI: 10.1371/journal.pcbi.1009218.].

Split scar sign to predict complete response in rectal cancer after neoadjuvant chemoradiotherapy: systematic review and meta-analysis.

OBJECTIVES: Magnetic resonance imaging (MRI) is the modality of choice for rectal cancer initial staging and restaging after neoadjuvant chemoradiation. Our objective was to perform a meta-analysis of the diagnostic performance of the split scar sign (SSS) on rectal MRI in predicting complete response after neoadjuvant therapy. METHODS: MEDLINE, EMBASE, and Cochrane databases were searched for relevant published studies through June 2023. Primary studies met eligibility criteria if they evaluated the diagnostic performance of the SSS to predict complete response on pathology or clinical follow-up in patients undergoing neoadjuvant chemoradiation. A meta-analysis with a random-effects model was used to estimate pooled sensitivity and specificity, area under the curve (AUC), and diagnostic odds ratio (DOR) of the SSS. RESULTS: A total of 4 studies comprising 377 patients met the inclusion criteria. The prevalence of complete response in the studies was 21.7-52.5%. The pooled sensitivity and specificity of the SSS to predict complete response were 62.0% (95% CI, 43.5-78.5%) and 91.9% (95% CI, 78.9-97.2%), respectively. The estimated AUC for SSS was 0.83 (95% CI, 0.56-0.94) with a DOR of 18.8 (95% CI, 3.65-96.5). CONCLUSION: The presence of SSS on rectal MRI demonstrated high specificity for complete response in patients with rectal cancer after neoadjuvant chemoradiation. This imaging pattern can be a valuable tool to identify potential candidates for organ-sparing treatment and surveillance. CLINICAL RELEVANCE STATEMENT: SSS presents high specificity for complete response post-neoadjuvant. This MRI finding enhances rectal cancer treatment assessment and aids clinicians and patients in choosing watch-and-wait over immediate surgery, which can potentially reduce costs and associated morbidity. KEY POINTS: •Fifteen to 50% of rectal cancer patients achieve complete response after neoadjuvant chemoradiation and may be eligible for a watch-and-wait strategy. •The split scar sign has high specificity for a complete response. •This imaging finding is valuable to select candidates for organ-sparing management.

Fresh-frozen allogeneic bone blocks grafts for alveolar ridge augmentation: Biological and clinical aspects.

The possibilities for oral bone regeneration procedures vary depending on the type of bone defect to be treated, which in turn dictate the type of graft to be used. Atrophic alveolar ridges are non-contained defects and pose a challenging defect morphology for bone regeneration/augmentation. Successful results are regularly obtained with the use of particulate grafts in combination with barrier membranes. In cases of very narrow ridges with need of larger amount of bone augmentation, block grafts are often used. Fresh-frozen allogeneic bone block grafts have been proposed as an alternative to autogenous (AT) bone blocks. Based on a systematic appraisal of pre-clinical in vivo studies and clinical trials including a direct comparison of fresh-frozen bone (FFB) blocks versus AT bone blocks it can be concluded that a FFB block graft: (a) cannot be considered as a reliable replacement of a AT bone block, and (b) should only be considered in cases where the amount of necessary augmentation-in a lateral direction-is relatively limited, so that the main portion of the body of the implant lies within the inner (i.e., the vital) aspect of the block.

The Brazilian Atlantic Bushmaster Lachesis (Linnaeus, 1766) Mitogenome With Insights On Snake Evolution And Divergence (Serpentes: Viperidae: Crotalinae).

This study presents the first complete mitogenome of the Brazilian Atlantic bushmaster Lachesis with insights into snake evolution. The total length was 17,177 bp, consisting of 13 PCGs, 22 tRNAs, two rRNAs and a duplicate control region (CRs). Almost all genes were encoded by the heavy-strand, except for the ND6 gene and eight tRNAs (tRNA-Gln, Ala, Asn, Cys, Tyr, Ser[TGA anticodon], Glu, Pro). Only ATG, ATA, and ATC were starting codons for protein-coding sequences. Stop codons mainly were TAA, AGA, AGG, and TAG; whereas ND1, ND3, and CYTB terminated with incomplete stop codons. Phylogeny retrieved Lachesis within the Crotalinae as the sister group of Agkistrodon; and the Lachesis+Agkistrodon clade as the sister group of (Sistrurus+Crotalus)+Bothrops. The tree supports Crotalinae, Viperinae, and Azemiopinae in the Viperidae family, being sister taxa of Colubridae+(Elapidae+Psammophiidae). The mean genetic distance across 15 snake families and 57 nucleotide sequences was 0.37. The overall mean value of genetic distance across the Crotalinae was 0.23, with Lachesis muta exhibiting the shortest distance of 0.2 with Agkistrodon piscivorus, Protobothrops dabieshanensis and P. flavoviridis and the greatest 0.25 with Gloydius blomhoffii, Trimeresurus albolabris, S. miliarius, and Deinagkistrodon acutus. The complete Atlantic L. muta mitogenome presented herein is only the third annotated mitogenome from more than 430 described Brazilian snake species.

First report of Epicoccum nigrum causing leaf spots on Campomanesia guazumifolia (Camb.) Berg in Brazil.

Campomanesia guazumifolia is a native tree that produces fruit that can be consumed fresh or used by industry (Donadio et al., 2002). In February 2022, in the experimental area of the Universidade Tecnológica Federal do Paraná - Brazil, disease was observed in 22 trees, with 50% to 80% severity in crown leaves. Symptoms were small, irregular, or circular-shaped, dark-brown lesions with yellow halos (Figure S1). As the disease progressed, the lesions increased in size, without distinction between mature and young tissues, causing complete leaf wilting. Twenty symptomatic leaves from 11 trees grown in the same orchard line were collected. For fungal isolation, the leaf surfaces were disinfected with 0.5% NaOCl solution for 1 min, rinsed in sterile distilled water, and dried on sterile filter paper. Five fragments of diseased leaf tissue were placed on a potato dextrose agar medium. The morphological characteristics of the colony, such as filamentous mycelium and golden yellow on the upper part, with the presence of circular to ovoid and multicellular conidia (mean 21.00 µm x 24.45 µm, n = 30) of the nine isolates, coincided with the description of the fungus of the genus Epicoccum (Valenzuela-Lopez et al., 2018). Further identification of one of these nine isolates was confirmed by amplifying and sequencing three loci (ITS, ß-tubulin, and RPB2) using the ITS1/ITS4, Bt2a/Bt2b, and 5F2/7cR primer pairs, respectively (White et al., 1990, Glass and Donaldson, 1995, O'Donnell et al., 2007). A single representative isolate (Cgen01) was analyzed and submitted to GenBank (OR020968, OR079879, and OR079878). The Bayesian Inference was used to reconstruct the phylogenetic trees (Figure S2), starting from random trees for 5,000,000 generations, using MrBayes v. 3.2.1 (Ronquist et al., 2012). The isolate clustered together with the isolate of Epicoccum nigrum (Chen et al., 2017) with a high posterior probability (0.98). For the pathogenicity tests, four young, healthy branches containing 20 leaves were spray-inoculated with 1.5 mL of conidia suspension of Cgen01 (106 conidia mL-1), covered with perforated transparent plastic bags, and moistened with distilled water in the orchard. The air temperature ranged from 14ºC to 25ºC. Sterile distilled water was used as a control. Three replicates (pathogen and control) on different trees were evaluated. After five days, the fungus was re-isolated from the symptomatic lesion, showing morphological characteristics similar to those of Cgen01. Control branches did not show fungal growth. The inoculation test was conducted twice and similar symptoms were observed. This is the first report of leaf spots caused by E. nigrum on C. guazumifolia in Brazil. E. nigrum, an endophytic fungus described as a mycoparasite, showed phytopathogenic behavior in this study, causing spots and loss of leaves in C. guazumifolia, drastically reducing the production of photoassimilates and affecting the quality of the fruits.

Effect of a gel containing green tea extract and hyaluronic acid on palate pain scores and wound healing after free gingival graft: a quasi-randomized controlled clinical trial.

OBJECTIVES: The aim of this study was to evaluate the effect of a gel containing green tea extract and hyaluronic acid (HA) on pain scores and wound healing in donor sites after free gingival graft (FGG). MATERIALS AND METHODS: Forty-two patients requiring FGG were included in three groups: (1) control group (n = 14), no material was placed in the donor area; only the clot was kept in position by sutures; (2) placebo group (n = 14), vehicle gel applied 3 times a day for 7 days; and (3) test group (n = 14), gel containing green tea extract and HA applied 3 times a day for 7 days. The wound size by clinical measurement (WS-CM) and photographic image (WS-PI), complete wound epithelialization (CWE), and palatal mucosa color were evaluated after 3 days and 1, 2, and 4 weeks postoperatively. The visual analog scale (VAS) for pain and analgesic consumption were used to assess participant's perception in the same postoperative periods. RESULTS: A similar progressive reduction in the wound size, associated with an improvement in the color pattern, was observed in all groups (p > 0.05). No significant differences were found for CWE and pain assessment between the examined groups (p > 0.05). CONCLUSION: The gel containing green tea extract and HA application in palatal wounds after FGG removal does not provide clinical healing benefits using this investigated protocol. CLINICAL RELEVANCE: This is the first clinical study evaluating the effect of gel containing green tea extract and HA on the palate postoperative pain control and wound healing after FGG. TRIAL REGISTRATION: http://clinicaltrials.gov : NCT05270161.

Digital planning and guided dual technique in esthetic crown lengthening: a randomized controlled clinical trial.

OBJECTIVES: The present study aimed to compare the guided dual technique with the conventional technique in esthetic crown lengthening (ECL). MATERIALS AND METHODS: The trial registration number is NCT04922086. Twenty-four participants diagnosed with altered passive eruption (APE) type I subcategory B were selected and allocated into two groups. In the control group (n = 12), the ECL procedure was planned by clinical examination and transgingival probing; in the test group (n = 12), the ECL procedure was carried out using digital planning and a double guide. Clinical parameters were assessed at baseline, immediately after the intervention (IAI), and at 4, 8, and 12 months of follow-up. RESULTS: The clinical crown length (CCL) mean at baseline was 8.09 mm (± 0.77) and increased significantly to 9.92 mm (± 0.62) IAI, with minimal significant reduction after 12 months (9.47 mm [± 0.60]) in the control group. Similarly, in the test group, the mean CCL at baseline was 8.04 mm (± 0.69) and increased significantly to 9.94 mm (± 0.57) IAI, with minimal reduction after 12 months (9.35 mm [± 0.80]). No differences were found between the mean CCL determined in the digital planning and after 12 months. No correlation was found between gingival thickness and gingival margin stability. High esthetic satisfaction was demonstrated by participants/specialists without differences between groups. CONCLUSIONS: In conclusion, the guided dual technique was as effective as the conventional technique for treatment of APE, with stable results after 12 months of follow-up. CLINICAL RELEVANCE: The guided dual technique aims to transfer both the gingival and bone resection planned position to facilitate the ECL surgical procedure and increase treatment predictability.

Feeding behaviour, locomotion rhythms and blood biochemistry of the neotropical red-tail catfish (Phractocephalus hemioliopterus).

The study evaluated the feeding behaviour of Phractocephalus hemioliopterus through the animals' ability to adapt to the self-feeding system, their preferred feeding times and locomotor activity, as well as the blood biochemistry of juveniles fed in a light/dark cycle. The study was carried out through two experiments, the first of which contained two phases. In experiment 1 - phase I, 24 juveniles (35.28 ± 0.62 g) were distributed in eight 48 l tanks. The tanks were equipped with a self-feeding system and the experiment consisted of evaluating whether the animals were able to adapt to the self-feeding system, as well as evaluating the preferred feeding times and locomotor activity of these animals. A feeding challenge to the animals was introduced in phase II, based on the results of phase I. The results of the first phase evidenced a nocturnal feeding preference. Thus, the feeding challenge consisted of measuring whether the animal would feed during the day and how long it would take to adapt. When the animals consumed 100% of the amount of feed provided daily, phase II was ended. In experiment 2, 24 juveniles of P. hemioliopterus (182.00 ± 14.03 g) were distributed in eight 96 l tanks. This experiment consisted of two treatments with four repetitions, one with exclusive feeding during the middle of the light cycle and another with exclusive feeding in the middle of the dark cycle. At the end, blood samples were collected from the animals for blood biochemistry evaluations. In experiment 1 - phase I, the results showed that the fish adapted very well to the self-feeding system and had a strictly nocturnal feeding behaviour and locomotor rhythm. When they were submitted to the feeding challenge in phase II, the feed intake was stabilized from the 17th day onwards, proportionally to the nocturnal consumption observed in the first phase, thus demonstrating feeding plasticity. In experiment 2, the feeding times influenced the animals' biochemical parameters. Animals fed during the night had higher values of cholesterol and triglycerides than animals fed during the day. It is concluded that P. hemioliopterus has fast adaptability to a self-feeding system, with strictly nocturnal feeding and locomotor behaviours. However, it has feeding plasticity, adapting its behaviour according to food availability. Blood biochemical parameters are influenced by the light/dark feeding cycle.

Feeding and locomotor rhythm of the ornamental cichlid fish Amatitlania sp. in different social groups.

The aim of this study was to evaluate the feeding activity and the rhythm of daily locomotor activity of the convict cichlid (Amatitlania sp.) kept in different social groups under a self-feeding system. A total of 120 animals was distributed among six repetitions of four social groups, as follows: group 1 with one male and one female per tank; group 2 with three males and three females per tank; group 3 with six males per tank; and group 4 with six females per tank. Feeding activity (FA) and locomotor activity (LA) were evaluated using photoelectric presence-sensors connected to automatic feeders. The fish were fed a commercial extruded diet (46% crude protein and 3600 kcal kg-1 of digestible energy). Animal growth was evaluated for all groups. After 30 days of experimentation, the fish stabilized their demands by adjusting their consumption. Amatitlania sp. showed predominantly diurnal FA and LA. All groups showed a peak of activity when the light was turned on and when it was turned off. In summary, FA and LA of Amatitlania sp. are predominantly diurnal and independent of social group. Pairs and groups of males and females together consume less food in relation to groups of one sex or the other due to reproductive behaviour. On the other hand, groups of only males or females consume more food because they lack reproductive stimuli and thus prioritize growth. These results may support good feeding management practices for this ornamental cichlid. Studies relating feeding behaviour with different social groups are of great importance for determining effective feeding strategies for this species in captivity. Thus, such a study assists in a more efficient production of Amatitlania sp.