Differentiation and Maturation of Muscle and Fat Cells in Cultivated Seafood: Lessons from Developmental Biology.

Cultivated meat, also known as cultured or cell-based meat, is meat produced directly from cultured animal cells rather than from a whole animal. Cultivated meat and seafood have been proposed as a means of mitigating the substantial harms associated with current production methods, including damage to the environment, antibiotic resistance, food security challenges, poor animal welfare, and-in the case of seafood-overfishing and ecological damage associated with fishing and aquaculture. Because biomedical tissue engineering research, from which cultivated meat draws a great deal of inspiration, has thus far been conducted almost exclusively in mammals, cultivated seafood suffers from a lack of established protocols for producing complex tissues in vitro. At the same time, fish such as the zebrafish Danio rerio have been widely used as model organisms in developmental biology. Therefore, many of the mechanisms and signaling pathways involved in the formation of muscle, fat, and other relevant tissue are relatively well understood for this species. The same processes are understood to a lesser degree in aquatic invertebrates. This review discusses the differentiation and maturation of meat-relevant cell types in aquatic species and makes recommendations for future research aimed at recapitulating these processes to produce cultivated fish and shellfish.

3D Bioprinting of Novel κ-Carrageenan Bioinks: An Algae-Derived Polysaccharide.

Novel green materials not sourced from animals and with low environmental impact are becoming increasingly appealing for biomedical and cellular agriculture applications. Marine biomaterials are a rich source of structurally diverse compounds with various biological activities. Kappa-carrageenan (κ-c) is a potential candidate for tissue engineering applications due to its gelation properties, mechanical strength, and similar structural composition of glycosaminoglycans (GAGs), possessing several advantages when compared to other algae-based materials typically used in bioprinting such as alginate. For those reasons, this material was selected as the main polysaccharide component of the bioinks developed herein. In this work, pristine κ-carrageenan bioinks were successfully formulated for the first time and used to fabricate 3D scaffolds by bioprinting. Ink formulation and printing parameters were optimized, allowing for the manufacturing of complex 3D structures. Mechanical compression tests and dry weight determination revealed young's modulus between 24.26 and 99.90 kPa and water contents above 97%. Biocompatibility assays, using a mouse fibroblast cell line, showed high cell viability and attachment. The bioprinted cells were spread throughout the scaffolds with cells exhibiting a typical fibroblast-like morphology similar to controls. The 3D bio-/printed structures remained stable under cell culture conditions for up to 11 days, preserving high cell viability values. Overall, we established a strategy to manufacture 3D bio-/printed scaffolds through the formulation of novel bioinks with potential applications in tissue engineering and cellular agriculture.

Gel and thermoluminescence dosimetry for dose verifications of a real anatomy simulated prostate conformal radiation treatment in the presence of metallic femoral prosthesis.

This study aims to verify the dose delivery of prostate radiotherapy treatments in an adult pelvic phantom with two metallic hip and femur prosthesis using a four-field box technique. The prostate planned target volume (PTV) tridimensional (3D) dose distribution was evaluated using gel dosimetry, and thermoluminescent dosimeters (TLD) were used for point-dose measurements outside it. Both results were compared to the treatment planning system (TPS) dose calculation without using heterogeneity corrections to evaluate the influence of the metal in the dose distribution. MAGIC-f gel dosimeter (Methacrylic and Ascorbic acid in Gelatin Initiated by Copper with Formaldehyde) associated with magnetic resonance imaging was used. TLD were positioned at several points at the bone metal interface and the sacrum region. The comparison of the gel measured and the TPS calculated dose distributions were done using gamma analysis (3%/3 mm), and a pass rate of 93% was achieved. The TLD dose values at the bone-metal interface showed variations from the planned dose. However, at the sacrum region, where the beams did not intercept the prosthesis, there was a good agreement between TPS planning and TLD measurements. Our results show how the combination of 3D dosimetry and measurements at specific points in the phantom allowed a comprehensive view of the dose distribution and identified that care must also be paid to regions outside the PTV.

Biomechanical Assessment of Bicuspid Aortic Valve Phenotypes: A Fluid-Structure Interaction Modelling Approach.

PURPOSE: Bicuspid aortic valve (BAV) is a congenital heart malformation with phenotypic heterogeneity. There is no prior computational study that assesses the haemodynamic and valve mechanics associated with BAV type 2 against a healthy tricuspid aortic valve (TAV) and other BAV categories. METHODS: A proof-of-concept study incorporating three-dimensional fluid-structure interaction (FSI) models with idealised geometries (one TAV and six BAVs, namely type 0 with lateral and anterior-posterior orientations, type 1 with R-L, N-R and N-L leaflet fusion and type 2) has been developed. Transient physiological boundary conditions have been applied and simulations were run using an Arbitrary Lagrangian-Eulerian formulation. RESULTS: Our results showed the presence of abnormal haemodynamics in the aorta and abnormal valve mechanics: type 0 BAVs yielded the best haemodynamical and mechanical outcomes, but cusp stress distribution varied with valve orifice orientation, which can be linked to different cusp calcification location onset; type 1 BAVs gave rise to similar haemodynamics and valve mechanics, regardless of raphe position, but this position altered the location of abnormal haemodynamic features; finally, type 2 BAV constricted the majority of blood flow, exhibiting the most damaging haemodynamic and mechanical repercussions when compared to other BAV phenotypes. CONCLUSION: The findings of this proof-of-concept work suggest that there are specific differences across haemodynamics and valve mechanics associated with BAV phenotypes, which may be critical to subsequent processes associated with their pathophysiology processes.

A newly developed Escherichia coli isolate panel from a cross section of U.S. animal production systems reveals geographic and commodity-based differences in antibiotic resistance gene carriage.

There are limited numbers of Escherichia coli isolate panels that represent United States food animal production. The majority of existing Escherichia coli isolate panels are typically designed: (i) to optimize genetic and/or phenotypic diversity; or (ii) focus on human isolates. To address this shortfall in agriculturally-related resources, we have assembled a publicly-available isolate panel (AgEc) from the four major animal production commodities in the United States, including beef, dairy, poultry, and swine, as well as isolates from agriculturally-impacted environments, and other commodity groups. Diversity analyses by phylotyping and Pulsed-field Gel Electrophoresis revealed a highly diverse composition, with the 300 isolates clustered into 71 PFGE sub-types based upon an 80% similarity cutoff. To demonstrate the panel's utility, tetracycline and sulfonamide resistance genes were assayed, which identified 131 isolates harboring genes involved in tetracycline resistance, and 41 isolates containing sulfonamide resistance genes. There was strong overlap in the two pools of isolates, 38 of the 41 isolates harboring sulfonamide resistance genes also contained tetracycline resistance genes. Analysis of antimicrobial resistance gene patterns revealed significant differences along commodity and geographical lines. This panel therefore provides the research community an E. coli isolate panel for study of issues pertinent to U.S. food animal production.

Glycosaminoglycans and vitamin C affect broiler bone parameters.

The purpose of this study was to determine if in ovo feeding and rearing with glycosaminoglycans and vitamin C could influence bone and cartilage macroscopy, mineral composition, mineral density and surface area, bone breaking strength, and bone histology in broilers. Fertile eggs from breeders (Cobb) were either uninjected or injected with 4 µg of additive/100 µL water on day 4 of incubation. Every 100 g of in ovo additive contained 30 g of chondroitin sulfate, 30 g of glucosamine, and 5 g of vitamin C. After hatching, the chicks from both incubation treatments were submitted to additional treatments during the growth phase from 1 to 42 D of age (diet without and with 0.74 g of additive/kg of feed). Every 100 kg of feed contained 30 g of glucosamine sulfate, 24 g of chondroitin sulfate, and 20 g of vitamin C. A completely randomized factorial design (2 × 2) was applied. The data were submitted to variance analysis using the general linear model procedure of SAS (SAS Institute). In ovo feeding with 4 µg of additive plus dietary supplementation with 0.74 g of additive/kg of feed resulted in the highest cartilage weight of the femur proximal epiphysis in birds (P = 0.0098). The highest ash, phosphorus and calcium percentage, mineral density and mineral composition were identified for femur and tibia in the following treatments: in ovo feeding plus diet without additive during rearing, or uninjected eggs plus dietary supplementation during rearing. In ovo feeding with 4 µg of the additive reduced (P = 0.0008) the number of chondrocytes in the proximal epiphysis of the tibia cartilage and increased (P < 0.0001) the number of osteocytes in the tibia diaphysis of broilers. We conclude that in ovo feeding or dietary supplementation during broiler rearing with glycosaminoglycans (chondroitin sulfate and glucosamine sulfate) and vitamin C benefits the development of bird bones and cartilage, and may represent a solution to bone problems in broilers.

Effect of the addition of glycosaminoglycans on bone and cartilaginous development of broiler chickens.

Locomotion issues in broiler production may decrease performance (carcass yield and traits) and lead to high financial losses. This study evaluates the addition of glucosaminoglycans in broiler diets to minimize the lack of proper bone development and joint weakening. The experiment was conducted using 2,160 broilers randomly distributed in a factorial pattern (3 × 3) using 3 levels of glucosamine sulfate (0, 0.12, and 0.24%) and 3 levels of chondroitin sulfate addition (0, 0.08, and 0.16%). Eight repetitions were used for each treatment, distributed in 72 pens with 30 broilers each. There was a quadratic effect on feed conversion for broilers from 1 to 42 d old (P = 0.0123) for the addition of chondroitin, and better feed conversion was obtained by adding 0.08% of chondroitin. The relative tibia weight, the width of the proximal epiphysis and diaphysis presented a linear increased effect in broilers at 42 d old. An interaction was found between the amount of chondroitin × glucosamine and the number of chondrocytes in the proximal cartilage of the tibia (P = 0.0072). There was a quadratic effect of glucosamine levels (P = 0.0107) in the birds that had received the 0.16% addition of chondroitin, and the presence of 0.18% glucosamine increased the number chondrocytes in the cartilage of broilers. These results provide the first evidence that broilers may benefit from increased dietary chondroitin sulfate. These results indicate that the addition of glucosamine and chondroitin sulfates in broiler feed rations might alleviate leg conditions and decrease financial losses in the broiler industry.

Burden of rotavirus infections in Liguria, Northern Italy: hospitalisations and potential savings by vaccination.

We evaluated the rates of gastroenteritis admissions to the emergency department and of rotavirus-related hospitalisations in children ≤5 years of age in 2006 at an Italian paediatric hospital. We calculated the number of rotavirus cases avoidable through the universal vaccination of children. Epidemiological data were extracted from the Data Elaboration Centre. To calculate the hospitalisation rate due to rotavirus, the virus was sought in the faeces of children hospitalised for acute gastroenteritis by means of rapid immunochromatographic assay. Emergency department admissions due to gastroenteritis numbered 2,396 (11.58% of the total admissions). Of these, 276 children (11.52%) were examined and then sent home, 1,286 (53.67%) were kept in short observation and 776 (32.38%) were hospitalised. In 27.83% of hospitalised cases, the rotavirus test proved positive. The rotavirus hospitalisation rate was 55 per 10,000 children ≤5 years of age in Genoa in 2006. In 85.6% of hospitalised patients with community-acquired rotavirus infection, the disease was severe. The number of avoidable cases confirmed that the vaccination of children ≤1 year of age could reduce the burden of rotavirus infection, especially with regard to hospitalisation (45 per 10,000 children ≤5 years of age) and admissions to short observation (85 per 10,000), generating benefits for the Italian healthcare system.

Noninvasive ventilation with positive airway pressure in paediatric intensive care.

AIM: The aim of the present study is to retrospectively evaluate the effectiveness of noninvasive pressure ventilation in the 24-bed Pediatric Intensive Care Unit (PICU) of the G. Gaslini Institute during a 24-month period. METHODS: A retrospective analysis of the characteristics (pH, CO2, SpO2, respiratory rate, oxygen requirement) of patients treated with noninvasive mechanical ventilation for different acute pathologies has been performed. RESULTS: Twenty patients (mean age 7.4+/-0.28 years) with acute respiratory failure due to different pathologies were treated with noninvasive mechanical ventilation. They were divided into 2 groups: the hypoxic group, suffering from pulmonary diseases, and the hypercapnic group, presenting a failure of the mechanical strength or increased dead space. Modalities of ventilation were pressure assisted/controlled or pressure support, delivered through nasal or facial masks. Fifteen out of 20 patients presented a marked improvement of oxygenation and ventilation. Mean times of treatment were 69 and 200 h in the hypoxic and hypercapnic groups, respectively. Five patients required intubation. Two patients presented reversible skin lesions over the nasal bridge. CONCLUSIONS: Noninvasive ventilation can be used in PICU. Major advantages regard immunocompromised children and patients with exacerbations from chronic respiratory diseases, whereas the exact role of noninvasive positive pressure ventilation in patients affected by acute respiratory distress syndrome is still controversial.

[Investigation of an outbreak of norovirus gastroenteritis in a geriatric hospital]. / Investigation d'une épidémie de gastro-entérite à norovirus dans un hôpital gériatrique.

In aged-care facilities, gastroenteritis outbreaks are responsible for big trouble in the management of cares to the elderly. In November 2002, a gastroenteritis outbreak was observed in 5 of the 6 wards of the geriatric hospital La Charité, University Hospital of Saint-Etienne, France, with an attack rate of 38.5% in the elderly (70 infected from 182 patients) and of 26.0% in the nursing staff (40 infected from 154 agents). The outbreak lasted 30 days with a peak corresponding to 79.8% of the cases between the 11(th) and the 20(th) of November. The first cases were observed in the two short-term-care wards; then, the outbreak spread rapidly to 3 of the 4 long-term care units. Health care workers were contaminated later than the elderly (P < 0.001 by Kruskal-Wallis test). A self-administered questionnaire was documented by most of the nursing staff; the most frequently observed clinical symptoms in this population were nausea (82.5%), abdominal pain (80.0%), diarrhoea (70.5%), asthenia (67.5%) and vomiting (62.5%). Thirty-five percent of the health care workers ceased their work. The causative agent of the gastroenteritis was identified by RT-PCR in the stools of 5 aged persons as a norovirus close to the Lordsdale strain (genogroup II). These findings illustrate the respective role of elderly and health care workers in the spread of the gastroenteritis outbreak inside the geriatric hospital.

[Outbreaks due to respiratory syncytial virus and influenzavirus A/H3N in institutionalized aged. Role of immunological status to influenza vaccine and possible implication of caregivers in the transmission]. / Infections groupées à virus respiratoire syncytial et à influenzavirus A/H3N2 chez des sujets âgés en institution. Influence du statut vaccinal anti-grippal et implication possible des soignants dans la transmission.

OBJECTIVES: Report of epidemiological, clinical and virological data collected from the prospective surveillance of febrile episodes observed in aged residents of a long-stay care unit of 33 beds, at the University Hospital of Saint-Etienne, during the 1997-1998 winter season. METHODS: Systematic collection of clinical and biological data from febrile patients (> or = 38 degrees C) on a form, including virological findings obtained from a nasal swab and paired serum specimens. RESULTS: From 38 patients (37 of them having been vaccinated against influenza in October 1997), 18 febrile episodes were recorded in 16 subjects, including 3 respiratory syncytial virus infections and a late-occurring outbreak (March 1998) of influenza due to a A/H3N2 strain (15 cases, 14 of them virologically confirmed). No death was noted after the influenza outbreak. In 8 of the 9 tested patients with influenza, "protective" titres of antibodies directed towards the hemagglutinin of the vaccinal strain were present by radial hemolysis test three months before the beginning of the outbreak. During the influenza outbreak, the attack rate of symptomatic infection was 45.5% in elderly and 47.5% in healthcare workers (mainly unvaccinated). The occurrence of the first cases in the latter suggests their possible role in the transmission of the virus to the aged. CONCLUSION: This study underlines the epidemic circulation of multiple respiratory viruses during the same winter season in long-stay care facilities, the occurrence of clinical influenza infections in vaccinated patients exhibiting protective antibody titres and the role of unvaccinated healthcare workers in the propagation of influenza in institutionalised aged.

Phenotype/genotype correlation and cystic fibrosis related diabetes mellitus (Italian Multicenter Study).

BACKGROUND: A genotype/phenotype correlation between early onset cystic fibrosis related diabetes (CFRD) and the N1303K mutation of the CF gene was previously identified in a small series of 28 CFRD patients, out of 313 CF patients. PATIENTS AND METHODS: In order to confirm the observation, data of 141 CFRD patients out of 1,229 CF patients attending 14 Italian CF centers were collected. All patients were older than 10 years and had been genotyped. RESULTS: DeltaF508 was the most frequent mutation (147/282 alleles: 52%) and N1303K the second most frequent mutation (18/282 alleles: 6.3%) in CFRD patients, without significant difference as compared with CF patients without DM (52% vs 48.6% and 6.3% vs 5.1%, respectively). W1282X was the third most frequent mutation in CFRD patients, more frequent than in CF patients without DM (5.3% vs 2%; p<0.001). CONCLUSIONS: Unlike the previous study, we did not find a higher frequency of the N1303K mutation in CFRD patients; moreover, data from this large CF series showed a significant correlation between the W1282X mutation and CFRD.

Genetic history of cystic fibrosis mutations in Italy. I. Regional distribution.

Earlier analysis of the Italian population showed patterns of genetic differentiation that were interpreted as being the result of population settlements going back to pre-Roman times. DNA disease mutations may be a powerful tool in further testing this hypothesis since the analysis of diseased individuals can detect variants too rare to be resolved in normal individuals. We present data on the relative frequencies of 60 cystic fibrosis (CF) mutations in Italy and the geographical distribution of the 12 most frequent CF mutations screened in 3492 CF chromosomes originating in 13 Italian regions. The 12 most frequent mutations characterize about 73% of the Italian CF chromosomes. The most common mutation, delta F508, has an average frequency of 51%, followed by N1303K and G542X, both with average frequencies around 5%. Multivariate analyses show that the relative frequencies of CF mutations are heterogeneous among Italian regions, and that this heterogeneity is weakly correlated with the geographical pattern of non-DNA 'classical' genetic markers. The northern regions are well differentiated from the central-southern regions and within the former group the western and eastern regions are remarkably distinct. Moreover, Sardinia shows the presence of mutation T338I, which seems absent in any other European CF chromosome. The north-western regions of Italy, characterized by the mutation 1717-1G-->A, were under Celtic influence, while the north-east regions, characterized by the mutations R1162X, 2183AA-->G and 711 + 5G-->A, were under the influence of the Venetic culture.

[Confusion syndromes in hospitalized aged patients: polymorphism of symptoms and course. Prospective study of 183 patients]. / Les syndromes confusionnels du sujet âgé hospitalisé: polymorphisme sémiologique et évolutif. Etude prospective de 183 patients.

Using explicit criteria contained in the DSM III R, we collected in a prospective cohort study clinical features, outcome and risk factors from two cohorts of delirium in hospitalized elderly patients: 138 hospitalized in geriatric department and 45 patients admitted to an acute and comprehensive care hospital. The clinical features were assessed using a quantitative scale (developed by Derouesné). Delirium was unrecognized or misdiagnosed by physicians in 34% of the cases. The onset was known only two thirds of cases. The incidence of hyperactive type, prolonged hospital stay, poor outcomes (persistent delirium leading up to dementia) were highest in subjects admitted in comprehensive hospital. The etiology of delirium is complex and multifactorial. An underlying cause was identified in 80% of patients. The length or the worsening of delirium was significantly higher in patients with psychiatric or dementia comorbidity (OR: 0.2; IC 95%: 0.1-0.5). The prognosis was better in patients without psychoactive medications (OR: 0.2; IC 95%: 0.1-0.4) or with metabolic abnormalities or acute diseases and disorders (OR: 3.3; IC 95%: 1.5-7.6). The predisposing factors to the development of dementia were prior use of psychoactive medications and signs of prior cognitive impairment. This article suggests delirium in elderly patients is associated with several outcomes. The prognosis should be improved at admission by specific scale and an evaluation of predisposing and precipitating factors.