Comparative phenotypic and functional analyses of the effects of autologous plasma and recombinant human macrophage-colony stimulating factor (M-CSF) on porcine monocyte to macrophage differentiation

Abstract Porcine monocyte-derived macrophages (moMΦ) have been employed as a model cell in numerous studies of the porcine immune system. However, the lack of a standardized method for moMΦ differentiation hampers the comparison of results coming from the use of different laboratory protocols. In this study we compared the use of varying concentrations of autologous plasma (10, 20 and 30% v/v) or recombinant human macrophage-colony stimulating factor (hM-CSF; 50, 100, and 200 ng/ml) to differentiate porcine monocytes into macrophages. Changes in cell morphology and surface marker expression were assessed by confocal microscopy and flow cytometry. Macrophage differentiation was evaluated by analysing TNF-α response to LPS stimulation and determining cytokine secretion patterns under both basal conditions and after classical and alternative activation. The effects of the differentiation methods on metabolic activity and susceptibility to infection with the myelotropic African swine fever virus (ASFV) were also evaluated. Monocytes cultured using the different culture conditions tested augmented in dimension and cellular complexity, but increasing porcine plasma concentrations resulted in a dose dependent enhancement in granularity and a marked pleomorphism. As expected, CD163, MHC class II DR and CD203a expression were up-regulated in both hM-CSF (M-CSF-moMΦ) and autologous plasma cultured macrophages (AP-moMΦ), although a lower percentage of CD163+ cells were found following differentiation with high percentages of porcine plasma. We observed enhanced number of viable cells using high concentration of hM-CSF compared to porcine plasma, suggesting a proliferative effect. Irrespective of differentiation conditions, monocyte differentiation into macrophages resulted in an increased susceptibility to ASFV and yielded larger amounts of LPS-induced TNF-α. AP-moMΦ showed a higher basal release of IL-1RA compared to those cultured with hM-CSF and displayed a reduced ability to respond to classical activation, suggesting that the use of high percentages of porcine plasma led to the acquisition of a M2-like phenotype. We conclude that all the protocols tested in this study can be considered as suitable to produce porcine moMΦ, although the use of hM-CSF provides high responsiveness to M1 polarization. Since a higher phenotypic and functional inter-animal variability was observed in AP-moMΦ, we propose that the use of low concentration of hM-CSF should be adopted as the method of choice to provide a better reproducibility between experiments

Impact of environmental pollution and weather changes on the incidence of ST-elevation myocardial infarction

Background: Environmental pollution and weather changes unfavorably impact on cardiovascular disease. However, limited research has focused on ST-elevation myocardial infarction (STEMI), the most severe yet distinctive form of acute coronary syndrome. Methods and results: We appraised the impact of environmental and weather changes on the incidence of STEMI, analysing the bivariate and multivariable association between several environmental and atmospheric parameters and the daily incidence of STEMI in two large Italian urban areas. Specifically, we appraised: carbon monoxide (CO), nitrogen dioxide (NO2), nitric oxide (NOX), ozone, particulate matter smaller than 10 μm (PM10) and than 2.5 μm (PM2.5), temperature, atmospheric pressure, humidity and rainfall. A total of 4285 days at risk were appraised, with 3473 cases of STEMI. Specifically, no STEMI occurred in 1920 (44.8%) days, whereas one or more occurred in the remaining 2365 (55.2%) days. Multilevel modelling identified several pollution and weather predictors of STEMI. In particular, concentrations of CO (p=0.024), NOX (p=0.039), ozone (p=0.003), PM10 (p=0.033) and PM2.5 (p=0.042) predicted STEMI as early as three days before the event, as well as subsequently, and NO predicted STEMI one day before (p = 0.010), as well as on the same day. A similar predictive role was evident for temperature and atmospheric pressure (all p < 0.05). Conclusions: The risk of STEMI is strongly associated with pollution and weather features. While causation cannot yet be proven, environmental and weather changes could be exploited to predict STEMI risk in the following days

Association of kidney disease measures with risk of renal function worsening in patients with type 1 diabetes

Background: Albuminuria has been classically considered a marker of kidney damage progression in diabetic patients and it is routinely assessed to monitor kidney function. However, the role of a mild GFR reduction on the development of stage 653 CKD has been less explored in type 1 diabetes mellitus (T1DM) patients. Aim of the present study was to evaluate the prognostic role of kidney disease measures, namely albuminuria and reduced GFR, on the development of stage 653 CKD in a large cohort of patients affected by T1DM. Methods: A total of 4284 patients affected by T1DM followed-up at 76 diabetes centers participating to the Italian Association of Clinical Diabetologists (Associazione Medici Diabetologi, AMD) initiative constitutes the study population. Urinary albumin excretion (ACR) and estimated GFR (eGFR) were retrieved and analyzed. The incidence of stage 653 CKD (eGFR < 60 mL/min/1.73 m2) or eGFR reduction > 30% from baseline was evaluated. Results: The mean estimated GFR was 98 \ub1 17 mL/min/1.73m2 and the proportion of patients with albuminuria was 15.3% (n = 654) at baseline. About 8% (n = 337) of patients developed one of the two renal endpoints during the 4-year follow-up period. Age, albuminuria (micro or macro) and baseline eGFR < 90 ml/min/m2 were independent risk factors for stage 653 CKD and renal function worsening. When compared to patients with eGFR > 90 ml/min/1.73m2 and normoalbuminuria, those with albuminuria at baseline had a 1.69 greater risk of reaching stage 3 CKD, while patients with mild eGFR reduction (i.e. eGFR between 90 and 60 mL/min/1.73 m2) show a 3.81 greater risk that rose to 8.24 for those patients with albuminuria and mild eGFR reduction at baseline. Conclusions: Albuminuria and eGFR reduction represent independent risk factors for incident stage 653 CKD in T1DM patients. The simultaneous occurrence of reduced eGFR and albuminuria have a synergistic effect on renal function worsening

Arthroscopic transphyseal anterior cruciate ligament reconstructions in adolescent athletes

PURPOSE: To evaluate the 5-year outcome of arthroscopic transphyseal anterior cruciate ligament (ACL) reconstruction in 19 adolescent athletes. METHODS: 14 male and 5 female adolescent athletes aged 12 to 16 (mean, 13.9) years with Tanner stage 2 or 3 open physes underwent arthroscopic transphyseal ACL reconstruction by a single surgeon and were followed up for 5 years. Patients were evaluated using the numerical rating score (NRS) for pain, knee osteoarthritis outcome score (KOOS), International Knee Documentation Committee (IKDC) score, Tegner Activity Scale, and Lysholm Score, as well as the leg length discrepancy, femorotibial alignment, varus or valgus deformities, active and passive knee range of motion. RESULTS: At 5-year follow-up, physes were closed in all patients. The mean NRS for pain improved from 7.2 to 1.6; the KOOS improved from 55.3 to 88; the mean IKDC score improved from 34.5 to 84; the mean Tegner Activity Scale improved from 2.7 to 8.2 and was comparable with that before injury (8.4); and the mean Lysholm score improved from 36.3 to 84.6. All except 2 patients returned to their pre-injury level of sports activity after a mean of 25 weeks. The 2 exceptions had a 2+ Jerk test and a 3+ Lachman test; one of them also had positive signs for a lateral meniscal lesion. Both had sustained a second trauma not long before the 5-year follow-up. Two patients had reduced sensitivity in the anteromedial aspect of the proximal third of the tibia. One patient had leg length discrepancy of +1.5 cm owing to overgrowth response of the physis. CONCLUSION: Transphyseal ACL reconstruction is a viable option for skeletally immature patients, with high reproducibility, a high rate of return to sport, and a low incidence of growth disturbance. Early surgery can prevent the onset of meniscal lesions and early osteoarthritis

Isolated Subscapularis Tendon Tear in a Skeletally Immature Soccer Player.

Subscapularis injury in adolescents, usually associated to an avulsion fracture of the lesser humeral tuberosity, accounts for less than 2% of all fractures of the proximal humerus. Isolated tears of the subscapularis tendon without a history of dislocation and associated avulsion fractures are an even rarer occurrence, and treatment is controversial. This article describes a rare case of a 12-year-old suffering from an isolated subscapularis tear and discusses its management. The patient was evaluated at presentation, and at 1 to 2.5 months after he underwent a cuff tear arthroscopic repair with a single "all suture" anchor loaded with two wires, active/passive range of motion (A/PROM), Constant-Murley score, and American Shoulder and Elbow Surgeons (ASES) score were noted. Patient reported an excellent outcome, recovered the whole ROM, was pain free, and returned to the previous level of activity. Isolated avulsion of the subscapularis tendon requires a high index of suspicion for a proper diagnosis as early treatment is required for a good recovery. Arthroscopy reserves more advantages in proper hands, restoring the previous levels of function and activity. An increase in attention for this condition is mandatory in a society where many adolescents are getting more and more active in high levels of sport activities

Distalization and Medialization of Tibial Tuberosity for the Treatment of Potential Patellar Instability with Patella Alta.

Purpose  The aim of the study was to test the distalization and medialization of the tibial tuberosity (DMTT) for the treatment of patellar instability associated with patella alta, focusing on residual instability and pain. Methods  Twenty-four consecutive patients (26 knees) suffering from patellofemoral instability and patella alta were treated by DMTT. Two groups were identified, which differed for documented history of frank patella dislocation. The groups were named objective patellar instability (OPI) (history of dislocation) and potential patella instability (PPI) (no dislocation). Outcome was measured with visual analogue scale (VAS), Kujala score, and Tegner score. Comparison between groups was performed using Student's t -test, Wilcoxon rank score, and Fisher's exact test (significance at p  < 0.05). Results  At 50 ± 18 and 41 ± 18 months of follow-up, respectively, both PPI and OPI groups obtained a significant pain reduction and functional improvement. The PPI group showed a significant decrease of the subjective instability. No procedure-related complications were reported. Conclusion  This study suggests that DMTT is a viable option for PPI patients with patella alta. The outcome was comparable between PPI and OPI cases; however, decrease in subjective instability was significantly greater in PPI patients. Level of Evidence  Level III, retrospective comparative study

Relationship between magnetism and prion protein

The mechanism of conversion of the normal prion protein (PrPC) into aggregates of its pathological conformer (PrPSc) reamins unclear. The aim of this study was to evaluate the effects induced by exposure of biological samples containing PrPSC to a magnetic field induced prominent molecular changes of samples indicated by the IR spectra located in the region that contains contribution primarily from absorption of amides. This finding suggests the existence of a strong correlation between magnetism and PrPsc and supports a new hypothesis that explains the conversion of normal PrPc to abnormal isoform PrPsc