Accuracy of the London Atlas to estimate the age of legal majority in a sample of the Amazon Regio.

Background: Estimating the age of majority is a challenging task in forensic odontology, especially because the third molars are usually the only developing teeth between the ages of 16 and 21 years. The London Atlas emerged as an alternative to estimate age using dental development, eruption and deciduous root resorption as parameters. The method has performed well in young age categories, while its performance for age estimation via third molars has been disputed. The present study tested the performance of the London Atlas to estimate the age of legal majority in a sample of individuals from the Amazon Region. Material and Methods: The sample consisted of 1.256 panoramic radiographs of women (n = 694) and men (n = 562) between 16 and 22.9 years. The method was applied to the maxillary (#28) and mandibular (#38) left third molars. For comparative purposes, the sample was divided into seven age groups: 16├ 16.99; 17├ 17.99; 18├ 18.99; 19├ 19.99; 20├ 20.99; 21├ 21.99; and 22├ 22.99 years. Chronological and estimated ages were compared descriptively by means of mean absolute errors (MAE) and root mean squared errors (RMSE), as well as through Receiver Operating Characteristic (ROC) curves and their area under the curve (AUC). Results: The MAE of the age estimates using tooth #28 was 1.76 years for females and 1.52 years for males. When the tooth #38 was used, the MAE for the females and males were 1.68 and 1.51 years, respectively. The MAE and RMSE increased in ascending order between age categories. Tooth #28 led to 74% of correct classifications around the age of legal majority, while tooth #38 reached 77%. The area under the curve was 0.75 for tooth #28 and 0.73 for tooth #38. Conclusions: The London Atlas should be used carefully to estimate the age of legal majority and not as a single method when the age threshold is 18 years. Key words:Age determination by teeth, forensic dentistry, forensic science, third molar.

On the forensic relevance of tattoos: distinguishing black inks with energy dispersive spectroscopy and backscattered scanning electron microscopy.

PURPOSE: This study aimed to analyze black tattoo inks by means of energy dispersive spectroscopy and backscattered scanning electron microscopy. METHODS: The sample consisted of five types of commercial tattoo pigments of the black colour (Easy Glow™, Electric Ink™, Iron Works™, Master Ink™, and Viper™). An Energy Dispersive Spectroscopy (EDS) detector (Silicon Drift Detector - SDD - type) attached to a Scanning Electron Microscope (SEM) device (Tescan Vega3 LMU, Libusina, Czech Republic) was used. X-ray characteristic signs were detected for each tattoo ink in an interval between 0 and 2.5 keV. The electron acceleration potential in the microscope was 15 keV. Two regions were analyzed for each sample (n = 10). On each region, a micrography of backscattered electrons (BSE) was obtained. Means and standard deviations (SD) of the weight percentages (Wt%) were calculated. RESULTS: C and O were predominant, with a mean O/C ratio between 2.69 and 2.74 Wt%. Electric Ink and Master Ink were the most similar pigments, while Easy Glow was the most distinctive - with agglomerates of Al that had a concentration 25 × higher than other specimens. Other compounds detected in the sample were Cl and Cu. CONCLUSION: EDS and SEM were efficient to distinguish black tattoo inks. These are our preliminary outcomes on the use of EDS and SEM to analyze black tattoo inks. Thus, careful interpretation is necessary to avoid rash applications in human identification practice.

Binary decisions of artificial intelligence to classify third molar development around the legal age thresholds of 14, 16 and 18 years.

Third molar development is used for dental age estimation when all the other teeth are fully mature. In most medicolegal facilities, dental age estimation is an operator-dependent procedure. During the examination of unaccompanied and undocumented minors, this procedure may lead to binary decisions around age thresholds of legal interest, namely the ages of 14, 16 and 18 years. This study aimed to test the performance of artificial intelligence to classify individuals below and above the legal age thresholds of 14, 16 and 18 years using third molar development. The sample consisted of 11,640 panoramic radiographs (9680 used for training and 1960 used for validation) of males (n = 5400) and females (n = 6240) between 6 and 22.9 years. Computer-based image annotation was performed with V7 software (V7labs, London, UK). The region of interest was the mandibular left third molar (T38) outlined with a semi-automated contour. DenseNet121 was the Convolutional Neural Network (CNN) of choice and was used with Transfer Learning. After Receiver-operating characteristic curves, the area under the curve (AUC) was 0.87 and 0.86 to classify males and females below and above the age of 14, respectively. For the age threshold of 16, the AUC values were 0.88 (males) and 0.83 (females), while for the age of 18, AUC were 0.94 (males) and 0.83 (females). Specificity rates were always between 0.80 and 0.92. Artificial intelligence was able to classify male and females below and above the legal age thresholds of 14, 16 and 18 years with high accuracy.

Psiconeurodontologia, (bio)decodificação dental e os mil e um nomes para o charlatanismo odontológico no século XXI: uma revisão de escopo / Psychoneurodontology, dental (bio)decoding and a thousand terms for odontological charlatanism in the 21st century: a scoping review

Junto às tecnologias que surgem para aprimorar a Odontologia mundial, emergem temáticas que progressivamente prosperam no obscuro e na incerteza. Tais "áreas odontológicas" se caracterizam por uma roupagem prolixa e de alto apelo público (como por meio de inferências sobre o emocional e os mistérios do corpo humano). Em tempos de disseminação da psiconeurodontologia e (bio)decodificação dental, assim como da incerteza acerca de sua confiabilidade, faz-se imperativa a busca pela fundamentação científica que sustenta esta temática. Para isto, foi realizada uma revisão de escopo seguindo as recomendações do PRISMA-ScR e JBI. Uma ampla busca foi realizada em cinco bases de dados e em parte da literatura cinza. Não houve restrição quanto ao ano de publicação e nem quanto ao idioma. Os dados foram coletados e expostos na forma narrativa/descritiva. De maneira complementar, utilizou-se a ferramenta JBI para avaliação da qualidade metodológica. A busca resultou em 4.785 registros iniciais, dos quais somente dois foram classificados como elegíveis. Ambos os estudos foram detectados a partir da literatura cinzenta e consistiam em trabalhos peruanos de conclusão de curso em nível de graduação (Bacharelado) em Odontologia. Os trabalhos aferiam a ocorrência de lesão de cárie em quadrantes de dentes permanentes e decíduos e correlacionavam com um questionário psicoemocional. Os dois trabalhos concluíram em prol da aplicação da psiconeurodontologia e da (bio)decodificação dental como ferramentas adicionais ao armamentário diagnóstico do Cirurgião-dentista. Detectou-se alto risco de viés, em especial acerca das limitações metodológicas dos estudos elegíveis. Tendo em vista os métodos e resultados observados nos estudos elegíveis, pode-se concluir que não há fundamentação científica na psiconeurodontologia ou na (bio)decodificação dental, muito menos validade e confiabilidade para a sua aplicação na prática clínica odontológica atual

Along with the technologies that emerged to improve Dentistry worldwide, there are themes that progressively dive into darkness and uncertainty. Such "dental areas" are characterized by a verbose approach and strong public appeal (such as through inferences about the emotions and mysteries of the human body). In times of dissemination of psychoneurodontology and dental (bio)decoding, as well as uncertainty about their reliability, the search for alleged scientific foundations that support this theme is imperative. To this end, a scoping review was carried out following the recommendations of PRISMA-ScR and JBI. A broad search was carried out in five databases and part of the gray literature. There was no restriction on the year of publication or language. Data were collected and presented in narrative/descriptive form. Complementarily, the JBI tool was used to assess methodological quality. The search resulted in 4.785 initial records, of which only two were classified as eligible. Both studies were detected from the gray literature and consisted of Peruvian studies at Bachelor's degree in Dentistry. The studies measured the occurrence of caries lesions in quadrants of permanent and deciduous teeth and correlated them with a psychoemotional questionnaire. The two studies concluded in favor of the application of psychoneurodontology and dental (bio)decoding as additional tools to the dental surgeon's diagnostic armamentarium. A high risk of bias was detected, especially regarding the methodological limitations of the eligible studies. Considering the methods and results observed in the eligible studies, it can be concluded that there is no scientific basis in psychoneurodontology or dental (bio)decoding, as well as lack of validity and reliability for its application in current clinical dental practice

Microbial Lipid Based Biorefinery Concepts: A Review of Status and Prospects.

The use of lignocellulosic biomass as a raw material for the production of lipids has gained increasing attention, especially in recent years when the use of food in the production of biofuels has become a current technology. Thus, the competition for raw materials for both uses has brought the need to create technological alternatives to reduce this competition that could generate a reduction in the volume of food offered and a consequent commercial increase in the value of food. Furthermore, the use of microbial oils has been studied in many industrial branches, from the generation of renewable energy to the obtainment of several value-added products in the pharmaceutical and food industries. Thus, this review provides an overview of the feasibility and challenges observed in the production of microbial lipids through the use of lignocellulosic biomass in a biorefinery. Topics covered include biorefining technology, the microbial oil market, oily microorganisms, mechanisms involved in lipid-producing microbial metabolism, strain development, processes, lignocellulosic lipids, technical drawbacks, and lipid recovery.

Dendrimers and Derivatives as Multifunctional Nanotherapeutics for Alzheimer's Disease.

Alzheimer's disease (AD) is the most prevalent form of dementia. It affects more than 30 million people worldwide and costs over US$ 1.3 trillion annually. AD is characterized by the brain accumulation of amyloid ß peptide in fibrillar structures and the accumulation of hyperphosphorylated tau aggregates in neurons, both leading to toxicity and neuronal death. At present, there are only seven drugs approved for the treatment of AD, of which only two can slow down cognitive decline. Moreover, their use is only recommended for the early stages of AD, meaning that the major portion of AD patients still have no disease-modifying treatment options. Therefore, there is an urgent need to develop efficient therapies for AD. In this context, nanobiomaterials, and dendrimers in particular, offer the possibility of developing multifunctional and multitargeted therapies. Due to their intrinsic characteristics, dendrimers are first-in-class macromolecules for drug delivery. They have a globular, well-defined, and hyperbranched structure, controllable nanosize and multivalency, which allows them to act as efficient and versatile nanocarriers of different therapeutic molecules. In addition, different types of dendrimers display antioxidant, anti-inflammatory, anti-bacterial, anti-viral, anti-prion, and most importantly for the AD field, anti-amyloidogenic properties. Therefore, dendrimers can not only be excellent nanocarriers, but also be used as drugs per se. Here, the outstanding properties of dendrimers and derivatives that make them excellent AD nanotherapeutics are reviewed and critically discussed. The biological properties of several dendritic structures (dendrimers, derivatives, and dendrimer-like polymers) that enable them to be used as drugs for AD treatment will be pointed out and the chemical and structural characteristics behind those properties will be analysed. The reported use of these nanomaterials as nanocarriers in AD preclinical research is also presented. Finally, future perspectives and challenges that need to be overcome to make their use in the clinic a reality are discussed.

Cupuassu juice fermentation by Lactiplantibacillus plantarum Lp62 produces an antioxidant enriched probiotic beverage.

The present work aimed to produce a cupuassu juice (Theobroma grandiflorum) fermented by the probiotic bacterium Lactiplantibacillus plantarum Lp62 and to analyze its antioxidant potential, antimicrobial activity, and resistance to biological barriers. The fermented beverage showed an increase in the content of phenolics, flavonoids, and antioxidant potential. The culture showed antagonistic activity against pathogens, but this result was not observed when the juice was tested. The probiotic strain remained viable under refrigeration, even in an acidified environment, and survived simulated gastrointestinal transit in vitro. L. plantarum Lp62 showed 30% adherence to HT-29 intestinal cells and proved to be safe in terms of antibiotic resistance and production of virulence factors. Fermentation increased the functional characteristics of cupuassu juice. This drink proved to be a good vehicle for the delivery of the probiotic bacteria L. plantarum Lp62.

Efficacy and adverse effects of ketamine versus electroconvulsive therapy for major depressive disorder: A systematic review and meta-analysis.

BACKGROUND: ECT is considered the fastest and most effective treatment for TRD. Ketamine seems to be an attractive alternative due to its rapid-onset antidepressant effects and impact on suicidal thoughts. This study aimed to compare efficacy and tolerability of ECT and ketamine for different depression outcomes (PROSPERO/CRD42022349220). METHODS: We searched MEDLINE, Web of Science, Embase, PsycINFO, Google Scholar, Cochrane Library and trial registries, which were the ClinicalTrials.gov and the World Health Organization's International Clinical Trials Registry Platform, without restrictions on publication date. SELECTION CRITERIA: randomized controlled trials or cohorts comparing ketamine versus ECT in patients with TRD. RESULTS: Eight studies met the inclusion criteria (of 2875 retrieved). Random-effects models comparing ketamine and ECT regarding the following outcomes were conducted: a) reduction of depressive symptoms severity through scales, g = -0.12, p = 0.68; b) response to therapy, RR = 0.89, p = 0.51; c) reported side-effects: dissociative symptoms, RR = 5.41, p = 0.06; nausea, RR = 0.73, p = 0.47; muscle pain, RR = 0.25, p = 0.02; and headache, RR = 0.39, p = 0.08. Influential & subgroup analyses were performed. LIMITATIONS: Methodological issues with high risk of bias in some of the source material, reduced number of eligible studies with high in-between heterogeneity and small sample sizes. CONCLUSION: Our study showed no evidence to support the superiority of ketamine over ECT for severity of depressive symptoms and response to therapy. Regarding side effects, there was a statistically significant decreased risk of muscle pain in patients treated with ketamine compared to ECT.

Aspects related to biofilm production and antifungal susceptibility of clinically relevant yeasts of the genus Trichosporon.

Trichosporonosis corresponds to a systemic fungal disease that leads to high mortality rates and is frequently associated with medical devices. It affects immunosuppressed patients in particular and is strongly linked to acquired human immunodeficiency, organ and tissue transplants, and malignant hematologic diseases such as leukemia and lymphomas. Trichosporon infections have been increasingly reported worldwide; however, little information is available either about their characteristics or the causative microorganism. Thus, the aims of the present study were: to investigate 59 yeasts of the genus Trichosporon by verifying the biofilm formation capacity of isolates; to analyze the susceptibility patterns of planktonic cells against the antifungals fluconazole, itraconazole, amphotericin-B, voriconazole, and caspofungin by comparing European Committee for Antimicrobial Susceptibility Testing (EUCAST) broth microdilution technique with the commercial method Etest; and to assess the susceptibility patterns of biofilm cells (sessile) against the same antifungals through broth microdilution. The ability to form biofilm on the surface of polystyrene plates was noted for all isolates, and 54.3% of samples were considered strong producers. Comparison between the antifungal susceptibility techniques evidenced that Etest showed higher and discordant minimum inhibitory concentrations (MICs) from those obtained by the microdilution method, especially for fluconazole, itraconazole, and caspofungin. Considering the susceptibility of biofilms, most species had high MIC50 and MIC90 against the tested antifungals, showing 4-to-66-fold higher concentrations for amphotericin B and 2-to-33-fold greater concentrations for caspofungin. These results highlight the importance of further studies with Trichosporon spp. for comparison between laboratory findings and in vivo response, considering both the susceptibility tests and the behavior of biofilm cells against drugs.

This study investigated 59 isolates of the medically important yeast Trichosporon in relation to their ability to form biofilms and the susceptibility of biofilms to antifungal agents. All isolates were able to produce biofilms and biofilms showed lower antifungal susceptibility.

Agricultural practices in Brazilian organic farms and microbiological characteristics of samples collected along the production chain.

AIMS: To gather data on agricultural practices in organic farms in Sao Paulo, Brazil, and evaluate their relationship with the microbiological characteristics of samples collected along the production chain. METHODS AND RESULTS: Practices data were based on field observations and interviews with farmers in 10 selected organic lettuce producing farms. Counts of Enterobacteriaceae and surveys for Salmonella were performed in samples of lettuce (before and after washing), fertilizers, irrigation and washing water, all collected in the same farm. Water samples were also tested for total coliforms and generic Escherichia coli. Isolated Enterobacteriaceae were identified by MALDI-TOF MS. Contamination of lettuce was influenced by some agricultural practices: chicken manure-based fertilization resulted in higher Enterobacteriaceae counts in lettuce when compared to other types of manure, whereas pre-washed lettuces presented lower microbial counts than non-pre-washed samples. Salmonella was detected in one lettuce sample by qPCR. Escherichia coli was detected in all irrigation water samples. All sample types contained Enterobacteriaceae species commonly reported as opportunistic human pathogens. CONCLUSIONS: The data highlight the need for improvement in the good agricultural practices in the studied farms. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides information on agricultural practices and microbiological characteristics of organic lettuce, contributing to the development of more accurate risk assessments.

Production and characterization of nanoemulsion with low-calorie structured lipids and its potential to modulate biomarkers associated with obesity and comorbidities.

Structured lipids (SL) containing behenic acid have been produced in order to obtain low-calorie lipids for foods; however, the development of a high nutritional value and a stable nanoemulsion carrier system for these SL is an interesting breakthrough for this field of research, improving technologic and biological potential for food application. In this sense, the aim of this study was to evaluate the stability of a nanoemulsion containing SL NeSL (produced with olive oil, soybean oil and fully hydrogenated crambe oil), the behavior during in vitro digestion and the effects on biomarkers involved in the obesity in cell models. The samples showed good stability throughout storage (30 days) under refrigeration and room temperature and after the gastric digestion phase compared to the controls (nanoemulsion of olive and soybean oil). After the intestinal phase, there was an increase in oil droplet size and zeta potential, a characteristic of coalescence. In the lipid accumulation model in adipocytes, the highest concentration (50 µL/mL) of NeSL resulted in 42% less lipid accumulation, compared to the control. Furthermore, the sample was able to reduce inflammatory cytokines produced by macrophages provoked by LPS (lipopolysaccharide). The combination of the oils in NeSL resulted in a fatty acid profile with beneficial health properties, which may have contributed to less lipid accumulation and improved inflammatory parameters. This SL in the form of a nanoemulsion, may be used as a partial fat substitute in low-calorie food products.

Relation between CANDIDA species isolated from vaginal mucosa and lesions caused by high-risk human papillomavirus hpv for cervical cancer

This study characterized and related yeasts of the genus Candida isolated from vaginal mucous membranes of women with lesions caused by high-risk HPV for cervical cancer. Forty-two women treated at the Lower Genital Tract Pathology Clinic of the University of São Paulo Medical School Hospital of Clinics were examined, with 30 high-grade (G1) uterine lesions with a mean age of 36.5 years ± 11. 1 and 12 with low grade (G2) uterine lesions with a mean age of 34.7 years ± 15.5. Clinical conditions and laboratory data on HPV were collected from patients' medical records; the socio-demographic data obtained from an appropriate questionnaire. For the study of association between the variables, Odds Ratio analysis was used from the STATA 13.1 program. Patients G1 had a higher prevalence for diabetes and the results indicated 27% prevalence of Candida spp. in vaginal mucosa, in G2 this was 33% in vaginal mucosa. Among the species found in vaginal mucosa of patients, Candida albicans was the most isolated with 88%, followed by C. tropicalis (8%) and C. glabrata (4%). The strains of C. albicans isolated from mucosa presented sensitivity to all antifungal agents tested, unlike the C. tropicalis strain isolated in G2 in vaginal mucosa, which presented a resistance profile to fluconazole. Thus, monitoring and supervision through clinical and laboratory testing of HPV patients is important, reinforcing the need for care, treatment and prevention of HPV-related infections and Candida spp.

Soft tissue thickness in Brazilian adults of different skeletal classes and facial types: A cone beam CT - Study.

The aim in this study was to evaluate the influence of skeletal class, facial type, and sex on soft tissue thickness (STT) of the craniofacial midline in a Brazilian subpopulation. Thus, 121 cone beam computed tomography (CBCT) scans (54 males and 67 females, age 21 to 40 yrs) composed the study sample. Patients were classified according to skeletal class (class I, II, and III) and facial type (brachycephalic, mesocephalic, and dolichocephalic), and STT was measured (mm) in 10 landmarks in the craniofacial midline for each CBCT scan. Multivariate analysis of covariance evaluated facial STT with regard to multiple independent variables (sex, age - covariate, facial type, and skeletal class). TEM and rTEM assessed the intra-examiner agreement. STT was significantly greater in males than in females for all regions measured (p < 0.05), except for the pogonion-pogonion' landmark (p>0.05). In general, class III individuals had significantly thicker soft tissue in the maxilla - subspinale-subnasale', prosthion-labrale superius', and incision-stomion' regions, while class II subjects had thicker soft tissue in the infradentale-labrale inferius' mandibular landmark (p < 0.05). Regarding facial type, dolichocephalic individuals showed significantly thicker soft tissue in the supramentale-supramentale' mandibular landmark, whereas brachycephalic subjects had thicker soft tissue in maxillary regions - prosthion-labrale superius' and incision-stomion' (p < 0.05). rTEM values were below 5% for most landmarks, and all TEM values were below 1 mm. Skeletal class and facial type influence STT, showing a soft tissue compensation, with deeper soft tissue in areas with lower skeletal development, and/or where bone is positioned more posteriorly.

Anti-glycation effect and the α-amylase, lipase, and α-glycosidase inhibition properties of a polyphenolic fraction derived from citrus wastes.

The advanced glycation end products (AGEs) constitute a wide variety of substances synthesized from interactions between amino groups of proteins and reducing sugars, which excess induces pathogenesis of chronic diseases. Brazil is the major producer of citrus, a low-cost source of hesperidin, which is a polyphenol recognized for its capacity to inhibit AGEs formation. This is the first work to evaluate the effects of a polyphenolic fraction derived from citrus wastes on the antiglycation and on the inhibition properties of digestive enzymes on the possibility to process these wastes in high value-added products. At concentrations of 10, 15 and 20 mg/mL inhibition of AGEs was higher than 60%. The extracts were able to inhibit by 76% the activity of pancreatic lipase and by 98% the activity of α-glucosidase. For the α-amylase the inhibition capacity was lower than 50%. Strong correlation was obtained among anti-glycation with polyphenolic content and antioxidant capacity.

Role of neuroendocrine modulation and biochemistry in the sepsis in Piaractus mesopotamicus.

Sepsis is a systemic process with multifactorial pathophysiology that affects most animal species. It is responsible for high rates of morbidity and mortality. This work aimed to study the biochemical and neuroendocrine changes of the sepsis process in Piaractus mesopotamicus after Aeromonas hydrophila inoculation analyzing changes in blood leukocyte and differences in neuroendocrine-biochemical modulation using RNA-seq. Fish showed hypercortisolemia, inhibition of glucose absorption, followed by hypocortisolemia and then hyperglycemia. Thyroid hormones (T3 and T4) showed immediate decrease in serum and T4 increased 6 h post-inoculation (HPI). Sepsis-induced hormonal alterations triggered changes in the metabolic pathways increasing protein and lipid catabolism, use of transient anaerobic glycolysis and liver injury. A reference transcriptome was constructed based on blood leukocytes from P. mesopotamicus. The assembly resulted in total 266,272 contigs with a N50 of 2786 bp. There was a reorganization of plasma membrane of leukocytes at the beginning of the septic process with increased expression of neuroendocrine receptors and with continuous flow of neurotransmitters, hormones and solutes with compensatory regulation at 6 HPI. Three and nine HPI seemed to be critical, the expression of a number of transcription factors was increased, including the modulatory DEGs related to glucocorticoid and thyroid hormones induced and suppressed (FDR < 0.05). Neuroendocrine modulation can regulate leukocytes and biochemical parameters of peripheral blood, being important sources for the study of the pathophysiology of sepsis. These finding highlights the importance of further studies focusing on biochemical-neuroendocrine changes in blood leukocytes and systemic sepsis.

Multicenter study of Candida species in oral mucosa of different patients: analysis of 711 strains and literature review

This study aimed to identify Candida spp. collected from oral mucosa and maintained in culture collections, correlating the findings with the medical history of patients and comparing with data from the literature over the past five years. Seven hundred and eleven oral Candida spp. isolates, collected between 2013 and 2017, were selected and identified using traditional and molecular methods. In addition, a literature review was performed with the key words: "Oral", "Candida" and "Yeast". Seven species of the genus Candida: were identified C. albicans(73.3%); C. tropicalis (9.3%); C. parapsilosis (8.2%); C. glabrata (3.9%); C. guilliermondii(2.8%); C. krusei (1.7%) and C. lusitaniae (0.3%). The strains identified as C. albicans were submitted to molecular methods using specific primers and of these, 5.8% were identified as C. dubliniensis strains. The greatest diversity of strains was found in patients presenting no systemic diseases or HIV +, while the highest percentage of strains of Candidanon-albicanswere observed in cancer patients. This study reports a representative distribution of Candidaspecies among individuals exhibiting distinct clinical conditions, in order to contribute to the design of future research on details of aspects involved in the infections caused by these microorganisms. The correct identification of oral Candida strains contributes to a realistic epidemiological approach and future clinical protocols against these pathogens

PAMAM dendrimers: blood-brain barrier transport and neuronal uptake after focal brain ischemia.

Drug delivery to the central nervous system is restricted by the blood-brain barrier (BBB). However, with the onset of stroke, the BBB becomes leaky, providing a window of opportunity to passively target the brain. Here, cationic poly(amido amine) (PAMAM) dendrimers of different generations were functionalized with poly(ethylene glycol) (PEG) to reduce cytotoxicity and prolong blood circulation half-life, aiming for a safe in vivo drug delivery system in a stroke scenario. Rhodamine B isothiocyanate (RITC) was covalently tethered to the dendrimer backbone and used as a small surrogate drug as well as for tracking purposes. The biocompatibility of PAMAM was markedly increased by PEGylation as a function of dendrimer generation and degree of functionalization. The PEGylated RITC-modified dendrimers did not affect the integrity of an in vitro BBB model. Additionally, the functionalized dendrimers remained safe when in contact with the bEnd.3 cells and rat primary astrocytes composing the in vitro BBB model after hypoxia induced by oxygen-glucose deprivation. Modification with PEG also decreased the interaction and uptake by endothelial cells of PAMAM, indicating that the transport across a leaky BBB due to focal brain ischemia would be facilitated. Next, the functionalized dendrimers were tested in contact with red blood cells showing no haemolysis for the PEGylated PAMAM, in contrast to the unmodified dendrimer. Interestingly, the PEG-modified dendrimers reduced blood clotting, which may be an added beneficial function in the context of stroke. The optimized PAMAM formulation was intravenously administered in mice after inducing permanent focal brain ischemia. Twenty-four hours after administration, dendrimers could be detected in the brain, including in neurons of the ischemic cortex. Our results suggest that the proposed formulation has the potential for becoming a successful delivery vector for therapeutic application to the injured brain after stroke reaching the ischemic neurons.