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This study set out to investigate whether US Heritage Spanish features a more streamlined verbal paradigm in psych verb constructions compared to standard varieties of Spanish, where HS speakers find an invariable third-person singular form acceptable with both singular and plural grammatical subjects. In standard Spanish, the semantic subjects of psych verbs are typically pre-verbal experiencers cast as oblique arguments in inverse predicates such as in me encantan los buhos ‘I love owls’. The translation of this sentence shows that equivalent English predicates are typically direct constructions. The data were gathered using an acceptability judgement questionnaire that was distributed to participants that fit into one of three groups: early bilingual heritage speakers of Spanish from California, advanced Spanish as L2 speakers, and non-bilingual native speakers of Spanish who had learnt English as an L2 as adults. The Heritage Spanish speakers in this group often patterned differently from both other groups, who surprisingly patterned together. We argue that this is due to L2 speakers’ mode of acquisition (formal and subject to prescriptive grammar), in comparison with Heritage Spanish speakers’ naturalistic acquisition. Specifically, we find evidence for a streamlining of the Spanish verbal paradigm not immediately attributed to English interference, and that in psych verb constructions, Heritage Spanish speakers more readily accept a third-person singular invariable verbal form. This differentiation of the verbal paradigm from standard Spanish use should be considered a bona fide linguistic change, but not proof of either incomplete acquisition or language attrition. Since Heritage Spanish speakers are, after all, native speakers of Spanish, this study shows that Heritage Spanish should be considered and studied as any other dialect of Spanish, with its distinctive grammatical features, and subject to variability and change.

This study explores anxiety and depression among young individuals in Kosovo, considering socio-political and economic influences. Introducing Positive Youth Development (PYD) as a framework, the study explores its integration with Internal Cohesion Psychotherapy (ICP) to address youth mental health challenges. Empirical evidence underscores ICP’s positive impact on mental health outcomes, emphasizing developmental assets. Thematic analysis of participant narratives reveals six key themes, showcasing ICP’s transformative nature, the critical role of social support networks, values and beliefs, reflections on personal growth, and therapy’s extended impact on relationships. Participants’ stories highlight ICP’s profound influence on personal growth, social relationships, and mental health. The study examines internal and external assets, emphasizing social networks, values, beliefs, and cultural competence. Insights into ICP’s intrapersonal and professional systems reveal developmental assets fostering growth, motivation, and engagement. External assets like empowerment, safety, and service to others, alongside internal assets like commitment to learning, positive values, integrity, and responsibility, contribute to an understanding of factors influencing Kosovo’s young population. In conclusion, the integration of PYD and ICP offers a powerful synergy to address mental health challenges, providing valuable insights into specific developmental assets crucial for optimizing interventions and promoting mental health among young Kosovars.

The Italian Association for Laboratory Animal Sciences (AISAL) held its annual meeting in September 2022. The main topic of the meeting was the section ‘Quality Control of the Components in the Study’ of the PREPARE guidelines. Among the audience, all stakeholders of AISAL were present: animal caretakers, technicians, facility managers, veterinarians, scientists, and animal welfare officers. One session was also dedicated to the round table on education and training, with the aim to discuss the legislative requirements as per Italian Decree 5 August 2021.

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Low-temperature treatments can be applied to grapes or must before alcoholic fermentation to enhance the wine’s sensory characteristics. Several studies have shown that such practices have a positive effect on the polyphenol profile of the wine, but only a few surveys have examined the effect of these treatments on the yeast microbiota of grapes and wine. Therefore, this study aimed to evaluate how cryoextraction (freezing the grape with liquid nitrogen) and cold pre-fermentative maceration (at 5 °C for 48 h) affect the Saccharomyces and non-Saccharomyces populations during the winemaking process of red grapes, cv Sangiovese, conducted at two temperatures (20 and 30 °C). This research analyzed the concentration of various yeast species, their fermentation abilities, and the resulting wine’s aromatic profile. The Principal Component Analysis performed on yeast concentrations during the fermentations of various wines did not group the experimental wines based on treatment. However, the same groupings were highlighted when the concentrations of the volatile compounds, quantified in the experimental wines, were processed using the same statistical approach. Therefore, cryoextraction and cold pre-fermentative maceration seem to contribute less to the aromatic profile than the yeasts involved in the fermentation process.

Sulphur dioxide (SO2) is usually used in winemaking due to its effectiveness as an antimicrobial and antioxidant agent. However, there is growing interest in finding alternatives to SO2 because of its adverse effects on human health. Therefore, in this work, a low-SO2-producing Saccharomyces cerevisiae strain was selected and the AIR-MIXINGTM M.ITM (A.M.) system, capable of determining a high extraction of polyphenols, was used to produce a red wine without added sulphites. A tank managed with the traditional pumping-over technique was used as a comparison. Microbiological and chemical monitoring of both fermentations performed by plate counts and HPLC analyses, respectively, did not indicate significant differences as regards the yeast growth kinetics and the degradation of the sugars, while it highlighted a faster extraction kinetics of colour and total polyphenols in the fermentation carried out with the A.M. system. Both experimental wines showed a total SO2 content 2-producing yeasts in combination with the A.M. system could be a suitable approach to produce wines without sulphites added.

Recent interest in the special beer category has encouraged the search for novel brewing materials, including new ingredients and novel yeast strains, in order to differentiate the finished products. The aim of this work was to select non-brewing S. cerevisiae strains for the production of a fruit beer with raspberry. The in vitro tests and the wort fermentations allowed the selection of two sourdough S. cerevisiae strains, showing high maltose and maltotriose consumption, high ethanol production, and high viability. Fruit beers (FB) and control beers (CB) without raspberries were prepared. Fruit addition accelerated sugar consumption (7 days compared to 13 days) and increased ethanol and glycerol production by yeasts. Raspberry addition and the inoculated yeast strongly affected the aroma profile of beers. FB samples showed a higher amount of volatile organic compounds (VOCs); the most represented classes were alcohols, followed by esters and acids. FB inoculated by the selected S. cerevisiae SD12 showed the highest VOCs concentration (507.33 mg/L). Results highlighted the possible application of sourdough yeast strains for the brewing process, which, combined with raspberry addition, can be exploited for the production of beers with enhanced aromatic features and suitable chemical properties.

Background: Measles infection in the hospital setting is a major issue. Despite the availability of an effective vaccine, measles outbreaks continue to occur in some European countries. We aimed to evaluate the immunological status of medical students attending the Tor Vergata Polyclinic (PTV). Methods: Measles antibodies titers were assessed by venipuncture on a sample of 2717 medical students who underwent annual health surveillance visits from January 2021 to March 2023. Subjects showing serum IgG values above 1.0 S/CO were considered serologically protected. Personal data, country of origin, and main demographic characteristic were also collected. Results: 66.7% (1467 Italian and 346 foreign) of medical students showed protective IgG antibodies levels. Female students were serologically immune more frequently than males (68.6% vs. 63.3%; p < 0.01 at Chi2). The mean antibody titer was 1.72 S/CO, significantly higher in females than males (1.67 vs. 1.75, respectively; p < 0.05), and significantly related to age (p < 0.01). Albanian students, who were the largest foreign population in our study, showed a low serological protection rate (40/90: 44.4%). Conclusions: The proportion of serologically non-immune students is high, raising concerns about the possible risk of hospital transmission. Substantial differences in the rate of immunity have been found between subjects coming from different parts of Europe and the world. Pre-training assessment of all medical students and vaccination of susceptible individuals is highly recommended, particularly for those from low immunization rate countries.

The use of earthenware amphorae in winemaking can give wines unique attributes enhancing their typicity. Therefore, in this study, spontaneous and inoculated in-amphora fermentations of Trebbiano Toscano grape must were monitored to assess the Saccharomyces cerevisiae strains occurring in each fermentation as well as the chemical characteristics of the wines. Strain typing via Interdelta analyses pointed out that the commercial starters did not dominate, showing 24% and 13% implantation percentages, and that 20 indigenous strains were present at different percentages, ranging from 2 to 20%, in inoculated and spontaneous fermentations. The assessment of the technical characteristics of the indigenous strains via fermentations at lab and pilot scale (20 L amphorae) and the sensory analysis of the experimental wines allowed for the selection of two indigenous strains to be used as starter cultures in comparison to a commercial strain in 300-L-amphorae vinifications in the cellar. The observed fermentative performances and sensory analysis of the experimental wines highlighted that one indigenous S. cerevisiae strain dominated the process and conferred distinctive sensory characteristics to the Trebbiano Toscano wine, demonstrating its effectiveness in managing the in-amphora fermentations. In addition, the results demonstrated the ability of amphorae to protect the polyphenolic compounds from oxidation during wine ageing. Indeed, the concentration of both hydroxycinnamic acids and flavonols decreased, with an average reduction of 30% and 14%, respectively, while hydroxybenzoic acids remained unchanged.

Millet is the sixth-highest yielding grain in the world and a staple crop for millions of people. Fermentation was applied in this study to improve the nutritional properties of pearl millet. Three microorganism combinations were tested: Saccharomyces boulardii (FPM1), Saccharomyces cerevisiae plus Campanilactobacillus paralimentarius (FPM2) and Hanseniaspora uvarum plus Fructilactobacillus sanfranciscensis (FPM3). All the fermentation processes led to an increase in minerals. An increase was observed for calcium: 254 ppm in FPM1, 282 ppm in FPM2 and 156 ppm in the unfermented sample. Iron increased in FPM2 and FPM3 (approx. 100 ppm) with respect the unfermented sample (71 ppm). FPM2 and FPM3 resulted in richer total phenols (up to 2.74 mg/g) compared to the unfermented sample (2.24 mg/g). Depending on the microorganisms, it was possible to obtain different oligopeptides with a mass cut off ≤10 kDalton that was not detected in the unfermented sample. FPM2 showed the highest resistant starch content (9.83 g/100 g) and a prebiotic activity on Bifidobacterium breve B632, showing a significant growth at 48 h and 72 h compared to glucose (p < 0.05). Millet fermented with Saccharomyces cerevisiae plus Campanilactobacillus paralimentarius can be proposed as a new food with improved nutritional properties to increase the quality of the diet of people who already use millet as a staple food.

Compounding is the final processing step for quality adjustment and control before recycled thermoplastic polymer material can be introduced into production processes. Motivated by the need for higher recyclate shares, the research question is which quality problems recycling compounders are encountered in practice, where they occur, and which mitigation options might be reasonable. Therefore, an online survey with 20 recycling compounders based in Germany was conducted asking about typical processing steps and processed materials, test procedures for quality assurance, quality problems, and possibilities for reducing quality problems. Results show that compounders mainly name impurities and contaminations of the input material as challenging and the reason for quality problems. The study shows that the problems are not dependent on the material input type. Quality problems occur along the entire secondary value chain, with companies manufacturing components themselves being particularly affected. The composition determination of the input materials helps to minimize quality problems.

Probiotic milk is a class of fermented milk that possesses health-promoting effects, not only due to the lactic acid bacteria (LAB) presence but potentially also to yeast activity. Hence, the aim of this work was to isolate and select yeasts from spontaneous milk fermentations to be used as inoculum, together with LAB, for manufacturing a potentially probiotic acidic low-alcohol fermented milk. Six yeast species were detected from the spontaneous milk fermentation. A screening of 13 yeast strains and 14 previously isolated LAB strains, based on the resistance to bile salts and to acidic conditions, was carried out. The best performing strains were successively tested for in vitro gastrointestinal tolerance. A strain of Kluyveromyces marxianus and a strain of Lactococcus lactis were selected for the manufacturing of two different fermented milk. The values of the main technological and microbiological parameters (pH, organic acids, ethanol, and microbial concentrations) of the experimental milk were in the range of those reported for this category of products. The evaluation of microorganism survival in fermented milk samples subjected to simulated gastrointestinal conditions highlighted a high resistance of both strains. In conclusion, the selected microbial starter culture enabled the setting up of potential probiotic fermented milk.

Since its publication, the 3Rs principle has provided a cornerstone for more ethical and humane biomedical and regulatory research. In Europe, the 3Rs principle has been incorporated into the European Directive 63/2010/EU, with the ultimate aim of fully replacing the procedures on live animals for scientific and educational purposes as soon as it is scientifically possible to do so. Thus, a critical shift in the discussion on animal use in biomedical and regulatory research is undergoing in Europe, a discussion where satisfying the “replacement” principle is becoming more and more defined as a scientific rather than ethical need. 3Rs Centres have been established in recent years across Europe. To date, Ireland has no 3Rs Centre, and the uptake of the 3Rs principle, and in particular of the “replacement” aspect, has been slow. In this Commentary, we present the Irish context of the use of animal models in biomedical and regulatory research, and urge for what, in the authors’ opinion, are the most critical actions that Ireland must undertake to align its biomedical (basic, applied and translational) research with the European 3Rs strategy.

The inoculation of grape juice with Saccharomyces cerevisiae strains selected from indigenous yeast populations can be a suitable tool to control alcoholic fermentation, contributing to producing wines with typical flavor and aroma and, hence, the demand for native starter cultures is increasing. However, since low amounts of indigenous yeast biomasses are usually required for local winemaking, the industrial production of these yeasts can be expensive. Therefore, in this study, after selecting an indigenous S. cerevisiae strain based on relevant oenological and technological features, a pilot-plant for easy and rapid production of fresh yeast biomass directly in a winery located in Tuscany, was exploited. The selected yeast strain was used as a starter to carry out 25 and 100 hL fermentations and its enological performance was compared with that of the commercial starter normally used in the winery. Chemical and sensory analysis of the resulting wines showed that they differentiated according to the used yeast strain, with the wines produced by the indigenous S. cerevisiae strain being characterized by a distinctive aromatic and sensory profile. In conclusion, the pilot-plant effectively resulted in producing fresh yeast starter cultures in the winery to be successfully used to carry out alcoholic fermentations.

Functional and nutritional properties of baked goods can be enhanced by the use of sourdough fermentation, which is defined as a mixture of water and flour fermented by lactobacilli (LAB) and yeasts. Previous data highlighted the ability of sourdoughs obtained with selected LAB strains and commercial flour to produce bioactive peptides equipped with antioxidant and anti-inflammatory properties. More recently, it has been proven that choosing the most suitable combination of selected LAB and high-functional-value flours greatly increase the amount of low-molecular-weight antioxidant compounds responsible for improving the nutritional value of the products. This work aimed to isolate bioactive peptides both from sourdoughs and baked breads obtained with pigmented wheat and barley and a selected combination of LABs. Soluble water extracts were obtained, and low-molecular-weight peptides were isolated. Antioxidant activity was detected by assaying the intracellular ROS production in stressed cultured macrophages, treated with peptides. Moreover, anti-inflammatory activity, highlighted by NFkB pathway inhibition and by COX2 reduction in stressed cells, was demonstrated for peptides obtained from breads. The results allowed the conclusion that the combination of flours and LAB used in the present manuscript led to the production of bakery products with beneficial effects on oxidative and inflammatory status.

Aminoglycoside or colistin therapy may alter the renal tubular function without decreasing the glomerular filtration rate. This association has never been extensively investigated. We conducted a systematic review of the literature following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses recommendations. Databases searched included United States National Library of Medicine, Excerpta Medica, and Web of Science. For the final analysis, we evaluated 46 reports, published after 1960, describing 82 cases. A total of 286 electrolyte and acid-base disorders were reported. Hypomagnesemia, hypokalemia, and hypocalcemia were reported in more than three quarter of cases. Further disorders were, in decreasing order of frequency, metabolic alkalosis, hyponatremia, hypophosphatemia, hypouricemia, hypernatremia, and metabolic acidosis. Six electrolyte and acid-base disorders were reported in seven cases, five in 12 cases, four in 16 cases, three in 31 cases, two in 11 cases, and one in five cases. Laboratory features consistent with a loop of Henle/distal tubular dysfunction were noted in 56 (68%), with a proximal tubular dysfunction in three (3.7%), and with a mixed dysfunction in five (6.1%) cases. The laboratory abnormality was unclassified in the remaining 18 (22%) cases. Treatment with aminoglycosides or colistin may trigger a proximal tubular or, more frequently, a loop of Henle/distal tubular dysfunction.

Leukocytoclastic small-vessel vasculitis of the skin (with or without systemic involvement) is often preceded by infections such as common cold, tonsillopharyngitis, or otitis media. Our purpose was to document pediatric (≤18 years) cases preceded by a symptomatic disease caused by an atypical bacterial pathogen. We performed a literature search following the Preferred Reporting of Systematic Reviews and Meta-Analyses guidelines. We retained 19 reports including 22 cases (13 females and 9 males, 1.0 to 17, median 6.3 years of age) associated with a Mycoplasma pneumoniae infection. We did not find any case linked to Chlamydophila pneumoniae, Chlamydophila psittaci, Coxiella burnetii, Francisella tularensis, or Legionella pneumophila. Patients with a systemic vasculitis (N = 14) and with a skin-limited (N = 8) vasculitis did not significantly differ with respect to gender and age. The time to recovery was ≤12 weeks in all patients with this information. In conclusion, a cutaneous small-vessel vasculitis with or without systemic involvement may occur in childhood after an infection caused by the atypical bacterial pathogen Mycoplasma pneumoniae. The clinical picture and the course of cases preceded by recognized triggers and by this atypical pathogen are indistinguishable.

Steroids, providing information regarding several biological patterns including stress and sexual behavior, have been investigated in different matrices in laboratory mice. Data regarding hair quantification, indicative of longer timespans when compared to blood and saliva, are lacking. The aim of the work was to analyze the hormonal hair profile of laboratory male mice and to investigate potential relationships with age and housing, as a potential tool for welfare assessment. Fifty-six adult male C57BL/6J and C57BL/6OlaHsd substrain mice were included in the study, housed in pairs or groups. Testosterone (T) and dehydroepiandrosterone (DHEA) were quantified by radioimmunoassay, corticosterone (CORT) by ELISA. Mean hormone levels were 6.42 pg/mg for T, 23.16 pg/mg for DHEA and 502.1 pg/mg for CORT. Age influenced all hormones by significantly increasing T and DHEA levels and decreasing CORT; only DHEA, significantly higher in grouped mice, was influenced by housing conditions. The influence of age indicates the need for accurate age-related reference intervals, while the higher levels of DHEA in grouped animals suggests that such housing practice may be beneficial for social interactions. In conclusion, it seems that hair hormones quantification may be a good tool for welfare assessment in laboratory mice and may help in refining husbandry.

The bacterial community profile of cricket powder highlighted the presence of four main genera: Bacteroides spp., Parabacteroides spp., Lactococcus spp., and Enterococcus spp. The spontaneous fermentation of cricket powder allowed for the isolation and characterization of seven lactic acid bacteria strains belonging to six species: Latilactobacillus curvatus, Lactiplantibacillus plantarum, Latilactobacillus sakei, Lactococcus garvieae, Weissella confusa, and Enterococcus durans. The strains were characterized and selected according to different technological properties. L. plantarum CR L1 and L. curvatus CR L13 showed the best performance in terms of general aminopeptidase activity, acidification, and growth rate in MRS broth and in dough with cricket powder and wheat flour, as well as robustness during consecutive backslopping. Thus, they were used as starter-mixed to produce sourdough to manufacture bread fortified with 20% cricket powder. The addition of cricket powder led to a significant increase of protein (up to 94%) and lipid content, from 0.7 up to 6 g/100 g of bread. Spontaneous fermentation represents a source of microbial diversity that can be exploited in order to obtain potential starters for food with innovative ingredients. Edible insects powder can be successfully added in leavened baked goods to enhance their nutritional value.

The use of sourdough fermentation and whole grain flours in baked goods manufacturing are known to enhance their functional and nutritional features. In this context, it is necessary to select the most suitable lactic acid bacteria strains and flour combination to achieve this goal. A characterization of 70 lactobacilli strains based on pro-technological and nutritional properties was carried out. The screening allowed the selection of 10 strains that were used to ferment sourdoughs made with two varieties of common wheat, the conventional red-grained cv Aubusson, a blue-grained variety rich in anthocyanins cv Skorpion, and a hull-less barley variety, cv Rondo. From each fermented sourdough, a water soluble extract was obtained and evaluated for its antioxidant activity performed on cultured cells (RAW 264.7 murine macrophage) by assaying Reactive Oxygen Species (ROS) content. Sourdoughs made with pigmented wheat and barley, had an antioxidant activity greater than that recovered in those made with conventional wheat flour, in spite they have been inoculated with the same LAB strains. Results highlighted the interdependence between flour and the inoculated lactic acid bacteria that has to be taken into account for the development of healthy breads exploiting high functional value cereals through biotechnological processes.

γ-Aminobutyric acid (GABA) is the primary inhibitory neurotransmitter of the central nervous system and possesses various physiological functions. GABA production can be obtained thanks to lactic acid bacteria activity in different foods such as sourdoughs. Recently, breads made from blends of pseudocereals and wheat flours have attracted much attention. Amaranth is especially interesting because of its high nutritional value, having a high protein content and containing different antioxidant compounds. Therefore, this study aimed to obtain sourdough breads enriched with GABA thanks to bacterial activity and to investigate the effect of amaranth flour addition on the antioxidant and sensorial properties of bread. Eighteen lactobacilli strains were assayed for GABA production in amaranth and wheat flour liquid sourdoughs. Two strains, Lactobacillus brevis A7 and Lactobacillus farciminis A11, demonstrated high GABA producing capability; hence, they were used to prepare breads containing 20% amaranth flour. The results confirmed the capability of the two strains to increase GABA concentrations (up to 39 mg/kg) in breads. Samples with amaranth addition showed a significantly higher total phenolic content compared to the control bread (more than 15 mg GAE 100 g−1 dwb); sensory analysis showed that breads with amaranth were moderately acceptable. Nevertheless, their general liking evaluation was significantly lower compared to the control bread. The addition of pseudocereal to traditional wheat sourdough and selection of lactobacilli allowed the production of baked goods with enhanced GABA content and antioxidant capacity, but recipes have to be developed to increase the organoleptic acceptability of the final products.

Seronegative latent carriers (SNLCs) are animals that carry the virus without detectable antibodies and pose a risk for disease transmission and diagnostic challenges, suggesting the importance of consideration of marker vaccines in managing them. Therefore, in this study, we evaluated two modified live infectious bovine rhinotracheitis (IBR) marker vaccines (single and double deletions) for their ability to generate SNLC calves. These vaccines were administered to four groups (n = 3 in each group) of three-month-old calves in the presence or absence of passive immunity. Three hundred days after the first vaccination and after confirming the IBR seronegativity of all animals, dexamethasone was administered intravenously for five consecutive days. Only animals immunized with the modified live IBR marker vaccine (single deletion) in the absence of passive immunity exhibited a more enduring immune response than those vaccinated in the presence of passive immunity. Moreover, the administration of a modified live IBR marker vaccine (double deletion) to calves with passive immunity generated SNLC. These findings underscore the potential of live IBR marker vaccine (double-deletions) to aid serological diagnostic tools and develop vaccination protocols in achieving the desired immune response, particularly in the context of latent carrier status, offering valuable insights into optimizing vaccination strategies for effective IBR control.

Catheter-based revascularization procedures were developed as an alternative to systemic thrombolysis for patients with intermediate-high- and high-risk pulmonary embolisms. USAT IH-PE is a retrospective and prospective multicenter registry of such patients treated with ultrasound-facilitated, catheter-directed thrombolysis, whose preliminary results are presented in this study. The primary endpoint was the incidence of pulmonary hypertension (PH) at follow-up. Secondary endpoints were short- and mid-term changes in the echocardiographic parameters of right ventricle (RV) function, in-hospital and all-cause mortality, and procedure-related bleeding events. Between March 2018 and July 2023, 102 patients were included. The majority were at intermediate-high-risk PE (86%), were mostly female (57%), and had a mean age of 63.7 ± 14.5 years, and 28.4% had active cancer. Echocardiographic follow-up was available for 70 patients, and in only one, the diagnosis of PH was confirmed by right heart catheterization, resulting in an incidence of 1.43% (CI 95%, 0.036-7.7). RV echocardiographic parameters improved both at 24 h and at follow-up. In-hospital mortality was 3.9% (CI 95%, 1.08-9.74), while all-cause mortality was 11% (CI 95%, 5.4-19.2). Only 12% had bleeding complications, of whom 4.9% were BARC ≥ 3. Preliminary results from the USAT IH-PE registry showed a low incidence of PH, improvement in RV function, and a safe profile.

Different types of vaccines against Infectious Bovine Rhinotracheitis (IBR) are commercially available. Among these, inactivated glycoprotein E (gE)-deleted marker vaccines are commonly used, but their ability to induce passive immunity is poorly known. Here, we evaluated the passive immunity transferred from dams immunised with commercial inactivated gE-deleted marker vaccines to calves. We vaccinated 12 pregnant cattle devoid of neutralising antibodies against Bovine alphaherpesvirus 1 (BoHV-1) and divided them into two groups with 6 animals each. Both groups were injected with a different inactivated gE-deleted marker vaccine administrated via intranasal or intramuscular routes. An additional 6 pregnant cattle served as the unvaccinated control group. After calving, the number of animals in each group was increased by the newborn calves. In the dams, the humoral immune response was evaluated before calving and, subsequently, at different times until post-calving day 180 (PCD180). In addition, the antibodies in colostrum, milk, and in serum samples from newborn calves were evaluated at different times until PCD180. The results indicated that inactivated glycoprotein E (gE)-deleted marker vaccines are safe and produce a good humoral immune response in pregnant cattle until calving and PCD180. Moreover, results showed that, in calf serum, passive immunity persists until PCD180.

A multicenter pilot program for universal newborn screening of Sickle cell disease (SCD) was conducted in two centres of Northern Italy (Padova and Monza). High Performance Liquid Chromatography (HPLC) was performed as the first test on samples collected on Guthrie cards and molecular analysis of the β-globin gene ( HBB ) was the confirmatory test performed on the HPLC-positive or indeterminate samples. 5466 samples of newborns were evaluated. Of these, 5439/5466 were submitted to HPLC analysis and the molecular analysis always confirmed in all the alteration detected in HPLC (62/5439 newborns); 4/5439 (0.07%) were SCD affected, 37/5439 (0.68%) were HbAS carriers and 21/5439 (0.40%) showed other hemoglobinopathies. Stored dried blood spots were adequate for HPLC and β-globin gene molecular analysis. Samples were suitable for analysis until sixteen months old. A cut-off of A 1 percentage, in order to avoid false negative or unnecessary confirmation tests, was identified. Our experience showed that several technical issues need to be addressed and resolved while developing a multicenter NBS program for SCD in a country where there is no national neonatal screening (NBS) program for SCD and NBS programs occur on a regional basis., Competing Interests: Conflicts of InterestThe authors declare no conflicts of interest., (© 2018 by the authors.)