Promiscuous Roles of Autophagy and Proteasome in Neurodegenerative Proteinopathies.

Alterations in autophagy and the ubiquitin proteasome system (UPS) are commonly implicated in protein aggregation and toxicity which manifest in a number of neurological disorders. In fact, both UPS and autophagy alterations are bound to the aggregation, spreading and toxicity of the so-called prionoid proteins, including alpha synuclein (α-syn), amyloid-beta (Aβ), tau, huntingtin, superoxide dismutase-1 (SOD-1), TAR-DNA-binding protein of 43 kDa (TDP-43) and fused in sarcoma (FUS). Recent biochemical and morphological studies add to this scenario, focusing on the coordinated, either synergistic or compensatory, interplay that occurs between autophagy and the UPS. In fact, a number of biochemical pathways such as mammalian target of rapamycin (mTOR), transcription factor EB (TFEB), Bcl2-associated athanogene 1/3 (BAG3/1) and glycogen synthase kinase beta (GSk3β), which are widely explored as potential targets in neurodegenerative proteinopathies, operate at the crossroad between autophagy and UPS. These biochemical steps are key in orchestrating the specificity and magnitude of the two degradation systems for effective protein homeostasis, while intermingling with intracellular secretory/trafficking and inflammatory pathways. The findings discussed in the present manuscript are supposed to add novel viewpoints which may further enrich our insight on the complex interactions occurring between cell-clearing systems, protein misfolding and propagation. Discovering novel mechanisms enabling a cross-talk between the UPS and autophagy is expected to provide novel potential molecular targets in proteinopathies

Treatment of esophageal achalasia in children: Today and tomorrow

Esophageal achalasia (EA) is a rare esophageal motility disorder in children. Laparoscopic Heller myotomy (LHM) represents the treatment of choice in young patients. Peroral endoscopic myotomy (POEM) is becoming an alternative to LHM. The aim of this study is to evaluate the effectiveness, safety, and outcomes of POEM vs LHM in treatment of children with EA. Data of pediatric patients with EA, who underwent LHM and POEM from February 2009 to December 2013 in two centers, were collected. Eighteen patients (9 male, mean age: 11.6 years; range: 2-17 years) were included. Nine patients (6 male, mean age: 10.7 years; range: 2-16 years) underwent LHM, and the other 9 (3 males, mean age: 12.2 years; range: 6-17 years) underwent POEM procedure. Mean operation time was shorter in POEM group compared with LHM group (62/149 minutes). Myotomy was longer in POEM group than in LHM group (11/7 cm). One major complication occurred after LHM (esophageal perforation). No clinical and manometric differences were observed between LHM and POEM in follow-up. The incidence of iatrogenic gastroesophageal reflux disease was low (1 patient in both groups). Results of a midterm follow-up show that LHM and POEM are safe and effective treatments also in children. Besides, POEM is a mini-invasive technique with an inferior execution timing compared to LHM. A skilled endoscopic team is mandatory to perform this procedur

Quantitative ultrastructural morphometry and gene expression of mTOR-related mitochondriogenesis within glioblastoma cells

In glioblastoma (GBM) cells, an impairment of mitochondrial activity along with autophagy suppression occurs. Autophagy suppression in GBM promotes stemness, invasion, and poor prognosis. The autophagy deficit seems to be due, at least in part, to an abnormal up-regulation of the mammalian target of rapamycin (mTOR), which may be counteracted by pharmacological mTORC1 inhibition. Since autophagy activation is tightly bound to increased mitochondriogenesis, a defect in the synthesis of novel mitochondria is expected to occur in GBM cells. In an effort to measure a baseline deficit in mitochondria and promote mitochondriogenesis, the present study used two different GBM cell lines, both featuring mTOR hyperactivity. mTORC1 inhibition increases the expression of genes and proteins related to autophagy, mitophagy, and mitochondriogenesis. Autophagy activation was counted by RT-PCR of autophagy genes, LC3-immune-fluorescent puncta and immune-gold, as well as specific mitophagy-dependent BNIP3 stoichiometric increase in situ, within mitochondria. The activation of autophagy-related molecules and organelles after rapamycin exposure occurs concomitantly with progression of autophagosomes towards lysosomes. Remarkably, mitochondrial biogenesis and plasticity (increased mitochondrial number, integrity, and density as well as decreased mitochondrial area) was long-lasting for weeks following rapamycin withdrawal

Monteggia fracture associated with ipsilateral intercondylar distal humeral fracture with posterior interosseous nerve palsy: case report, medico-legal implications, and methodological assessment analysis

BACKGROUND: The Monteggia fracture, defined as a fracture of the proximal third of the ulnar shaft associated with an anterior or posterior dislocation of the proximal radial epiphysis, is a serious injury accounting for 0.7% of all elbow fractures and dislocations in adults. For adult patients, good results can only be obtained through early diagnosis and adequate surgical treatment. Monteggia fracture-dislocations associated with distal humeral fracture are extremely rare injuries in adults and there are few cases described in the literature. Medico-legal implications arising from such conditions have a host of complexities that cannot be discounted.CASE REPORT: This case report revolves around a patient affected by a type I Monteggia fracture-dislocation, according to the Bado classification, associated with an ipsilateral intercondylar distal humeral fracture. To our knowledge, this combination of lesions has never been reported before in adult patients. A positive result was obtained due to early diagnosis, achievement of anatomical reduction, and optimal stabilization with internal fixation which made it possible to achieve early functional recovery.CONCLUSIONS: Monteggia fracture -dislocations associated with ipsilateral intercondylar distal humeral fracture are extremely rare in adults. In the case herein reported, a favorable outcome was obtained due to early diagnosis, achievement of anatomical reduction and management with internal fixation with plate and screws, as well as early functional training. Misdiagnosis makes such lesions risky in terms of potentially delayed treatment, increasing the need for surgical interventions and the possibility of high-risk complications and disabling sequelae, with possible medico -legal implications. In the case of unrecognized injuries under urgent circumstances, the injuries may become chronic, making the treatment more complex. The ultimate outcomes of a misdiagnosed Monteggia lesion can lead to very serious functional and aesthetic damage

Morpho-chemical observations of human deciduous teeth enamel in response to biomimetic toothpastes treatment

Today, biomaterial research on biomimetic mineralization strategies represents a new challenge in the prevention and cure of enamel mineral loss on delicate deciduous teeth. Distinctive assumptions about the origin, the growth, and the functionalization on the biomimetic materials have been recently proposed by scientific research studies in evaluating the different clinical aspects of treating the deciduous tooth. Therefore, appropriate morpho-chemical observations on delivering specific biomaterials to enamel teeth is the most important factor for controlling biomineralization processes. Detailed morpho-chemical investigations of the treated enamel layer using three commercial toothpastes (Biorepair, F1400, and F500) were performed through variable pressure scanning electron microscopy (VP-SEM) and energy dispersive X-ray spectroscopy (EDS) on deciduous teeth in their native state. A new microscopy methodology allowed us to determine the behaviors of silicate, phosphate, and calcium contents from the early stage, as commercially available toothpastes, to the final stage of delivered diffusion, occurring within the enamel layer together with their penetration depth properties. The reported results represent a valuable background towards full comprehension of the role of organic-inorganic biomaterials for developing a controlled biomimetic toothpaste in biofluid media

Spontaneous calcification process in primary renal cells from a medullary sponge kidney patient harbouring a GDNF mutation.

Medullary nephrocalcinosis is a hallmark of medullary sponge kidney (MSK). We had the opportunity to study a spontaneous calcification process in vitro by utilizing the renal cells of a patient with MSK who was heterozygous for the c.-27 + 18G>A variant in the GDNF gene encoding glial cell-derived neurotrophic factor. The cells were obtained by collagenase digestion of papillary tissues from the MSK patient and from two patients who had no MSK or nephrocalcinosis. These cells were typed by immunocytochemistry, and the presence of mineral deposits was studied using von Kossa staining, scanning electron microscopy analysis and an ALP assay. Osteoblastic lineage markers were studied using immunocytochemistry and RT-PCR. Staminality markers were also analysed using flow cytometry, magnetic cell separation technology, immunocytochemistry and RT-PCR. Starting from p2, MSK and control cells formed nodules with a behaviour similar to that of calcifying pericytes; however, Ca2PO4 was only found in the MSK cultures. The MSK cells had morphologies and immunophenotypes resembling those of pericytes or stromal stem cells and were positive for vimentin, ZO1, aSMA and CD146. In addition, the MSK cells expressed osteocalcin and osteonectin, indicating an osteoblast-like phenotype. In contrast to the control cells, GDNF was down-regulated in the MSK cells. Stable GDNF knockdown was established in the HK2 cell line and was found to promote Ca2PO4 deposition when the cells were incubated with calcifying medium by regulating the osteonectin/osteopontin ratio in favour of osteonectin. Our data indicate that the human papilla may be a perivascular niche in which pericyte/stromal-like cells can undergo osteogenic differentiation under particular conditions and suggest that GDNF down-regulation may have influenced the observed phenomenon

Risk factors for abnormally invasive placenta: a systematic review and meta-analysis.

Purpose of the article. To explore the strength of association between different maternal and pregnancy characteristics and the occurrence of abnormally invasive placenta (AIP). MATERIALS AND METHODS: Pubmed, Embase, CINAHL databases were searched. The risk factors for AIP explored were: obesity, age > 35 years, smoking before or during pregnancy, placenta previa, prior cesarean section (CS), placenta previa and prior CS, prior uterine surgery, abortion and uterine curettage, in vitro fertilization (IVF) pregnancy and interval between a previous CS and a subsequent pregnancy. Random-effect head-to-head meta-analyses were used to analyze the data. RESULTS: Forty-six were included in the systematic review. Maternal obesity (Odd ratio, OR: 1.4, 95% CI 1.0-1.8), advanced maternal age (OR: 3.1, 95% CI 1.4-7.0) and parity (OR: 2.5, 95% CI 1.7-3.6), but not smoking were associated with a higher risk of AIP. The presence of placenta previa in women with at least a prior CS was associated with a higher risk of AIP compared to controls, with an OR of 12.0, 95% CI 1.6-88.0. Furthermore, the risk of AIP increased with the number of prior CS (OR of 2.6, 95% CI 1.6-4.4 and 5.4, 95% CI 1.7-17.4 for two and three prior CS respectively). Finally, IVF pregnancies were associated with a high risk of AIP, with an OR of 2.8 (95% CI 1.2-6.8). CONCLUSION: A prior CS and placenta previa are among the strongest risk factors for the occurrence of AIP