Validation of the SkinEthic HCE Time-to-Toxicity test method for eye hazard classification of chemicals according to UN GHS.

This study describes the within- and between-laboratory reproducibility (WLR and BLR) of a Time-to-Toxicity (TTT) approach for chemicals based on the SkinEthic™ HCE tissue construct, capable to distinguish chemicals that do not require classification for serious eye damage/eye irritation (No Cat.) from chemicals that require classification for eye irritation (Cat. 2), and serious eye damage (Cat. 1). The WLR and BLR was assessed with three participating laboratories. Each laboratory tested 40 coded chemicals in three independent runs. The predictive capacity of the method was assessed on a larger set of 150 chemicals (70 liquids and 80 solids) by combining the results of this study with the results of the test method developer. The WLR for the 20 liquids ranged from 85% to 95% with a BLR of 90%. For the 20 solids, a WLR and BLR of 100% was obtained. The test method developer obtained a WLR of 80% and 95%, based on 50 liquids and 48 solids tested in three independent runs, respectively. Regarding the predictive capacity, the SkinEthic™ HCE TTT test method identified 80.8% Cat. 1, 69.2% Cat. 2, and 74.9% No Cat. correctly. An independent peer review panel concluded that based on all available data, the relevance and reliability of the SkinEthic™ HCE TTT has been demonstrated for discriminating the three UN GHS eye hazard categories.

Medical Legal Aspects of Telemedicine in Italy: Application Fields, Professional Liability and Focus on Care Services During the COVID-19 Health Emergency.

Telemedicine services can be classified into the macro-categories of specialist Telemedicine, Tele-health and Tele-assistance. From a regulatory perspective, in Italy, the first provision dedicated to the implementation of Telemedicine services is represented by the Agreement between the Government and the Regions on the document bearing "Telemedicine-National guidelines," approved by the General Assembly of the Superior Health Council in the session of 10th July 2012 and by the State Regions Conference in the session of 20th February 2014. Scientifically, several studies in the literature state that information and communication technologies have great potential to reduce the costs of health care services in terms of planning and making appropriate decisions that provide timely tools to patients. Another clear benefit is the equity of access to health care. The evolution of telemedicine poses a series of legal problems ranging from the profiles on the subject of authorization and accreditation to those concerning the protection of patient confidentiality, the definition and solution of which, in the absence of specific regulatory provisions, is mainly left to the assessment of compatibility of the practices adopted so far, with the general regulatory framework. In terms of professional liability, it is necessary to first clarify that the telemedicine service is comparable to any diagnostic-therapeutic health service considering that the telemedicine service does not replace the traditional health service, but integrates the latter to improve its effectiveness, efficiency and appropriateness.

CON4EI: SkinEthic™ Human Corneal Epithelium Eye Irritation Test (SkinEthic™ HCE EIT) for hazard identification and labelling of eye irritating chemicals.

Assessment of ocular irritancy is an international regulatory requirement and a necessary step in the safety evaluation of industrial and consumer products. Although a number of in vitro ocular irritation assays exist, none are capable of fully categorizing chemicals as a stand-alone assay. Therefore, the CEFIC-LRI-AIMT6-VITO CON4EI (CONsortium for in vitro Eye Irritation testing strategy) project was developed with the goal of assessing the reliability of eight in vitro/alternative test methods as well as establishing an optimal tiered-testing strategy. One of the in vitro assays selected was the validated SkinEthic™ Human Corneal Epithelium Eye Irritation Test method (SkinEthic™ HCE EIT). The SkinEthic™ HCE EIT has already demonstrated its capacity to correctly identify chemicals (both substances and mixtures) not requiring classification and labelling for eye irritation or serious eye damage (No Category). The goal of this study was to evaluate the performance of the SkinEthic™ HCE EIT test method in terms of the important in vivo drivers of classification. For the performance with respect to the drivers all in vivo Cat 1 and No Cat chemicals were 100% correctly identified. For Cat 2 chemicals the liquids and the solids had a sensitivity of 100% and 85.7%, respectively. For the SkinEthic™ HCE EIT test method, 100% concordance in predictions (No Cat versus No prediction can be made) between the two participating laboratories was obtained. The accuracy of the SkinEthic™ HCE EIT was 97.5% with 100% sensitivity and 96.9% specificity. The SkinEthic™ HCE EIT confirms its excellent results of the validation studies.

Multi-laboratory evaluation of SkinEthic HCE test method for testing serious eye damage/eye irritation using solid chemicals and overall performance of the test method with regard to solid and liquid chemicals testing.

A prospective multicentre study of the reconstructed human corneal epithelial tissue-based in vitro test method (SkinEthic™ HCE) was conducted to evaluate its usefulness to identify chemicals as either not classified for serious eye damage/eye irritation (No Cat.) or as classified (Cat. 1/Cat. 2) within UN GHS. The aim of this study was to demonstrate the transferability and reproducibility of the SkinEthic™ HCE EITS protocol for solids and define its predictive capacity. Briefly, 60 chemicals were three times tested (double blinded) in 3 laboratories and 35 additional chemicals were tested three times in one laboratory. Good within laboratory reproducibility was achieved of at least 95% (57/60) and 96.8% (92/95) for the extended data set. Furthermore, the overall concordance between the laboratories was 96.7% (58/60). The accuracy of the SkinEthic™ HCE EITS for the extended dataset, based on bootstrap resampling, was 81.0% (95% CI: 78.9% to 83.2%) with a sensitivity of 90.5% (95% CI: 88.1% to 92.9%) and specificity of 73.6% (95% CI: 71.7% to 75.5%). Overall, 200 chemicals were tested (105 liquids (EITL protocol) and 95 solids (EITS protocol)) resulting in a sensitivity of 95.2%, specificity of 72.1% and accuracy of 83.7%, thereby meeting all acceptance criteria for predictive capacity.

Multi-laboratory validation of SkinEthic HCE test method for testing serious eye damage/eye irritation using liquid chemicals.

A prospective multicentric study of the reconstructed human corneal epithelial tissue-based in vitro test method (SkinEthic™ HCE) was conducted to evaluate its usefulness to identify chemicals as either not classified for serious eye damage/eye irritation (No Cat.) or as classified (Cat. 1/Cat. 2) within UN GHS. The aim of this study was to demonstrate the transferability and reproducibility of the SkinEthic™ HCE EITL protocol for liquids and define its predictive capacity. Briefly, 60 chemicals were three times tested (double blinded) in 3 laboratories and 45 additional chemicals were tested three times in one laboratory. Good within laboratory reproducibility was achieved of at least 88.3% (53/60) and 92.4% (97/105) for the extended data set. Furthermore, the overall concordance between the laboratories was 93.3% (56/60). The accuracy of the SkinEthic™ HCE EITL for the extended dataset, based on bootstrap resampling, was 84.4% (95% CI: 81.9% to 87.6%) with a sensitivity of 99.0% (95% CI: 96.4% to 100%) and specificity of 68.5% (95% CI: 64.0% to 74.0%), thereby meeting all acceptance criteria for predictive capacity. This efficient transferable and reproducible assay is a promising tool to be integrated within a battery of assays to perform an eye irritation risk assessment.

Prospective multicentre study of the U-SENS test method for skin sensitization testing.

The U-SENS™ is a test method based on the human myeloid U937 cell line to assess the skin sensitisation potential of substances. To demonstrate its robustness, a multicentre validation study with four laboratories testing 24 coded substances has been conducted according to internationally agreed principles. The primary objective of the study was to enlarge the U-SENS™'s reproducibility database. Secondary objectives were to provide additional evidence on its transferability and its predictive capability. Reproducibility within laboratories was approximately 92%, while the reproducibility between laboratories was 87.5%. Predictivity for the 24 validation substances was high, with sensitivity, specificity and accuracy being on average at least 93.8%. Similar performances are obtained for 38 substances when combining the study results with those of an earlier multicentre study, as well as with an automated version of the U-SENS™. With reliability and relevance similar to comparable non-animal skin sensitisation test methods, which have achieved regulatory acceptance, it is concluded that the U-SENS™ is a well reproducible and predictive test method. This profiles the U-SENS™ as a valuable addition to the suite of non-animal testing methods for skin sensitisation with the potential to significantly contribute to the development of integrated testing strategies.

Effects of acute hypercapnia with and without acidosis on lung inflammation and apoptosis in experimental acute lung injury.

We investigated the effects of acute hypercapnic acidosis and buffered hypercapnia on lung inflammation and apoptosis in experimental acute lung injury (ALI). Twenty-four hours after paraquat injection, 28 Wistar rats were randomized into four groups (n=7/group): (1) normocapnia (NC, PaCO2=35-45 mmHg), ventilated with 0.03%CO2+21%O2+balancedN2; (2) hypercapnic acidosis (HC, PaCO2=60-70 mmHg), ventilated with 5%CO2+21%O2+balancedN2; and (3) buffered hypercapnic acidosis (BHC), ventilated with 5%CO2+21%O2+balancedN2 and treated with sodium bicarbonate (8.4%). The remaining seven animals were not mechanically ventilated (NV). The mRNA expression of interleukin (IL)-6 (p=0.003), IL-1ß (p<0.001), and type III procollagen (PCIII) (p=0.001) in lung tissue was more reduced in the HC group in comparison with NC, with no significant differences between HC and BHC. Lung and kidney cell apoptosis was reduced in HC and BHC in comparison with NC and NV. In conclusion, in this experimental ALI model, hypercapnia, regardless of acidosis, reduced lung inflammation and lung and kidney cell apoptosis.

Alveolar fibroblasts in acute lung injury: biological behaviour and clinical relevance.

Although fibroblasts are key cells in the lung repair/fibrosis process, their characteristics are poorly studied in acute lung injury (ALI) and acute respiratory distress syndrome (ARDS). The aims of our study were to: 1) determine the biological behaviour of alveolar fibroblasts during ALI; and 2) to evaluate the clinical relevance of positive alveolar fibroblast culture from patients with ALI/ARDS. Cells were cultured from bronchoalveolar lavage (BAL) obtained from 68 critically ill, ventilated patients: ALI n = 17; ARDS n = 31; and ventilated controls n = 20. Patients were followed for 28 days and clinical data was recorded. We studied proliferation, migration and collagen-1 synthesis capacities of fibroblasts. Cells expressing fibroblast markers were cultured from BAL obtained in six (35%) ALI patients and six (19%) ARDS patients, but never from ventilated controls. Alveolar fibroblasts exhibited a persistent activated phenotype with enhanced migratory and collagen-1 production capacities, with hyporesponsiveness to prostaglandin E(2) compared to normal lung fibroblasts (p< or =0.04). Positive fibroblast culture was associated with both an increased collagen-1 concentration and monocyte/macrophage percentage in BAL fluid (p< or =0.01), and with a reduced duration of mechanical ventilation (p<0.001). We conclude that activated alveolar fibroblasts can be cultured either in ALI or ARDS and that their presence might reflect the initiation of the organising phase of ALI.

Role of duplex scan in endoleak detection after endoluminal abdominal aortic aneurysm repair.

OBJECTIVE: to validate the role of duplex scan in endoleak detection in postoperative surveillance of endoluminal abdominal aneurysm repair (EAAR). PATIENTS AND METHODS: between April 1997 and March 1999, 103 patients were eligible for duplex and computed tomography (CT) scan after EAAR. Mean follow-up was 8 months (range 1-24 months). The study protocol comprised concurrent examination with colour-duplex and CT scan at 1, 6, and 12 months after EAAR, for a total of 198 concurrent examinations. All duplex scan examinations were performed by two vascular surgeons with the same machine (ATL HDI 3000). Interobserver agreement in endoleak detection (kappa=1) and in type of endoleak (kappa=0.7) was evaluated in 50 random duplex examinations. Endoleak detection was examined comparatively in duplex and CT scan, the latter being the gold standard. Sensitivity and specificity tests together with negative- and positive-predictive values (NPV and PPV) were calculated. RESULTS: duplex scan was not feasible in one patient. On CT scan the endoleak rate was 4% at one month, 3% at 6 months, and 4% at one year. Overall, CT scan detected 12 endoleaks. With respect to endoleak detection, duplex scan revealed a great ability in ruling out false-negative results (sensitivity 91.7%, NPV 99.4%), but overestimated the presence of endoleak (specificity 98.4%, PPV 78. 6%). Regarding type of endoleak, the ability of duplex scan to identify the source of endoleak was low (sensitivity 66.7%). CONCLUSIONS: duplex scan, if validated, appears to be a reliable means for excluding the presence of endoleak after EAAR.

10-years experience with a 28,800 roller bottle plant for FMD vaccine production.

The roller bottle plant at the Istituto Zooprofilattico of Brescia is described and the following topics are discussed: (1) results of ten years cell and virus culture; (2) critical approach to advantages and disadvantages of applied technology, and (3) FMD vaccine requirements and cell substrate characterization.

Infectious bursal disease vaccine: some remarks on production and control.

A vaccine against infectious bursal disease (IBD) is prepared with a strain of IBD virus attenuated in chicken embryos. Safety and potency tests are carried out on embryos and chickens susceptible to IBD virus.

Data on a FMD virus of type "A" isolated in Italy in 1975.

A strain of FMD virus type A was isolated in a disease outbreak in Alessandria, Italy, in December 1975. Affected animals were young unvaccinated bovines and pigs of a very restricted region. No further spread has been observed. The strain was compared with some subtype strains of type A, including the Italian vaccinal strain. This Italy 1975 strain differs from all the other viruses included in the performed tests.

The use of suspension cultures for FMD vaccine production. Criteria for the evaluation of cells, virus and vaccine.

A survey of problems connected with the cell suspension culture for the production of FMD vaccine is presented, in order to stimulate a discussion. Introductory remarks are made on the following topics: 1) suspension cell lines used for FMD vaccine production, 2) terminology, 3) criteria for the characterization of a cell culture, 4) evaluation of FMD virus produced according to the cell suspension technique and correlation existing between in vitro tests and antigenicity in domestic animals, 5) criteria which must be met by FMD vaccine produced from virus cultivated on cell suspension.

Vaccination against Newcastle disease: efficacy of an oil emulsion vaccine.

Vaccination using injectable Newcastle vaccine (inactivated) oil emulsion at 21 days of age stimulated high and persistent H.I. antibody levels. Vaccine prepared on an industrial scale contained not less then 100 PD50 in 0.5 ml. High levels of maternal antibodies had a negative influence on vaccination of chickens when carried out during the first 3 weeks of age, but not when performed later. Vaccine stored in a refrigerator (at 8 degrees to 4 degrees C) or at room temperature (20 degrees C) retained its potency for at least 1 year.