Autophagy is the major mechanism involved in degradation and recycling of intracellular components, and its alterations have been proposed to cause beta cell dysfunction. In this study, we explored ...the effects of autophagy modulation in human islets under conditions associated to endoplasmic reticulum (ER) stress. Human pancreatic islets were isolated by enzymatic digestion and density gradient purification from pancreatic samples of non-diabetic (ND;
= 17; age 65 ± 21 years; gender: 5 M/12 F; BMI 23.4 ± 3.3 kg/m
) and T2D (
= 9; age 76 ± 6 years; 4 M/5 F; gender: BMI 25.4 ± 3.7 kg/m
) organ donors. Nine ND organ donors were treated for hypertension and 1 for both hypertension and hypercholesterolemia. T2D organ donors were treated with metformin (1), oral hypoglycemic agents (2), diet + oral hypoglycemic agents (3), insulin (3) or insulin plus metformin (3) as for antidiabetic therapy and, of these, 3 were treated also for hypertension and 6 for both hypertension and hypercholesterolemia. Two days after isolation, they were cultured for 1-5 days with 10 ng/ml rapamycin (autophagy inducer), 5 mM 3-methyladenine or 1.0 nM concanamycin-A (autophagy blockers), either in the presence or not of metabolic (0.5 mM palmitate) or chemical (0.1 ng/ml brefeldin A) ER stressors. In ND islets palmitate exposure induced a 4 to 5-fold increase of beta cell apoptosis, which was significantly prevented by rapamycin and exacerbated by 3-MA. Similar results were observed with brefeldin treatment. Glucose-stimulated insulin secretion from ND islets was reduced by palmitate (-40 to 50%) and brefeldin (-60 to 70%), and rapamycin counteracted palmitate, but not brefeldin, cytotoxic actions. Both palmitate and brefeldin induced PERK, CHOP and BiP gene expression, which was partially, but significantly prevented by rapamycin. With T2D islets, rapamycin alone reduced the amount of p62, an autophagy receptor that accumulates in cells when macroautophagy is inhibited. Compared to untreated T2D cells, rapamycin-exposed diabetic islets showed improved insulin secretion, reduced proportion of beta cells showing signs of apoptosis and better preserved insulin granules, mitochondria and ER ultrastructure; this was associated with significant reduction of PERK, CHOP and BiP gene expression. This study emphasizes the importance of autophagy modulation in human beta cell function and survival, particularly in situations of ER stress. Tuning autophagy could be a tool for beta cell protection.
Summary
Visceral leishmaniasis is an important parasitic disease that affects humans and animals. The response against the protozoan involves the interaction of both innate and adaptive branches of ...the immune system, and an important immune sensor is represented by the toll‐like receptor (TLR) family. Here, we investigated the pattern of TLR‐2, TLR‐4 and TLR‐9 gene expression in different compartments (brain, choroid plexus, spleen and lymph node) of dogs naturally infected with Leishmania infantum. Gene expression of the TLRs varied according to the compartment evaluated. In the brain, there was only an upregulation of TLR‐2, whereas in the choroid plexus, TLR‐2 and TLR‐9 were both upregulated. Further, the peripheral lymphoid organs (spleen and lymph nodes) showed increased TLR‐2 and TLR‐4 expression. This study provides the first insight about TLR expression in the central nervous system of infected dogs, and gives additional evidence of the compartmentalization of the immune response during visceral leishmaniasis.
Summary
Visceral leishmaniasis is a complex disease caused by Leishmania infantum, and in dogs, besides the classical symptoms, there are descriptions of inflammatory alterations in the brain. Brain ...inflammation is a strictly controlled process, and as the brain counts on the efficiency of the blood–brain barrier (BBB), we aimed to assess BBB integrity in dogs with spontaneous visceral leishmaniasis. Therefore, we evaluated markers in the cerebrospinal fluid (CSF) and in brain tissue related to BBB disruption and brain inflammation. Elevated albumin quota revealed BBB breakdown, corroborated by increased concentrations of anti‐Leishmania antibodies in the CSF. In the brain, albumin and IgG staining formed halos around blood vessels, a classical indicator of BBB leakage. Soluble IgG was also detected in the choroid plexus and ependyma, and in these structures, IgG stained random resident cells. IgG+ cells and Fcγ‐RI+ cells were identified in the choroid plexus, ependyma and perivascular in the brain parenchyma. The data support the occurrence of BBB disruption in dogs with spontaneous visceral leishmaniasis, and IgG as a key molecule that is capable of initiating and/or maintaining the inflammatory stimuli in the nervous milieu and the CSF as an important disseminator of inflammatory stimuli within the CNS.
Abstract
Purpose:
the purpose of the present paper is to present the short-term results of a “detachment-free” (DF) anterolateral approach for primary total hip replacement (THR) performed in a large ...series of patients.
Methods:
two hundred patients submitted to primary THR were retrospectively reviewed for the present study. In all cases, the surgery was performed using a minimally invasive DF anterolateral approach, which entails no disconnection of tendons and no muscle damage. The study population consisted of 96 men (48%) and 104 women (52%), with an average age of 69.4 years (range 38-75). Clinical and radiographic follow-up was performed after 12 months.
Results:
the clinical results, evaluated using the Harris Hip Score, were excellent in 95% of the cases and good in 5%; no cases had fair or poor results. X-rays taken at 3, 6 and 12 months after surgery did not show heterotopic ossification, mobilization of the prosthetic components, or hip dislocation. No infections, deep vein thrombosis, or failure of the gluteal muscles were reported.
Conclusions:
the DF anterolateral approach for THR proved safe and provided effective results at shortterm follow-up.
Level of evidence:
Level IV, therapeutic case series.
If increasing practitioners’ diagnostic accuracy, medical artificial intelligence (AI) may lead to better treatment decisions at lower costs, while uncertainty remains around the resulting ...cost-effectiveness. In the present study, we assessed how enlarging the data set used for training an AI for caries detection on bitewings affects cost-effectiveness and also determined the value of information by reducing the uncertainty around other input parameters (namely, the costs of AI and the population’s caries risk profile). We employed a convolutional neural network and trained it on 10%, 25%, 50%, or 100% of a labeled data set containing 29,011 teeth without and 19,760 teeth with caries lesions stemming from bitewing radiographs. We employed an established health economic modeling and analytical framework to quantify cost-effectiveness and value of information. We adopted a mixed public–private payer perspective in German health care; the health outcome was tooth retention years. A Markov model, allowing to follow posterior teeth over the lifetime of an initially 12-y-old individual, and Monte Carlo microsimulations were employed. With an increasing amount of data used to train the AI sensitivity and specificity increased nonlinearly, increasing the data set from 10% to 25% had the largest impact on accuracy and, consequently, cost-effectiveness. In the base-case scenario, AI was more effective (tooth retention for a mean 2.5%–97.5% 62.8 59.2–65.5 y) and less costly (378 284–499 euros) than dentists without AI (60.4 55.8–64.4 y; 419 270–593 euros), with considerable uncertainty. The economic value of reducing the uncertainty around AI’s accuracy or costs was limited, while information on the population’s risk profile was more relevant. When developing dental AI, informed choices about the data set size may be recommended, and research toward individualized application of AI for caries detection seems warranted to optimize cost-effectiveness.
This open access edited book brings together a closer examination of European and Asian responses to the escalating rivalry between the US and China. As the new Cold War has surfaced as a perceivable ...reality in the post-COVID era, the topic itself is of great importance to policymakers, academic researchers, and the interested public. Furthermore, this manuscript makes a valuable contribution to an under-studied and increasingly important phenomenon in international relations: the impact of the growing strategic competition between the United States and China on third parties, such as small and middle powers in the two arguably most affected regions of the world: Europe and East Asia. The European side has been under-studied and explicitly comparative work on Europe and East Asia is extremely rare. Given that the manuscript focuses heavily on recent developments—and because many of these developments have been quite dramatic—there are very few publications that cover the same topics.
Aim
Anorectal malformations (ARMs) are rare congenital colorectal anomalies with long lasting consequences, among which faecal incontinence is one of the most relevant since it may strongly affect ...patients’ health‐related quality of life (HRQoL). Although a growing body of literature supports the importance of self‐efficacy in chronic disease health outcomes, only few studies have focused on self‐efficacy in ARMs and in faecal incontinence. The purpose of the present study is to examine the mediational role of self‐efficacy in the path between faecal incontinence and HRQoL in patients born with ARMs.
Method
Ninety‐eight adult patients from the Italian Association for Anorectal Malformations (AIMAR) responded to measures of faecal incontinence, self‐efficacy for managing ARM consequences, and physical and mental HRQoL (SF‐36). Data were analysed by means of structural equation models.
Results
The tested model provides support for the guiding hypothesis. Fit indices indicate that the model fits the data well (χ2 = 33.48, df = 23, P = 0.07; comparative fit index CFI = 0.97; root mean square error of approximation RMSEA = 0.07; standardized root mean square residual SRMR = 0.05). Faecal incontinence has negative effects on both physical and mental HRQoL, as well on self‐efficacy. In turn, self‐efficacy has a positive and direct effect on mental HRQoL.
Conclusion
Faecal incontinence is the most relevant and negative factor influencing HRQoL; in addition, self‐efficacy contributes in reducing emotional distress and in improving mental health outcomes. Longitudinal and controlled studies may be helpful to evaluate the effectiveness of self‐efficacy interventions in improving mental HRQoL in patients with faecal incontinence.
Abstract Visceral leishmaniasis is a chronic disease caused by Leishmania infantum . We aimed to detect the parasite in the brain of fifteen naturally-infected dogs using in situ hybridization and ...immunohistochemistry, and the gene expression of selected chemokines by RT-qPCR. We detected no parasite in the brain, but perivascular deposition of parasite DNA and IgG in the choroid plexus. We noticed up-regulation of CCL-3, CCL-4 and CCL-5, coherent with T lymphocyte accumulation, stating the brain as a pro-inflammatory environment. Indeed, not necessarily the parasite itself, but rather its DNA seems to act as a trigger to promote brain inflammation during visceral leishmaniasis.
Summary
In this study, we investigated the bone cell activity in patients with osteogenesis imperfecta (OI) treated and untreated with neridronate. We demonstrated the key role of Dickkopf-1 (DKK1), ...receptor activator of nuclear factor-κB ligand (RANKL), and tumor necrosis factor alpha (TNF-α) in regulating bone cell of untreated and treated OI subjects. These cytokines could represent new pharmacological targets for OI.
Introduction
Bisphosphonates are widely used in the treatment of children with osteogenesis imperfecta (OI) with the objective of reducing the risk of fractures. Although bisphosphonates increase bone mineral density in OI subjects, the effects on fracture incidence are conflicting. The aim of this study was to investigate the mechanisms underlying bone cell activity in subjects with mild untreated forms of OI and in a group of subjects with severe OI treated with cycles of intravenous neridronate.
Methods
Sclerostin, DKK1, TNF-α, RANKL, osteoprotegerin (OPG), and bone turnover markers were quantified in serum of 18 OI patients (12 females, mean age 8.86 ± 3.90), 8 of which were receiving cyclic intravenous neridronate, and 21 sex- and age-matched controls. The effects on osteoblastogenesis and OPG expression of media conditioned by the serum of OI patients and anti-DKK1 neutralizing antibody were evaluated. Osteoclastogenesis was assessed in cultures from patients and controls.
Results
DKK1 and RANKL levels were significantly increased both in untreated and in treated OI subjects with respect to controls. The serum from patients with high DKK1 levels inhibited both osteoblast differentiation and OPG expression in vitro. High RANKL and low OPG messenger RNA (mRNA) levels were found in lymphomonocytes from patients. High amounts of TNF-α were expressed by monocytes, and an elevated percentage of circulating CD11b-CD51/CD61+ osteoclast precursors was observed in patients.
Conclusions
Our study demonstrated the key role of DKK1, RANKL, and TNF-α in regulating bone cell activity of subjects with OI untreated and treated with bisphosphonates. These cytokines could represent new pharmacological targets for OI patients.
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