A critical review of the mechanisms involved in the occurrence of growth-related abnormalities affecting broiler chicken breast muscles

In the past decade, the poultry industry has faced the occurrence of growth-related muscular abnormalities that mainly affect, with a high incidence rate, the Pectoralis major of the fast-growing genotypes selected for their production performances (high growth rate and breast yield). These myopathies are termed as White Striping, Wooden Breast, and Spaghetti Meat and exhibit distinctive phenotypes. A spatiotemporal distribution has been demonstrated for these disorders as in the early stage they primarily affect the superficial area in the cranial portion of the muscle and, as the birds grow older, involve the entire tissue. Aside from their distinctive phenotypes, these myopathies share common histological features. Thus, it might be speculated that common causative mechanisms might be responsible for the physiological and structural perturbations in the muscle associated with these conditions and might underpin their occurrence. The present review paper aims to represent a critical survey of the outcomes of all the histologic and ultrastructural observations carried out on White Striping, Wooden Breast, and Spaghetti Meat affected muscles. Our analysis has been performed by combining these outcomes with the findings of the genetic studies, trying to identify possible initial causative mechanisms triggering the onset and the time-series of the events ultimately resulting in the development and progression of the growth-related myopathies currently affecting broilers Pectoralis major muscles. Several evidences support the hypothesis that sarcoplasmic reticulum stress, primarily induced an accumulation of misfolded proteins (but also driven by other factors including altered calcium homeostasis and accumulation of fatty acids), may be responsible for the onset of these growth-related myopathies in broilers. At the same time, the development of hypoxic conditions, as a direct consequence of an inadequate vascularization, triggers a time-series sequence of events (i.e., phlebitis, oxidative stress, etc.) resulting in the activation of response mechanisms (i.e., modifications in the energetic metabolism, inflammation, degeneration, and regeneration) which are all strictly related to the progression of these myopathic disorders

Prevalence of alcohol and drugs in urine of patients involved in road accidents

Objective. Road injuries are the leading injury-related cause of death among people aged 15-44. A clear dose-effect relationship has been demonstrated for drug and alcohol use and road traffic accidents. The objective of our study was to estimate the prevalence of drug and/or alcohol use in subjects admitted for road traffic accidents to an Emergency Department. Methods. In this study, conducted between January and April 2006, 100 patients of both sexes (age 18-65) examined after road traffic accidents were consecutively enrolled. A commercial rapid urine test was used to detect drugs by the Emergency Department staff. The alcohol concentration was determined from a blood sample at the central laboratory. Results. Most of the patients were drivers under 35 years of age. 67/100 road traffic accidents occurred at the weekend (Friday- Sunday), nearly 60% between 24:00-09:00 hrs; on non-weekend days about 80% of road traffic accidents were recorded between 14:00-24:00 hrs (p inf. 0.0001). With the alcoholemia test and urine test for drugs detection 43/100 patients showed a single or multiple positivity. Alcohol and drug presence is relevant during the weekend (37/43 cases), in contrast with non weekend (6/43 cases) [OR 3.04 (95% CI 1.43; 6.46)]. Alcohol was the most frequently detected abuse substance (72%), followed by benzodiazepines (42%), tetrahydrocannabinol (21%) and cocaine (14%). Discussion. 43% of patients examined were under the influence of psychotropic substances (alcohol, drugs or both). The greater part of road traffic accidents in positive test patients occurred during the week-end, in particular during the late night/early morning hours, probably after recreational time. The high incidence of alcohol and/or drug abuse may have caused physical and/or psychological problems, therefore the high number of road traffic accidents, especially if taken in combination. Conclusion. The rapid urine test used cannot represent a diagnosis, and requires a confirmation test. It can be used for medical purposes as an easy and fast preliminary response which enables a faster diagnostic and therapeutic guideline, but it cannot be used for sanctions. Further studies are advisable with an increase of number of patients, in a wider temporal range, including control subjects, and using confirmation tests

Tailoring the CRISPR system to transactivate coagulation gene promoters in normal and mutated contexts

Engineered transcription factors (TF)have expanded our ability to modulate gene expression and hold great promise as bio-therapeutics. The first-generation TF, based on Zinc Fingers or Transcription-Activator-like Effectors (TALE), required complex and time-consuming assembly protocols, and were indeed replaced in recent years by the CRISPR activation (CRISPRa)technology. Here, with coagulation F7/F8 gene promoters as models, we exploited a CRISPRa system based on deactivated (d)Cas9, fused with a transcriptional activator (VPR), which is driven to its target by a single guide (sg)RNA. Reporter gene assays in hepatoma cells identified a sgRNA (sgRNA F7.5 )triggering a ~35-fold increase in the activity of F7 promoter, either wild-type, or defective due to the c.-61T>G mutation. The effect was higher (~15-fold)than that of an engineered TALE-TF (TF4)targeting the same promoter region. Noticeably, when challenged on the endogenous F7 gene, the dCas9-VPR/sgRNA F7.5 combination was more efficient (~6.5-fold)in promoting factor VII (FVII)protein secretion/activity than TF4 (~3.8-fold). The approach was translated to the promoter of F8, whose reduced expression causes hemophilia A. Reporter gene assays in hepatic and endothelial cells identified sgRNAs that, respectively, appreciably increased F8 promoter activity (sgRNA F8.1 , ~8-fold and 3-fold; sgRNA F8.2 , ~19-fold and 2-fold)with synergistic effects (~38-fold and 2.7-fold). Since modest increases in F7/F8 expression would ameliorate patients' phenotype, the CRISPRa-mediated transactivation extent might approach the low therapeutic threshold. Through this pioneer study we demonstrated that the CRISPRa system is easily tailorable to increase expression, or rescue disease-causing mutations, of different promoters, with potential intriguing implications for human disease models

Factors affecting the development of Bovine Respiratory Disease: a cross-sectional study in beef steers shipped from France to Italy

Bovine respiratory disease (BRD) is a complex, multifactorial syndrome and one of the major welfare and economical concerns for the cattle industry. This 1-year cross-sectional study was aimed at documenting the prevalence of BRD-related pathogens and clinical signs before and after a long journey and at identifying possible predisposition factors. Male Limousine beef steers (n = 169) traveling from France to Italy were health checked and sampled with Deep Nasopharyngeal Swabs (DNS) at loading (T0) and 4 days after arrival (T1). Real-time quantitative PCR was used to quantify the presence of bovine viral diarrhea virus (BVDV), bovine respiratory syncytial virus (BRSV), bovine alphaherpesvirus 1 (BoHV-1), bovine coronavirus (BCoV), bovine adenovirus (BAdV), bovine parainfluenza virus 3 (BPIV-3), Histophilus somni, Mannheimia haemolytica, Mycoplasma bovis, and Pasteurella multocida. Weather conditions at departure and arrival were recorded, and the travel conditions were taken from the travel documentation. At T0, even if no animals displayed clinical signs, some of them were already positive for one or more pathogens. At T1, the number of animals displaying clinical signs and positive for BCoV, BAdV, BRSV, H. somni, M. haemolytica, M. bovis, and P. multocida increased dramatically (p < 0.001). Transport also significantly increased co-infection passing from 16.0% at T0 to 82.8% at T1 (p < 0.001). An extra stop during the journey seemed to favor BRSV, M. haemolytica, and P. multocida (p < 0.05). Weather conditions, in particular sudden climate changes from departure to arrival and daily temperature variance, were found to be predisposing factors for many of the pathogens. The farm of arrival also played a role for BRSV, BAdV, and H. somni (p < 0.05). BCoV increased dramatically, but no associations were found confirming that it spreads easily during transport phases. Our findings increased our understanding of factors increasing the likelihood of BRD-related pathogens shedding and can be useful to minimize the incidence of BRD and to implement animal transport regulations

The Use of Infrared Thermography (IRT) as Stress Indicator in Horses Trained for Endurance: A Pilot Study

The aim of this pilot study was to document the effects of endurance training at different intensities on heart rate (HR), blood count, serum cortisol, and maximal temperatures of different body locations, namely eye, crown, pastern pasterns, gluteus and longissimus dorsi muscle (mm), measured by infrared thermography technique (IRT) in horses trained for endurance. Possible associations among the studied parameters were also investigated. Our hypothesis was that temperature, measured by IRT after endurance training of different intensities would vary depending on the intensity and would be positively correlated with HR and serum cortisol. Eight horses were tested before and after training of different intensities (low, moderate, and high). The results partially supported our hypothesis; all the studied parameters increased after training (p < 0.05), eye temperature (ET) correlated positively with HR (p < 0.01), and crown temperature (CT) correlated positively with cortisol (p < 0.01). However, only HR and white blood cells increased with the intensity of the exercise (p = 0.0016 and p = 0.0142, respectively). Our findings suggest the evaluation of ET and CT may become a useful non-invasive tool to detect physiological stress during training and to evaluate how the horses cope with the training. Infrared thermography technique may also become a useful tool for the early identification of horses that are not fit to compete or to continue the competition. However, further studies should be conducted on a larger number of horses and during competitions to ascertain our preliminary findings

A Cretaceous carbonate delta drift in the Montagna della Maiella, Italy

The Upper Cretaceous (Campanian\u2013Maastrichtian) bioclastic wedge of the Orfento Formation in the Montagna della Maiella, Italy, is compared to newly discovered contourite drifts in the Maldives. Like the drift deposits in the Maldives, the Orfento Formation fills a channel and builds a Miocene delta-shaped and mounded sedimentary body in the basin that is similar in size to the approximately 350 km 2 large coarse-grained bioclastic Miocene delta drifts in the Maldives. The composition of the bioclastic wedge of the Orfento Formation is also exclusively bioclastic debris sourced from the shallow-water areas and reworked clasts of the Orfento Formation itself. In the near mud-free succession, age-diagnostic fossils are sparse. The depositional textures vary from wackestone to float-rudstone and breccia/conglomerates, but rocks with grainstone and rudstone textures are the most common facies. In the channel, lensoid convex-upward breccias, cross-cutting channelized beds and thick grainstone lobes with abundant scours indicate alternating erosion and deposition from a high-energy current. In the basin, the mounded sedimentary body contains lobes with a divergent progradational geometry. The lobes are built by decametre thick composite megabeds consisting of sigmoidal clinoforms that typically have a channelized topset, a grainy foreset and a fine-grained bottomset with abundant irregular angular clasts. Up to 30 m thick channels filled with intraformational breccias and coarse grainstones pinch out downslope between the megabeds. In the distal portion of the wedge, stacked grainstone beds with foresets and reworked intraclasts document continuous sediment reworking and migration. The bioclastic wedge of the Orfento Formation has been variously interpreted as a succession of sea-level controlled slope deposits, a shoaling shoreface complex, or a carbonate tidal delta. Current-controlled delta drifts in the Maldives, however, offer a new interpretation because of their similarity in architecture and composition. These similarities include: (i) a feeder channel opening into the basin; (ii) an excavation moat at the exit of the channel; (iii) an overall mounded geometry with an apex that is in shallower water depth than the source channel; (iv) progradation of stacked lobes; (v) channels that pinch out in a basinward direction; and (vi) smaller channelized intervals that are arranged in a radial pattern. As a result, the Upper Cretaceous (Campanian\u2013Maastrichtian) bioclastic wedge of the Orfento Formation in the Montagna della Maiella, Italy, is here interpreted as a carbonate delta drift

U and Th content in the Central Apennines continental crust: a contribution to the determination of the geo-neutrinos flux at LNGS

The regional contribution to the geo-neutrino signal at Gran Sasso National Laboratory (LNGS) was determined based on a detailed geological, geochemical and geophysical study of the region. U and Th abundances of more than 50 samples representative of the main lithotypes belonging to the Mesozoic and Cenozoic sedimentary cover were analyzed. Sedimentary rocks were grouped into four main "Reservoirs" based on similar paleogeographic conditions and mineralogy. Basement rocks do not outcrop in the area. Thus U and Th in the Upper and Lower Crust of Valsugana and Ivrea-Verbano areas were analyzed. Based on geological and geophysical properties, relative abundances of the various reservoirs were calculated and used to obtain the weighted U and Th abundances for each of the three geological layers (Sedimentary Cover, Upper and Lower Crust). Using the available seismic profile as well as the stratigraphic records from a number of exploration wells, a 3D modelling was developed over an area of 2^{\circ}x2^{\circ} down to the Moho depth, for a total volume of about 1.2x10^6 km^3. This model allowed us to determine the volume of the various geological layers and eventually integrate the Th and U contents of the whole crust beneath LNGS. On this base the local contribution to the geo-neutrino flux (S) was calculated and added to the contribution given by the rest of the world, yielding a Refined Reference Model prediction for the geo-neutrino signal in the Borexino detector at LNGS: S(U) = (28.7 \pm 3.9) TNU and S(Th) = (7.5 \pm 1.0) TNU. An excess over the total flux of about 4 TNU was previously obtained by Mantovani et al. (2004) who calculated, based on general worldwide assumptions, a signal of 40.5 TNU. The considerable thickness of the sedimentary rocks, almost predominantly represented by U- and Th- poor carbonatic rocks in the area near LNGS, is responsible for this difference.Comment: 45 pages, 5 figures, 12 tables; accepted for publication in GC

Proteomic analysis of urine in medication-overuse headache patients: possible relation with renal damages

Medication-overuse headache (MOH) is a chronic disorder associated with overuse of analgesic drugs, triptans, non-steroidal anti-inflammatory drugs (NSAIDs) or other acute headache compounds. Various epidemiologic investigations proved that different drug types could cause nephrotoxicity, particularly in chronic patients. The aim of the present work was to analyze, by a proteomic approach, the urinary protein profiles of MOH patients focusing on daily use of NSAIDs, mixtures and triptans that could reasonably be related to potential renal damage. We selected 43 MOH patients overusing triptans (n = 18), NSAIDs (n = 11), and mixtures (n = 14), for 2–30 years with a mean daily analgesic intake of 1.5 ± 0.9 doses, and a control group composed of 16 healthy volunteers. Urine proteins were analyzed by mono-dimensional gel electrophoresis and identified by mass spectrometry analysis. Comparing the proteomic profiles of patients and controls, we found a significantly different protein expression, especially in the NSAIDs group, in which seven proteins resulted over-secreted from kidney (OR = 49, 95% CI 2.53–948.67 vs. controls; OR = 11.6, 95% CI 0.92–147.57 vs. triptans and mixtures groups). Six of these proteins (uromodulin, α-1-microglobulin, zinc-α-2-glycoprotein, cystatin C, Ig-kappa-chain, and inter-α-trypsin heavy chain H4) were strongly correlated with various forms of kidney disorders. Otherwise, in mixtures and in triptans abusers, only three proteins were potentially associated to pathological conditions (OR = 4.2, 95% CI 0.33–53.12, vs. controls). In conclusion, this preliminary proteomic study allowed us to define the urinary protein pattern of MOH patients that is related to the abused drug. According with the obtained results, we believe that the risk of nephrotoxicity should be considered particularly in MOH patients who abuse of NSAIDs

The morphology and biochemistry of nanostructures provide evidence for synthesis and signaling functions in human cerebrospinal fluid

<p>Abstract</p> <p>Background</p> <p>Cerebrospinal fluid (CSF) contacts many brain regions and may mediate humoral signaling distinct from synaptic neurotransmission. However, synthesis and transport mechanisms for such signaling are not defined. The purpose of this study was to investigate whether human CSF contains discrete structures that may enable the regulation of humoral transmission.</p> <p>Methods</p> <p>Lumbar CSF was collected prospectively from 17 participants: with no neurological or psychiatric disease, with Alzheimer's disease, multiple sclerosis, or migraine; and ventricular CSF from two cognitively healthy participants with long-standing shunts for congenital hydrocephalus. Cell-free CSF was subjected to ultracentrifugation to yield supernatants and pellets that were examined by transmission electron microscopy, shotgun protein sequencing, electrophoresis, western blotting, lipid analysis, enzymatic activity assay, and immuno-electron microscopy.</p> <p>Results</p> <p>Over 3,600 CSF proteins were identified from repeated shotgun sequencing of cell-free CSF from two individuals with Alzheimer's disease: 25% of these proteins are normally present in membranes. Abundant nanometer-scaled structures were observed in ultracentrifuged pellets of CSF from all 16 participants examined. The most common structures included synaptic vesicle and exosome components in 30-200 nm spheres and irregular blobs. Much less abundant nanostructures were present that derived from cellular debris. Nanostructure fractions had a unique composition compared to CSF supernatant, richer in omega-3 and phosphoinositide lipids, active prostanoid enzymes, and fibronectin.</p> <p>Conclusion</p> <p>Unique morphology and biochemistry features of abundant and discrete membrane-bound CSF nanostructures are described. Prostaglandin H synthase activity, essential for prostanoid production and previously unknown in CSF, is localized to nanospheres. Considering CSF bulk flow and its circulatory dynamics, we propose that these nanostructures provide signaling mechanisms <it>via </it>volume transmission within the nervous system that are for slower, more diffuse, and of longer duration than synaptic transmission.</p