Analysis of prognostic factors in COVID-19 hospitalized patients: an Italian single-center case-control study.

OBJECTIVE: COVID-19 clinical presentation ranges from asymptomatic infection to an inflammatory cytokine storm with multi-organ failure and fatal outcomes. The identification of high-risk patients for severe disease is crucial to plan an early treatment and intensive follow-up. We aimed to investigate negative prognostic factors in a group of patients hospitalized for COVID-19. PATIENTS AND METHODS: 181 patients (90 men and 91 women, mean age 66.56 ± 13.53 years) were enrolled. Each patient received a work-up including medical history, clinical examination, arterial blood gas analysis, laboratory blood tests, feasible ventilatory support required during hospital stay, intensive care setting required, duration of illness and length of hospital stay (>or<25 days). For the assessment of the severity of COVID-19, three main indicators were considered: 1) the intensive care unit (ICU) admission 2) the hospitalization length >25 days; 3) the need of non-invasive ventilation (NIV). RESULTS: The independent risk factor associated with the ICU admission were lactic dehydrogenase elevation (p=0.046), C reactive protein elevation (p=0.014) at hospital admission and direct oral anticoagulant home therapy (p=0.048); for hospital length >25 days: early corticosteroid therapy (p=0.035); for NIV treatment: ferritin elevation at hospital admission (p=0.006). CONCLUSIONS: The presence of the above factors may be useful to identify patients at high risk of developing a severe COVID-19 that need an early treatment and intensive follow-up.

Anti-gastric parietal cell antibodies for autoimmune gastritis screening in juvenile autoimmune thyroid disease.

OBJECTIVE: Patients with autoimmune thyroid disease (ATD) have a higher prevalence of autoimmune gastritis (AIG) compared with the general population. The association between ATD and AIG is poorly characterized in the pediatric age. We reviewed the prevalence of anti-gastric parietal cell antibodies (PCA) in young patients with ATD to evaluate its usefulness as a marker for AIG screening. METHODS: We evaluated 220 children and adolescents (11.28 ± 6.37 years) with ATD (186 with autoimmune thyroiditis (AT) and 34 with Graves' disease (GD). At ATD diagnosis and annually thereafter, blood counts and PCA levels were measured. In patients positive for PCA, plasma gastrin, chromogranin A, vitamin B12, iron and ferritin levels and H. pylori antigen were measured. PCA-positive patients > 18 years were invited to undergo a gastroscopic exam. RESULTS: PCA positivity was detected in ten (4.5%) subjects (5F/5M; 12.6 ± 3.4 years). The prevalence of PCA positivity was not significantly different in the comparison of GD and AT patients (p = 0.9). PCA positivity was detected after 2.7 ± 2.7 years of follow-up in AT and 4.4 ± 4.0 years in GD (p = 0.4). Autoantibody positivity was more prevalent in female patients, in both AT and GD (p = 0.02 and p = 0.03, respectively). At detection of PCA positivity, five out of ten PCA-positive patients had iron deficiency, four vitamin B12 deficiency, two anemia, three hypergastrinemia and two elevated chromogranin values. Two patients had H. pylori infection. Gastroscopy was performed in the five ATD patients and in all patients, AIG was confirmed. CONCLUSION: In the juvenile population, ATD and AIG may also be associated. PCA screening is useful to detect subjects at risk for this condition. Due to the longer life expectancy of the pediatric population and considering the relatively high risk of malignant transformation, early surveillance monitoring is mandatory for children and adolescents with ATD.

[Effect of treatment with selective serotonin reuptake inhibitors on lipid profile: state of the art]. / Effetto del trattamento con inibitori selettivi del reuptake della serotonina sul profilo lipidico: stato dell'arte.

Selective Serotonin Reuptake Inhibitors (SSRI) are an effective treatment for depressive disorder. Nevertheless, there is evidence suggesting a negative effect of these drugs on the lipid profile of the patients. We carried out a systematic review of the literature evaluating the influence of therapy with SSRI on lipid profile. Data source was MEDLINE. Clinical trials, prospective studies, retrospective studies and reviews published until November 2011 were considered. We identified twelve studies published from 1994 to 2011, of which four were randomized clinical trials, six were prospective studies and two were retrospective studies. Sertraline and Paroxetine seemed to have negative effects on the serum levels of Total and LDL Cholesterol. Citalopram did not demonstrate any influence on Total and LDL Cholesterol blood levels, being conversely associated with a slight increase of the HDL Cholesterol levels. Few data were found about the effects of Fluoxetina e Fluvoxamina on lipid profile and no data were found about Escitalopram. Sertaline and Paroxetine, two effective and widely used drugs for the treatment of major depression, seem to have a negative effect on the lipid profile; Citalopram, with its neutral or positive effect on lipid profile, should be considered the treatment of choice for depressive patients affected by dyslipidemia.

[Evaluation of the working load in subjects using equipment fitted with VDU]. / Valutazione del carico lavorativo in soggetti che utilizzano apparecchiature munite di videoterminali.

In Italy the health and safety of VDU workers is regulated by section VI of Legislative Decree 626/94 (with later modifications). In compliance with this law, employers shall be obliged to analyse workstations, assess and reduce risks; they shall also identify any worker who habitually uses display screen equipment as significant part of his normal work (20 hours per week). Our study presents a new method, named WODSE (Workers of Display Set Equipment), for the objective evaluation of VDU workers effort. It is a software created for measurement of VDT usage time, able to calculate weekly use for each worker. We applied this method to tellers of a great national services agency, obtaining an improvement of VDU risk assessment and, consequently, management.

[Health surveillance in health care workers]. / La sorveglianza sanitaria degli operatori sanitari: esperienza in un Policlinico universitario.

In a complex reality as the hospital, occupational physician plays a key role in risk prevention through health surveillance. The medical examination is intended to highlight any possible deterioration of workers' health and to detect workers' hypersusceptibility to occupational hazards. We report biostatistic data resulted from health surveillance conducted on health care workers in 2005 and 2006 in a universitary hospital, with particular regard to the judgement of the fitness to work and the reasons that has determined it. Our report, in agreement with data available in literature, shows that manual patient lifting is one of the most common professional hazards within the hospital and occupational physician must pay a special attention to it, promoting an integrated answer.

[Biomechanical risk assessment of manual material handling in vegetables and fruit departments of supermarkets]. / Valutazione del rischio da movimentazione manuale dei carichi nel reparto ortofrutta in un esercizio commerciale di medie dimensioni.

There is little in the literature about the risks of manual handling of material in supermarkets and what there is refers solely to storehouse work. This contrasts with the substantial number of studies of the risk of repeated arm movements among supermarket cash-desk staff. The scarcity of information is partly due to the difficulties of applying widely employed, standardized evaluation methods in this sector. One of the conditions limiting the application of the NIOSH protocol in this retail sector is that lifting tasks are so often closely tied to transport. The biomechanical analysis method we used brought to light considerable risks in many of the steps investigated: unpacking the pallet, unloading the crates from the pallet to the ground, lifting them from the floor onto display stands, and filling the boxes on the stands with goods before the shop opens. Images acquired on site were analyzed in the laboratory. We selected the most indicative images, which were then studied as regards posture and biomechanics using Apalys 3.0 software (ILMCAD GmbH, Ilmenau, Germany). Biomechemical analysis was done on the following movements: unloading crates from the pallet, positioning them on fruit and vegetable department display stands, and filling the boxes on the stands. We obtained a prediction of 2720 to 5472 N for the load at the lumbosacral junction (L5-S1). Simulation of the NIOSH index gave a value of 2.69 in the only case where the Waters protocol could be applied.

[Evaluation and risk management of the risk of robbery]. / Valutazione e gestione del rischio rapina.

The carried out work have permitted to make a technical evaluation of the risk robbery in a big Company of services and to formulate a strategy in order to prevent the criminal event and its effects on the workers' health. At this point an algorithm of prevision of risk has been formulated, taking account the elements of probability of criminal act and those ones referring to the relating damage according to the following formula: RISK(R) = PROBABILITY(P) x DAMAGE(D). The use of this method has proved useful to obtain the right evaluation risk robbery and it has permitted to elaborate a document of Health Protection to use towards the workers who are involved in the event with the active participation of the Company doctor.

[Role of the occupational health physician in the evaluation of biological risk and cancer in a hospital setting]. / Ruolo del medico competente nella valutazione del rapporto tra rischio biologico e cancro in ambiente sanitario.

OBJECTIVE: Elaborating and proposing a model of behaviour which is useful for any occupational doctor of a hospital in the management of the people exposed to biological risk, with the aim of preventing or early diagnosing neoplasias caused by cancerogenic infective agents. MATERIALS AND METHODS: This study was conducted analyzing the literature data regarding biohazard work accidents which happened in health environment and the scientific evidence of the causal relationship between infective agents and development of neoplasias. RESULTS: Data in literature show that the biohazard work accidents are very numerous and there is high percentage of sub-communication of them; many infective agents that the health workers can get in contact with, after biohazard biological accidents, are cancerogenic: HBV, HCV, HIV, HP belong to group 1 of IARC classification. CONCLUSION: Health workers exposed to biological risk and in particular those who got in contact with infective biological liquids can be considered at risk of neoplasia development; for this reason, we propose a three phases behaviour model: 1. Biological risk accidents prevention; 2. Prevention of infective disease after an accident; 3. Neoplasia prevention and/or early diagnosis after the development of the infective disease.

Occupational risk factors for the sporadic form of Creutzfeldt-Jakob disease.

Some case reports among European farmers and a few case-control studies suggested the hypothesis of an increased risk of the sporadic form of CJD (sCJD) associated with livestock farming or work as a butcher. Also, the discovery of the possibility of transmission of the disease via blood or by contact following corneal or dura madre transplant suggested that health occupations might also run higher sCJD risks. However, a meta-analysis of three case-control studies and a multicentre European study did not find any positive association between sCJD and health-related jobs or occupational contact with livestock, such as cattle and sheep, or animal products. To explore possible occupational risk factors for Creutzfeldt-Jakob disease (CJD), we used a publicly available US database including about 6 million deaths in 24 states during 1984-95. Cases were 636 deaths (300 men and 336 women) with CJD (ICD-9 code 046.1) as the underlying cause of death. Controls were 3,180 deaths randomly selected from among those who died from all other diseases except those affecting the central nervous system. CJD cases represented a wide variety of occupations (159) and industries (147). Among occupations and industries, for which previous reports suggested potential exposure to a transmissible spongiform encephalopathy (TSE) agent, the OR for CJD was significantly increased among butchers (OR = 6.8, 95% C.I. 1.5, 30.1, based on 4 cases and 3 controls), and persons working in offices of physicians (OR = 4.6, 95% C.I. 1.2, 17.6 based on 5 cases and 4 controls). Nine other occupations and seven other industries, for which no previous suggestion existed in the literature, also showed significant associations. Overall, our results suggest that occupational exposures are not an important source of sCJD infection. However, as the excess among butchers and some workers in health occupations was consistent with previous reports, more indepth research is warranted to address the hypothesis.

[Presentation forms of occupational psychologic disorders: experience at a national-level call center]. / Forme di presentazione del disagio psicologico a genesi occupazionale: esperienza presso il call center di una azienda di rilevanza nazionale.

Often in Occupational Medicine doctors misunderstand or underestimate the occupational psychologic discomfort forms deriving from "transverse risks" of organizational kind. In this communication is shortly described a our study with in a "call center". We explain the several kinds of psychologic discomfort which we have here recognized and studied. We show, at last, some of job's organization modifications rose from the first outcome of our study.

Acoso moral (mobbing): un nuevo riesgo en la medicina del trabajo / Moral harassment: a new risk in occupational medicine

Los riesgos psicosociales se están constituyendo en una de las principales causas de alteración de la salud en los puestos de trabajo. En los últimos años, el "riesgo relacional o interpersonal" - mobbing - se ha ido incrementando debido a los cambios macroeconómicos y por el cambio en la tipología del trabajo y en los riesgos laborales derivados. Cada trabajador, independientemente de las características de su propia personalidad y del propio carácter, puede ser objeto de acoso moral. Los primeros efectos derivados del mobbing son observables sobre la salud de la víctima que, casi siempre, después de un intervalo variable, se altera con manifestaciones en la esfera neuropsíquica, Las consecuancias sociales pueden ser devastadoras. El costo del mobbing no se limita a los aspectos individuales, sino que se refleja generalmente a nivel de la empresa. La gestión del fenómeno de mobbing es multidisciplinaria. A nivel asistencial, el rol del médico del trabajo, del psiquiatra y del psicólogo son interdependientes y deben por lo tanto ser integrados en una estructura funcional unitaria...

Acoso moral (mobbing): un nuevo riesgo en la medicina del trabajo / Moral harassment: a new risk in occupational medicine

Los riesgos psicosociales se están constituyendo en una de las principales causas de alteración de la salud en los puestos de trabajo. En los últimos años, el "riesgo relacional o interpersonal" - mobbing - se ha ido incrementando debido a los cambios macroeconómicos y por el cambio en la tipología del trabajo y en los riesgos laborales derivados. Cada trabajador, independientemente de las características de su propia personalidad y del propio carácter, puede ser objeto de acoso moral. Los primeros efectos derivados del mobbing son observables sobre la salud de la víctima que, casi siempre, después de un intervalo variable, se altera con manifestaciones en la esfera neuropsíquica, Las consecuancias sociales pueden ser devastadoras. El costo del mobbing no se limita a los aspectos individuales, sino que se refleja generalmente a nivel de la empresa. La gestión del fenómeno de mobbing es multidisciplinaria. A nivel asistencial, el rol del médico del trabajo, del psiquiatra y del psicólogo son interdependientes y deben por lo tanto ser integrados en una estructura funcional unitaria...(AU)

Pro-sequence assisted folding and disulfide bond formation of human nerve growth factor.

Nerve growth factor (NGF) is a member of the neurotrophin family. These growth factors support neuronal survival and differentiation. Neurotrophins are synthesized as pre-pro-proteins. Whereas the pre-sequences mediate secretion, the function of the pro-peptides is largely unknown. To test the role of the pro-sequence as a folding enhancer, recombinant human pro-NGF (rh-pro-NGF) was produced in Escherichia coli. The oxidative refolding of rh-pro-NGF and rh-NGF was studied using electrospray mass spectrometry (ESIMS) time-course analysis. This analysis permitted both the identification and quantification of intermediates present during the process. The disulfide bonds formed at different times of the refolding processes were characterized by proteolytic digestion followed by matrix assisted laser desorption ionization mass spectrometry (MALDIMS) analysis. Folding yields and kinetics of rh-pro-NGF were significantly enhanced when compared to the in vitro refolding of mature rh-NGF. These results suggest that the pro-sequence of NGF promotes folding of the mature part.

Contribution of individual disulfide bonds to the oxidative folding of ribonuclease A.

The eight cysteine residues of ribonuclease A form four disulfide bonds in the native protein. We have analyzed the folding of three double RNase A mutants (C65A/C72A, C58A/C110A, and C26A/C84A, lacking the C65-C72, C58-C110, and C26-C84 disulfide bonds, respectively) and two single mutants (C110A and C26A), in which a single cysteine is replaced with an alanine and the paired cysteine is present in the reduced form. The folding of these mutants was carried out in the presence of oxidized and reduced glutathione, which constitute the main redox agents present within the ER. The use of mass spectrometry in the analysis of the folding processes allowed us (i) to follow the formation of intermediates and thus the pathway of folding of the RNase A mutants, (ii) to quantitate the intermediates that formed, and (iii) to compare the rates of formation of intermediates. By comparison of the folding kinetics of the mutants with that of wild-type RNase A, the contribution of each disulfide bond to the folding process has been evaluated. In particular, we have found that the folding of the C65A/C72A mutant occurs on the same time scale as that of the wild-type protein, thus suggesting that the removal of the C65-C72 disulfide bond has no effect on the kinetics of RNase A folding. Conversely, the C58A/C110A and C26A/C84A mutants fold much more slowly than the wild-type protein. The removal of the C58-C110 and C26-C84 disulfide bonds has a dramatic effect on the kinetics of RNase A folding. Results described in this paper provide specific information about conformational folding events in the regions involving the mutated cysteine residues, thus contributing to a better understanding of the complex mechanism of oxidative folding.

Mammalian FKBP-25 and its associated proteins.

Soluble proteins from porcine brain were divided into two packs: (1) proteins which pass freely through CM52-cellulose, and (2) proteins retained on CM52. Each of these two packs of proteins was fractionated on preparative flat-bed isoelectrofocusing gel in the range of pH 2-12. Native FKBP-25 and its truncated forms were found among other proteins retained on CM52-cellulose. Immunoblotting with anti-FKBP-25 showed two bands in the range 27-30 kDa, one due to unmodified FKBP-25 and other due to FKBP-25 mixed with high-mobility group II protein (HMG-II). Selective immunostaining with anti-FKBP-25 antibodies of proteins which were not retained on CM52-cellulose showed several bands within the range of pI 7-5 and mass of 23 +/- 2 kDa. These fractions of proteins were next resolved on two-dimensional gels and immunostained with anti-FKBP-25 antibodies. Six proteins in the pI range 7-5 were detected. Edman degradation of alpha-chymotrypsin digests of the major spot suggests that it contains the GTP-binding protein Rab5 co-migrating with guanylyl kinase, whereas MALDI-TOF showed that a residual content of FKBP-25 may be also associated with these two proteins. A residual quantity of FKBP-25 was also associated with the phosphatidylethanolamine-binding protein which is abundant in the brain.

Investigation of the DsbA mechanism through the synthesis and analysis of an irreversible enzyme-ligand complex.

Approaching the molecular mechanism of some enzymes is hindered by the difficulty of obtaining suitable protein-ligand complexes for structural characterization. DsbA, the major disulfide oxidase in the bacterial periplasm, is such an enzyme. Its structure has been well characterized in both its oxidized and its reduced states, but structural data about DsbA-peptide complexes are still missing. We report herein an original, straightforward, and versatile strategy for making a stable covalent complex with a cysteine-homoalanine thioether bond instead of the labile cystine disulfide bond which normally forms between the enzyme and polypeptides during the catalytic cycle of DsbA. We substituted a bromohomoalanine for the cysteine in a model 14-mer peptide derived from DsbB (PID-Br), the membrane partner of DsbA. When incubated in the presence of the enzyme, a selective nucleophilic substitution of the bromine by the thiolate of the DsbA Cys(30) occurred. The major advantage of this strategy is that it enables the direct use of the wild-type form of the enzyme, which is the most relevant to obtain unbiased information on the enzymatic mechanism. Numerous intermolecular NOEs between DsbA and PID could be observed by NMR, indicating the presence of preferential noncovalent interactions between the two partners. The thermodynamic properties of the DsbA-PID complex were measured by differential scanning calorimetry. In the complex, the values for both denaturation temperature and variation in enthalpy associated with thermal unfolding were between those of oxidized and reduced forms of DsbA. This progressive increase in stability along the DsbA catalytic pathway strongly supports the model of a thermodynamically driven mechanism.

Estimation of lead intake from crystalware under conditions of consumer use.

The purpose of this study was to estimate the lead intake from crystalware resulting from short-term contacts with beverages, under conditions that are likely to occur to a consumer. The extraction ability of different kinds of beverages was estimated by comparison with 4% acetic acid under conditions of continuous contact for 3 h. It was found that lead release increased in the following order: cola drink > HAc > whisky > white wine. Under conditions of repeated use under different scenarios, lead release showed a steep decrease with increasing number of contacts, for both wine and cola drink. The maximum lead intake resulted from the cola drink, corresponding to an ingestion of 14.5 micrograms Pb for consumption of 350 ml beverage. Assuming a fixed contribution from the diet of 71 micrograms/day, in the six scenarios taken into consideration, total daily lead intake levels ranged from a minimum of about 76 micrograms up to a maximum of 86 micrograms lead. As these values, converted on a weekly basis, would correspond to 35% and 40% PTWI respectively, significant health risks resulting from the ingestion of beverages in contact with crystalware can be excluded. Finally it was found that the use of a dishwater did not affect significantly the release of lead into wine, while release into cola drink was slightly but significantly increased after the third cycle.

Aspartate aminotransferase from the Antarctic bacterium Pseudoalteromonas haloplanktis TAC 125. Cloning, expression, properties, and molecular modelling.

The gene encoding aspartate aminotransferase from the psychrophilic bacterium Pseudoalteromonas haloplanktis TAC 125 was cloned, sequenced and overexpressed in Escherichia coli. The recombinant protein (PhAspAT) was characterized both at the structural and functional level in comparison with the E. coli enzyme (EcAspAT), which is the most closely related (52% sequence identity) bacterial counterpart. PhAspAT is rapidly inactivated at 50 degrees C (half-life = 6.8 min), whereas at this temperature EcAspAT is stable for at least 3 h. The optimal temperature for PhAspAT activity is approximately 64 degrees C, which is some 11 degrees C below that of EcAspAT. The protein thermal stability was investigated by following changes in both tryptophan fluorescence and amide ellipticity; this clearly suggested that a first structural transition occurs at approximately 50 degrees C for PhAspAT. These results agree with the expected thermolability of a psychrophilic enzyme, although the observed stability is much higher than generally found for enzymes isolated from cold-loving organisms. Furthermore, in contrast with the higher efficiency exhibited by several extracellular psychrophilic enzymes, both kcat and kcat/Km of PhAspAT are significantly lower than those of EcAspAT over the whole temperature range. This behaviour possibly suggests that the adaptation of this class of endocellular enzymes to a cold environment may have only made them less stable and not more efficient. The affinity of PhAspAT for both amino-acid and 2-oxo-acid substrates decreases with increasing temperature. However, binding of maleate and 2-methyl-L-aspartate, which both inhibit the initial steps of catalysis, does not change over the temperature range tested. Therefore, the observed temperature effect may occur at any of the steps of the catalytic mechanism after the formation of the external aldimine. A molecular model of PhAspAT was constructed on the basis of sequence homology with other AspATs. Interestingly, it shows no insertion or extension of loops, but some cavities and a decrease in side chain packing can be observed.

Early intermediates in the PDI-assisted folding of ribonuclease A.

The oxidative refolding of ribonuclease A has been investigated in several experimental conditions using a variety of redox systems. All these studies agree that the formation of disulfide bonds during the process occurs through a nonrandom mechanism with a preferential coupling of certain cysteine residues. We have previously demonstrated that in the presence of glutathione the refolding process occurs through the reiteration of two sequential reactions: a mixed disulfide with glutathione is produced first which evolves to form an intramolecular S-S bond. In the same experimental conditions, protein disulfide isomerase (PDI) was shown to catalyze formation and reduction of mixed disulfides with glutathione as well as formation of intramolecular S-S bonds. This paper reports the structural characterization of the one-disulfide intermediate population during the oxidative refolding of Ribonuclease A under the presence of PDI and glutathione with the aim of defining the role of the enzyme at the early stages of the reaction. The one-disulfide intermediate population occurring at the early stages of both the uncatalyzed and the PDI-catalyzed refolding was purified and structurally characterized by proteolytic digestion followed by MALDI-MS and LC/ESIMS analyses. In the uncatalyzed refolding, a total of 12 disulfide bonds out of the 28 theoretical possible cysteine couplings was observed, confirming a nonrandom distribution of native and nonnative disulfide bonds. Under the presence of PDI, only two additional nonnative disulfides were detected. Semiquantitative LC/ESIMS analysis of the distribution of the S-S bridged peptides showed that the most abundant species were equally populated in both the uncatalyzed and the catalyzed process. This paper shows the first structural characterization of the one-disulfide intermediate population formed transiently during the refolding of ribonuclease A in quasi-physiological conditions that mimic those present in the ER lumen. At the early stages of the process, three of the four native disulfides are detected, whereas the Cys26-Cys84 pairing is absent. Most of the nonnative disulfide bonds identified are formed by nearest-neighboring cysteines. The presence of PDI does not significantly alter the distribution of S-S bonds, suggesting that the ensemble of single-disulfide species is formed under thermodynamic control.