Characterization of Philadelphia-like Pre-B Acute Lymphoblastic Leukemia: Experiences in Mexican Pediatric Patients.

Ph-like subtypes with CRLF2 abnormalities are frequent among Hispano-Latino children with pre-B ALL. Therefore, there is solid ground to suggest that this subtype is frequent in Mexican patients. The genomic complexity of Ph-like subtype constitutes a challenge for diagnosis, as it requires diverse genomic methodologies that are not widely available in diagnostic centers in Mexico. Here, we propose a diagnostic strategy for Ph-like ALL in accordance with our local capacity. Pre-B ALL patients without recurrent gene fusions (104) were classified using a gene-expression profile based on Ph-like signature genes analyzed by qRT-PCR. The expressions of the CRLF2 transcript and protein were determined by qRT-PCR and flow cytometry. The P2RY8::CRLF2, IGH::CRLF2, ABL1/2 rearrangements, and Ik6 isoform were screened using RT-PCR and FISH. Surrogate markers of Jak2-Stat5/Abl/Ras pathways were analyzed by phosphoflow. Mutations in relevant kinases/transcription factors genes in Ph-like were assessed by target-specific NGS. A total of 40 patients (38.5%) were classified as Ph-like; of these, 36 had abnormalities associated with Jak2-Stat5 and 4 had Abl. The rearrangements IGH::CRLF2,P2RY8::CRLF2, and iAMP21 were particularly frequent. We propose a strategy for the detection of Ph-like patients, by analyzing the overexpression/genetic lesions of CRLF2, the Abl phosphorylation of surrogate markers confirmed by gene rearrangements, and Sanger sequencing.

Effect of surface treatment and aging on bond strength of composite resin onlays.

STATEMENT OF PROBLEM: Additional polymerization of indirect composite resins enhances their physical properties but lessens the potential for chemical bonding. PURPOSE: The purpose of this in vitro study was to evaluate the influence of different surface treatments and 6-month water storage on the microtensile bond strength (µTBS) of composite resin onlays. MATERIAL AND METHODS: Composite resin onlays (Filtek Z250) randomly received 6 different surface treatments: (1) airborne-particle abrasion with 27-µm alumina particles+Adper Scotchbond 1XT adhesive application, (2) airborne-particle abrasion with alumina particles+silane application (ESPE SIL)+Adper Scotchbond 1XT, (3) airborne-particle abrasion with alumina particles+Scotchbond Universal adhesive, (4) tribochemical silica coating with 30-µm particles (CoJet Sand)+Adper Scotchbond 1XT adhesive, (5) tribochemical silica coating+silane application+Adper Scotchbond 1XT, and (6) tribochemical silica coating+Scotchbond Universal adhesive. Onlays were luted to fresh composite resin specimens with RelyX Ultimate resin cement. Bonded assemblies were stored in water for 24 hours or 6 months at 37°C and subjected to the µTBS test. Additional surface-treated composite resin onlays were analyzed with a contact profilometer to determine average roughness, and micromorphologic changes were analyzed with scanning electron microscopy. RESULTS: Airborne-particle abrasion with alumina followed by Adper Scotchbond 1XT or Scotchbond Universal adhesive application provided the highest bond strength values at 24 hours. Lower values were obtained after tribochemical silica coating. After 6 months of artificial aging, airborne-particle abrasion with alumina or silica-coated alumina particles followed by Scotchbond Universal application yielded the greatest bond strength results. Airborne-particle abrasion with alumina produced the highest roughness values and a more irregular surface. CONCLUSION: Adhesive selection seems to be relevant to the µTBS of luted composite resin onlays after 6 months of water aging, as specimens treated with Scotchbond Universal, after alumina airborne-particle abrasion or tribochemical silica coating, yielded the highest values and better aging stability.

3D Graphene/silk fibroin scaffolds enhance dental pulp stem cell osteo/odontogenic differentiation.

OBJECTIVES: The current in vitro study aims to evaluate silk fibroin with and without the addition of graphene as a potential scaffold material for regenerative endodontics. MATERIAL AND METHODS: Silk fibroin (SF), Silk fibroin/graphene oxide (SF/GO) and silk fibroin coated with reduced graphene oxide (SF/rGO) scaffolds were prepared (n = 30). The microarchitectures and mechanical properties of scaffolds were evaluated using field emission scanning electron microscopy (FESEM), pore size and water uptake, attenuated total reflectance fourier transformed infrared spectroscopy (ATR-FTIR), Raman spectroscopy and mechanical compression tests. Next, the study analyzed the influence of these scaffolds on human dental pulp stem cell (hDPSC) viability, apoptosis or necrosis, cell adhesion, odontogenic differentiation marker expression and mineralized matrix deposition. The data were analyzed with ANOVA complemented with the Tukey post-hoc test (p < 0.005). RESULTS: SEM analysis revealed abundant pores with a size greater than 50 nm on the surface of tested scaffolds, primarily between 50 nm and 600 µm. The average value of water uptake obtained in pure fibroin scaffolds was statistically higher than that of those containing GO or rGO (p < 0.05). ATR-FTIR evidenced that the secondary structures did not present differences between pure fibroin and fibroin coated with graphene oxide, with a similar infrared spectrum in all tested scaffolds. Raman spectroscopy showed a greater number of defects in the links in SF/rGO scaffolds due to the reduction of graphene. In addition, adequate mechanical properties were exhibited by the tested scaffolds. Regarding biological properties, hDPSCs attached to scaffolds were capable of proliferating at a rate similar to the control, without affecting their viability over time. A significant upregulation of ALP, ON and DSPP markers was observed with SF/rGO and SF/GO groups. Finally, SF/GO and SF/rGO promoted a significantly higher mineralization than the control at 21 days. SIGNIFICANCE: Data obtained suggested that SF/GO and SF/rGO scaffolds promote hDPSC differentiation at a genetic level, increasing the expression of key osteo/odontogenic markers, and supports the mineralization of the extracellular matrix. However, results from this study are to be interpreted with caution, requiring further in vivo studies to confirm the potential of these scaffolds.

Long-Term In Vitro Adhesive Properties of Two Universal Adhesives to Dentin.

The composition of universal adhesives, as well as the adhesive strategy, may influence bonding effectiveness and durability. This study aimed to evaluate the microtensile bond strength (µTBS) and nanoleakage (NL), immediately and after 6-month aging, and in situ degree of conversion (DC), of two universal adhesives (Scotchbond Universal Adhesive, SBU; Xeno Select, XEN) applied with etch-and-rinse (ER) and self-etch (SE) strategies, in comparison with a two-step SE adhesive (Clearfil SE Bond, CSE). Dentin surfaces of fifty human third molars were randomly assigned to the following adhesives: two universal adhesives, SBU and XEN, applied in ER or SE mode and CSE, used as control. Teeth were sectioned into beams to be tested under µTBS, half of them after 24 h, and the rest after 6 months of water aging. Selected beams from each tooth were used for NL evaluation and in situ DC quantification. SBU and CSE showed significantly higher mean µTBS and lower nanoleakage than XEN, regardless of the evaluation time and adhesion strategy. XEN-SE yielded the lowest degree of conversion. Therefore, adhesive properties of universal adhesives to dentin are material dependent, regardless of the adhesion strategy, exhibiting XEN a significantly worse performance than SBU.

Exploring the therapeutic potential of an antinociceptive and anti-inflammatory peptide from wasp venom.

Animal venoms are rich sources of neuroactive compounds, including anti-inflammatory, antiepileptic, and antinociceptive molecules. Our study identified a protonectin peptide from the wasp Parachartergus fraternus' venom using mass spectrometry and cDNA library construction. Using this peptide as a template, we designed a new peptide, protonectin-F, which exhibited higher antinociceptive activity and less motor impairment compared to protonectin. In drug interaction experiments with naloxone and AM251, Protonectin-F's activity was decreased by opioid and cannabinoid antagonism, two critical antinociception pathways. Further experiments revealed that this effect is most likely not induced by direct action on receptors but by activation of the descending pain control pathway. We noted that protonectin-F induced less tolerance in mice after repeated administration than morphine. Protonectin-F was also able to decrease TNF-α production in vitro and modulate the inflammatory response, which can further contribute to its antinociceptive activity. These findings suggest that protonectin-F may be a potential molecule for developing drugs to treat pain disorders with fewer adverse effects. Our results reinforce the biotechnological importance of animal venom for developing new molecules of clinical interest.

Dopamine-loaded nanoparticle systems circumvent the blood-brain barrier restoring motor function in mouse model for Parkinson's Disease.

Parkinson's disease (PD) is a progressive and chronic neurodegenerative disease of the central nervous system. Early treatment for PD is efficient; however, long-term systemic medication commonly leads to deleterious side-effects. Strategies that enable more selective drug delivery to the brain using smaller dosages, while crossing the complex brain-blood barrier (BBB), are highly desirable to ensure treatment efficacy and decrease/avoid unwanted outcomes. Our goal was to design and test the neurotherapeutic potential of a forefront nanoparticle-based technology composed of albumin/PLGA nanosystems loaded with dopamine (ALNP-DA) in 6-OHDA PD mice model. ALNP-DA effectively crossed the BBB, replenishing dopamine at the nigrostriatal pathway, resulting in significant motor symptom improvement when compared to Lesioned and L-DOPA groups. Notably, ALNP-DA (20 mg/animal dose) additionally up-regulated and restored motor coordination, balance, and sensorimotor performance to non-lesioned (Sham) animal level. Overall, ALNPs represent an innovative, non-invasive nano-therapeutical strategy for PD, considering its efficacy to circumvent the BBB and ultimately deliver the drug of interest to the brain.

Animal venoms: therapeutic tools for tackling Parkinson's disease.

Parkinson's disease (PD) is a neurodegenerative pathology of the central nervous system, mainly involving the selective and progressive loss of dopaminergic neurons from the substantia nigra, resulting in motor and non-motor symptoms. PD remains an incurable ailment; thus, treatments are limited to symptom alleviation. With long-term use, conventional treatments can become inefficient, often triggering possible side effects. Considering these drawbacks, drug discovery constantly turns to nature as a source of efficient therapeutics. Thus, this review explores animal venoms as a rich source of bioactive compounds with potent neuropharmacological profiles for the development of effective adjuvant treatments with fewer side effects, ultimately aiming for the neuroprotection of dopaminergic neurons and the symptomatic relief of PD.

The Müller-Lyer illusion as a tool for schizophrenia screening.

The perceptual deficit hypothesis for schizophrenia is based on more general models of normal human visual perception, which have traditionally postulated that objects must compete for attention and processing space in the visual system. Recent evidence suggests that susceptibility of schizophrenics to the Müller-Lyer (ML) illusion may be a marker of vulnerability, detectable in prodromic patients, but disappearing with the progression of the illness. This illusion consists of overestimating the length of a straight line with converging arrowheads at the ends, while underestimating those with diverging arrowheads. Although the ML illusion has been shown to occur in touch as well as vision, it is not known whether abnormal contextual suppression extends to other sensory modalities in schizophrenics. Another challenge consists in verifying whether different visual parameters of the illusion which favor the magnocellular and parvocellular systems would have diverse ML illusion effects in schizophrenia. In this review we present data showing the degree of illusion in capuchin monkeys (Cebus spp.), a possible animal model for schizophrenia. To this end, a computer program was developed to conduct experiments in humans and non-human primates, allowing the display of illusory figures, manipulation of the stimuli's exposure time, interval between stimuli and number of trials. In the non-primate experiments, the visual illusion test based on achromatic ML illusion figures indicated the presence of the ML illusory effect in 10 capuchin monkeys. These results suggest that Cebus might be a good model for the experimental study of schizophrenia.

Arthropod toxins and their antinociceptive properties: From venoms to painkillers.

The complex process of pain control commonly involves the use of systemic analgesics; however, in many cases, a more potent and effective polypharmacological approach is needed to promote clinically significant improvement. Additionally, considering side effects caused by current painkillers, drug discovery is once more turning to nature as a source of more efficient therapeutic alternatives. In this context, arthropod venoms contain a vast array of bioactive substances that have evolved to selectively bind to specific pharmacological targets involved in the pain signaling pathway, playing an important role as pain activators or modulators, the latter serving as promising analgesic agents. The current review explores how the pain pathway works and surveys neuroactive compounds obtained from arthropods' toxins, which function as pain modulators through their interaction with specific ion channels and membrane receptors, emerging as promising candidates for drug design and development.

Perception of the Müller-Lyer illusion in capuchin monkeys (Cebus apella).

Visual illusions are formed by differences between the perception of one figure and its real physical characteristics. The Müller-Lyer illusion is the best known and most studied geometric illusion, consisting in the subject's judgment between two parallel lines that have the same size, one flanked with outward-pointing arrowheads, and the other with inward-pointing arrowheads. These arrowheads act as inductors that make the lines to be perceived as having different sizes, inward-pointing stimuli being estimated as longer. This study aimed to investigate the Müller-Lyer illusion in capuchin monkeys (Cebus apella), a New World primate not yet investigated for this illusion. For this purpose, stimuli were presented on a touch screen monitor. Ten adult subjects (five females and five males) were used. Before the tests, they were trained to discriminate between two physically different lines with and without arrowheads. The longer lines were always the positive (rewarded) stimuli. Regarding the Müller-Lyer Illusion test, all monkeys, unrespective of gender, demonstrated susceptibility to the illusion, by choosing preferentially the line with inward-pointing arrowheads. In order to determine the degree of the illusion, a point of subjective equality test (PSE) was performed. The PSE without arrowheads values were lower than the PSE with arrowheads. Thus, it was demonstrated that capuchin monkeys were susceptible to the Müller-Lyer illusion, once the perception of the lines' size was influenced by the presence of the arrowheads and by their orientation.

Mechanical characterization of resin cements used for luting fiber posts by nanoindentation.

OBJECTIVES: To evaluate the mechanical behavior of resin cements used for luting fiber post. The influence of the curing mode is analysed. METHODS: Nanoindentation techniques have been applied to determine hardness and Young's modulus of disc-shaped specimens of three types of cements: chemical- , photo- and dual-cured, provided by Ivoclar-Vivadent. Results obtained have been compared with measurements performed inside the post-cement-dentin system. Mechanical properties are evaluated together with scanning electron micrographs showing the post-cement and dentin-cement interfaces. RESULTS AND SIGNIFICANCE: Differences have been detected between mechanical measurements performed inside the post-cement-dentin system and those carried out in laboratory disc-shaped specimens. The close presence of post and dentin boundaries has a lot of influence on the cement behavior. The nanoindentation measurements indicate that the photo-cured cement exhibits a high hardness and stiffness, but with a more marked tendency to brittle failure. The chemically cured cement is the material with higher ability to bear deformation without damage, although its hardness and elastic modulus are significantly lower. Dual-cured cements present the best combination of properties.

Photodynamic therapy mediated by acai oil (Euterpe oleracea Martius) in nanoemulsion: A potential treatment for melanoma.

Melanoma is the most aggressive and lethal form of skin cancer, responsible for >80% of deaths. Standard treatments for late-stage melanoma usually present poor results, leading to life-threatening side effects and low overall survival. Thus, it is necessary to rethink treatment strategies and design new tools for the treatment of this disease. On that ground, we hereby report the use of acai oil in nanoemulsion (NanoA) as a novel photosensitizer for photodynamic therapy (PDT) used to treat melanoma in in vitro and in vivo experimental models. NIH/3T3 normal cells and B16F10 melanoma cell lines were treated with PDT and presented 85% cell death for melanoma cells, while maintaining high viability in normal cells. Flow cytometry indicated that cell death occurred by late apoptosis/necrosis. Tumor bearing C57BL/6 mice treated five times with PDT using acai oil in nanoemulsion showed tumor volume reduction of 82% in comparison to control/tumor group. Necrotic tissue per tumor area reached its highest value in PDT-treated mice, supporting PDT efficacy. Overall, acai oil in nanoemulsion was an effective photosensitizer, representing a promising source of new photosensitizing molecules for PDT treatment of melanoma, a tumor with an inherent tendency to be refractory for this type of therapy.

Pharmacological Alternatives for the Treatment of Neurodegenerative Disorders: Wasp and Bee Venoms and Their Components as New Neuroactive Tools.

Neurodegenerative diseases are relentlessly progressive, severely impacting affected patients, families and society as a whole. Increased life expectancy has made these diseases more common worldwide. Unfortunately, available drugs have insufficient therapeutic effects on many subtypes of these intractable diseases, and adverse effects hamper continued treatment. Wasp and bee venoms and their components are potential means of managing or reducing these effects and provide new alternatives for the control of neurodegenerative diseases. These venoms and their components are well-known and irrefutable sources of neuroprotectors or neuromodulators. In this respect, the present study reviews our current understanding of the mechanisms of action and future prospects regarding the use of new drugs derived from wasp and bee venom in the treatment of major neurodegenerative disorders, including Alzheimer's Disease, Parkinson's Disease, Epilepsy, Multiple Sclerosis and Amyotrophic Lateral Sclerosis.

Morphological Analysis of Reticuloendothelial System in Capuchin Monkeys (Sapajus spp.) after Meso-2,3-Dimercaptosuccinic Acid (DMSA) Coated Magnetic Nanoparticles Administration.

Magnetic nanoparticles can be used for numerous in vitro and in vivo applications. However, since uptake by the reticuloendothelial system represents an obstacle for the achievement of nanoparticle diagnostic and therapeutic goals, the aim of the present study was to evaluate the uptake of dimercaptosuccinic acid coated magnetic nanoparticles by reticuloendothelial system phagocytic cells present in lymph nodes, spleen, and liver tissue and how the presence of these particles could have an impact on the morphology of these organs in capuchin monkeys (Sapajus spp.). Animals were intravenously injected with dimercaptosuccinic acid coated magnetic nanoparticles and euthanized 12 hours and 90 days post-injection. Organs were processed by transmission electron microscopy and histological techniques. Samples of spleen and lymph nodes showed no morphological changes. Nevertheless, liver samples collected 90 days post-administration showed slight morphological alteration in space of Disse. Moreover, morphometrical analysis of hepatic mitochondria was performed, suggesting a clear positive correlation between mitochondrial area and dimercaptosuccinic acid coated magnetic nanoparticles administration time. The present results are directly relevant to current safety considerations in clinical diagnostic and therapeutic uses of magnetic nanoparticles.

Neuroactive compounds obtained from arthropod venoms as new therapeutic platforms for the treatment of neurological disorders.

The impact of neurological disorders in society is growing with alarming estimations for an incidence increase in the next decades. These disorders are generally chronic and can affect individuals early during productive life, imposing real limitations on the performance of their social roles. Patients can have their independence, autonomy, freedom, self-image, and self-confidence affected. In spite of their availability, drugs for the treatment of these disorders are commonly associated with side effects, which can vary in frequency and severity. Currently, no effective cure is known. Nowadays, the biopharmaceutical research community widely recognizes arthropod venoms as a rich source of bioactive compounds, providing a plethora of possibilities for the discovery of new neuroactive compounds, opening up novel and attractive opportunities in this field. Several identified molecules with a neuropharmacological profile can act in the central nervous system on different neuronal targets, rendering them useful tools for the study of neurological disorders. In this context, this review aims to describe the current main compounds extracted from arthropod venoms for the treatment of five major existing neurological disorders: stroke, Alzheimer's disease, epilepsy, Parkinson's disease, and pathological anxiety.

Microhardness of superficial and deep sound human dentin.

Our purpose in this study was to determine the microhardness of superficial and deep dentin by means of two indentation methods (Knoop and Vickers) under two different applied loads. Twelve dentin discs approximately 2-mm thick were obtained from both superficial and deep dentin by transversally sectioning the crowns of sound, extracted human third molars with a diamond blade under water irrigation. Dentin surfaces were sequentially polished, and indentations (n = 20 per surface) were performed with either Vickers indentor at loads of 300 and 500 g, respectively, or Knoop indentor at loads of 50 and 100 g, respectively. Average Vickers hardness number (VHN) and Knoop hardness number (KHN) were calculated and treated with two-way analysis of variance (ANOVA) and Student's t test. Microhardness of dentin was not influenced by the different loads applied for both indentation methods. Knoop hardness was significantly higher for superficial than for deep dentin (p < 0.05). Conversely, Vickers hardness was not significantly different for both substrates (p > 0.05). Differences in dentin hardness as a function of depth exist, but they might not be relevant, and no alteration of the distribution of stresses along the adhesive interface is expected.

Tensile strength and microhardness of treated human dentin.

OBJECTIVES: To determine the ultimate tensile strength and Knoop hardness of mineralized, EDTA-treated, sodium hypochlorite (NaOCl)-treated, EDTA-treated resin-infiltrated, and NaOCl-treated resin-infiltrated dentin. METHODS: Dumbell-shaped specimens with a cross-sectional area of 0.5 mm2 were prepared from the crowns of extracted human third molars. Specimens were randomly assigned to the following experimental groups: (1) mineralized dentin; (2) 0.5 M EDTA-demineralized dentin, pH 7/5 days; (3) 5% NaOCl-deproteinized dentin/2 days; (4) EDTA-treated, Single Bond resin-infiltrated dentin; (5) NaOCl-treated, Single Bond resin-infiltrated dentin. All specimens were tested in tension in a Vitrodyne testing machine at 0.6 mm/min. Knoop microhardness was measured on the fractured edges of specimens in groups 1, 3, 4, and 5. Results were analyzed by ANOVA and SNK tests (p < 0.05). RESULTS: Both EDTA and NaOCl treatments caused significant reductions in the tensile strength and microhardness of mineralized dentin (p < 0.05) with the largest reductions observed after NaOCl treatment (p < 0.05) Resin infiltration of treated dentin resulted in moderate increase of its tensile strength and microhardness, however, the original mineralized values were not recovered (p < 0.05). SIGNIFICANCE: Whenever dentin surfaces are treated with EDTA or NaOCl prior to a clinical bonding procedure, clinicians must be aware that a weak layer may be present at the interface, which may lead to premature failures of resin/dentin bonds.

Sorption and solubility of resin-based restorative dental materials.

OBJECTIVES: To measure the water sorption and solubility of different resin-based restorative dental materials. METHODS: Eight commercial restorative materials were selected: two resin composites (Z100 and Prodigy), four polyacid-modified resin composites (Compoglass, Compoglass F, Dyract and Dyract AP), and two light-cured glass ionomers (Vitremer and Fuji II LC). Five disc specimens were prepared of each material, following the manufacturer's instructions, and were grounded wet with silicon carbide paper. Water sorption and solubility of the different materials were calculated by means of weighting the samples before and after water immersion and desiccation. Data were analyzed by one-way ANOVA and Student-Newman-Keuls tests (P<0.05). RESULTS: Compoglass and Compoglass F showed the lowest values of water sorption and solubility, while Vitremer and Fuji II LC displayed the highest values. Solubility values of Prodigy, Z100, Dyract and Dyract AP did not show significant differences among them, while their water sorption values attained some differences and were lower for Prodigy followed by Dyract and Z100. CONCLUSIONS: The attained water sorption and solubility values are mainly influenced by the generic type of material and variations occurring between materials of the same type may result from differences in resin matrix compositions.

Perspectives on the application of nanotechnology in photodynamic therapy for the treatment of melanoma.

Malignant melanoma is the most aggressive form of skin cancer and has been traditionally considered difficult to treat. The worldwide incidence of melanoma has been increasing faster than any other type of cancer. Early detection, surgery, and adjuvant therapy enable improved outcomes; nonetheless, the prognosis of metastatic melanoma remains poor. Several therapies have been investigated for the treatment of melanoma; however, current treatment options for patients with metastatic disease are limited and non-curative in the majority of cases. Photodynamic therapy (PDT) has been proposed as a promising minimally invasive therapeutic procedure that employs three essential elements to induce cell death: a photosensitizer, light of a specific wavelength, and molecular oxygen. However, classical PDT has shown some drawbacks that limit its clinical application. In view of this, the use of nanotechnology has been considered since it provides many tools that can be applied to PDT to circumvent these limitations and bring new perspectives for the application of this therapy for different types of diseases. On that ground, this review focuses on the potential use of developing nanotechnologies able to bring significant benefits for anticancer PDT, aiming to reach higher efficacy and safety for patients with malignant melanoma.