Monoclonal antibodies against human CD34 antigens do not cross-react with ovine umbilical cord blood cells

CD34 is a cell surface glycoprotein expressed by hematopoietic progenitors and endothelial cells. It is widely used in the clinic for isolation of human hematopoietic stem cells. In recent years large animals are gaining increasing importance in biomedical research for the study and therapy of human diseases. Sheep has proved to be an useful experimental model for preclinical trials in transplantation procedures. Unfortunately, the lack of specie-specific monoclonal antibodies (MABS) recognizing hemopoietic progenitor cells hampers the use of this animal in experimental hematology. The aim of this paper was to determine whether commercial monoclonal antibodies specific for human CD34 molecule could cross-react with hematopoietic progenitor cells (HPC) present in sheep umbilical cord blood (UCB). Six anti-human CD34 MABS, recognizing the three different epitope classes, were tested in flow cytometry on purified mononuclear cells (MNC) isolated from cord blood of both species. None of the MABS used in this trial seemed to be able to identify HPC from sheep UCB. These data suggest that the panel of monoclonal antibodies used for cross reactivity detection has to be expanded with recently produced reagents. Further studies should be directed towards the production of ovine specific anti CD34 MABS

Use of a glucomannan polymer to reduce the effects of mycotoxin-contaminated diets in finishing pigs

The use of feed additives with mycotoxin adsorption capacity is a common strategy for controlling negative effects of mycotoxins in swine production systems. However, adsorbents that may results very effective under experimental conditions, i.e. when feed contamination level is rather high, do not necessarily retain their efficacy when tested under field conditions feed with generally low mycotoxin contamination. In this study, the effects of diets artificially contaminated with aflatoxin B1 or ochratoxin A on fattening performance and serum chemistry of fattening pigs are investigated. Moreover, the ability of a commercial glucomannan polymer (Gm polimer) to reduce or eliminate the effects of the contaminated feeds is tested. Thirty heavy pigs (BW = 110±10.6 kg) were fed 6 diets (n = 5 pigs/diet) for 4 weeks until slaughtering. Diets were: control without toxin added (C); added with 0.02 ppm of aflatoxin B1 (AFB1); added with 0.05 ppm of ochratoxin A (OTA); other three diets as the previous but the addition of 2.0 g/kg of Gm polymer (C-GM, AFB1-GM, OTA-GM). Daily weight gain (ADG) and Feed efficiency ratio (FE) were measured every two weeks. Data were analyzed with a two-way ANOVA that included the fixed effect of diet, time and their interaction. After the first 2 weeks the ADG did not differ significantly between the diets, even if the ADG of AFB1 diet was about 20% lower than AFB1-Gm or C. In the last 2 weeks the ADG of AFB1 diet was significantly lover than the other diets (P<0.01) and was about one-half of the values reported for the same group in the first period. The contamination with ochratoxin A did not affect fattening performance of pigs during the whole experimental period. No damages were found in kidneys of all diets. Moreover, no evidence of association between observed liver damages and different diets was found. Finally, no differences between experimental diets were evidenced for the haematological parameters

Stress responses in lambs castrated with three different methods

The present work was conducted to evaluate the animal response to stress in lambs caused by three different castration techniques. Forty-six male lambs aged 4-5 months were randomly allocated to one of four groups including Burdizzo (B), scrotal ablation (SA), orchiectomy (OR) and control handling (H). Local anaesthesia (lidocaine 2%) was administered in both spermatic cords and the scrotal neck of lambs before each treatment. Blood samples were collected at -30, -10, +1, +20, +40, +60, +120, and +180 minutes. Serum cortisol concentrations were determined using a competitive immunoassay and the area under the curve (AUC) was calculated for each lamb. The following biochemical parameters were assayed for each animal at each time point: alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), creatine kinase (CK) and glucose (GLU). The time needed for total lesion resolution and weight gain of each animal was recorded. Orchiectomy elicits the greatest cortisol response, significantly greater than that seen in similarly handled controls (P≤0.01), Burdizzo and scrotal ablation groups (P≤0.05). The serum cortisol AUC was higher in the scrotal ablation group (P≤0.05) than controls, but lower than in the orchiectomy group (P≤0.05). The Burdizzo group didn’t differ from controls. Serum glucose levels of the castrated lambs differed significantly from the control group, following a trend similar to cortisol. No change was seen in ALT, AST, LDH or CK. No difference in weight gain was seen among the groups. Our results suggest that use of the Burdizzo is the preferable castration technique for adult lambs, while scrotal ablation is a valid surgical alternative to orchiectomy and permits more rapid wound healing that is ideal for extensive management where flocks are not under close observation

Machine milking management and milk nitrogen fractions in primiparous ewes

Little work has been carried out on management techniques to reduce stress of first machine milking in ewes. The aim of this work was to evaluate the effect of milking parlour training before weaning on sheep milk yield and milk protein fractions. One week before weaning, an experimental group (EG) was introduced into the machine milking parlour and hand-milked once a day, to adapt them to the new environment and noise of the milking machine, while a control group (CG) was hand-milked once a day in a traditional fold parlour until weaning. After weaning, both groups were machine milked, separately, twice a day in the milking parlour. Milk yield and nitrogen fractions were recorded for the first 10 days of machine milking. Differences between samplings within group were observed for all parameters considered. Only on the first day of machine milking, differences between groups occurred for total nitrogen, total protein and casein. Higher content of nitrogen fractions in the milk of EG was due to its lower milk production. In fact, daily production of nitrogen fractions was higher in CG than in EG. High variability for most parameters was observed during the first 5 days of machine milking in both groups. A week of training to machine milking parlour was not enough to reduce the negative effects of weaning and machine milking stresses on primiparous ewes. (interaction?

Sostanze indesiderate negli alimenti zootecnici: effetti sugli ovini e riflessi nel latte e derivati

Negli ovini da latte l’ingestione di alimenti contaminati da sostanza indesiderate rappresenta un rischio sia per animali al pascolo che per quelli confinati. L’assunzione di composti tossici negli ovini da latte comporta effetti negativi a carico dell’animale oltre che costituire un potenziale elemento di rischio per il consumatore finale nel caso in cui il composto tossico permanga nel latte o nelle carni. I metalli pesanti, le diossine e le micotossine rappresentano le sostanze indesiderate negli alimenti per animali di maggiore interesse per la filiera di produzione degli ovini da latte. La presenza di residui di contaminanti nel latte ovino e nei derivati caseari costituisce un importante indicatore della igienicità della filiera produttiva e allo stesso tempo è ritenuto un marcatore indiretto della salubrità dell’ambiente di produzione. Questo lavoro riassume gli aspetti di maggiore rilievo circa gli effetti dei metalli pesanti, delle diossine e di alcune micotossine sugli ovini, con particolare riferimento al loro trasferimento nel latte e nei derivati

Value at Risk and Expected Shortfall based on Gram-Charlier-like expansions

This paper offers a new approach to modeling the distribution of a portfolio composed of either asset returns or insurance losses. To capture the leptokurtosis, which is inherent in most financial series, data are modeled by using Gram-Charlier (GC) expansions. Since we are interested in operating with several series simultaneously, the distribution of the sum of GC random variables is derived. This latter turns out to be a tail-sensitive density, suitable for modeling the distribution of a portfolio return-losses and, accordingly, can be conveniently adopted for computing risk measures such as the value at risk and the expected shortfall as well as some performance measures based on its partial moments. The closed form expressions of these risk measures are derived for cases when the density of a portfolio is the sum of GC expansions, either with the same or different kurtosis. An empirical application of this approach to a portfolio of financial asset indexes provides evidence of the comparative effectiveness of this technique in computing risk measures, both in and out of the sample period

Functionalized multiwalled carbon nanotubes as ultrasound contrast agents

Ultrasonography is a fundamental diagnostic imaging tool in everyday clinical practice. Here, we are unique in describing the use of functionalized multiwalled carbon nanotubes (MWCNTs) as hyperechogenic material, suggesting their potential application as ultrasound contrast agents. Initially, we carried out a thorough investigation to assess the echogenic property of the nanotubes in vitro. We demonstrated their long-lasting ultrasound contrast properties. We also showed that ultrasound signal of functionalized MWCNTs is higher than graphene oxide, pristine MWCNTs, and functionalized single-walled CNTs. Qualitatively, the ultrasound signal of CNTs was equal to that of sulfur hexafluoride (SonoVue), a commercially available contrast agent. Then, we found that MWCNTs were highly echogenic in liver and heart through ex vivo experiments using pig as an animal model. In contrast to the majority of ultrasound contrast agents, we observed in a phantom bladder that the tubes can be visualized within a wide variety of frequencies (i.e., 5.5–10 MHz) and 12.5 MHz using tissue harmonic imaging modality. Finally, we demonstrated in vivo in the pig bladder that MWCNTs can be observed at low frequencies, which are appropriate for abdominal organs. Importantly, we did not report any toxicity of CNTs after 7 d from the injection by animal autopsy, organ histology and immunostaining, blood count, and chemical profile. Our results reveal the enormous potential of CNTs as ultrasound contrast agents, giving support for their future applications as theranostic nanoparticles, combining diagnostic and therapeutic modalities

Effect of extruded linseed supplementation on blood metabolic profile and milk performance of Saanen goats

This study assessed the effects of dietary supplementation with extruded linseed on milk yield and composition, milk fatty acid (FA) profile and renal and hepatic metabolism of grazing goats in mid-lactation. Forty Saanen goats were divided into two isoproductive groups: one group was fed the control diet (CON) composed of hay and pelleted concentrate and the other group was supplemented with additional 180 g/day of extruded linseed (LIN; dry matter basis), which supplied 70 g/day of fat per head for 9 weeks. Animals grazed on pasture for ∼3 h/day after the first of the 2 daily milkings. Milk samples were collected weekly and analyzed for fat, protein, lactose, milk urea nitrogen (MUN) and somatic cell count. Blood samples were collected every 2 weeks and analyzed for total bilirubin, creatinine, aspartate transaminase (AST), alanine transaminase (ALT), gamma glutamyl transpeptidase, alkaline phosphatase, total protein and urea nitrogen. Milk yield was higher in the LIN than in the CON group (2369 v. 2052 g/day). LIN group had higher milk fat (37.7 v. 33.4 g/kg) and protein (30.7 v. 29.1 g/kg) concentration and lower MUN (35.0 v. 43.3 mg/dl) than CON group. Goats fed LIN had greater proportions of 18:1 trans11, 18:2 cis9trans11 and total polyunsatured fatty acids n-3 in milk fat, because of higher 18:3n-3 and 20:5n-3 FA, and lower proportions of short- and medium-chain FAs than goats fed CON. All kidney and liver function biomarkers in serum did not differ between dietary groups, except for AST and ALT, which tended to differ. Extruded linseed supplementation to grazing mid-lactating goats for 2 months can enhance the milk performance and nutritional profile of milk lipids, without altering the general hepatic and renal metabolism

Characterization of the interaction of African swine fever virus with monocytes and derived macrophage subsets.

Abstract African swine fever (ASF) is a devastating disease for which there is no vaccine available. The ASF virus (ASFV) primarily infects cells of the myeloid lineage and this tropism is thought to be crucial for disease pathogenesis. A detailed in vitro characterization of the interactions of a virulent Sardinian isolate (22653/14) and a tissue culture adapted avirulent strain (BA71V) of ASFV with porcine monocytes, un-activated (moMΦ), classically (moM1) and alternatively (moM2) activated monocyte-derived macrophages was conducted in an attempt to better understand this relationship. Using a multiplicity-of-infection (MOI) of 1, both viruses were able to infect monocytes and macrophage subsets, but BA71V presented a reduced ability to infect moM1 compared to 22653/14, with higher expression of early compared to late proteins. Using an MOI of 0.01, only 22653/14 was able to replicate in all the macrophage subsets, with initially lowest in moM1 and moM2. No differences were observed in the expression of CD163 between ASFV infected and uninfected bystander cells. ASFV down-regulated CD16 expression but did not modulate MHC class II levels in monocytes and macrophage subsets. BA71V-infected but not 22653/14-infected moMΦ and moM2 presented with a reduced expression of MHC class I compared to the mock-infected controls. Higher levels of IL-18, IL1-β and IL-1α were released from moM1 after infection with BA71V compared to 22653/14 or mock-infected control. These results revealed differences between these ASFV strains, suggesting that virulent isolates have evolved mechanisms to counteract activated macrophages responses, promoting their survival, dissemination in the host and so ASF pathogenesis

A simplified genomic profiling approach predicts outcome in metastatic colorectal cancer

The response of metastatic colorectal cancer (mCRC) to the first-line conventional combination therapy is highly variable, reflecting the elevated heterogeneity of the disease. The genetic alterations underlying this heterogeneity have been thoroughly characterized through omic approaches requiring elevated efforts and costs. In order to translate the knowledge of CRC molecular heterogeneity into a practical clinical approach, we utilized a simplified Next Generation Sequencing (NGS) based platform to screen a cohort of 77 patients treated with first-line conventional therapy. Samples were sequenced using a panel of hotspots and targeted regions of 22 genes commonly involved in CRC. This revealed 51 patients carrying actionable gene mutations, 22 of which carried druggable alterations. These mutations were frequently associated with additional genetic alterations. To take into account this molecular complexity and assisted by an unbiased bioinformatic analysis, we defined three subgroups of patients carrying distinct molecular patterns. We demonstrated these three molecular subgroups are associated with a different response to first-line conventional combination therapies. The best outcome was achieved in patients exclusively carrying mutations on TP53 and/or RAS genes. By contrast, in patients carrying mutations in any of the other genes, alone or associated with mutations of TP53/RAS, the expected response is much worse compared to patients with exclusive TP53/RAS mutations. Additionally, our data indicate that the standard approach has limited efficacy in patients without any mutations in the genes included in the panel. In conclusion, we identified a reliable and easy-to-use approach for a simplified molecular-based stratification of mCRC patients that predicts the efficacy of the first-line conventional combination therapy