Modelling the long-term health impact of COVID-19 using Graphical Chain Models brief heading: long COVID prediction by graphical chain models.

BACKGROUND: Long-term sequelae of SARS-CoV-2 infection, namely long COVID syndrome, affect about 10% of severe COVID-19 survivors. This condition includes several physical symptoms and objective measures of organ dysfunction resulting from a complex interaction between individual predisposing factors and the acute manifestation of disease. We aimed at describing the complexity of the relationship between long COVID symptoms and their predictors in a population of survivors of hospitalization for severe COVID-19-related pneumonia using a Graphical Chain Model (GCM). METHODS: 96 patients with severe COVID-19 hospitalized in a non-intensive ward at the "Santa Maria" University Hospital, Terni, Italy, were followed up at 3-6 months. Data regarding present and previous clinical status, drug treatment, findings recorded during the in-hospital phase, presence of symptoms and signs of organ damage at follow-up were collected. Static and dynamic cardiac and respiratory parameters were evaluated by resting pulmonary function test, echocardiography, high-resolution chest tomography (HRCT) and cardiopulmonary exercise testing (CPET). RESULTS: Twelve clinically most relevant factors were identified and partitioned into four ordered blocks in the GCM: block 1 - gender, smoking, age and body mass index (BMI); block 2 - admission to the intensive care unit (ICU) and length of follow-up in days; block 3 - peak oxygen consumption (VO2), forced expiratory volume at first second (FEV1), D-dimer levels, depression score and presence of fatigue; block 4 - HRCT pathological findings. Higher BMI and smoking had a significant impact on the probability of a patient's admission to ICU. VO2 showed dependency on length of follow-up. FEV1 was related to the self-assessed indicator of fatigue, and, in turn, fatigue was significantly associated with the depression score. Notably, neither fatigue nor depression depended on variables in block 2, including length of follow-up. CONCLUSIONS: The biological plausibility of the relationships between variables demonstrated by the GCM validates the efficacy of this approach as a valuable statistical tool for elucidating structural features, such as conditional dependencies and associations. This promising method holds potential for exploring the long-term health repercussions of COVID-19 by identifying predictive factors and establishing suitable therapeutic strategies.

Breaking bad news: an active learning method for medical students.

BACKGROUND: Breaking bad news is one of the most difficult aspects of communication in medicine. The objective of this study was to assess the relevance of a novel active learning course on breaking bad news for fifth-year students. METHODS: Students were divided into two groups: Group 1, the intervention group, participated in a multidisciplinary formative discussion workshop on breaking bad news with videos, discussions with a pluri-professional team, and concluding with the development of a guide on good practice in breaking bad news through collective intelligence; Group 2, the control group, received no additional training besides conventional university course. The relevance of discussion-group-based active training was assessed in a summative objective structured clinical examination (OSCE) station particularly through the students' communication skills. RESULTS: Thirty-one students were included: 17 in Group 1 and 14 in Group 2. The mean (range) score in the OSCE was significantly higher in Group 1 than in Group 2 (10.49 out of 15 (7; 13) vs. 7.80 (4.75; 12.5), respectively; p = 0.0007). The proportion of students assessed by the evaluator to have received additional training in breaking bad news was 88.2% (15 of the 17) in Group 1 and 21.4% (3 of the 14) in Group 2 (p = 0.001). The intergroup differences in the Rosenberg Self-Esteem Scale and Jefferson Scale of Empathy scores were not significant, and both scores were not correlated with the students' self-assessed score for success in the OSCE. CONCLUSION: Compared to the conventional course, this new active learning method for breaking bad news was associated with a significantly higher score in a summative OSCE. A longer-term validation study is needed to confirm these exploratory data.

Superoxide dismutase content and microsomal lipid composition of tumours with different growth rates.

The content of cytosolic superoxide dismutase has been determined in Morris hepatomas 3924A (fast-growing) and 44 (slow-growing) and in ascites tumour cells (Novikoff hepatoma and Ehrlich-Lettré). The enzyme is decreased in all the tumours examined. The lowest amounts were found in the tumours with the fastest growth rates. Measurements of the lipid composition and fluidity of microsomal membranes isolated from Morris hepatomas show that also these parameters are changed in relation to the growth rate. The lipid to protein ratio and the degree of fatty acid unsaturation decrease gradually from rat liver to hepatoma 44 and 3924A microsomes. The different lipid composition is reflected also by differences in the physical environment of the bilayer, as indicated by data obtained with spin-labeled fatty acids. It is proposed that the changes in the membrane lipid composition and organization are consequent to the decrease in the protective effect of cytosolic superoxide dismutase against the O2- induced lipid peroxidation.

Initiating activity of 1,1,2,2-tetrachloroethane in two-stage BALB/c 3T3 cell transformation.

By using in vitro two-stage BALB/c 3T3 cell transformation assay, we have tested the effect of promoting treatment with tetradecanoylphorbol acetate (TPA) on transformation induced by 1,1,2,2-tetrachloroethane (1,1,2,2-TTCE). Cells were treated with subeffective or transforming concentrations of 1,1,2,2-TTCE in the presence of an S9-mix activating system, followed by TPA promoting treatment. The transforming activity of 1,1,2,2-TTCE is evident only by reseeding confluent cells and allowing additional rounds of cell replications in the amplification test. Treatment with TPA leads to a marked transformation yield in all plates scored even at the lowest assayed dosage of 1,1,2,2-TTCE, without performing amplification of transformation.

Benzene adducts with rat nucleic acids and proteins: dose-response relationship after treatment in vivo.

The dose-response relationship of the benzene covalent interaction with biological macromolecules from rat organs was studied. The administered dose range was 3.6 x 10(7) starting from the highest dosage employed, 486 mg/kg, which is oncogenic for rodents, and included low and very low dosages. The present study was initially performed with tritium-labeled benzene, administered by IP injection. In order to exclude the possibility that part of the detected radioactivity was due to tritium incorporated into DNA from metabolic processes, 14C-benzene was then also used following a similar experimental design. By HPLC analysis, a single adduct from benzene-treated DNA was detected; adduct identification will be attempted in the near future. Linear dose-response relationship was observed within most of the range of explored doses. Linearity was particularly evident within low and very low dosages. Saturation of benzene metabolism did occur at the highest dosages for most of the assayed macromolecules and organs, especially in rat liver. This finding could be considered as indicative of the dose-response relationship of tumor induction and could be used in risk assessment.

In vivo and in vitro binding of 1,2-dibromoethane and 1,2-dichloroethane to macromolecules in rat and mouse organs.

The comparative interaction of equimolar amounts of 1,2-dichloroethane and 1,2-dibromoethane with rat and mouse nucleic acids was studied in both in vivo (liver, lung, kidney and stomach) and in vitro (liver microsomal and/or cytosolic fractions) systems. In vivo, liver and kidney DNA showed the highest labeling, whereas the binding to lung DNA was barely detectable. Dibromoethane was more highly reactive than dichloroethane in both species. With dichloroethane, mouse DNA labeling was higher than rat DNA labeling whatever the organ considered: the opposite was seen for the bioactivation of dibromoethane. RNA and protein labelings were higher than DNA labeling, with no particular pattern in terms of organ or species involvement. In vitro, in addition to a low chemical reactivity towards nucleic acids shown by haloethanes per se, both compounds were bioactivated by either liver microsomes and cytosolic fractions to reactive forms capable of binding to DNA and polynucleotides. UV irradiation did not photoactivate dibromoethane and dichloroethane. The in vitro interaction with DNA mediated by enzymatic fractions was PB-inducible (one order of magnitude, using rat microsomes). In vitro bioactivation of haloethanes was mainly performed by microsomes in the case of dichloroethane and by cytosolic fractions in the case of dibromoethane. When microsomes plus cytosol were used, rat enzymes were more efficient than mouse enzymes in inducing a dibromoethane-DNA interaction: the opposite situation occurred for dichloroethane-DNA interaction, and this is in agreement with the in vivo pattern. In the presence of both metabolic pathways, addition or synergism occurred. Dibromoethane was always more reactive than dichloroethane. An indication of the presence of a microsomal GSH transferase was achieved for the activation of dibromoethane. No preferential binding in vitro to a specific polynucleotide was found. Polynucleotide labeling was higher than (or equal to) DNA binding. The labeling of microsomal RNA and proteins and of cytosolic proteins was many times lower than that of DNA or polynucleotides. The in vivo and in vitro data reported above give an unequivocal indication of the relative reactivity of the haloethanes examined with liver macromolecules from the two species and agree, on the whole, with the relative genotoxicity (DNA repair induction ability, mutagenicity and carcinogenicity) of the chemicals.

Retrograde cerebral perfusion during profound hypothermia and circulatory arrest in pigs.

The purpose of this study was to evaluate the use of retrograde cerebral perfusion via the superior vena cava during profound hypothermia and circulatory arrest (CA) in pigs. In three groups of 5 pigs each, group A (control) underwent cardiopulmonary bypass and normothermic CA for 1 hour, group B underwent cardiopulmonary bypass, profound hypothermia, and CA (15 degrees C nasopharyngeal) for 1 hour, and group C underwent the same procedure as group B plus retrograde cerebral perfusion. In group A none awoke. In group B, 2 of 5 did not awake and 3 of 5 awoke unable to stand, 2 with perceptive hind limb movement and 1 moving all extremities. In group C all awoke, 4 of 5 able to stand and 1 of 5 unable to stand but moving all limbs. In neurologic evaluation group B had significantly lower Tarlov scores than group C (p = 0.0090). Group B mean wake-up time, plus or minus standard error of the mean, was 124.6 +/- 4.6 minutes versus 29.2 +/- 5.1 in group C (p = 0.0090). In group B late phase CA cerebral blood flow dropped 30.9% +/- 4.8%, but in group C it rose 24.7% +/- 9.3% (p = 0.0007, pooled variance t test, two-tailed). In group B late phase CA brain oxygenation decreased 46.0% +/- 13.9% but it increased 26.1% +/- 5.4% in group C (p = 0.0013). This difference was reduced somewhat during rewarming (B, -21.2% +/- 14.9%; C, 16.4% +/- 4.7%; p = 0.043). Group B rewarming jugular venous O2 saturation was 30.8% +/- 2.5% versus 56.0% +/- 4.4% in group C (p = 0.0011). We conclude that in pigs retrograde cerebral perfusion combined with profound hypothermia during CA significantly reduces neurologic dysfunction, providing superior brain protection.

Effects of the protease inhibitor antipain on cell malignant transformation.

BACKGROUND: Several natural products have been found to exhibit a chemopreventive activity both in in vivo and in vitro experimental systems. Among them, protease inhibitors seem to play a key role in the regulation of growth and phenotypic expression of transformed cells as well as in the regulation of the late events of carcinogenesis. We evaluated the effect of antipain (AP), a natural protease inhibitor, on chemically induced BALB/c 3T3 cell transformation, on invasion and chemotactic motility of transformed cells and on their gelatinase expression. METHODS: BALB/c 3T3 cells were plated and exposed to 2.5 micrograms/ml 3-MCA or 50 micrograms/ml, 1,2-DBE. The effect of a non-cytotoxic dosage of AP (10 microM) was studied by: a) pretreating cells with AP for 48 hours before the carcinogen exposure; b) adding AP simultaneously to the carcinogen treatment; c) chronic addition of AP at each medium change throughout the experimental duration. The effectiveness of the treatment was analysed as the ability to reduce or inhibit the occurrence of transformed foci. Modulation of the invasive phenotype by anti-transforming dosages of AP was evaluated by in vitro Matrigel invasion assay. Gelatin zymography was performed in order to assess AP regulation of proteolytic enzymes, such as metalloproteases, involved in invasion and metastasis. RESULTS: AP treatment can reduce the transformation rate both in 3-MCA- and 1,2-DBE-initiated cells. Its effectiveness depends on the administration schedule, and chronic addition seems to be the most effective treatment. The concentration of AP, which is effective in the antitransformation assay, is not able to significantly affect the migration and invasion of chemically transformed cells or their gelatinase activity. CONCLUSIONS: AP can suppress chemically induced BALB/c 3T3 cell transformation through mechanisms which do not involve modulation of the invasive phenotype.

The covalent interaction of 1,4-dibromobenzene with rat and mouse nucleic acids: in vivo and in vitro studies.

1,4-Dibromobenzene (1,4-DBB) was covalently bound to DNA from liver, kidney, lung and stomach of mice after intraperitoneal administration. The covalent binding index (CBI) value (23 in mouse liver) was typical of weak initiators. On the contrary, no interaction with DNA from rat organs was observed (CBI detection limit: 1.3-2.6). The in vitro interaction of 1,4-DBB with calf thymus DNA was mediated mainly by microsomes, especially those from liver of both species and from mouse lung. Mouse subcellular fractions were more active then rat subcellular fractions. Unlike liver cytosol, subcellular cytosolic fractions from lung, kidney and stomach were capable of bioactivating 1,4-DBB, although to a lesser extent than liver microsomes. Both cytochrome P-450 and GSH-transferases are involved in 1,4-DBB bioactivation.

In vivo and in vitro interaction of 1,2-dichlorobenzene with nucleic acids and proteins of mice and rats.

Twenty-two hours after i.p. injection into male Wistar rats and BALB/c mice, 1,2-dichlorobenzene (1,2-DCB) was covalently bound to DNA, RNA, and proteins of liver, kidney, lung and stomach. The covalent binding index to liver DNA was typical of carcinogens classified as weak initiators. The enzyme-mediated in vitro interaction of 1,2-DCB with calf thymus DNA of synthetic polyribonucleotides was carried out by a microsomal mixed-function oxidase system and microsomal GSH-transferases, which seemed to be effective only in liver and lung of rat and mouse. Cytosolic GSH-transferases played a minor role in 1,2-DCB bioactivation. The latter finding provides the first evidence of 1,2-DCB genotoxicity in mammalian cells. The type of halide, the number of halosubstituents and their spatial disposition on the benzene ring are the major determinants of halobenzenes activability to intermediate(s) capable of interacting covalently with DNA and other macromolecules in biologic systems.

The different genotoxicity of p-dichlorobenzene in mouse and rat: measurement of the in vivo and in vitro covalent interaction with nucleic acids.

Twenty-two hours after i.p. injection to male Wistar rats and BALB/c mice para-dichlorobenzene (p-DCB) is bound covalently to DNA from liver, kidney, lung and stomach of mice but not of rats. DNA adducts in mouse liver are repaired in seventy-two hours. The covalent binding index value, calculated on the labelling of mouse liver DNA, classifies p-DCB as a weak initiator with an oncogenic activity lower than that of chlorobenzene. The labelling of RNA and proteins from the different organs of both species is, however, low. In vitro interaction with calf thymus DNA mediated by mouse and rat microsomes from liver and lung did occur. Binding extent was strongly reduced by addition of 2-diethylaminoethyl-2,2-diphenylvalerate hydrochloride (SKF 525-A) to the microsomal standard incubation mixture, whereas it was enhanced by adding GSH. Cytosolic fractions from kidney and lung were able to induce binding of p-DCB to DNA to a lower extent with respect to microsome-mediated binding. These results indicate that microsomal mixed function oxidase system and microsomal GSH-transferases can be involved in overall activating metabolism whereas cytosolic GSH-transferases play a minor role. This study, which is a part of a structure-activity relationship approach on benzene and its haloderivatives, provides the first evidence of genotoxicity of p-DCB in mammalian cell. It allows to partly explain variations of susceptibility of different species to hepatocarcinogenesis and of hepatotoxicity of different isomers.

Chloroform bioactivation leading to nucleic acids binding.

Chloroform was bound covalently to DNA, RNA and proteins of rat and mouse organs in vivo after i.p. injection. Covalent Binding Index values of rat and mouse liver DNA classify chloroform as a weak initiator. Labelings of RNA and proteins from various organs of both species were higher than that of DNA. In an in vitro cell-free system, chloroform was bioactivated by cytochrome P450-dependent microsomal fractions, by cytosolic GSH-transferases from rat and mouse liver, and particularly by the latter enzymes from mouse lung. This observation suggests that GSH plays a role in the binding of chloroform metabolites to DNA. The presence of both microsomal and cytosolic enzymatic systems in the standard incubation mixture generally led to an additive or synergistic bioactivating effect for rat and mouse, respectively.

Dipyridamole echocardiography and 99mTc-MIBI spect dipyridamole scintigraphy for cardiac evaluation prior to peripheral vascular surgery.

Coronary artery disease accounts for most of the early and late mortality and morbidity associated with vascular surgery. Cardiac pre-operative evaluation is mandatory for the assessment of cardiac risk. The aim of this study is to compare dipyridamole scintigraphy with 99mTc-MIBI (MIBI-dipy) and dipyridamole echocardiography (ECHO-dipy) and to evaluate their capability in identifying cardiac risk for strong events such as death, unstable angina or myocardial infarction. METHODS. Sixty consecutive patients (mean age 67 +/- 7) were enrolled. 52 performed ECHO-dipy, 51 MIBI-dipy. 40 patients went to aorto-femoral or aorto-iliac graft replacement and 15 to vascular angioplasty. Five patients did not undergo surgery. RESULTS. Eighteen patients (30%) had stress defects and 9 patients also rest defects with MIBI-dipy. Six patients new asinergic areas at ECHO-dipy. Three pts died in the first year follow-up for a cerebrovascular event, a myocardial infarction and a sudden death respectively. Sensitivity and specificity, positive and negative predictive value were 100%, 69%, 16%, 100% for MIBI-dipy and 66%, 94%, 40%, 98% for ECHO-dipy. CONCLUSIONS. As other authors reported, stress scintigraphy is a pre-operative test showing high sensitivity but with no satisfying specificity. Stress echocardiography, in our population, can produce a good negative predictive value. It is a less expensive and widespread clinical tool useful in the evaluation of preoperative patients. Its positive predictive power is not satisfying but it is shared with all non-invasive pre-operative tests available now.

[Surgical day hospital: technical possibilities and organizational model]. / Day hospital in chirurgia: possibilità techniche e modello organizzativo.

The Authors propose an organizational model for a surgical day hospital program, which is being used for a pilot day surgery unit in the I Department of Surgery of the Rome University "La Sapienza". The program requires little capital investment, as it is closely linked, geographically and administratively, to the main surgical unit, and uses the present staff, facilities and support services. The model is based on a computerized LAN (Local Area Network), providing fast recording, scheduling, management and trannsfer of medical data for each patient. The present situation is reported in detail. Data from the authors' outpatient department for 1988, have been recorded and elaborated. The results show a low use of surgical day care, limited to minor surgical procedures, and with not a single operation performed under general anesthesia. The authors hope to see a growth in the use of day surgery and a more selective use of inpatient care.

[Primary hepatic leiomyoma]. / Leiomioma epatico primitivo.

A young woman of 34 years old had an asymptomatic intrahepatic mass. The histological diagnosis, performed after a right lobe hepatectomy, was of leiomyoma of the liver.

Palmitoylethanolamide controls reactive gliosis and exerts neuroprotective functions in a rat model of Alzheimer's disease.

Given the complex heterogeneity of pathological changes occurring in Alzheimer's disease (AD), any therapeutic effort absolutely requires a multi-targeted approach, because attempts addressing only a single event may result ineffective. Palmitoylethanolamide (PEA), a naturally occurring lipid amide between palmitic acid and ethanolamine, seems to be a compound able to fulfill the criteria of a multi-factorial therapeutic approach. Here, we describe the anti-inflammatory and neuroprotective activities of systemic administration of PEA in adult male rats given intrahippocampal injection of beta amyloid 1-42 (Aß 1-42). Moreover, to investigate the molecular mechanisms responsible for the effects induced by PEA, we co-administered PEA with the GW6471, an antagonist of peroxisome proliferator-activated receptor-α (PPAR-α). We found that Aß 1-42 infusion results in severe changes of biochemical markers related to reactive gliosis, amyloidogenesis, and tau protein hyperphosphorylation. Interestingly, PEA was able to restore the Aß 1-42-induced alterations through PPAR-α involvement. In addition, results from the Morris water maze task highlighted a mild cognitive deficit during the reversal learning phase of the behavioral study. Similarly to the biochemical data, also mnestic deficits were reduced by PEA treatment. These data disclose novel findings about the therapeutic potential of PEA, and suggest novel strategies that hopefully could have the potential not just to alleviate the symptoms but also to modify disease progression.