Effect of induced ruminal acidosis on blood variables in heifers.

BACKGROUND: Ruminal acidosis is responsible for the onset of different pathologies in dairy and feedlot cattle, but there are major difficulties in the diagnosis. This study modelled the data obtained from various blood variables to identify those that could indicate the severity of ruminal acidosis. Six heifers were fed three experimental rations throughout three periods. The diets were characterised by different starch levels: high starch (HS), medium starch (MS) and low starch, as the control diet (CT). Ruminal pH values were continuously measured using wireless sensors and compared with pH measurements obtained by rumenocentesis. Blood samples were analysed for complete blood count, biochemical profile, venous blood gas, blood lipopolysaccharide (LPS) and LPS-binding proteins (LBP). RESULTS: The regression coefficient comparing the ruminal pH values, obtained using the two methods, was 0.56 (P = 0.040). Feeding the CT, MS and HS led to differences in the time spent below the 5.8, 5.5 and 5.0 pH thresholds and in several variables, including dry matter intake (7.7 vs. 6.9 vs. 5.1 kg/d; P = 0.002), ruminal nadir pH (5.69 vs. 5.47 vs. 5.44; P = 0.042), mean ruminal pH (6.50 vs. 6.34 vs. 6.31; P = 0.012), haemoglobin level (11.1 vs. 10.9 vs. 11.4 g/dL; P = 0.010), platelet count (506 vs. 481 vs. 601; P = 0.008), HCO3(-) (31.8 vs. 31.3 vs. 30.6 mmol/L; P = 0.071) and LBP (5.9 vs. 9.5 vs. 10.5 µg/mL; P < 0.001). A canonical discriminant analysis (CDA) was used to classify the animals into four ruminal pH classes (normal, risk of acidosis, subacute ruminal acidosis and acute ruminal acidosis) using haemoglobin, mean platelet volume, ß-hydroxybutyrate, glucose and reduced haemoglobin. CONCLUSIONS: Although additional studies are necessary to confirm the reliability of these discriminant functions, the use of plasma variables in a multifactorial model appeared to be useful for the evaluation of ruminal acidosis severity.

The effect of crate height on the behavior of female turkeys during commercial pre-slaughter transportation.

Limited information is available on suitable height of transport crates for turkeys. We compared behaviors and physiological indicators of four groups of 10 female turkeys each confined in either conventional (38.5 cm height) or experimental (77 cm height) crates during six commercial pre-slaughter transportations for 86 km (76 ± 4 min) along two tracts with one-lane streets, crossroads, bends, roundabouts (S1 and S2) and a highway tract (H) between S1 and S2. Only 36% of birds in the higher crates maintained a standing position. In conventional versus experimental crates, the frequency of rising attempts was five/bird/hour versus less than one/bird/hour, while wing flapping was seven/bird/hour versus 20/bird/hour, and balance loss was one versus four/bird/hour. The behaviors of both groups differed significantly according to the route tract, with a lower frequency of stress-related behaviors at H. No scratches, fractures or hematomas were detected in any birds after transportation. Crate height had no significant effect on hemato-biochemical markers. These results suggest that crates enabling a standing position may increase potentially dangerous behaviors. Moreover, busy and curvy routes should be avoided, as they may contribute to increasing the frequency of stress-related behaviors.

The degree of acceptability of swine blood values at increasing levels of hemolysis evaluated through visual inspection versus automated quantification.

The pronounced fragility that characterizes swine erythrocytes is likely to produce a variable degree of hemolysis during blood sampling, and the free hemoglobin may then unpredictably bias the quantification of several analytes. The aim of this study was to evaluate the degree of acceptability of values obtained for several biochemical parameters at different levels of hemolysis. Progressively increased degrees of physical hemolysis were induced in 3 aliquots of 30 nonhemolytic sera, and the relative effects on the test results were assessed. To define the level of hemolysis, we used both visual estimation (on a scale of 0 to 3+) and analytical assessment (hemolytic index) and identified the best analytical cutoff values for discriminating the visual levels of hemolysis. Hemolysis led to a variable and dose-dependent effect on the test results that was specific for each analyte tested. In mildly hemolyzed specimens, C-reactive protein, haptoglobin, ß1-globulin, ß2-globulin, α1-globulin, γ-globulin, sodium, calcium, and alkaline phosphatase were not significantly biased, whereas α2-globulin, albumin, urea, creatinine, glucose, total cholesterol, aspartate aminotransferase, alanine aminotransferase, gamma-glutamyl transferase, nonesterified fatty acids, bilirubin, phosphorus, magnesium, iron, zinc, copper, lipase, triglycerides, lactate dehydrogenase, unbound iron-binding capacity, and uric acid were significantly biased. Chloride and total protein were unbiased even in markedly hemolyzed samples. Analytical interference was hypothesized to be the main source of this bias, leading to a nonlinear trend that confirmed the difficulty in establishing reliable coefficients of correction for adjusting the test results.

Tail docking and the rearing of heavy pigs: the role played by gender and the presence of straw in the control of tail biting. Blood parameters, behaviour and skin lesions.

This study evaluated whether the specific heavy pig rearing context allowed the fattening of undocked pigs without an outbreak of tail biting. At the same time, gender and straw availability (small amounts) were considered to understand their possible interactions with tail presence in the display of tail biting. A 2 × 2 × 2 factorial design was adopted to test the effects of these factors on blood parameters, behaviour and tail/ear lesions. Few interactions among factors were detected. Undocked pigs showed lower cortisol (P<0.02), lying behaviour (P<0.001), and higher risk of tail/ear biting (weeks 3 and 9), but lower risk of tail lesions (week 14). Straw increased the motivation for exploring (P<0.001), reduced serum haptoglobin (P<0.001) and the risk for tail biting (weeks 3, 9, 18) and ear biting (weeks 3, 9). Results highlight the importance of straw as an environmental enrichment and seem to indicate that fattening undocked heavy pigs is possible.

Continuous straw provision reduces prevalence of oesophago-gastric ulcer in pigs slaughtered at 170 kg (heavy pigs).

Adopting a 2 × 2 × 2 factorial design, this study evaluated whether continuous straw provision by racks, tail docking and gender (barrows vs. females) have an effect on the prevalence of lung lesions and oesophago-gastric ulcer (OGU) visually scored at slaughter in 635 Italian heavy pigs (169 ± 4 kg). The lung lesions were very low (72% of pigs with score 0), and were not significantly different among the experimental groups. Overall, OGU was diagnosed in 47% of the pigs. The consumption of small amounts of straw (70 g/day/pig) represented a protective factor against the onset of OGU (OR: 0.27). Barrows were more likely than females to have OGU (OR: 1.52), while no significant differences between docked and undocked pigs were detected. Nevertheless, the presence of straw acted as a protective factor particularly in undocked pigs (OR: 0.16), suggesting that in this group the absence of rooting material may have a stronger effect on welfare.

Versatility of gene therapy vectors through viruses.

Several viruses have been engineered for gene therapy applications, and the specific properties of each viral vector have been exploited to target a variety of inherited and acquired diseases. Preclinical and clinical studies demonstrated that viral vectors are highly versatile tools capable of efficient transfer of foreign genetic information into almost all cell types and tissues. Gene therapy applications depend on vector characteristics, such as host range, cell- or tissue-specific targeting, genome integration, efficiency and duration of transgene expression, packaging capacity, and suitability for scale-up production. This review discusses the advances in the development of viral vectors, with particular emphasis on how knowledge of virus biology has been exploited to design a variety of vectors with improved safety characteristics and efficiency, potentially suitable for a large number of gene therapy applications.

Loss of growth hormone secretagogue receptor 1a and overexpression of type 1b receptor transcripts in human adrenocortical tumors.

OBJECTIVE AND METHODS: Quantitative analysis of mRNA expression of ghrelin and its receptors GHS-R1a and -R1b in a large series of normal and neoplastic human adrenocortical tissues. Evaluation of the effects of ghrelin on GHS-R expression and proliferation of human adrenocortical carcinoma (ACC) cell lines. RESULTS: Ghrelin and GHS-R transcripts are expressed in normal adrenal cortex, with GHS-R1b mRNA levels being 5- to 10-fold higher than GHS-R1amRNA. A significant increase in ghrelin expression was observed in adrenocortical adenomas, but not in carcinomas. GHS-R1a was undetectable in about 60% of both benign and malignant tumor samples, except for cortisol-producing adenomas, which showed increased receptor expression. At variance, GHS-R1b was overexpressed in both benign and malignant adrenocortical tumors. In vitro studies in human ACC cell lines demonstrated that GHS-R1a is downregulated and GHS-R1bmRNA expression is upregulated by ghrelin, while inhibiting cell proliferation. CONCLUSION: Downregulation ofGHS-R1a in adrenal tumors and the presence of high levels of GHS-R1b transcripts in adrenocortical tissue suggest a role for these receptors in adrenal function and growth. In this regard, ghrelin inhibits cell proliferation and modulates GHS-R expression in ACC cells in vitro.

Systemic efficacy of combined suicide/cytokine gene therapy in a murine model of hepatocellular carcinoma.

BACKGROUND/AIMS: Gene therapy might be a promising therapeutic approach for advanced hepatocellular carcinoma (HCC) not amenable to any effective traditional treatment. The aim of the study was to evaluate the in vitro and in vivo efficacy of combined gene therapy of HCC with two different MoMLV-derived retroviral vectors, an MFG- and a LXSN-derived vector, both containing HSV-TK and hIL-2. RESULTS: In vitro experiments on HCC cells showed efficient killing of transduced cells and efficient bystander effect after ganciclovir (GCV) treatment, with higher antitumor activity when the MFG-based vector was used. In vivo studies in a murine syngenic model of HCC demonstrated that treatment with GCV led to complete regression of tumors composed of transduced cells and regression of distant non-transduced tumors. Tumor transduction and efficacy of treatment was also demonstrated after in vivo delivery of vectors. Microarray analysis of tumor samples in mice receiving gene therapy showed up-regulation of genes involved in immune response and signal transduction. CONCLUSIONS: We demonstrated the in vitro and in vivo efficacy of combined retroviral-mediated gene therapy for HCC, with significant systemic therapeutic efficacy in vivo.

Retrovirus-mediated herpes simplex virus thymidine kinase gene transfer in pancreatic cancer cell lines: an incomplete antitumor effect.

INTRODUCTION: The transfer of drug-susceptible (suicide) genes to tumor cells by retroviral or adenoviral vectors is a novel approach to the treatment of human tumors. AIMS: To ascertain the antitumor effect of retroviral transduction of the pancreatic cancer cell lines MIA PaCa 2, CAPAN-1, PANC1, and PSN1 with the herpes simplex virus thymidine kinase (HSV-TK) gene. METHODOLOGY: The vector carried a neoselectable marker gene, the human interleukin-2 gene, an internal ribosome entry coding site, and the region coding HSV-TK. RESULTS: Twenty micromoles or less of ganciclovir did not modify nontransduced TK- cell growth, whereas > or =100 micromol completely inhibited TK- cell growth, indicating that this dosage is cytotoxic per se. The 4 TK- and the 4 transduced cell lines were treated daily with 0.001, 0.01, 0.1, 1, 10, and 20 micromol of ganciclovir for 13 days. CAPAN-1 cell growth was completely inhibited by 0.1 micromol of ganciclovir; higher doses were required to kill PANC1 (10 micromol) and PSN1 (20 micromol). MIA PaCa 2 cell growth decreased following a 20-micromol ganciclovir dosing. The bystander effect was great in the CAPAN-1 cell line and moderate in PANC1; no bystander effect was recorded in MIA PaCa 2 and PSN1 cell lines. CONCLUSION: Gene therapy with HSV-TK for pancreatic cancer seems effective in only a limited number of tumor-derived cell lines, and this limits its application in vivo.