Targeted nanotherapeutics for the treatment of Helicobacter pylori infection.

Helicobacter pylori infection is involved in gastric diseases such as peptic ulcer and adenocarcinoma. Approved antibiotherapies still fail in 10 to 40% of the infected patients and, in this scenario, targeted nanotherapeutics emerged as powerful allies for H. pylori eradication. Nano/microparticles conjugated with H. pylori binding molecules were developed to eliminate H. pylori by either (i) blocking essential mechanisms of infection, such as adhesion to gastric mucosa or (ii) binding and killing H. pylori through the release of drugs within the bacteria or at the site of infection. Glycan antigens (as Lewis B and sialyl-Lewis X), pectins, lectins, phosphatidylethanolamine and epithelial cell membranes were conjugated with nano/microparticles to successfully block H. pylori adhesion. Urea-coated nanoparticles were used to improve drug delivery inside bacteria through H. pylori UreI channel. Moreover, nanoparticles coated with antibodies against H. pylori and loaded with sono/photosensitizers, were promising for their application as targeted sono/photodynamic therapies. Further, non-specific H. pylori nano/microparticles, but only active in the acidic gastric environment, coated with binders to bacterial membrane, extracellular polymeric substances or to high temperature requirement A protease, were evaluated. In this review, an overview of the existing nanotherapeutics targeting H. pylori will be given and their rational, potential to counteract infection, as well as level of development will be presented and discussed.

Engineering resveratrol-loaded chitosan nanoparticles for potential use against Helicobacter pylori infection.

Helicobacter pylori (H. pylori) is a microorganism directly linked to severe clinical conditions affecting the stomach. The virulence factors and its ability to form biofilms increase resistance to conventional antibiotics, growing the need for new substances and strategies for the treatment of H. pylori infection. The trans-resveratrol (RESV), a bioactive polyphenol from natural sources, has a potential activity against this gastric pathogen. Here, Chitosan nanoparticles (NP) containing RESV (RESV-NP) were developed for H. pylori management. The RESV-NP were prepared using the ionic gelation method and characterized by Dynamic Light Scattering (DLS), Nanoparticle Tracking Analysis (NTA) and, Cryogenic Transmission Electron Microscopy (Cryo - TEM). The encapsulation efficiency (EE) and in vitro release rate of RESV were quantified using high-performance liquid chromatography (HPLC). RESV-NP performance against H. pylori was evaluated by the quantification of the minimum inhibitory/bactericidal concentrations (MIC/MBC), time to kill, alterations in H. pylori morphology in its planktonic form, effects against H. pylori biofilm and in an in vitro infection model. RESV-NP cytotoxicity was evaluated against AGS and MKN-74 cell lines and by hemolysis assay. Acute toxicity was tested using Galleria mellonella model assays. RESV-NP showed a spherical shape, size of 145.3 ± 24.7 nm, polydispersity index (PDI) of 0.28 ± 0.008, and zeta potential (ZP) of + 16.9 ± 1.81 mV in DLS, while particle concentration was 3.12 x 1011 NP/mL (NTA). RESV-NP EE was 72 %, with full release within the first 5 min. In microbiological assays, RESV-NP presented a MIC/MBC of 3.9 µg/mL, a time to kill of 24 h for complete eradication of H. pylori. At a concentration of 2xMIC (7.8 µg/mL), RESV-NP completely eradicated the H. pylori biofilm, and in an in vitro infection model, RESV-NP (4xMIC - 15.6 µg/mL) showed a significant decrease in bacterial load (1 Log10CFU/mL) when compared to the H. pylori J99 control. In addition, they did not demonstrate a toxic character at MIC concentration for both cell lines. The use of the RESV-NP with mucoadhesion profile is an interesting strategy for oral administration of substances targeting gastric disorders, linked to H. pylori infections.

One-Pot Microfluidics to Engineer Chitosan Nanoparticles Conjugated with Antimicrobial Peptides Using "Photoclick" Chemistry: Validation Using the Gastric Bacterium Helicobacter pylori.

Surface bioconjugation of antimicrobial peptides (AMP) onto nanoparticles (AMP-NP) is a complex, multistep, and time-consuming task. Herein, a microfluidic system for the one-pot production of AMP-NP was developed. Norbornene-modified chitosan was used for NP production (NorChit-NP), and thiolated-AMP was grafted on their surface via thiol-norbornene "photoclick" chemistry over exposure of two parallel UV LEDs. The MSI-78A was the AMP selected due to its high activity against a high priority (level 2) antibiotic-resistant gastric pathogen: Helicobacter pylori (H. pylori). AMP-NP (113 ± 43 nm; zeta potential 14.3 ± 7 mV) were stable in gastric settings without a cross-linker (up to 5 days in pH 1.2) and bactericidal against two highly pathogenic H. pylori strains (1011 NP/mL with 96 µg/mL MSI-78A). Eradication was faster for H. pylori 26695 (30 min) than for H. pylori J99 (24 h), which was explained by the lower minimum bactericidal concentration of soluble MSI-78A for H. pylori 26695 (32 µg/mL) than for H. pylori J99 (128 µg/mL). AMP-NP was bactericidal by inducing H. pylori cell membrane alterations, intracellular reorganization, generation of extracellular vesicles, and leakage of cytoplasmic contents (transmission electron microscopy). Moreover, NP were not cytotoxic against two gastric cell lines (AGS and MKN74, ATCC) at bactericidal concentrations. Overall, the designed microfluidic setup is a greener, simpler, and faster approach than the conventional methods to obtain AMP-NP. This technology can be further explored for the bioconjugation of other thiolated-compounds.

Infected Spinal Hematoma After Accidental Exteriorization of an Epidural Catheter: A Case Report.

Optimal treatment and outcome after spinal hematoma remain unclear. Close neurological monitoring is the key to successful treatment. Here, we present a case of an infected spinal hematoma management. We report the case of a 51-year-old male patient, American Society of Anesthesiologists physical status I, admitted to a level I hospital due to right lower limb necrotizing fasciitis. A lumbar epidural catheter was placed for pain control without complications. On the 26th day of hospitalization, three hours after the accidental exteriorization of the epidural catheter, the patient developed severe back pain not relieved by systemic analgesia. Prophylactic low-molecular-weight heparin had been administered less than six hours before. The patient had inflammatory signs and digital pressure pain at the catheter insertion site and a positive Brudzinski sign. Sensory-motor deficits were never felt. An urgent dorsolumbar MRI exhibited a significant hematic collection at the subdural and anterior epidural level, and an infected epidural hematoma was assumed. Empirical antibiotics and hourly monitoring of sensory-motor deficits, meningeal signs, and fever were initiated. The patient remained on absolute bed rest. Low-molecular-weight heparin was stopped. The pain disappeared on the third day after the MRI. Citrobacter koseri was isolated. A re-evaluation CT scan did not show spinal compression. The patient was discharged on the 27th day after an MRI in an asymptomatic condition and was referred to acute pain consultation. Neurological deficits are usually expected at the time of spinal hematoma diagnosis. However, the classic triad of pain, sensory deficits, and motor deficits is only seen in less than half of patients. Our patient only developed severe lumbar pain. In selected cases, diagnostic MRI associated with tight monitoring and conservative management can be applied.

Impact of Ketamine on Quality of Recovery after Laparoscopic Surgery: A Single-Centre Single-Blinded Trial Using the QoR-15 Questionnaire.

Background: The quality of recovery is a cluster of patient-related outcomes that emphasise not only pain but different physical and emotional dimensions. Traditionally, ketamine is used to improve postoperative analgesia and avoid opioid consumption and opioid-related side effects. Objective: The present study sought to evaluate if intraoperative ketamine administration (as a part of multimodal analgesia) influences the quality of recovery after laparoscopic surgery. Design: A prospective two-armed, single-blinded trial. Settings. Tertiary single-centre trial between July 2021 and January 2022. Patients. From the 146 patients initially admitted to the study, 127 patients were enrolled, 60 in the ketamine group (group K) and 67 in the control group (group NK). Intervention. Both groups received a rigid intraoperative anaesthesia protocol; furthermore, in group K, 0.5 mg/kg of the ideal body weight of ketamine was administered. Main Outcome Measures. The primary outcome was to evaluate the effect of ketamine administration on the postoperative quality of recovery using the Portuguese version of the Quality of Recovery-15 (QoR-15) Questionnaire 24 h after surgery. The total score and minimal clinically significant difference (MCID) of the QoR-15 were compared. Other variables were also assessed such as the presence of emergence delirium (ED), the Numeric Rating Scale (NRS) for pain, and the presence of postoperative nausea and vomiting (PONV). Results: A total of 127 patients were allocated to the study groups, 60 in group K and 67 in group NK. Regarding the primary outcome, no differences were found in individual categories (15 items) and in the total score of QoR-15 (p=0.214). Concerning improvement (MCID ≥ 8) or worsening (MCID ≤ 8) in quality of recovery, no difference was found between the groups (24 vs. 32 and 6 vs. 6; p=0.776). Finally, no difference was found in secondary postoperative outcomes including ED (p=0.55), NRS (p=0.401), and PONV (p=0.55). Conclusion: In this study, the administration of ketamine in laparoscopic surgery had no impact on the quality of recovery 24 h after surgery. This trial is registered with NCT03724019.

Grafting MSI-78A onto chitosan microspheres enhances its antimicrobial activity.

MSI-78A (Pexiganan A) is one of the few antimicrobial peptides (AMPs) able to kill Helicobacter pylori, a pathogenic bacterium that colonizes the gastric mucosa of half of the world's population. Antibiotics fail in 20-40% of H. pylori-infected patients, reinforcing the need for alternative treatments. Herein, a bioengineered approach was developed. MSI-78A with a C-terminal cysteine was grafted onto chitosan microspheres (AMP-ChMic) by thiol-maleimide (Michael-addition) chemistry using a long heterobifunctional spacer (NHS-PEG113-MAL). Microspheres with ∼4 µm diameter (near H. pylori length) and stable at low pH were produced by spray drying using a chitosan solution with an incomplete genipin crosslinking. A 3 × 10-5 µg AMP/microsphere grafting was estimated/confirmed by UV/Vis and FTIR spectroscopies. AMP-ChMic were bactericidal against H. pylori J99 (highly pathogenic human strain) at lower concentrations than the free peptide (∼277 µg grafted MSI-78A-SH/mL vs 512 µg free MSI-78A-SH/mL), even after pre-incubation in simulated gastric conditions with pepsin. AMP-ChMic killed H. pylori by membrane destabilization and cytoplasm release in a ratio of ∼10 bacteria/microsphere. This can be attributed to H. pylori attraction to chitosan, facilitating the interaction of grafted AMP with bacterium membrane. Overall, it was demonstrated that the peptide-microsphere conjugation chemistry did not compromise the MSI-78A antimicrobial activity, instead it boosted its bactericidal performance against H. pylori. STATEMENT OF SIGNIFICANCE: Half of the world's population is infected with Helicobacter pylori, a gastric bacterium that is responsible for 90% of non-cardia gastric cancers. Therefore, H. pylori eradication is now advocated in all infected individuals. However, available antibiotic therapies fail in up to 40% patients. Antimicrobial peptides (AMPs) are appealing alternatives to antibiotics, but their high susceptibility in vivo limits their clinical translation. AMP immobilization onto biomaterials surface will overcome this problem. Herein, we demonstrate that immobilization of MSI-78A (one of the few AMPs with activity against H. pylori) onto chitosan microspheres (AMP-ChMic) enhances its anti-H. pylori activity even at acidic pH (gastric settings). These results highlight the strong potential of AMP-ChMic as an antibiotic alternative for H. pylori eradication.

Extracellular matrix electrospun membranes for mimicking natural renal filtration barriers.

Kidney diseases are recognized as a major health problem, which affect 10% of the population. Because currently available therapies have many limitations, some tissue engineering strategies have been emerging as promising approaches in this field. In this work, porcine kidneys were decellularized to obtain decellularized kidney extracellular matrix (dKECM).1 Our results demonstrate a successful protocol of decellularization characterized by the removal of nucleic acid material and preservation of collagen and glycosaminoglycans. Blends of polycaprolactone (PCL)2 and dKECM were prepared by electrospinning and characterized. The biological performance of the membranes was tested with a human kidney cell line (HK-2) for 7 days. It was observed that cellular metabolic activity, proliferation and protein content increased with an increase in dKECM concentrations (30, 50 and 70%). Additionally, the expression of zona occludens-1 was revealed on dKECM-containing membranes but not on pure PCL membranes. To the best of our knowledge this is the first time that natural extracellular matrix is used to mimic the kidney basement membrane as an in vitro model. This could be a valuable tool for regenerative nephrology and may have an impact on the development of kidney advanced therapies in the future.

Development of non-orthogonal 3D-printed scaffolds to enhance their osteogenic performance.

Three-dimensional (3D)-printed polycaprolactone (PCL)-based scaffolds have been extensively proposed for Tissue Engineering (TE) applications. Currently, the majority of the scaffolds produced are not representative of the complex arrangement of natural structures, since the internal morphologies follow an orthogonal and regular pattern. In order to produce scaffolds that more closely replicate the structure of the extracellular matrix (ECM) of tissues, herein both circular and sinusoidal scaffolds were fabricated and compared to their conventional orthogonal counterparts. This is an innovative, versatile and efficient strategy to 3D print PCL scaffolds with unique curved geometries. The morphology and the mechanical behavior of the scaffolds were assessed. The biological response was analyzed by evaluating the cell seeding efficiency, cell adhesion, proliferation, and osteogenic activity of an osteoblastic-like cell line seeded in these scaffolds. The scaffolds were designed and produced to have a similar porosity of about 56%. The non-orthogonal structures demonstrated lead to higher values of Young's modulus, both under dry conditions and when immersed in PBS. Moreover, the biological data corroborate that non-orthogonal scaffolds influence the cellular responses in a positive manner, namely in the osteogenic activity when compared with the orthogonal controls. These results suggest that introducing less orthogonal elements, which better mimic the tissue ECM and architecture, may have a positive influence on the cellular behavior, being a potential strategy to address bone tissue engineering applications.

Cardiac channelopathies: The role of sodium channel mutations. / Canalopatias cardíacas: o papel das mutações nos canais de sódio.

INTRODUCTION AND OBJECTIVES: The importance of sodium channels for the normal electrical activity of the heart is emphasized by the fact that mutations (inherited or de novo) in genes that encode for these channels or their associated proteins cause arrhythmogenic syndromes such as the Brugada syndrome and the long QT syndrome (LQTS). The aim of this study is to conduct a review of the literature on the mutations in the sodium channel complex responsible for heart disease and the implications of a close relationship between genetics and the clinical aspects of the main cardiac channelopathies, namely at the level of diagnosis, risk stratification, prognosis, screening of family members and treatment. METHODS: The online Pubmed® database was used to search for articles published in this field in indexed journals. The MeSH database was used to define the following query: "Mutation [Mesh] AND Sodium Channels [Mesh] AND Heart Diseases [Mesh]", and articles published in the last 15 years, written in English or Portuguese and referring to research in human beings were included. CONCLUSIONS: In the past few years, significant advances have been made to clarify the genetic and molecular basis of these syndromes. A greater understanding of the underlying pathophysiological mechanisms showed the importance of the relationship between genotype and phenotype and led to progress in the clinical approach to these patients. However, it is still necessary to improve diagnostic capacity, optimize risk stratification, and develop new specific treatments according to the genotype-phenotype binomial.

Convertirse en madre durante la adolescencia: activación del rol materno en el control prenatal / Become a mother during adolescence: activation of the maternal role during prenatal control

Introducción: Las adolescentes gestantes constituyen un grupo poblacional vulnerable, con limitación en la construcción de la identidad personal frente a la materna e imposibilidad de constitución de una familia estable que afectan su calidad de vida y la del hijo por nacer. El objetivo del artículo es dar a conocer el alcance que tiene la aplicación de una estrategia de cuidado de enfermería que promovió el cuidado integral de la adolescente gestante en la activación del rol materno durante el control prenatal. Materiales y Métodos: Este artículo es el resultado de la aplicación de una estrategia de cuidado de enfermería: programa de atención integral a la adolescente gestante durante el control prenatal, con base en la teoría de Mercer, y estructurado desde el marco lógico; la estrategia contó con la participación de diez adolescentes en gestación y el equipo interdisciplinario. Resultados: Adopción e implementación de la estrategia y apertura Institucional del centro de apoyo y cuidado para adolescentes gestantes “Creciendo Juntos”, que promovió el cuidado integral y permitió la activación del rol materno durante el control prenatal. Discusión: El convertirse en madre adolescente, no se activa sin la influencia del cuidado estructurado brindado por enfermería quien lidera el equipo de salud, quienes permiten a las gestantes, disminuir sus miedos, conocer y vivir el proceso de gestación y aprender las tareas para trascender hacia la activación del rol materno. Conclusiones: La Estrategia valida la visibilidad del cuidado sustentado en una teoría de enfermería, al dar respuesta a las necesidades humanas.

Introduction: Pregnant adolescents are a vulnerable population group, with limitations on the construction of personal identity against maternal and inability to establish a stable family that affect their quality of life and the unborn child. The objective of this article is to present the scope of the application of a strategy for nursing care that promoted the comprehensive care of pregnant adolescent activation of the maternal role during prenatal care. Materials and Methods: This article is the result of the implementation of a strategy for nursing care: comprehensive care program for pregnant adolescent during antenatal care, based on the theory of Mercer, and structured from the logical framework; the strategy counted with the participation of ten adolescent pregnancy and the interdisciplinary team. Results: Adoption and implementation of the strategy and center opening Institutional support and care for pregnant adolescents "Growing Together", which promoted the comprehensive care and allowed the activation of the maternal role during prenatal care. Discussion: Becoming a teenage mother, is not activated without the influence of careful structured provided by nurses who heads the health team, who allow pregnant women to reduce their fears, knowing and living the gestation and learn the tasks to transcend to the activation of the maternal role. Conclusions: The Strategy validates the visibility of sustained care in a nursing theory, to respond to human needs.

Assessment of pandemic preparedness in a socially vulnerable community in south Texas.

The purpose of this research was to obtain information about general and pandemic preparedness efforts of residents within San Patricio County in South Texas, as well as to identify the most effective means of communicating the risks posed by pandemic influenza. The population of San Patricio County is socially vulnerable to a variety of disasters, including influenza pandemics due to the unique demographic profile of the county as well as its location on the Gulf Coast. The goals of this study were to help with pandemic planning efforts and to provide recommendations that could serve as a foundation for building more resilient communities within San Patricio County. Clearly the various governmental levels must work together to assist communities prepare for pandemic preparedness but broad, inclusive community participation is also necessary to strengthen community resilience.

Dermatofibroma in a black tattoo: report of a case.

Tattooing has been associated with a variety of complications including inflammatory and granulomatous reactions, transmission of infections, and neoplasms. We report a case of a 24-year-old male who presented with a 2-month history of an erythematous nodule involving a newly made tattoo on the right leg. An excisional biopsy was performed and the histopathological evaluation was consistent with dermatofibroma. Only three cases of dermatofibroma associated with tatooing were reported in literature. We report an additional case and review the literature regarding cutaneous reactions to tattoos.

Dermatofibroma in a black tattoo: report of a case / Dermatofibroma sob pigmento preto de tatuagem: relato de um caso

Tattooing has been associated with a variety of complications including inflammatory and granulomatous reactions, transmission of infections, and neoplasms. We report a case of a 24-year-old male who presented with a 2-month history of an erythematous nodule involving a newly made tattoo on the right leg. An excisional biopsy was performed and the histopathological evaluation was consistent with dermatofibroma. Only three cases of dermatofibroma associated with tatooing were reported in litetature. We report an additional case and review the literature regarding cutaneous reactions to tattoos.

Tatuagens têm sido associadas com uma variedade de complicações incluindo reações inflamatórias e granulomatosas, transmissão de infecções e neoplasias. Relatamos um caso de homem com 24 anos de idade que apresentava há dois meses nódulo eritematoso sob pigmento preto de uma tatuagem na coxa direita. A biópsia excisional foi realizada e a avaliação histológica foi consistente com dermatofibroma. Apenas três casos da associação dermatofibroma e tatuagem foram relatados na literatura. Nós reportamos um caso adicional e revisamos a literatura sobre reações cutâneas em tatuagens.

Multiple cavitary pulmonary nodules in association with pyoderma gangrenosum: case report.

Pyoderma gangrenosum is a rare neutrophilic disease of unknown origin that is associated with systemic diseases in 50% of cases. It is characterized by erythematous-violaceous nodular lesions that quickly progress to painful ulcers, with undermined edges, necrotic-hemorrhagic, varying in size and depth, located mainly in the lower limbs. Extracutaneous locations of pyoderma gangrenosum are rare, usually involving the lungs; the main differential diagnosis in these cases is Wegener granulomatosis. We report a case of pyoderma gangrenosum, which showed multiple cavitary lung nodules, with good response to high doses of steroids. Once excluded the possibility of Wegener granulomatosis, the authors concluded that it was the manifestation of systemic pyoderma gangrenosum with pulmonary involvement.

Multiple cavitary pulmonary nodules in association with pyoderma gangrenosum: case report / Múltiplos nódulos pulmonares cavitados em associação com pioderma gangrenoso: relato de caso

Pyoderma gangrenosum is a rare neutrophilic disease of unknown origin that is associated with systemic diseases in 50% of cases. It is characterized by erythematous-violaceous nodular lesions that quickly progress to painful ulcers, with undermined edges, necrotic-hemorrhagic, varying in size and depth, located mainly in the lower limbs. Extracutaneous locations of pyoderma gangrenosum are rare, usually involving the lungs; the main differential diagnosis in these cases is Wegener granulomatosis. We report a case of pyoderma gangrenosum, which showed multiple cavitary lung nodules, with good response to high doses of steroids. Once excluded the possibility of Wegener granulomatosis, the authors concluded that it was the manifestation of systemic pyoderma gangrenosum with pulmonary involvement.

Pioderma gangrenoso é doença neutrofílica rara de etiologia desconhecida, que se associa a doenças sistêmicas em 50% dos casos. Caracteriza-se clinicamente por lesão nodular eritematoviolácea ou pústula que progride rapidamente para úlcera dolorosa, de bordas irregulares, fundo necrohemorrágico e localização preferencial nos membros inferiores. Manifestações sistêmicas do Pioderma gangrenoso são raras, envolvem geralmente os pulmões e o principal diagnóstico diferencial nestes casos é a granulomatose de Wegener. Relatamos um caso de paciente portador de pioderma gangrenoso que apresentava múltiplos nódulos pulmonares cavitados, com boa resposta a altas doses de corticoterapia. Uma vez excluída a possibilidade de granulomatose de Wegener, os autores concluíram tratar-se da manifestação sistêmica do pioderma gangrenoso.

Do you know this syndrome? Xeroderma pigmentosum (XP).

Xeroderma pigmentosum is a rare genetic disease characterized by clinical and cellular hypersensitivity to ultraviolet radiation and DNA repair defects. Patients with xeroderma pigmentosum experience sun-induced cutaneous and ocular abnormalities, including cancer. Some develop neurological disorders. We describe the case of a 2 year-old child with DeSanctis-Cacchione's syndrome, with severe neurological deterioration associated with schizencephaly. In the current clinical classification of xeroderma pigmentosum, the term is reserved for cases with severe neurological disorders linked to dwarfism and immature sexual development. The association of xeroderma pigmentosum with schizencephaly has not to date been reported in the literature.

Annular elastolytic giant cell granuloma--case report.

Annular elastolytic giant cell granuloma is a rare granulomatous skin disease characterized histologically by phagocytosis of elastic fibers by multinucleated giant cells and clinically by annular patches with elevated borders and atrophic centers found mainly on sun-exposed skin. The pathogenesis of the disease is controversial. We report the case of a 44-year-old woman with classical clinical and histological findings of the disease whose condition improved when hydroxychloroquine was administered over a four-week period.

Granuloma anular elastolítico gigantocelular: relato de caso / Annular elastolytic giant cell granuloma: case report

O granuloma anular elastolítico gigantocelular é dermatose granulomatosa rara, caracterizada, histologicamente, por fagocitose das fibras elásticas por células gigantes multinucleadas e, clinicamente, por placas anulares com bordas elevadas e centro atrófico, com preferência por áreas expostas. A patogênese é controversa. Descreve-se um caso de mulher de 44 anos, com achados clínicos e histológicos clássicos, que apresentou boa resposta à hidroxicloroquina, após o período de 1 mês.

Annular elastolytic giant cell granuloma is a rare granulomatous skin disease characterized histologically by phagocytosis of elastic fibers by multinucleated giant cells and clinically by annular patches with elevated borders and atrophic centers found mainly on sun-exposed skin. The pathogenesis of the disease is controversial. We report the case of a 44-year-old woman with classical clinical and histological findings of the disease whose condition improved when hydroxychloroquine was administered over a four-week period.