Systemic Beta-Hydroxybutyrate Affects BDNF and Autophagy into the Retina of Diabetic Mice

Background: Diabetic retinopathy (DR) is a neurovascular disease, characterized by a deficiency of brain-derived neurotrophic factor (BDNF), a regulator of autophagy. Beta-hydroxybutyrate (BHB), previously reported as a protective agent in DR, has been associated with BDNF promotion. Here, we investigated whether systemic BHB affects the retinal levels of BDNF and local autophagy in diabetic mice with retinopathy; Methods: C57BL/6J mice were administered with intraperitoneal (i.p.) streptozotocin (STZ) (75 mg/kg) injection to develop diabetes. After 2 weeks, they received i.p. injections of BHB (25-50-100 mg/kg) twice a week for 10 weeks. Retinal samples were collected in order to perform immunofluorescence, Western blotting, and ELISA analysis; Results: BHB 50 mg/kg and 100 mg/kg significantly improved retinal BDNF levels (p < 0.01) in diabetic mice. This improvement was negatively associated with autophagosome-lysosome formations (marked by LC3B and ATG14) and to higher levels of connexin 43 (p < 0.01), a marker of cell integrity. Moreover, BHB administration significantly reduced M1 microglial activation and autophagy (p < 0.01); Conclusions: The systemic administration of BHB in mice with DR improves the retinal levels of BDNF, with the consequent reduction of the abnormal microglial autophagy. This leads to retinal cell safety through connexin 43 restoration

IL-17A neutralizing antibody regulates monosodium urate crystal-induced gouty inflammation

Gout is a paradigm of acute, self-limiting inflammation caused by the deposition of monosodium urate (MSU) crystals within intra-and/or peri-articular areas, leading to excruciating pain, joint swelling and stiffness. The infiltration of leukocytes drives the inflammatory response and remains an attractive target for therapeutic intervention. In this context, emerging evidence supports the view that systemic differentiation of Th17 cells and their in situ infiltration as one of the potential mechanisms by which these cells, and their main product IL-17, causes damage to target tissues. To test if IL-17 was having a detrimental role in gouty onset and progression we targeted this cytokine, using a neutralizing antibody strategy, in an experimental model of gout. Joint inflammation was induced in CD-1 mice by the intra-articular (i.a.) administration of MSU crystals (200 μg/20 μl). Animals from IL-17Ab-treated groups received 1, 3 and 10 μg (i.a.) in 20 μl of neutralizing antibody after MSU crystals administration. Thereafter, joints were scored macroscopically, and knee joint oedema determined with a caliper. Histological analysis, myeloperoxidase assay and western blots analysis for COX-2/mPGEs-1/IL-17R pathway were conducted at 18 h (peak of inflammation) to evaluate leukocytes infiltration and activation, followed by the analysis, in situ, of pro/anti-inflammatory cytokines and chemokines. Flow cytometry was also used to evaluate the modulation of infiltrated inflammatory monocytes and systemic Th17 and Treg profile. Treatment with IL-17Ab revealed a dose-dependent reduction of joint inflammation scores with maximal inhibition at 10 μg. The neutralizing antibody was also able to significantly reduce leukocytes infiltration and MPO activity as well the expression of JE, IL-1α, IL-1β, IL-16, IL-17, C5a, BLC and, with a less extent IP-10, Rantes, KC, TIMP-1, SDF-1 and metalloproteinases in inflamed tissues. Biochemical analysis also revealed that IL-17Ab treatment modulated COX-2/mPGEs-1 pathway (and related PGE2 production) without interfering with IL-17R expression. Furthermore, flow cytometry analysis highlighted a selective modulation of infiltrating inflammatory monocytes (B220-/GR1hi-F480hi/CD115+) and circulating Th17, but not Treg, cells after IL-17Ab treatment. Collectively the results of this study report for the first time, that i.a. injection of MSU crystals stimulates in vivo production of Th17 cells and Th17-related inflammatory cyto-chemokines. In addition, we have demonstrated that the administration of a neutralizing antibody against IL-17 attenuates joint symptoms, swelling and leukocytes infiltration to the inflamed tissue, possibly providing a new strategy for the treatment of gouty inflammation and/or arthritis

Surface Related Multiple Elimination (SRME): an outline of the theory and application to a real data case

L’obiettivo principale delle prospezioni sismiche è ricostruire un’immagine delle strutture geologiche del sottosuolo grazie all’analisi degli echi generati da sorgenti artificiali. Una fonte di disturbo che condiziona il risultato finale è la presenza di riflessioni multiple che si originano a causa della riverberazione del segnale utile all’interno di strati geologici . Le “surface-related multiples” possono essere definite come le multiple che non esisterebbero se l’interfaccia aria-strato superficiale fosse trasparente all’energia sismica. Nelle acquisizioni marine questa tipologia di disturbi condiziona fortemente l’immagine sismica finale. La metodologia di rimozione delle multiple, oggetto di questo lavoro di tesi, è denominata “Surface-Related Multiple Elimination (S.R.M.E)” ed ha la peculiarità di ricavare dai dati stessi le informazioni necessarie per stimare le “surface-related multiples”. Nel dominio common shot le multiple sono predette per una coppia sorgente-ricevitore (xs, xr) attraverso la sommatoria dei prodotti di convoluzione tra le tracce del common shot gather (xs, xk) e le tracce del receiver gather corrispondente(xk, xr). Ripetendo l’operazione per ogni ricevitore (xr) si ottengono le multiple predette associate alla sorgente (xs) che dovranno essere rimosse attraverso una sottrazione adattativa dai dati.L’obiettivo dell’elaborato di tesi è comprendere le basi teoriche della metodologia ed applicarla ad un dato reale. Partendo da un modello sintetico 1-D, fino alla sua estensione 2-D, sono fornite le prove sviluppate grazie ad un algoritmo da me realizzato in linguaggio Matlab e sono discussi i limiti e le potenzialità del metodo. Il problema principale della metodologia è dovuto ai limiti logistici di acquisizione, come ad esempio l’impossibilità di registrare l’intero range di offset oppure la mancata coincidenza spaziale di una posizione di energizzazione con una posizione di ricezione (differenti shot interval e receiver interval). Sono quindi spiegate le motivazioni teoriche e le conseguenze pratiche di queste limitazioni, sono menzionate inoltre le metodologie per ridurne le conseguenze. La parte finale dell’elaborato fornisce i risultati dell’applicazione del metodo S.R.M.E ad un caso reale; questa parte del lavoro è stata eseguita grazie all’ausilio del software Promax della Società Landmark

Drug-resistant Streptococcus pyogenes: a case report of pyoderma and cellulitis

In the last years there has been worldwide an increase in the incidence of Streptococcus pyogenes infections and its sequelae. The purpose of the present study is to describe a clinical case of pyoderma and cellulitis by S. pyogenes with a pattern of resistance to penicillin, erythromycin and levofloxacin. Antimicrobial susceptibility was analysed by automated method of disk diffusion while E-test method was used to verify the minimum inhibitory concentration to penicillin, erythromycin and levofloxacin. Although penicillin remains the first-choice treatment for S. pyogenes infection, worldwide antibiotic resistance and the associated phenotypes are highly variable across countries. This case report strengthens the need for continued surveillance to obtain further insights into the forces governing resistance in S. pyogenes.</em

Reporting epidemiology of antibiotic resistance

The aim of antimicrobial resistance surveillance is to monitor temporal trends and provide clinicians with data to define empirical treatment protocols. The surveillance methods adopted in different settings can be significantly different and, therefore, no reference can be made to a single set of standards. This paper outlines the main features of analysis and reporting of antimicrobial resistance data according to the guidelines issued by the US Clinical and Laboratory Standards Institute, and the surveillance systems adopted in Europe. In this article the strengths and weaknesses of the various types of analyses will be discussed highlighting the critical aspects to be taken into account in surveillance data reporting

Diagnosis of Bacteriuria and Leukocyturia by Automated Flow Cytometry Compared with Urine Culture▿

Urinary tract infection (UTI) is a widespread disease, and thus, the most common samples tested in diagnostic microbiology laboratories are urine samples. The “gold standard” for diagnosis is still bacterial culture, but a large proportion of samples are negative. Unnecessary culture can be reduced by an effective screening test. We evaluated the performance of a new urine cytometer, the Sysmex UF-1000i (Dasit), on 703 urine samples submitted to our laboratory for culture. We compared bacteria and leukocyte (WBC) counts performed with the Sysmex UF-1000i to CFU-per-milliliter quantification on CPS agar to assess the best cutoff values. Different cutoff values of bacteria/ml and WBC/ml were compared to give the best discrimination. On the basis of the results obtained in this study, we suggest that when the Sysmex UF-1000i analyzer is used as a screening test for UTI the cutoff values should be 65 bacteria/ml and 100 WBC/ml. Diagnostic performance in terms of sensitivity (98.2%), specificity (62.1%), negative predictive value (98.7%), positive predictive value (53.7%), and diagnostic accuracy (73.3%) were satisfactory. Screening with the Sysmex UF-1000i is acceptable for routine use. In our laboratory, we have reduced the number of bacterial cultures by 43%, speeded up their reporting, and decreased the inappropriate use of antibiotics

Training in Vascular Microsurgery: The Ex Vivo Biological Model on Domestic Turkey Leg (Meleagris gallopavo)

: There are various models for practicing microsurgical anastomoses, from synthetic to ex vivo and in vivo biological ones. In this study, we present the domestic turkey (Meleagris gallopavo) as an ex vivo biological model in the practice of surgical anastomoses. In our opinion, it represents a model that is very similar to a human one, low cost, and easy to find. In fact, our study shows that the diameters of the arteries and veins used for anastomoses (tibial artery diameter: 2.5 ± 0.6 mm; tibial vein diameter: 3.5 ± 1.2 mm) are similar to those of human arteries and veins most frequently used in microsurgical free flaps. So, we believe that this animal model is a great model for microsurgical training for doctors who approach this difficult and long to learn discipline

Oral Administration of Vitamin D3 Prevents Corneal Damage in a Knock-Out Mouse Model of Sjögren’s Syndrome

Background: Vitamin D deficiency has been associated with dry eye development during Sjögren’s syndrome (SS). Here, we investigated whether repeated oral vitamin D3 supplementation could prevent the corneal epithelium damage in an SS mouse model. Methods: 30 female mouse knock-out for the thrombospondin 1 gene were randomized (six per group) in untreated mice euthanized at 6 weeks as negative control (C−) or at 12 weeks as the positive control for dry eye (C+). Other mice were sacrificed after 6 weeks of oral vitamin D3 supplementation in the drinking water (1000, 8000, and 20,000 IU/kg/week, respectively). Results: The C+ mice showed alterations in their corneal epithelial morphologies and thicknesses (p p < 0.01 vs. C+). Moreover, while the C+ mice exhibited high levels and activity of corneal tumor necrosis factor alpha converting enzyme (TACE), neovascularization and fibrosis markers; these were all reduced in the M and H mice. Conclusions: Oral vitamin D3 supplementation appeared to counteract the negative effect of TACE on corneal epithelium in a mouse model of SS-associated dry eye

Pre-Menopausal Breast Fat Density Might Predict MACE During 10 Years of Follow-Up: The BRECARD Study

Objectives: This study sought to determine whether the breast gland adipose tissue is associated with different rates of major adverse cardiac events (MACEs) in pre-menopausal women. Background: To our knowledge, no study investigated the impact of breast adipose tissue infiltration on MACEs in pre-menopausal women. Methods: Prospective multicenter cohort study conducted on pre-menopausal women &gt;40 years of age without cardiovascular disease and breast cancer at enrollment. The study started in January 2000 and ended in January 2009, and the end of the follow-up for the evaluation of MACEs was in January 2019. Participants underwent mammography to evaluate breast density and were divided into 4 groups according to their breast density. The primary endpoint was the probability of a MACE at 10 years of follow-up in patients staged for different breast deposition/adipose tissue deposition. Results: The propensity score matching divided the baseline population of 16,763 pre-menopausal women, leaving 3,272 women according to the category of breast density from A to D. These women were assigned to 4 groups of the study according to baseline breast density. At 10 years of follow-up, we had 160 MACEs in group 1, 62 MACEs in group 2, 27 MACEs in group 3, and 16 MACEs in group 4. MACEs were predicted by the initial diagnosis of lowest breast density (hazard ratio: 3.483; 95% confidence interval: 1.476-8.257). Further randomized clinical trials are needed to translate the results of the present study into clinical practice. The loss of ex vivo breast density models to study the cellular/molecular pathways implied in MACE is another study limitation. Conclusions: Among pre-menopausal women, a higher evidence of adipose tissue at the level of breast gland (lowest breast density, category A) versus higher breast density shows higher rates of MACEs. Therefore, the screening mammography could be proposed in overweight women to stage breast density and to predict MACEs. (Breast Density in Pre-menopausal Women Is Predictive of Cardiovascular Outcomes at 10 Years of Follow-Up [BRECARD]; NCT03779217)