Studies on ''corky rugose wood'' of grapevine and on the diagnosis of grapevine virus B

Vines affected by corky rugose wood (CRW), a field syndrome characterized by pronounced cork production by the scion of several grapevine varieties just above the graft union, contain a number of filamentous and isometric phloem-limited viruses, such as grapevine leafroll-associated virus 1, 2, and 3 (GLRaV-1, GLRaV-2, GLRaV-3), grapevine virus A and B (GVA and GVB), and grapevine fleck virus (GFkV). However, the same viruses, with the exception of GVB, are widely represented also in vines with rugose wood without excessive corkyness. Although GVB was found in all vines indexing positive in LN 33 for corky bark disease, iis occurrence in CRW-affected vines was not consistent enough to suggest that it may have a determining role in the induction of this syndrome. Monoclonal antibodies to GVB raised previously were characterized and their possible use for reliable detection of GVB in field-grown vines investigated in detail. A triple antibody sandwich ELISA protocol that under our experimental conditions afforded consistent and repeatable results, was based on the use of crude cortical scraping extracts from mature canes collected in autumn, antibodies from a polyclonal antiserum for plate coating (trapping) and a monoclonal antibody for antigen detection

Silence and Voicing Accumulations in Italian Primary School Teachers With and Without Voice Disorders

Summary: Objectives. The relationship between the silence and voicing accumulations of primary school teachers and the teachers' clinical status was examined to determine whether more voicing accumulations and fewer silence accumulations were measured for the vocally unhealthy subjects than for the healthy subjects, which would imply more vocal loading and fewer short-term recovery moments. Methods. Twenty-six Italian primary school teachers were allocated by clinicians to three groups: (1) with organic voice disorders, (2) with subjectively mild organic alteration or functional voice symptoms, and (3) normal voice quality and physiology. Continuous silence and voicing periods were measured with the APM3200 during the teachers' 4-hour workdays. The accumulations were grouped into seven time intervals, ranging from 0.03-0.9 to 3.16-10 seconds, according to Italian prosody. The effects of group on silence and voicing accumulations were evaluated. Results. Regarding silence accumulations, Group 1 accumulated higher values in intervals between 0.1 and 3.15 seconds than other groups, whereas Groups 2 and 3 did not differ from each other. Voicing accumulations between 0.17 and 3.15 seconds were higher for subjects with a structural disorder. A higher time dose was accumulated by these subjects (40.6%) than other subjects (Group 2, 31.9%; Group 3, 32.3%). Conclusions. Although previous research has suggested that a rest period of a few seconds may produce some vocal fatigue recovery, these results indicate that periods shorter than 3.16 seconds may not have an observable effect on recovery. The results provide insight into how vocal fatigue and vocal recovery may relate to voice disorders in occupational voice users

Embryo Culture, In Vitro Propagation, and Molecular Identification for Advanced Olive Breeding Programs

The high biodiversity of the olive tree is an important opportunity to develop sustainable plans to control Xylella fastidiosa (X) through breeding programs. Olive tree breeding activities have been limited due to various features of this species including the long time required for seed germination caused by the inhibition effect of the woody endocarp, the seed integument, and the endosperm. Starting from F1 seeds by cross-breeding, the embryo culture was compared with traditional seed germination, evaluating the effectiveness of in vitro multiplication of the plantlets for large-scale production. The isolated embryos were established on a new medium based on Rugini ‘84 macroelements, Murashige & Skoog ‘62 microelements, with Nitsch J. P. & Nitsch C. ‘69 vitamine and subcultured on Leva MSM modified. The results obtained confirmed that in vitro culture of olive embryos is a valid tool for increasing the percentage and speed of germination, helping to reduce the time of the olive breeding programs, offering the possibility to effectively propagate plantlets for further experiments

Determination of the LOQ in real-time PCR by receiver operating characteristic curve analysis: application to qPCR assays for Fusarium verticillioides and F. proliferatum

Real-time PCR (qPCR) is the principal technique for the quantification of pathogen biomass in host tissue, yet no generic methods exist for the determination of the limit of quantification (LOQ) and the limit of detection (LOD) in qPCR. We suggest using the Youden index in the context of the receiver operating characteristic (ROC) curve analysis for this purpose. The LOQ was defined as the amount of target DNA that maximizes the sum of sensitivity and specificity. The LOD was defined as the lowest amount of target DNA that was amplified with a false-negative rate below a given threshold. We applied this concept to qPCR assays for Fusarium verticillioides and Fusarium proliferatum DNA in maize kernels. Spiked matrix and field samples characterized by melting curve analysis of PCR products were used as the source of true positives and true negatives. On the basis of the analysis of sensitivity and specificity of the assays, we estimated the LOQ values as 0.11 pg of DNA for spiked matrix and 0.62 pg of DNA for field samples for F. verticillioides. The LOQ values for F. proliferatum were 0.03 pg for spiked matrix and 0.24 pg for field samples. The mean LOQ values correspond to approximately eight genomes for F. verticillioides and three genomes for F. proliferatum. We demonstrated that the ROC analysis concept, developed for qualitative diagnostics, can be used for the determination of performance parameters of quantitative PCR

Mapping and Assessment of forest Ecosystem and Their Services. Applications and guidance for decision making in the framework of MAES

The aim of this report is to illustrate by means of a series of case studies the implementation of mapping and assessment of forest ecosystem services in different contexts and geographical levels. Methodological aspects, data issues, approaches, limitations, gaps and further steps for improvement are analysed for providing good practices and decision making guidance. The EU initiative on Mapping and Assessment of Ecosystems and their Services (MAES), with the support of all Member States, contributes to improve the knowledge on ecosytem services. MAES is one of the building-block initiatives supporting the EU Biodiversity Strategy to 2000

Trends in chronic hepatitis B virus infection in Italy over a 10-year period: Clues from the nationwide PITER and MASTER cohorts toward elimination

Objectives: The study measures trends in the profile of patients with chronic hepatitis B virus linked to care in Italy. Methods: A cross-sectional, multicenter, observational cohort (PITER cohort) of consecutive patients with hepatitis B surface antigen (HBsAg) over the period 2019-2021 from 46 centers was evaluated. The reference was the MASTER cohort collected over the years 2012-2015. Standard statistical methods were used. Results: The PITER cohort enrolled 4583 patients, of whom 21.8% were non-Italian natives. Compared with those in MASTER, the patients were older and more often female. The prevalence of hepatitis B e antigen (HBeAg) declined (7.2% vs 12.3; P <0.0001) and that of anti-hepatitis D virus (HDV) remained stable (9.3% vs 8.3%). In both cohorts, about 25% of the patients had cirrhosis, and those in the PITER cohort were older. HBeAg-positive was 5.0% vs 12.6% (P <0.0001) and anti-HDV positive 24.8% vs 17.5% (P <0.0017). In the logistic model, the variables associated with cirrhosis were anti-HDV-positive (odds ratio = 10.08; confidence interval 7.63-13.43), age, sex, and body mass index; the likelihood of cirrhosis was reduced by 40% in the PITER cohort. Among non-Italians, 12.3% were HBeAg-positive (vs 23.4% in the MASTER cohort; P <0.0001), and 12.3% were anti-HDV-positive (vs 11.1%). Overall, the adherence to the European Association for the Study of the Liver recommendations for antiviral treatment increased over time. Conclusion: Chronic hepatitis B virus infection appears to be in the process of becoming under control in Italy; however, HDV infection is still a health concern in patients with cirrhosis and in migrants

The Mycotoxin Deoxynivalenol Potentiates Intestinal Inflammation by Salmonella Typhimurium in Porcine Ileal Loops

Background and Aims: Both deoxynivalenol (DON) and nontyphoidal salmonellosis are emerging threats with possible hazardous effects on both human and animal health. The objective of this study was to examine whether DON at low but relevant concentrations interacts with the intestinal inflammation induced by Salmonella Typhimurium. Methodology: By using a porcine intestinal ileal loop model, we investigated whether intake of low concentrations of DON interacts with the early intestinal inflammatory response induced by Salmonella Typhimurium. Results: A significant higher expression of IL-12 and TNF alpha and a clear potentiation of the expression of IL-1 beta, IL-8, MCP-1 and IL-6 was seen in loops co-exposed to 1 mu g/mL of DON and Salmonella Typhimurium compared to loops exposed to Salmonella Typhimurium alone. This potentiation coincided with a significantly enhanced Salmonella invasion in and translocation over the intestinal epithelial IPEC-J2 cells, exposed to non-cytotoxic concentrations of DON for 24 h. Exposure of Salmonella Typhimurium to 0.250 mu g/mL of DON affected the bacterial gene expression level of a limited number of genes, however none of these expression changes seemed to give an explanation for the increased invasion and translocation of Salmonella Typhimurium and the potentiated inflammatory response in combination with DON. Conclusion: These data imply that the intake of low and relevant concentrations of DON renders the intestinal epithelium more susceptible to Salmonella Typhimurium with a subsequent potentiation of the inflammatory response in the gut