Long non-coding RNAs are essential for Schistosoma mansoni pairing-dependent adult worm homeostasis and fertility.

The trematode parasite Schistosoma mansoni causes schistosomiasis, which affects over 200 million people worldwide. Schistosomes are dioecious, with egg laying depending on the females' obligatory pairing with males. Long non-coding RNAs (lncRNAs) are transcripts longer than 200 nucleotides with low or no protein-coding potential that have been involved in other species with reproduction, stem cell maintenance, and drug resistance. In S. mansoni, we recently showed that the knockdown of one lncRNA affects the pairing status of these parasites. Here, we re-analyzed public RNA-Seq data from paired and unpaired adult male and female worms and their gonads, obtained from mixed-sex or single-sex cercariae infections, and found thousands of differentially expressed pairing-dependent lncRNAs among the 23 biological samples that were compared. The expression levels of selected lncRNAs were validated by RT-qPCR using an in vitro unpairing model. In addition, the in vitro silencing of three selected lncRNAs showed that knockdown of these pairing-dependent lncRNAs reduced cell proliferation in adult worms and their gonads, and are essential for female vitellaria maintenance, reproduction, and/or egg development. Remarkably, in vivo silencing of each of the three selected lncRNAs significantly reduced worm burden in infected mice by 26 to 35%. Whole mount in situ hybridization experiments showed that these pairing-dependent lncRNAs are expressed in reproductive tissues. These results show that lncRNAs are key components intervening in S. mansoni adult worm homeostasis, which affects pairing status and survival in the mammalian host, thus presenting great potential as new therapeutic target candidates.

De novo assembly of the complete mitochondrial genome of Mycetophylax simplex Emery, 1888 through organelle targeting revels no substantial expansion of gene spacers, but rather some slightly shorter genes.

Mitochondria play a key role in cell biology and have their own genome, residing in a highly oxidative environment that induces faster changes than the nuclear genome. Because of this, mitochondrial markers have been exploited to reconstruct phylogenetic and phylogeographic relationships in studies of adaptation and molecular evolution. In this study, we determined the complete mitogenome of the fungus-farming ant Mycetophylax simplex (Hymenoptera, Formicidae) and conducted a comparative analysis among 29 myrmicine ant mitogenomes. Mycetophylax simplex is an endemic ant that inhabits sand dunes along the southern Atlantic coast. Specifically, the species occur in the ecosystem known as "restinga", within the Atlantic Forest biome. Due to habitat degradation, land use and decline of restinga habitats, the species is considered locally extinct in extremely urban beaches and is listed as vulnerable on the Brazilian Red List (ICMBio). We employed a mitochondrion-targeting approach to obtain the complete mitogenome through high-throughput DNA sequencing technology. This method allowed us to determine the mitogenome with high performance, coverage and low cost. The circular mitogenome has a length of 16,367 base pairs enclosing 37 genes (13 protein-coding genes, 22 tRNAs and 2 rRNAs) along with one control region (CR). All the protein-coding genes begin with a typical ATN codon and end with the canonical stop codons. All tRNAs formed the fully paired acceptor stems and fold into the typical cloverleaf-shaped secondary structures. The gene order is consistent with the shared Myrmicinae structure, and the A + T content of the majority strand is 81.51%. Long intergenic spacers were not found but some gene are slightly shorter. The phylogenetic relationships based on concatenated nucleotide and amino acid sequences of the 13 protein-coding genes, using Maximum Likelihood and Bayesian Inference methods, indicated that mitogenome sequences were useful in resolving higher-level relationship within Formicidae.

The complete mitochondrial genome of Sporophila nigricollis (Aves, Passeriformes).

BACKGROUND: Sporophila nigricollis, popularly known as Yellow-bellied Seedeater, is a commonly trafficked bird in Brazil. This study describes the complete mitogenome of the species and its phylogenetic position. METHODS AND RESULTS: DNA sample was sequenced on MiSeq (Illumina®) sequencer. Reads were assembled to reference using Geneious. A total of 11,740 paired sequence reads were used to produce the complete mitochondrial DNA sequence with average coverage of 176x. The mitogenome was found to be circular in shape and consisted of 16,777 base pairs. The mitochondrial genome was annotated with MITOS and verified with ExPASy. Its structure contains two ribosomal RNA (rRNA) genes, 22 transporter RNA (tRNA) genes, 13 protein-coding genes (PCGs) and one control region. Twelve of the 13 PCGs have ATG as start codon. Seven of the 13 PCGs contain a TAA stop codon. Most of the tRNA genes and PCGs are encoded on the heavy strand. Phylogenetic analyses were conducted with MEGA using the maximum likelihood method. Sporophila nigricollis grouped together with other Thraupidae species. CONCLUSION: This study presents the first complete mitogenome of Sporophila nigricollis and can be useful for research on evolution, ecology and conservation of this species.

Gene therapy targeting the blood-brain barrier improves neurological symptoms in a model of genetic MCT8 deficiency.

A genetic deficiency of the solute carrier monocarboxylate transporter 8 (MCT8), termed Allan-Herndon-Dudley syndrome, is an important cause of X-linked intellectual and motor disability. MCT8 transports thyroid hormones across cell membranes. While thyroid hormone analogues improve peripheral changes of MCT8 deficiency, no treatment of the neurological symptoms is available so far. Therefore, we tested a gene replacement therapy in Mct8- and Oatp1c1-deficient mice as a well-established model of the disease. Here, we report that targeting brain endothelial cells for Mct8 expression by intravenously injecting the vector AAV-BR1-Mct8 increased tri-iodothyronine (T3) levels in the brain and ameliorated morphological and functional parameters associated with the disease. Importantly, the therapy resulted in a long-lasting improvement in motor coordination. Thus, the data support the concept that MCT8 mediates the transport of thyroid hormones into the brain and indicate that a readily accessible vascular target can help overcome the consequences of the severe disability associated with MCT8 deficiency.

Experimental pastes containing niobophosphate and 45S5 bioactive glasses for treatment of dentin hypersensitivity: dentin permeability and tubule obliteration.

OBJECTIVES: This study tested the ability of bioactive pastes containing niobophosphate and 45S5 glasses to reduce dentin permeability and to obliterate dentinal tubules, as a mean of reducing human dentin hypersensitivity. MATERIALS AND METHODS: Experimental pastes with concentrations of 10, 20, and 30 wt% of two bioactive glasses (45S5 or niobophosphate [NbG]) were formulated. A paste without bioactive glass (placebo) and a commercial paste (Nano P, FGM) were used as controls. Forty dentin disc specimens were obtained from caries-free extracted third human molars and divided in 8 groups (n = 5). Percentage of permeability (%Lp) was assessed in a dental permeability machine considering hydraulic conductance, immediately after pastes application and at day 7, day 14, and day 21. The precipitates formed on the surface of the dentin discs (and dentinal tubules) were analyzed by SEM/EDS and micro-Raman spectra. Data of dentin permeability (%) 2-way repeated-measures (ANOVA) and Holm-Sidak post-tests (α = 0.05). Dentinal tubule obliteration was visually (and elemental) evaluated and descriptively reported. RESULTS: The experimental bioactive glass pastes containing NbG and 45S5, regardless of the concentration, reduced dentin permeability in comparison with pastes without bioactive glasses (P < 0.05). The formulated placebo and commercial paste did not reduce permeability over time (P < 0.05). SEM/EDS and micro-Raman analyses showed that both type of bioactive pastes (NbG or 45S5-based) presented mineral precipitates obliterating the dentinal tubules at day 21. NbG seems to offer a better initial effect than 45S5, while at 21 days there is no difference between both glasses. CONCLUSION: Experimental bioactive pastes containing NbG and 45S5 (at concentrations of 10%, 20%, or 30%) have potential to reduce dentin permeability (over time) in comparison with pastes without bioactive glasses; and this occurs on behalf of obliteration of dentinal tubules by microparticle and precipitate formation. CLINICAL RELEVANCE: Bioactive pastes containing NbG and 45S5 may benefit patients presenting dentin hypersensitivity, because these pastes can start acting fast after application and maintain their action up to 21 days.

Ceftriaxone pretreatment confers neuroprotection in rats with acute glaucoma and reduces the score of seizures induced by pentylenotetrazole in mice.

ß-Lactam antibiotics such as ceftriaxone, are potent stimulators of the expression of l-glutamate transporter GLT-1 and may exert neuroprotective effects when chronically used in rats and mice. In this study, we used two animal models to test the neurological effect of subchronic treatment with ceftriaxone: experimental acute glaucoma in Wistar rats and induction of acute seizures with pentylenetetrazole in mice. We also assessed the performance of mice in the rotarod to calculate therapeutic indexes and exploratory activity in the open field. Our results showed that subchronic use of ceftriaxone was neuroprotective in both models, reducing injury in acute ischemia and ischemia/reperfusion in specific layers of retina and leading to a decrease in the seizure severity score. In behavioral experiments, we observed that ceftriaxone increased hyperactivity followed by a decrease in exploratory behavior in the open field, and there was no motor impairment in the rotarod test. We conclude that ceftriaxone may be useful as a tool in the development of new neuroprotective drugs targeting diseases which present a possible dysfunction in the balance of glutamatergic neurotransmission.

Pneumosinus dilatans following dermal filler injections.

When an adverse event occurs after a dermal filler treatment, simple ultrasound can be of help for diagnosis. But sometimes a more sophisticated imaging technique as MRI is needed to find the true reason for a complication. As in the case of pneumosinus dilatans after injection of a dermal filler, we present. The relation of this rare disorder and filler injection is discussed.

Detecting acetyl-coenzyme a carboxylase resistance gene in rice (Oryza sativa L.).

Herbicides inhibiting acetyl-coenzyme A carboxylase (ACCase) are very effective in controlling grass weeds including weedy-rice in paddy rice production systems. The ACCase inhibitor affects the enzyme by blocking fatty acid biosynthesis resulting in plant death. The herbicide resistance in rice is conferred by a single point mutation with an amino acid substitution of the carboxyl transferase domain of the ACCase gene. An assay based on the tetra-primer ARMS-PCR method was developed to detect the SNP G2027T that causes a tryptophan-cysteine substitution in the gene encoding chloroplastic ACCase in rice. The protocol was tested in 453 rice samples from a segregant population for validation of the assay. This technique can be exploited to monitor resistant lines in rice breeding programs to detect homozygous or heterozygous resistant genotypes and homozygous susceptible genotypes. The presence of resistant ACCase allele(s) can be detected with rapidity, simplicity, at low cost and can be used in any molecular biology laboratory with minimal equipment.

Etoposide-Loaded Poly(Lactic-co-Glycolic Acid) Intravitreal Implants: In Vitro and In Vivo Evaluation.

Etoposide-loaded poly(lactic-co-glycolic acid) implants were developed for intravitreal application. Implants were prepared by a solvent-casting method and characterized in terms of content uniformity, morphology, drug-polymer interaction, stability, and sterility. In vitro drug release was investigated and the implant degradation was monitored by the percent of mass loss. Implants were inserted into the vitreous cavity of rabbits' eye and the in vivo etoposide release profile was determined. Clinical examination and the Hen Egg Test-Chorioallantoic Membrane (HET-CAM) method were performed to evaluate the implant tolerance. The original chemical structure of the etoposide was preserved after incorporation in the polymeric matrix, which the drug was dispersed uniformly. In vitro, implants promoted sustained release of the drug and approximately 57% of the etoposide was released in 50 days. In vivo, devices released approximately 63% of the loaded drug in 42 days. Ophthalmic examination and HET-CAM assay revealed no evidence of toxic effects of implants. These results tend to show that etoposide-loaded implants could be potentially useful as an intraocular etoposide delivery system in the future.

DNA-Mediated Proximity-Based Assembly Circuit for Actuation of Biochemical Reactions.

Smart nanodevices that integrate molecular recognition and signal production hold great promise for the point-of-care (POC) diagnostic applications. Herein, the development of a DNA-mediated proximity assembly of biochemical reactions, which was capable of sensing various bio-targets and reporting easy-to-read signals is reported. The circuit was composed of a DNA hairpin-locked catalytic cofactor with inhibited activity. Specific molecular inputs can trigger a conformational switch of the DNA locks through the mechanisms of toehold displacement and aptamer switching, exposing an active cofactor. The subsequent assembly of an enzyme/cofactor pair actuated a reaction to produce colorimetric or fluorescence signals for detecting target molecules. The developed system could be potentially applied to smart biosensing in molecular diagnostics and POC tests.

Lipid and protein fingerprinting for Fusarium oxysporum f. sp. cubense strain-level classification.

Banana is one of the most popular fruits in the world but has been substantially impaired by Panama disease in the last years. Fusarium oxysporum f. sp. cubense (Foc) is the causal agent and colonizes banana cultivars from many subgroups with different aggressiveness levels, often leading to plant death while compromising new crops in infested areas. This study has evaluated the ability of MALDI-MS protein and lipid fingerprinting to provide intraspecies classification of Foc isolates and to screen biomolecules related to host-pathogen relationship. The MS data, when inspected via partial least square discriminant analysis (PLS-DA), distinguished the isolates by aggressiveness as well as by specific location and host. Although both lipids and proteins show discriminating tendencies, these differences were more clearly perceived via the protein profiles. Considering that Cavendish cultivar is the more resistant option to endure Foc presence in the field, the lipids and proteins related to this subgroup might have an important role in pathogen adaptation. This study reports a new application of MALDI-MS for the analysis of a banana pathogen with intraspecies classification ability. Graphical abstract MALDI-MS classified Foc isolates by aggressiveness level on banana revealing the additional influence of location and host cultivar on the expression of lipids and proteins.

Isoforms of Hsp70-binding human LDL in adult Schistosoma mansoni worms.

Schistosoma mansoni is one of the most common parasites infecting humans. They are well adapted to the host, and this parasite's longevity is a consequence of effective escape from the host immune system. In the blood circulation, lipoproteins not only help to conceal the worm from attack by host antibodies but also act as a source of lipids for S. mansoni. Previous SEM studies showed that the low-density lipoprotein (LDL) particles present on the surface of adult S. mansoni worms decreased in size when the incubation time increased. In this study, immunocytochemical and proteomic analyses were used to locate and identify S. mansoni binding proteins to human plasma LDL. Ultrathin sections of adult worms were cut transversely from the anterior, medial and posterior regions of the parasite. Immunocytochemical experiments revealed particles of gold in the tegument, muscle region and spine in male worms and around vitelline cells in females. Immunoblotting and 2D-electrophoresis using incubations with human serum, anti-LDL antibodies and anti-chicken IgG peroxidase conjugate were performed to identify LDL-binding proteins in S. mansoni. Analysis of the binding proteins using LC-MS identified two isoforms of the Hsp70 chaperone in S. mansoni. Hsp70 is involved in the interaction with apoB in the cytoplasm and its transport to the endoplasmic reticulum. However, further studies are needed to clarify the functional role of Hsp70 in S. mansoni, mainly related to the interaction with human LDL.

Influence of pH change and water storage on the sealing ability of two resin-based root-filling materials.

AIM: To evaluate the Influence of pH change and water storage up to 90 days on the sealing ability of two resin-based root-filling materials. MATERIALS AND METHODS: Forty-four human mandibular single-rooted teeth were instrumented and filled with gutta-percha/ AH Plus or Resilon/Epiphany SE (n=20 per group). Two teeth each were used as positive and negative controls. Specimens were set for 7 days under 100% humidity at 37°C. They were allocated into two subgroups (n=10) according to whether they were tested immediately or stored for up to 90 days in water before testing. Sealing ability was evaluated by passive dye penetration. Absorbance at 630 nm (in µg/ml) was measured by spectrophotometry. The pH values were obtained in triplicate. Data were submitted to ANOVA by post-hoc Tukey's test (α=0.05). RESULTS: Specimens filled with Resilon/Epiphany SE exhibited more leakage than specimens filled with gutta-percha/AH Plus at the immediate time point (p<0.001). No differences were detected between the groups after storage, or between the materials with pH changes after 30, 60 and 90 days (p>0.05). CONCLUSION: Gutta-percha/AH Plus provided superior sealing at the immediate time point. Water storage and pH changes did not Influence the sealing ability of tested materials. CLINICAL SIGNIFICANCE: These results suggest that Resilon/ Epiphany SE sealer offered no apparent advantage over the more conventional gutta-percha/AH Plus sealer technique in terms of sealing ability.

The Effect of 3% Phosphate Ascorbyl Gel on Bond Strength of Composite Resin to Enamel treated with 35% Hydrogen Peroxide.

OBJECTIVE: To evaluate the effect of 3% phosphate ascorbyl gel (PA) in different times onto the microshear bond strength of composite resin (CR) to bovine enamel treated with 35% hydrogen peroxide (HP). MATERIALS AND METHODS: Thirty enamel blocks of bovine incisors were made and divided into 5 groups (n = 6) with three specimens per group (n = 18), according to treatment: G1= No bleaching + CR; G2 = HP + CR after 15d; G3 = HP + CR after 24 hours; G4 = HP + PA (15 min) + CR after 24 hours; G5 = HP + PA (2 hours) + CR after 24 hours. The resin cylinders were made by Tygon matrices. Microshear bond strength test was performed using universal testing machine with a 50N load at a speed of 0.5 mm/min. Fracture modes were assessed by a stereomicroscope 40 ×. Microshear bond strength values were submitted to the analysis of variance (ANOVA) one-way and Tukey test (p < 0.05). RESULTS: G1 had significant results when compared to G3 and G5 (p < 0.01). However, G2, G3, G4 and G5 have showed no significant differences among groups (p > 0.05). Failure modes were categorized into adhesive (90%) and mixed (10%). CONCLUSION: The use of 3% phosphate ascorbyl gel for 15 minutes was able to improve bond strength of composite resin to bleached bovine enamel, but when 3% phosphate ascorbyl gel was applied during 40 minutes it negatively interfered in the adhesion of the resin to bleached bovine enamel.

[Women in situation of violence by their intimate partner: making a decision to seek a specialized violence support service]. / Mulheres em situação de violência pelo parceiro íntimo: tomada de decisão por apoio institucional especializado.

OBJECTIVE: This study aimed to evaluate the factors involved in the decision making process of women in situation of interpersonal violence perpetrated by an intimate partner in search for domestic violence support services. METHODS: Qualitative, descriptive and exploratory research. Sixteen women were interviewed through individual semi-structured interviews, at a Women's Reference Center in Duque de Caxias, Rio de Janeiro, Brazil. Data analysis was performed using the technique of collective subject discourse was used. RESULTS: The results showed that these women sought psychosocial support when interpersonal violence reached their limit of tolerance. The group meetings organized by the Reference Center made it possible for these women to recover their self-esteem and confidence and to increase awareness of the need to change their lives and achieve economic autonomy. CONCLUSIONS: In this scenario, by collecting data for the research, the nurses contributed to the promotion of gender equality and women's empowerment, as recommended by the Millennium Development Goals.

Left Ventricular Pseudoaneurysm: A Bursting Threat.

A 55-year-old man submitted to emergency surgery due to cardiac perforation by stabbing. One month later, he presented with chest pain, and a transthoracic echocardiogram revealed moderate-severe mitral regurgitation. After 6 months, a new transthoracic echocardiogram showed a left ventricular pseudoaneurysm, being later submitted to pseudoaneurysm exclusion and mitral valvuloplasty.

The Role of Platelet-Rich Plasma Injection for Muscle Strains in Athletes.

Muscle tears/strains are among the most common musculoskeletal injuries, posing a serious challenge for sports medicine. Aiming to reduce the time to return to play and the rate of reinjuries, apart from the traditional conservative treatments and rehabilitation protocols, new and innovative therapeutic options have emerged, particularly platelet-rich plasma (PRP). This study aims to present the available evidence regarding PRP injection for the treatment of muscle strains in athletes. Two databases were searched for articles published between January 2012 and December 2022 in Portuguese or English. The query used for the PubMed database was ("Muscles/injuries"[Mesh]) AND ("Athletes"[Mesh] OR "Athletic Injuries"[Mesh]) AND "Platelet-Rich Plasma"[Mesh], while for the Web of Science database the search was performed for "Platelet-rich plasma" AND "Muscle injuries" AND ("Athletes" OR "Athletic injuries"). Eleven studies involving athletes diagnosed with muscle injuries who received treatment with PRP injection alone, or in combination with traditional conservative treatment, compared to a control group, were included. Four randomized controlled trials, four systematic reviews/meta-analyses, two retrospective studies, and one comparative study were included. Current evidence from the highest-quality studies does not support the hypothesis of reduction of time to return to play and the rate of reinjuries after PRP injection, even though some studies reported positive results. However, the available evidence suggests that PRP might have a beneficial effect on the pain perceived by athletes following an acute muscle strain. It is challenging to arrive at definitive conclusions and translate these findings into a clinical context for treating muscle strains in athletes. The existing trials present several inconsistencies and limitations, with a heterogeneous set of patients and injuries, as well as the use of different and inconsistent methods for preparing, administering, and measuring the effects of PRP. To achieve consistent outcomes, standardizing PRP administration procedures is essential.

MMP-8 in Periodontal Sites of Postpartum and without-Any-Pregnancy Women.

The hypothesis that physiological changes in women can affect periodontal tissues is the subject of this study, and inflammatory markers such as matrix metalloproteinase-8 can measure susceptibility to inflammation. The study aimed to analyze MMP-8 levels in periodontal sites of postpartum women and women without a history of pregnancy, comparing health parameters and periodontal disease. This is a case-control study with 40 participants, 20 cases (women in the postpartum period) and 20 controls (women without any pregnancy), who underwent clinical periodontal examination and the collection of crevicular gingival fluid. The ELISA test was used to detect MMP-8 levels. Postpartum women had worse periodontal parameters, such as bleeding index on probing, number of sites with CAL ≥ 3, and fewer teeth present. In the group of women without a history of pregnancy, a significantly lower MMP-8 level was observed in healthy sites and a higher one was observed in periodontal pockets (p < 0.01). In contrast, in postpartum women, MMP-8 levels were elevated in both healthy sites and periodontal pockets (p > 0.01). The MMP-8 levels in gingival fluid appear to be related to periodontal clinical parameters and may be a possible marker of enzymatic changes involved in periodontal tissue destruction in postpartum women.

Antiepileptic profile of Parawixin-11, purified from Parawixia bistriata spider venom (Araneae, Araneidae), in Wistar rats.

The pharmacological treatment of epilepsy is often complex due to the lack of efficacy in many patients and profound side effects from current drugs, including sedation, motor impairment, and teratogenesis. In the quest for new antiepileptic drugs, animal venoms offer a valuable source of neuroactive molecules targeting ion channels and neurotransmitter receptors. This study investigates the antiepileptic potential of compounds isolated from the venom of the Parawixia bistriata spider. One compound, designated Parawixin-11, demonstrated significant anticonvulsant effects when injected into the cerebral ventricle in a dose-response manner. It effectively countered seizures induced by bicuculline (ED50 0.16 µg/animal), pentylenetetrazole (ED50 0.08 µg/animal), strychnine (ED50 0.05 µg/animal), pilocarpine (ED50 0.10 µg/animal), and NMDA (ED50 0.008 µg/animal). We also assessed whether intracerebroventricular administration of Parawixin-11 caused motor or cognitive impairments in rats using the open field, rotarod, and Morris water maze tests. No differences in exploration or movement were observed with doses of 0.3, 0.2, or 0.1 µg of Parawixin-11. Although there was an increased latency to find the platform during the acquisition phase of the Morris water maze test, no differences in spatial memory retention were noted. Given Parawixin-11's potency against NMDA-induced seizures, we hypothesize that it may modulate the glutamatergic system, aligning with the mechanisms of several spider-derived polyamines.