SARS-CoV-2 infection: NLRP3 inflammasome as plausible target to prevent cardiopulmonary complications?

NLRP3 (NOD-, LRR- and pyrin domain-containing protein 3) inflammasome has recently become an intriguing target of several chronic and viral diseases. Here, we argue that targeting NLRP3 inflammasome could be a strategy to prevent cardiovascular outcomes [fulminant myocarditis, heart failure, venous thromboembolism (VTE)] and acute respiratory distress syndrome (ARDS) in patients with SARS-CoV-2 infection. We discuss the rationale for NLRP3 targeting in clinical trials as an effective therapeutic strategy aimed to improve prognosis of COVID-19, analyzing the potential of two therapeutic options (tranilast and OLT1177) currently available in clinical practice.

Quality of eggs from Lohmann Brown Classic laying hens fed black soldier fly meal as substitute for soya bean.

Soya bean is the main protein source in poultry feed but rising prices make an alternative protein source necessary. Insects, such as the black soldier fly (Hermetia illucens), may be an attractive solution for hens, although little information is available on their effect on egg quality. The present study aims to fill this gap by testing the effect of 100% replacement of soya bean with H. illucens larva meal in the diet of Lohmann Brown Classic laying hens for 21 weeks. At the end of the trial, the eggs were characterized for parameters such as weight, colour, proximate composition of albumen and yolk, and content of carotenoids, tocopherols and cholesterol. The fatty acid profile of yolks was also determined. Hens fed the insect-based diet produced eggs (HIM group) with a higher proportion of yolk than the group fed the soya bean-based diet (SBM group). HIM was associated with redder yolks (red index 5.63 v. 1.36) than SBM. HIM yolks were richer in γ-tocopherol (4.0 against 2.4 mg/kg), lutein (8.6 against 4.9 mg/kg), ß-carotene (0.33 against 0.19 mg/kg) and total carotenoids (15 against 10.5 mg/kg) than SBM yolks. The fatty acid composition of HIM yolks was almost identical to that of SBM yolks. Finally, HIM yolks contained 11% less cholesterol than SBM yolks. These results suggest that H. illucens larva meal is a suitable total substitute for soya bean meal in the diet of Lohmann Brown Classic laying hens. A sustainable alternative to the plant protein source therefore seems feasible.

Immunohistochemical analysis of dendritic cells in skin lesions: correlations with survival time.

The response to wounds until healing requires the activity of many cell types coordinate in space and time, so that the types of cells in a wound and their localization may be of help to date lesions with respect to death, which would be useful in forensic pathology. Cells reacting to injury include dendritic cells; the early reaction of these cells to skin wounding has not yet been investigated in humans, which was the aim of this study. Samples of wounded and control skin were taken at autopsy and analyzed by affinity histochemistry. Both epidermal and dermal MHC-II+ cells increased transiently in number within the first hour after wounding, then decreased. In the epidermis the increase affected also CD1a+ cells, i.e. well differentiated Langherhans cells, which however increased less, earlier and for a shorter time period than MHC-II+ cells. Dermal MHC-II+ cells became part of a perivascular mononuclear cell infiltrate visible in the subpapillary dermis by 60 min after wounding, which contained also mast cells. The immediately perivascular MHC-II+ cells were DC-SIGN- and CD11c-, while MHC-II+, DC-SIGN+, CD11c+ dendritic cells were predominantly located at the periphery of infiltrates and some were near the epidermis. Mast cells underwent degranulation, besides increase in number, in the first hours after wounding. The results suggest that skin dendritic cells, including Langerhans cells, participate to the early response to wounding in concert with mast cells, and that subpapillary blood vessels are primary sites of cell infiltration during that response in humans. The results show that the ratio between CD1a positive and MHC-II positive cells in the epidermis, the degranulation index of mast cells and the relative volume of MHC-II positive cells in the dermis can be added to the tools useful to distinguish vital from post mortem lesions and, the first two of them, to estimate the interval between a lesion and death.

Comparative genotoxicity of cobalt nanoparticles and ions on human peripheral leukocytes in vitro.

Owing to the increasing development of nanotechnology, there is a need to assess how engineered nanomaterials can interact with living cells. The main purpose of the present study was to assess whether metal cobalt nanoparticles (CoNP 100-500 nm) are genotoxic compared to cobalt ions (Co(2+)). Uptake experiments were carried out by incubating peripheral blood leukocytes (PBLs) with (57)Co(2+) (added to stable Co(2+) 10(-2) M to obtain concentrations in the range of 10(-5) to 10(-4) M) or with (60)CoNP for 24 and 48 h. Whereas intracellular Co(2+) showed slight or no variations over the baseline levels, CoNP were taken up efficiently leading to intracellular CoNP concentrations of 485 +/- 106.1 and 970 +/- 99 fg per cell after 24 and 48 h, respectively. The genotoxicity end points considered in this study were the frequency of binucleated micronucleated (BNMN) cells and the percentage of tail DNA (% Tail DNA) fragmentation by means of the comet assay. Genotoxic effects were evaluated by incubating PBLs of three healthy donors with subtoxic concentrations (10(-5) to 8 x 10(-5)M) of Co(2+) in the form of cobalt chloride, CoNP and 'washed' CoNP, the latter to exclude any interference by Co(2+). On a group basis, Co(2+) induced a clear trend in the increase of the BNMN frequency, whereas CoNP showed only minor changes. Moreover, we observed a high variability among donors in the induction of micronuclei. The comet assay showed a statistically significant dose-related increase in % Tail DNA for CoNP (P < 0,001), whereas Co(2+) did not induce significant changes over control values. These findings suggest that nanosized Co can be internalized by human leukocytes and can interact with DNA leading to the observed genotoxic effects, which are, however, modulated both by donor's characteristics and/or by Co(2+) release.

Early increase in TNF-alpha-containing mast cells in skin lesions.

We have evaluated the numbers and immunohistochemical positivity for tumor necrosis factor (TNF) alpha of the mast cells in (a) 40 skin samples collected at autopsy from subjects who had survived for a few seconds to 1 h, (b) 10 samples of post-mortem skin lesions and (c) 10 surgical biopsies of healthy skin. Sections were treated with fluoresceinated avidin, to tag mast cell granules, followed by indirect immunohistochemistry for TNF-alpha with polyclonal primary and rhodaminated secondary antibodies. We could confirm a progressive increase in mast cell numbers, which became significant 1 h after trauma. More important, the fraction of mast cells positive for TNF-alpha increased progressively in the same time period and became significantly higher than controls in specimens collected more than 15 min after trauma. Samples from post-mortem lesions had significantly fewer mast cells and fewer TNF-alpha-positive cells than any other group of samples. The results suggest that mast cells are quickly recruited to an injured site in response to trauma and upregulate their TNF-alpha content, which can play an early role in directing tissue response to injury. The forensic pathologist can take advantage from this behavior of mast cells for the evaluation of the timing of early vital lesions.

Affinity cytochemistry analysis of mast cells in skin lesions: a possible tool to assess the timing of lesions after death.

The histamine content in vital wounds is known to increase, with a zenith after 3 h, and then decrease until 24 h after wounding. We addressed whether this biochemical alteration has a morphological counterpart. Since the main source of skin histamine are mast cells, the distribution and number of these cells was assessed upon labeling with fluorescent avidin and with antibodies to the mast cell specific enzymes, chymase and tryptase. Analyses were performed on skin from 15 healthy controls (from surgical biopsies), from 15 post-mortem lesions and 75 vital lesions, obtained at autopsy from subjects who had survived from a few seconds to 24 h. The number of mast cells per unit area of section surface increased progressively with survival time, up to a maximum in subjects who survived 1-3 h ( p<0.01), and decreased thereafter becoming less than in the controls if lesions had occurred earlier than 6 h before death ( p<0.01). Samples from post-mortem lesions had significantly fewer mast cells than those of any other groups of samples ( p<0.01). We suggest that in association to other histological and circumstantial evidence the analysis of mast cells by affinity cytochemistry can help to discriminate vital from post-mortem lesions and to estimate survival time after lesions.

Immunohistochemical localization of mast cells as a tool for the discrimination of vital and postmortem lesions.

In wounds that are inflicted at least 60 min before death, histamine levels can increase up to 100%. This functional effect might have a morphometric counterpart. Mast cells play a crucial role in acute inflammatory reactions and in the healing process of wounds. Therefore, the density of these cells was immunohistochemically assessed in tissue from 20 healthy controls (Group 1), 20 vital skin lesions (Group 2) (age range: a few seconds to 1 h), and 20 postmortem lesions (Group 3). A piece of skin close to the vital lesion was also obtained from the homolateral part of the body (Group 4). Mast cell density was significantly higher at the level of the vital lesions (11.28+/-2.44) than elsewhere (healthy controls 7.66+/-1.27, postmortem lesions 4.13+/-1.46, skin close to the vital lesions 4.88+/-1.59). No differences were found between the values assessed in the skin samples close to the vital lesions and in those in the postmortem lesions. Therefore, mast cell richness in the vital lesions exhibited a proportional morphological correlation with previously detected histamine values in cutaneous vital lesions. These results suggest that the detection of mast cells with immunohistochemical techniques can lead to a high level of discrimination (based on statistical data) between antemortem and postmortem lesions. This method could also be used to ascertain the vitality of lesions.

Airbag and hearing loss: our experience.

The international literature reports sporadic cases of sudden sensorineural hearing loss associated with Airbag inflation in subjects involved in car accidents, who had never suffered from hearing loss and had no history of pathologies that could otherwise explain the hearing loss. The authors have studied the actual incidence of this possibility, analysing 25 subjects submitted to an airbag inflation. Only subjects who had never suffered from previous otological pathology were considered for the study. The examination was carried out 3 to 10 days after the accident. Otoscopical examination, tympanometry and audiometry were normal in all the examined subjects. The Author's opinion is that traumatic hearing loss following airbag inflation is a rare or exceptional incident that probably occurs in subjects with a particular susceptibility and sensitivity.

Neck motion evaluation after whiplash: a radiographic and kinematic protocol.

Evaluation of functional damage is the main aspect in the diagnosis of whiplash injury, therefore some type of instrumental testing is strongly needed. We describe an instrumental protocol to test subjects with whiplash, to support clinical evaluation, in order to solve the difficulty of evaluating the functional injury and the lack of fixed parameters capable of identifying this kind of injury. Eighty subjects suffering from after effects of whiplash and seventy five normal subjects undergo two instrumental tests: kinematic examination of neck mobility, and measurements of extension-flexion X-rays of the cervical spine. The results exhibited by the traumatized patients have been compared with the normal range of motion obtained from the data bank of the normal subjects. This is a report in progress of a larger research on whiplash evaluation in a clinical setting.

[The heterogeneity of myocardial sympathetic innervation in normal subjects: an assessment by iodine-123 metaiodobenzylguanidine scintigraphy]. / Eterogeneità dell'innervazione simpatica del miocardio in soggetti normali: valutazione mediante scintigrafia con iodio-123 metaiodobenzilguanidina.

123I-radiolabeled metaiodobenzylguanidine (123I-MIBG) cardiac imaging has been used to evaluate the distribution of sympathetic nervous system (SNS) in the heart. Different heart diseases have shown impaired cardiac SNS distribution as reflected by MIBG activity. The aim of this study was to assess the cardiac distribution of SNS in normal subjects, using MIBG imaging. Ten normal subjects (1 male and 9 females, mean age 46 +/- 9 years) with no cardiac abnormalities underwent myocardial 123I-MIBG scintigraphy, Tc-99m methoxyisobutylisonitrile (MIBI) cardiac perfusion imaging and equilibrium radionuclide angiography (RNA). Regional myocardial MIBG and MIBI activities were quantitatively evaluated using a region of interest analysis. For this purpose, the left ventricle was divided into 6 myocardial regions as anterior, apical, inferior, septum, lateral and posterolateral. In particular, myocardial MIBG and MIBI activities were measured as myocardium to mediastinum ratio. Regional left ventricular function was assessed by RNA. Myocardial MIBG uptake was homogeneous in anterior (2.2 +/- 0.5), inferior (2.5 +/- 0.7), septal (2.4 +/- 0.4), lateral (2.3 +/- 0.4), and posterolateral (2.3 +/- 0.4) regions. Conversely, MIBG uptake was significantly lower in the apical region (1.9 +/- 0.3) compared to all other left ventricular segments (p < 0.05). Regional myocardial perfusion, as measured by MIBI uptake, was homogeneous in all regions. No regional left ventricular wall motion abnormalities were observed by RNA. In conclusion, our data suggest that a decreased MIBG uptake may be observed in the left ventricular apical region of normal subjects reflecting reduced sympathetic innervation of the apex. This finding is not related to myocardial perfusion or wall motion abnormalities. The knowledge of cardiac sympathetic innervation in normal subjects may be helpful to assess SNS abnormalities in heart disease.

[Evaluation of left ventricular function in patients with mammary carcinoma after treatment with epidoxorubicin using a high-dose, short-term protocol]. / Valutazione della funzione ventricolare sinistra in pazienti con carcinoma mammario sottoposti a terapia con epidoxorubicina secondo il protocollo "high-dose short-term".

Anthracyclines are effective chemotherapeutic agents against various malignancies but their therapeutic value is limited by well-known dose-related cardiotoxicity, mainly induced by oxygen free radicals. Left ventricular diastolic and systolic functional abnormalities precede the clinical evidence of cardiotoxicity. The aim of this study was to evaluate the possible cardiotoxicity of epidoxorubicin administered as "high-dose short-term" protocol. Twenty patients (mean age 50.4 +/- 7.9 years) without cardiac disease, affected by advanced breast cancer were studied. All patients were treated with epidoxorubicin as neoadjuvant chemotherapy according to the new protocol "high-dose short-term" (cumulative dose 475.8 +/- 35.6 mg/m2, range 450-600 mg/m2, in 4-6 weeks). The effectiveness of cancer chemotherapy was monitored by clinical evaluation and mammography performed before and after treatment. All patients underwent color Doppler echocardiography and resting radionuclide angiocardiography in baseline condition and 30 +/- 10 days after the last cycle of chemotherapy. All patients showed a significant reduction of tumor lesion after chemotherapy. Left ventricular systolic and diastolic function parameters obtained by echocardiography (fractional shortening 33.1 +/- 4.5% vs 32.4 +/- 4.8%; ejection fraction 63.6 +/- 6.2% vs 62.9 +/- 5.7%; E/A ratio 1.73 +/- 0.64 vs 1.82 +/- 0.67; E wave deceleration time 204 +/- 24.6 ms vs 208.5 +/- 31.7 ms;isovolumetric relaxation time 79 +/- 15.7 ms vs 80 +/- 17.8 ms) and radionuclide angiocardiography (ejection fraction 62.4 +/- 7% vs 61.8 +/- 5.9; peak ejection rate 2.87 +/- 0.44 VTD/s vs 2.74 +/- 0.46 VTD/s; peak filling rate 2.72 +/- 0.54 VTD/s vs 2.6 +/- 0.58 VTD/s) did not show significant changes after treatment. In conclusion, our results suggest that epidoxorubicin administration using the "high-dose short-term" protocol in patients with breast cancer does not induce early significant abnormalities of left ventricular systolic and diastolic function.

[Medico-legal aspects of chronic respiratory insufficiency under long-term oxygen inhalation therapy]. / Aspetti medico-legali dell'insufficienza respiratoria cronica con ossigeno terapia a lungo termine.

Ninety-one subjects affected by severe pulmonary insufficiency necessitating continuous treatment with oxygen were studied. The social and professional realities and their status within the national health service were determined. The results of the research, obtained through the use of the questionnaires, showed that the lives of these subjects are very much influenced by their pathology. Among the factors considered were necessity for home assistance, scarce or absence of autonomy and contact with the outside world and extremely reduced work capacity. Elevated public spending for assistance was analyzed in reference to public health insurance as regards each single subject and the type of lung dysfunction from which the subject suffered.

[Chronic kidney failure: dialysis and kidney transplantation. The medicolegal and social considerations]. / Insufficienza renale cronica: dialisi e trapianto renale. Considerazioni medico-legali e sociali.

The authors carried out a research on the social and labour problems of the people affected by chronic renal failure, either on dialysis treatment or already transplanted since two years. It has been given a questionnaire based on questions about the person, the working position, the social insurance situation, the family and personal problems. The results show a good social reinsert and a good recover of the transplanted in opposition of the many problems had by those on dialysis; in particular it has been considered also the problem of the periodic absences from work. It has been also considered the italian law situation regarding the people affected by chronic renal failure in some different social insurance fields.

[4'-epidoxorubicin: its cardiotoxicity. Possible cardiac protection with gallopamil, a drug with calcium-antagonist action]. / La 4'-epidoxorubicina: sua cardiotossicità. Eventuale cardioprotezione con il gallopamil, farmaco ad azione calcioantagonista.

The 4'-EPI-doxorubicin (4'-EDX) has shown antitumoral activity against a broad spectrum of human tumors. The same clinical reports have shown therapeutic activity comparable to doxorubicin (adriamycin), but a lower cardiac toxicity. 4'-EDX cardiotoxicity appears after highest cumulative doses: 935 mg/m2 bs for 4'-EDX vs 550 mg/m2 bs for adriamycin. Moreover we have observed a protective action of the calcium antagonist gallopamil. We have found an improvement in ventricular performance after using gallopamil, and a reduction of 4'-EDX cardiotoxicity. Cardiac function was evaluated by EKS, STI, echography, myocardial scintigraphy course. Our result indicates the possibility of treating tumors without serious cardiotoxic by 4'-EDX and gallopamil.

Differential effects of beclobrate on lipid/lipoprotein distribution in normal and hypercholesterolemic rats.

Beclobrate, a new fibric acid derivative, displays remarkable lipid lowering activity in rodents. In order to evaluate changes in the distribution and liver handling of lipoproteins, beclobrate was tested in rats fed on a normal or hypercholesterolemic diet. On the normal diet, beclobrate lowered total plasma cholesterol by 22-33.4% (10-50 mg/kg); the cholesterol reduction occurred mainly in high density lipoproteins (HDL) (by 24-45% with the three tested doses). The metabolic clearance of 125I-labelled beta-very low density lipoproteins (beta-VLDL) injected into these animals almost doubled (0.20 1/h vs. 0.13 1/h in controls) after treatment with 20 mg/kg of beclobrate. In addition, beclobrate administration dramatically increased the activity of the high-affinity receptors for beta-VLDL in isolated liver membranes (Bmax: 208 +/- 17.6 vs. 146 +/- 2.6 ng/mg of protein for controls). On the hypercholesterolemic diet, beclobrate treatment (50 mg/kg) was associated with a 25% reduction in total cholesterol accompanied, however, by a 166% rise in HDL cholesterol. In these animals, the composition of VLDL, typically cholesterol-enriched, became close to normal. The increased HDL was characterized by a remarkable enrichment with particles containing apolipoprotein E (apo E), which is compatible with either an improved peripheral cholesterol removal or an enhanced direct secretion of apo E. The two models offer different opportunities for evaluating the mechanism of action of this new lipid lowering agent. Lipoprotein catabolism and receptor-mediated clearance were characteristically improved in normolipidemic rats whereas major effects on HDL metabolism could be demonstrated in hypercholesterolemic rats.