Search

Antibody therapies for Alzheimer's Disease (AD) hold promise but have been limited by the inability of these proteins to migrate efficiently across the blood brain barrier (BBB). Central nervous system (CNS) gene transfer by vectors like adeno-associated virus (AAV) overcome this barrier by allowing the bodies' own cells to produce the therapeutic protein, but previous studies using this method to target amyloid-β have shown success only with truncated single chain antibodies (Abs) lacking an Fc domain. The Fc region mediates effector function and enhances antigen clearance from the brain by neonatal Fc receptor (FcRn)-mediated reverse transcytosis and is therefore desirable to include for such treatments. Here, we show that single chain Abs fused to an Fc domain retaining FcRn binding, but lacking Fc gamma receptor (FcγR) binding, termed a silent scFv-IgG, can be expressed and released into the CNS following gene transfer with AAV. While expression of canonical IgG in the brain led to signs of neurotoxicity, this modified Ab was efficiently secreted from neuronal cells and retained target specificity. Steady state levels in the brain exceeded peak levels obtained by intravenous injection of IgG. AAV-mediated expression of this scFv-IgG reduced cortical and hippocampal plaque load in a transgenic mouse model of progressive β-amyloid plaque accumulation. These findings suggest that CNS gene delivery of a silent anti-Aβ scFv-IgG was well-tolerated, durably expressed and functional in a relevant disease model, demonstrating the potential of this modality for the treatment of Alzheimer's disease.

The nose-to-brain delivery route is used to bypass the blood–brain barrier and deliver drugs directly into the brain. Over the years, significant signs of progress have been made in developing nano-drug delivery systems to address the very low drug transfer levels seen with conventional formulations (e.g., nasal solutions). In this paper, sericin nanoparticles were prepared using crocetin as a new bioactive natural cross-linker (NPc) and compared to sericin nanoparticles prepared with glutaraldehyde (NPg). The mean diameter of NPc and NPg was about 248 and 225 nm, respectively, and suitable for nose-to-brain delivery. The morphological investigation revealed that NPc are spherical-like particles with a smooth surface, whereas NPg seem small and rough. NPc remained stable at 4 °C for 28 days, and when freeze-dried with 0.1% w/v of trehalose, the aggregation was prevented. The use of crocetin as a natural cross-linker significantly improved the in vitro ROS-scavenging ability of NPc with respect to NPg. Both formulations were cytocompatible at all the concentrations tested on human fibroblasts and Caco-2 cells and protected them against oxidative stress damage. In detail, for NPc, the concentration of 400 µg/mL resulted in the most promising to maintain the cell metabolic activity of fibroblasts higher than 90%. Overall, the results reported in this paper support the employment of NPc as a nose-to-brain drug delivery system, as the brain targeting of antioxidants is a potential tool for the therapy of neurological diseases.

This review aims to provide the state of the art on polymeric and lipid nanoparticles, used or suggested to approach pediatric diseases’ problems and needs, and to inspire new researches in this field. Several drugs are currently not available in formulations suitable for pediatric patients. The United States Pediatric Formulation Initiative suggested applying new technologies to pediatric drug formulations, for instance, nanotechnology. The literature analysis showed that polymeric and lipid nanoparticles have been widely studied to treat pediatric diseases, and albumin nanoparticles and liposomes are already used in clinical practice. Nevertheless, these studies are focused almost exclusively on pediatric cancer treatment. Although nanomedicine may solve many needs of pediatric diseases and medicines, the unavailability of data on pharmacokinetics, safety and efficacy of both drugs and nanoparticles in pediatric patients limits the development of new pediatric medicines based on nanoparticles. Therefore, nanomedicine applied in pediatrics remains a significant challenge in the near future.

Genistein (GEN) is a soy-derived isoflavone that exhibits several biological effects, such as neuroprotective activity and the prevention of several types of cancer and cardiovascular disease. However, due to its poor water solubility and the extensive first-pass metabolism, the oral bioavailability of GEN is limited. In this work, solid lipid nanoparticles (SLN) were developed to preferentially reach the intestinal lymphatic vessels, avoiding the first-pass metabolism of GEN. GEN-loaded SLN were obtained by a hot homogenization process, and the formulation parameters were chosen based on already formulated studies. The nanoparticles were characterized, and the preliminary in vitro chylomicron formation was evaluated. The cell uptake of selected nanocarriers was studied on the Caco-2 cell line and intestinal mucosa. The SLN, characterized by a spherical shape, showed an average diameter (about 280 nm) suitable for an intestinal lymphatic uptake, good stability during the testing time, and high drug loading capacity. Furthermore, the intestinal mucosa and Caco-2 cells were found to uptake SLN. The approximately two-fold increase in particle size suggested a possible interaction between SLN and the lipid components of chylomicrons like phospholipid; therefore, the results may support the potential for these SLN to improve oral GEN bioavailability via intestinal lymphatic absorption.

The role of mycobacterial efflux pumps in drug-resistant tuberculosis has been widely reported. Recently, a new compound, named SS13, has been synthesized, and its activity as a potential efflux inhibitor has been demonstrated. In this work, the chemical–physical properties of the SS13 were investigated; furthermore, a formulative study aimed to develop a formulation suitable for oral administration was performed. SS13 shows nonintrinsic antitubercular activity, but it increases the antitubercular activity of all the tested drugs on several strains. SS13 is insoluble in different simulated gastrointestinal media; thus, its oral absorption could be limited. Solid lipid nanoparticles (SLNs) were, therefore, developed by using two different lipids, Witepsol and/or Gelucire. Nanoparticles, having a particle size (range of 200–450 nm with regards to the formulation composition) suitable for intestinal absorption, are able to load SS13 and to improve its permeation through the intestinal mucosa compared to the pure compound. The cytotoxicity is influenced by the concentration of nanoparticles administered. These promising results support the potential application of these nanocarriers for increasing the oral permeation of SS13 in multidrug-resistant tuberculosis management.

Biomedical applications of nanoparticles (NPs) have reached an increasing development in recent years. Recently, we demonstrated that newly synthesized poly (ethyl 2-cyanoacrylate) nanoparticles (PECA-NPs) are possible antitumor agents due to their cytotoxicity for cancer cells. Indocyanine green (ICG), an amphiphilic tricarbocyanine fluorescent dye, is widely used for the detection of tumoral extension in different organs during clinical surgery. Moreover, this fluorescent agent is unstable and it has a rapid clearance in physiological conditions in vivo. In this study, ICG was charged in PECA-NPs to improve its aqueous stability and make easier its use for the identification of tumor cells. Microscopic and ultrastructural aspects concerning the related in vitro interactions between ICG-loaded NPs and tumor cell culture were investigated. Obtained results showed an effective stabilization of ICG; furthermore, color inclusions inside the cells treated with ICG-loaded NPs demonstrated the internalization of NPs with associated ICG. Transmission electron microscopy (TEM) analysis demonstrated the cytoplasmic presence of coated vesicles (Ø ≤ 100 nm), hypothesizing their involvement in the mechanism of endocytosis. Therefore, ICG-loaded NPs could be proposed as agents for tumor diagnosis, hypothesizing also in the future a specific therapeutic treatment.

The National Union for the Fight Against Illiteracy, which was founded in Rome in December 1947, pursued the main aim of contributing to the recovery of illiterate adults, who was numerous in southern Italy, to form them as socially and politically aware citizens. The choice to keep equal distance from all political organizations was the basis on which the Union founded its educational work. To develop its work the Union created the Centers of Popular Culture, and as long as the Cpc were under the control of the Union, the principle of party equidistance was respected. But in June 1970, the Union gave life to the Regional Committees of the Cpc, which arose in Basilicata, Calabria, Campania, Lazio, Puglia and Sardinia. The Committees characterized in a political-ideological way their courses and programs, including training for Cpc animators. This, at least, is what emerges from the activity of the Regional Committee of the Sardinian CPC, which is the subject of this paper.

Genistein has been reported to have antioxidant and neuroprotective activity. Despite encouraging in vitro and in vivo results, several disadvantages such as poor water solubility, rapid metabolism, and low oral bioavailability limit the clinical application of genistein. The aim of this study was to design and characterize genistein-loaded chitosan nanoparticles for intranasal drug delivery, prepared by the ionic gelation technique by using sodium hexametaphosphate. Nanoparticles were characterized in vitro and their cytotoxicity was tested on PC12 cells. Genistein-loaded nanoparticles were prepared, and sodium hexametaphosphate was used as a valid alternative to well-known cross-linkers. Nanoparticle characteristics as well as their physical stability were affected by formulation composition and manufacturing. Small (mean diameters of 200–300 nm) and homogeneous nanoparticles were obtained and were able to improve genistein penetration through the nasal mucosa as compared to pure genistein. Nanoparticle dispersions showed a pH consistent with the nasal fluid and preserved PC12 cell vitality.

Early pancreas development, given its complexity, is generally considered as a paradigm for branching morphogenesis and for the development of two organs in one: the Langherans islets, programmed to secrete hormones into the bloodstream, and the exocrine pancreas compartment, composed of two major cell types, acinar and ductal cells, devoid to secrete digestive enzymes into the duodenum through a branched network of ducts. Exocrine and endocrine pancreas are generally presumed to originate from a common multi-lineage progenitor cell (MPC), emerging within the definitive endoderm surrounding the posterior foregut. Bipotential precursors committed to the pancreatic fate originate the MPC, that are considered the progenitors of all pancreatic cells operating in the mature pancreas, including acinar, ductal, endocrine and stromal cell types. Pluripotent stem cells (PSCs) are able to differentiate into several cell types, including acinary cells, duct cells and islet cells, depending on certain transcription factors, which function in a coordinated way during pancreas development. The epidemiological entity of pancreatic diseases such as diabetes mellitus and issues regarding the management of the diabetic patient have constantly stimulated the great current interest aimed at regenerative pancreatic medicine. Several studies in rats have demonstrated the existence of stem/progenitor cells in the adult pancreas and have clarified the mechanism by which pancreatic stem cells differentiate into acinar, ductal and endocrine cells. In this context, the cellular microenvironment called “niche” plays a major role in inducing differentiation of stem/progenitor cells by adequate cellular signals. Within the niche, undifferentiated pluripotent cells give rise to asymmetrically dividing daughter cells. The main purpose of this work was to identify stem cells and progenitor cells in the human pancreas during intrauterine development in relation to what is already known in the adult pancreas and in experimental models. Proceedings of the 2nd International Course on Perinatal Pathology (part of the 11th International Workshop on Neonatology · October 26th-31st, 2015) · Cagliari (Italy) · October 31st, 2015 · Stem cells: present and future Guest Editors: Gavino Faa, Vassilios Fanos, Antonio Giordano

The human adrenal glands arise around the 4th week of gestation and during the intrauterine life produce many substances that are responsible for the maintenance of fetal homeostasis and organ maturation. Stem cell niches represent the microenvironment suitable for life and replication of adrenal stem cells. Adrenal gland stem cells have the capacity to self-renew and generate functional differentiated daughter cells that replenish lost cells. Morphologically the adrenal stem cells appeared as small, polymorphic cells, closed together, with basophilic nucleus, located between adrenal and renal capsules. This study was mainly based on a morphological and immunohistochemical approach, particularly on characterization and localization of the multiple stem/progenitor cells that contribute to the development of the human adrenal gland. Proceedings of the 2nd International Course on Perinatal Pathology (part of the 11th International Workshop on Neonatology · October 26th-31st, 2015) · Cagliari (Italy) · October 31st, 2015 · Stem cells: present and future Guest Editors: Gavino Faa, Vassilios Fanos, Antonio Giordano

In the first part of this review a brief summary of the embryology and histology of the gastrointestinal tract is provided. In the second part intestinal stem cells (ISCs) are discussed. Several signaling pathways play a crucial role in the crypt base in the regulation of ISC proliferation and self-renewal; Wnt, Notch, BMP, Ephrin, JAK/STAT1, PTEN, AKT, PI3K and many more. Numerous investigators are involved in studying the location, number, and behavior of ISCs within the base of the intestinal crypts. Several markers are espressed by ISCs. Among these, Leucine-rich-repeat-containing G-protein-coupled receptor-5 (Lgr5), Sox9, Prominin-1, DCAMKL-1, EphB2, p-PTEN, p-AKT, Fgfr3, m-TER, and CD44. Stem cell therapy has shown promise for the treatment of some diseases characterized by tissue damage with ischemic and inflammatory lesions like inflammatory bowel disease (IBD) and necrotizing enterocolitis (NEC). Proceedings of the 2nd International Course on Perinatal Pathology (part of the 11th International Workshop on Neonatology · October 26th-31st, 2015) · Cagliari (Italy) · October 31st, 2015 · Stem cells: present and future Guest Editors: Gavino Faa, Vassilios Fanos, Antonio Giordano

We report a sampling strategy based on Mendelian Breeding Units (MBUs), representing an interbreeding group of individuals sharing a common gene pool. The identification of MBUs is crucial for case-control experimental design in association studies. The aim of this work was to evaluate the possible existence of bias in terms of genetic variability and haplogroup frequencies in the MBU sample, due to severe sample selection. In order to reach this goal, the MBU sampling strategy was compared to a standard selection of individuals according to their surname and place of birth. We analysed mitochondrial DNA variation (first hypervariable segment and coding region) in unrelated healthy subjects from two different areas of Sardinia: the area around the town of Cabras and the western Campidano area. No statistically significant differences were observed when the two sampling methods were compared, indicating that the stringent sample selection needed to establish a MBU does not alter original genetic variability and haplogroup distribution. Therefore, the MBU sampling strategy can be considered a useful tool in association studies of complex traits.

Wilms’ Tumor 1 (WT1) is a transcription factor involved in the development of the urogenital system. The purpose of this study was to analyze the immunoreactivity for WT1 protein in different tissues and organs in human fetuses in early phases of gestation. To this end, samples from multiple organs were obtained from 4 human fetuses, ranging from 7 up to 12 weeks of gestation. Each sample was formalin-fixed, paraffin embedded and immunostained for WT1. Our data show that WT1 is involved in development of multiple human organs in a more vast series of cells types than previously reported. Immunostaining for WT1 was characterized by a predominant cytoplasmic reactivity in the vast majority of cell types. Mesenchimal progenitors in the fetal lung, ductal plate progenitors in fetal liver, cap mesenchimal cells in the developing kidney, fetal zone cells in adrenal glands, atrial and ventricular cardiomyocytes in the fetal heart, radial glial cells in the fetal cerebral cortex and skeletal muscle cell precursors showed the highest levels of WT1 immunoreactivity. Future studies will be needed to detect differences in the expression of WT1 in various organs at different gestational ages, in order to better evaluate the role of WT1 in cell proliferation and differentiation during intrauterine human development.

Perinatal asphyxia is one of the leading causes of morbidity and mortality in the perinatal period, with 4 million neonates suffering annually from birth asphyxia. Tissue hypoxia can cause several pathological changes in multiple organs, hypoxic-ischemic encephalopathy (HIE) representing one of the most severe consequences in the newborn, occasionally leading to the insurgence of the multi-organ dysfunction syndrome. The pathological diagnosis of neonatal asphyxia is complex, histological markers of tissue hypoxia often overlapping with pathological changes due to other etiologies. This work is aimed at summarizing the most important pathological markers of asphyxia occurring in a newborn in the different organs. The endothelial lesions (swelling, apoptosis, detachment and loss of the endothelial barrier) in our experience, represent the most relevant pathological changes induced by hypoxia in all the organs. The finding of increased hepatic hemopoiesis represents one of the most important markers of chronic tissue hypoxia. In conclusion, the accurate histological study of all the organs in every case of perinatal asphyxia may allow, in expert hands, perinatal pathologists to give important data to neonatologists for reaching, together, a complex clinical/pathological diagnosis, able to explain the clinical course in the majority of asphyxiated newborns undergoing multiple organ dysfunction. Proceedings of the International Course on Perinatal Pathology (part of the 10th International Workshop on Neonatology · October 22nd-25th, 2014) · Cagliari (Italy) · October 25th, 2014 · The role of the clinical pathological dialogue in problem solving Guest Editors: Gavino Faa, Vassilios Fanos, Peter Van Eyken

One of the most significant causes of neonatal morbidity and mortality is represented by neonatal sepsis that often manifests itself as a systemic inflammatory response syndrome (SIRS). The progression of SIRS usually leads to multiple organ dysfunction, occasionally culminating in multiple organ failure (MOF). The loss of endothelial barrier represents the unifying lesion of multiple organs in newborns affected by sepsis and the most important pathological change responsible for the evolution toward MOF in neonates. The aim of this study is to present the most important pathological changes occurring in neonatal sepsis. Proceedings of the International Course on Perinatal Pathology (part of the 10th International Workshop on Neonatology · October 22nd-25th, 2014) · Cagliari (Italy) · October 25th, 2014 · The role of the clinical pathological dialogue in problem solving Guest Editors: Gavino Faa, Vassilios Fanos, Peter Van Eyken

One of the most severe events occurring in critically ill patients admitted to a neonatal intensive care unit (NICU) center is represented by the multiple organ failure (MOF), a systemic inflammatory response leading to a progressive organ dysfunction and mortality in newborns. MOF may occur in newborns primarily affected by multiple single organ diseases, including respiratory distress syndrome neonatal sepsis with acute kidney injury, post-asphyxial hypoxic-ischemic encephalopathy and pandemic influenza A (H1N1) infection. In a previous article from our group, based on the histological examination of all organs at autopsy of newborns affected by MOF, all organs studied did not escape to be damaged, including thymus and pancreas normally not mentioned in the literature of MOF. The aim of this article is to review the most important pathological changes pathologists should look for in every case of MOF occurring in the perinatal period, with particular attention to systemic endothelial changes occurring in blood vessels in all organs and sytems. On the basis of our experience, matching data during the last phases of the clinicopathological diagnosis represents a useful method, much more productive as compared to the method based on giving pathological answers to the clinical questions prospected before autopsy. As for the pathological features observed in neonatal MOF, one of them deserves a particular attention: the vascular lesions, and in particular the multiple changes occurring during MOF development in endothelial cells, ending with the loss of the endothelial barrier, probably the most relevant histological lesion followed by the insurgence of interstitial edema and disseminated intravascular coagulation. Small vessels should be observed at high power, with particular attention to the size and shape of endothelial nuclei, in order to evidence endothelial swelling, probably the initial modification of the endothelial cells leading to their death. Finally, only the clinical pathological discussion may lead to a good diagnosis, correlating the morphological evidences with the clinical history and the sequence of clinical events that, at the best of our experience, are always different in a new case of MOF. Proceedings of the International Course on Perinatal Pathology (part of the 10th International Workshop on Neonatology · October 22nd-25th, 2014) · Cagliari (Italy) · October 25th, 2014 · The role of the clinical pathological dialogue in problem solving Guest Editors: Gavino Faa, Vassilios Fanos, Peter Van Eyken

Mitochondrial DNA (mtDNA) sequence variation was examined in Finns, Swedes and Tuscans by PCR amplification and restriction analysis. About 99% of the mtDNAs were subsumed within 10 mtDNA haplogroups (H, I, J, K, M, T, U, V, W, and X) suggesting that the identified haplogroups could encompass virtually all European mtDNAs. Because both hypervariable segments of the mtDNA control region were previously sequenced in the Tuscan samples, the mtDNA haplogroups and control region sequences could be compared. Using a combination of haplogroup-specific restriction site changes and control region nucleotide substitutions, the distribution of the haplogroups was surveyed through the published restriction site polymorphism and control region sequence data of Caucasoids. This supported the conclusion that most haplogroups observed in Europe are Caucasoid-specific, and that at least some of them occur at varying frequencies in different Caucasoid populations. The classification of almost all European mtDNA variation in a number of well defined haplogroups could provide additional insights about the origin and relationships of Caucasoid populations and the process of human colonization of Europe, and is valuable for the definition of the role played by mtDNA backgrounds in the expression of pathological mtDNA mutations