Effects of diets containing grape seed, linseed, or both on milk production traits, liver and kidney activities, and immunity of lactating dairy ewes.

This study aimed to evaluate the effects of the dietary inclusion of grape seed, alone or in combination with linseed, on milk production traits, immune response, and liver and kidney metabolic activity of lactating ewes. Twenty-four Sarda dairy ewes were randomly assigned to 4 dietary treatments consisting of a control diet (CON), a diet containing 300 g/d per head of grape seed (GS), a diet containing 220 g/d per head of extruded linseed (LIN), and a diet containing a mix of 300 g/d per head of grape seed and 220 g/d per head of extruded linseed (MIX). The study lasted 10 wk, with 2 wk of adaptation period and 8 wk of experimental period. Milk yield was measured and samples were collected weekly and analyzed for fat, protein, casein, lactose, pH, milk urea nitrogen, and somatic cell count. Blood samples were collected every 2 wk by jugular vein puncture and analyzed for hematological parameters, for albumin, alkaline phosphatase, bilirubin, creatinine, gamma glutamyltransferase, aspartate aminotransferase, alanine aminotransferase, protein, blood urea nitrogen, and for anti-albumin IgG, IL-6, and lymphocyte T-helper (CD4(+)) and lymphocyte T-cytotoxic (CD8(+)) cells. On d 0, 45, and 60 of the trial, lymphocyte response to phytohemagglutinin was determined in vivo on each animal by measuring skin-fold thickness (SFT) at the site of phytohemagglutinin injection. Humoral response to chicken egg albumin was stimulated by a subcutaneous injection with albumin. Dietary treatments did not affect milk yield and composition. Milk urea nitrogen and lactose were affected by diet × period. Diets did not influence hematological, kidney, and liver parameters, except for blood urea nitrogen, which decreased in LIN and increased in MIX compared with CON and GS. Dietary treatments did not alter CD4(+), CD8(+), and CD4(+)-to-CD8(+) ratio. The SFT was reduced in GS and MIX and increased in LIN compared with CON. The IgG and IL-6 were affected by diet × period. The reduction in IgG on d 60 and SFT in ewes fed GS suggests an immunomodulatory effect of this residue. The limited variation in milk and hematological and metabolic parameters suggests that GS and LIN can be included, alone or in combination, in the diet of dairy ewes without adverse effects on milk production and health status.

Changes in the gene expression profile of gastric cancer cells in response to ibuprofen: a gene pathway analysis.

Nonsteroidal anti-inflammatory drugs possess antiproliferative activities that can affect cancer cells. The aim of this study was to examine the antiproliferative effects of ibuprofen on the MKN-45 cell line. Cells were treated with ibuprofen for 24, 48 or 72 h, and cell proliferation was evaluated by cell counting and [(3)H]-thymidine incorporation. Using microarray technology, we studied changes in the gene expression profiles over time after ibuprofen treatment. Ibuprofen induced a dose- and time-dependent reduction in cell number without altering cell viability. Genes involved in the 'biological oxidation' and 'G(1)/S checkpoint' pathways were the most significantly represented at 24 h, whereas genes involved in the 'cell cycle' and 'DNA replication' pathways were represented at 48 and 72 h. Genes associated with the 'apoptosis' pathway were also significantly represented at 72 h. Modulation of the expression of p53 and p53-induced genes (CDKN1A/p21 and GADD45), which are involved in the G(1)/S transition, suggested an effect of ibuprofen on cell-cycle progression. Using flow cytometry, we observed an early block in the G(1) phase of the cell cycle after ibuprofen treatment. In addition, P450 family transcripts were upregulated and intracellular reactive oxygen species (ROS) was increased following 12 h of ibuprofen treatment. Ibuprofen induced ROS, which resulted in cellular alterations that promoted a p53-dependent G(1) blockade. These findings suggest that ibuprofen exerts its antiproliferative actions through cell-cycle control and the induction of apoptosis. Both of these mechanisms appear to be independent of ibuprofen's anti-inflammatory effects.

Basic butylated methacrylate copolymer/kappa-carrageenan interpolyelectrolyte complex: preparation, characterization and drug release behaviour.

The formation of a novel interpolyelectrolyte complex (IPEC) between basic butylated methacrylate copolymer and kappa-carrageenan was investigated and the product formed was characterized. Turbidity measurements and elemental analyses pointed to a 1:1 interaction of the repeating units. These results and FT-IR confirmed IPEC formation. Electronic microscopy images, particle size determination by image analysis and N(2) (77K) adsorption measurements were consistent with a porous material. This IPEC formed presented very good flowability and compactibility. Two maxima were observed in the swelling behaviour as a function of pH. The performance of the IPEC as a matrix for controlled release of drugs was evaluated, using ibuprofen as a model drug. Release profiles were properly represented by a mathematical model, which indicates that the system releases ibuprofen in a zero-order manner. These profiles could be controlled by conveniently modifying the proportion of the IPEC in the tablets.

tBid induces alterations of mitochondrial fatty acid oxidation flux by malonyl-CoA-independent inhibition of carnitine palmitoyltransferase-1.

Recent studies suggest a close relationship between cell metabolism and apoptosis. We have evaluated changes in lipid metabolism on permeabilized hepatocytes treated with truncated Bid (tBid) in the presence of caspase inhibitors and exogenous cytochrome c. The measurement of beta-oxidation flux by labeled palmitate demonstrates that tBid inhibits beta-oxidation, thereby resulting in the accumulation of palmitoyl-coenzyme A (CoA) and depletion of acetyl-carnitine and acylcarnitines, which is pathognomonic for inhibition of carnitine palmitoyltransferase-1 (CPT-1). We also show that tBid decreases CPT-1 activity by a mechanism independent of both malonyl-CoA, the key inhibitory molecule of CPT-1, and Bak and/or Bax, but dependent on cardiolipin decrease. Overexpression of Bcl-2, which is able to interact with CPT-1, counteracts the effects exerted by tBid on beta-oxidation. The unexpected role of tBid in the regulation of lipid beta-oxidation suggests a model in which tBid-induced metabolic decline leads to the accumulation of toxic lipid metabolites such as palmitoyl-CoA, which might become participants in the apoptotic pathway.

PROPAT: a study to improve the quality and reduce the cost of diabetes care.

OBJECTIVE: In PROPAT we implemented an integrated approach to diabetes care designed to improve the quality and reduce the cost of care. STUDY DESIGN AND METHODS: PROPAT was a case-control study matching patients by age and gender (diabetes:control ratio 1:2) within IOMA, a public employment-based health maintenance organization (HMO) of the Province of Buenos Aires, Argentina. Costs were evaluated using prevalence data from an HMO perspective. We currently report clinical and biochemical data and costs from the first 297 patients enrolled who completed 1 year in PROPAT, and compare them with those derived from control patients. RESULTS: All recommended practices recorded as care provided at baseline increased significantly 1 year after implementing PROPAT, with a parallel significant improvement in several clinical and biochemical parameters, and markedly lower total annual per capita costs. CONCLUSIONS: These results demonstrate that the implementation of a comprehensive diabetes care program can simultaneously improve quality while reducing costs.

Nitrate uptake improvement by modified activated carbons developed from two species of pine cones.

Activated carbons from two species of pine cones (Pinus canariensis and Cupressus sempervirens) were prepared by phosphoric acid activation and tested for the removal of nitrate ions from aqueous solution. To investigate the feasibility of improving their nitrate adsorption capacity, two different post-treatments­a thermal treatment and a treatment with saturated urea solution­were also applied to the prepared activated carbons. Comparison of the treated and untreated activated carbons showed that both post-treatments improved the nitrate adsorption performance more than twice. The maximum adsorption capacity, as evaluated from determination of the adsorption isotherms for the P. canariensis based carbons, and their proper representation by the Langmuir model, demonstrated that the post-treatment with the urea solution led to activated carbons with increased nitrate removal effectiveness, even superior to other reported results. Enhancements in their adsorption capacity could be mainly ascribed to higher contents of nitrogen and basic functional groups, whereas porous structure of the activated carbons did not seem to play a key role in the nitrate uptake.

Development and characterization of activated hydrochars from orange peels as potential adsorbents for emerging organic contaminants.

Activated hydrochars obtained from the hydrothermal carbonization of orange peels (Citrus sinensis) followed by various thermochemical processing were assessed as adsorbents for emerging contaminants in water. Thermal activation under flows of CO2 or air as well as chemical activation with phosphoric acid were applied to the hydrochars. Their characteristics were analyzed and related to their ability to uptake three pharmaceuticals (diclofenac sodium, salicylic acid and flurbiprofen) considered as emerging contaminants. The hydrothermal carbonization and subsequent activations promoted substantial chemical transformations which affected the surface properties of the activated hydrochars; they exhibited specific surface areas ranging from 300 to ∼620 m(2)/g. Morphological characterization showed the development of coral-like microspheres dominating the surface of most hydrochars. Their ability to adsorb the three pharmaceuticals selected was found largely dependent on whether the molecules were ionized or in their neutral form and on the porosity developed by the new adsorbents.

Responses of hematological parameters, beta-endorphin, cortisol, reactive oxygen metabolites, and biological antioxidant potential in horses participating in a traditional tournament.

Several concerns have been raised over the health of animals used in equestrian games that have their origins in historical or religious events and are currently held in many countries. This study investigated physiological stress response and health status of horses participating in the Sartiglia, a historical horse tournament held in the city of Oristano, Italy, which is principally based on the attempts of masked horsemen at a gallop to run a sword through a hole in a suspended silver star. Blood samples were collected from 21 horses the day before the tournament (D0), during the tournament (D1), and the day after the tournament (D2). Samples were analyzed for complete blood count and biochemical, hormonal, and oxidative stress assays. Data were analyzed using the mixed effect model with sampling session as one of the fixed effects. On the whole, blood parameters evidenced an optimal health status of horses at D0. Significant dehydration and increase of circulating glucose, enzymes, cortisol, and ß-endorphin were registered at D1 (P < 0.001) with a complete recovery of physiological values just at D2. The reactive oxygen metabolites (d-ROM), from which the prooxidant activity can be evaluated, showed an increase from D0 to D1 and D2. Concentration of biological antioxidant potential, which measured the antioxidant capacity, was characterized by the maximum level registered during the tournament and counteracted the simultaneous increase of d-ROM. It can be hypothesized that the tournament played an important role in causing high levels of oxidant markers not only because of the physical exercise represented by the gallop but also because the emotional stressors. In conclusion, the tournament caused significant changes of most parameters, which rapidly recovered to baseline values within the day after. These data will certainly be useful for a future implementation of tests in equine medicine and for the improvements of knowledge of changes of blood parameters and health of horses in similar tournaments.

Oxaliplatin (L-OHP) treatment of human myeloma cells induces in vitro growth inhibition and apoptotic cell death.

Oxaliplatin (L-OHP), a diaminocyclohexane platinum derivative, is an active and well tolerated anticancer drug which is presently used in the treatment of gastrointestinal tumours. Since the efficacy of L-OHP in the treatment of multiple myeloma (MM) has not yet been evaluated, we studied the antiproliferative activity of this compound in vitro in a panel of MM cell lines (XG1, XG1a, U266 and IM-9). We found that L-OHP inhibited the growth of MM cells at therapeutically achievable concentrations (IC(50): 5-10 microM after 24 h of exposure) and was more active than Cisplatin (CDDP) or Carboplatin (CBDCA). The activity of L-OHP was apparently not affected by interleukin-6 (IL-6), the major growth and survival factor of MM cells. We also found that L-OHP induced apoptotic cell death. We demonstrated that the combination of L-OHP with Dexamethasone (Dex) resulted in the enhancement of the anti-myeloma effects. L-OHP and Dex both induced poly adenosine diphosphate (ADP)-ribose polymerase (PARP) cleavage and this induction was enhanced by the combined treatment. L-OHP-induced apoptosis correlated with caspase-3 cleavage, but this correlation could not be demonstrated in Dex-treated cells. Taken together, these in vitro results provide a rationale for the experimental use of L-OHP in the treatment of MM patients and suggest therapeutic combinations of potential value.

A novel monoclonal antibody recognizing human thymocytes and B-cell chronic lymphocytic leukemia cells.

This paper describes a new murine monoclonal antibody, UN5, raised against human thymocytes. This antibody recognizes a molecule of approximately 45 kDa on thymocytes. Flow cytometric analysis reveals a high intensity of labeling with the majority of thymocytes, whereas only CD20+ cells from peripheral whole-blood samples are weakly stained. Peripheral T cells, granulocytes, platelets and red blood cells do not express this antigen, while monocytes are only weakly labeled by UN5. Furthermore, the UN5 antibody discriminates between different types of B-cell malignancies, reacting with a subgroup of B-cell chronic lymphocytic leukemias and hairy cell leukemias, but not with the other kinds of hematopoietic malignancies tested. Antibody UN5 should prove a useful tool for the study of T-cell precursors and for analysis of both normal and neoplastic B cells.

Analysis of peripheral blood normal and malignant cells with the novel murine monoclonal antibody UN2.

The monoclonal antibody (mAb) UN2 was generated upon immunization of a Balb/c mouse with human thymocytes. mAb UN2 recognized an antigen expressed by a subpopulation of human thymocytes and peripheral blood lymphocytes. In thymus, mAb UN2 recognized cortical cells; its expression was higher on CD3bright than on CD3dim thymocytes. This antigen was also detected on peripheral blood granulocytes, monocytes, platelets and on cell lines MOLT4, U937 and KG1. mAb UN2 was submitted to the 5th International Workshop and Conference on Human Leukocyte Differentiation Antigens, Boston, MA, 1993, and was assigned to the CD31. Expression of the UN2-recognized antigen in malignant lymphoid cells from 57 cases of B-cell chronic lymphoproliferative disease and 4 of B-cell acute lymphoblastic leukemia was analysed in flow cytometry. Among the 57 cases of B-cell chronic lymphoproliferative malignancies studied, 49 were classified as B-cell chronic lymphocytic leukemia. These showed high (86 +/- 8%) UN2 antigen expression. In 8 cases of hairy-cell leukemia the percentage of cells reacting with mAb UN2 was 42 +/- 4%; the fluorescence intensity of labelled cells was lower than that displayed by cells of B-cell chronic lymphocytic leukemia and comparable to that of normal lymphoid cells. mAb UN2 could prove useful in analysis of the lymphoid development and diagnostics of B-cell chronic lymphoproliferative disorders.

Breast cancer estrogen and progesterone receptors.

Data from 2933 consecutive cases of primary breast carcinoma, observed in our Institute from 1984 to 1994, having documented estrogen (ER) and progesterone (PR) receptor levels, were obtained from the Institute's Hospital Tumor Registry and analysed after being categorised as follows: age, less than or equal to 60 vs. >60; menopausal status, pre-menopausal vs. post-menopausal; histology, ductal vs. lobular vs. others; tumor size, T-1 vs. T-2, T-3, T-4; nodal status, N-0, vs. N+; histologic grade, 1-2 vs. 3; focality, unifocal vs. multifocal; ER status, <10 fmol/mg protein vs. greater than or equal to 15. At multivariate analysis, using a logistic model including age, histology, tumor size, nodal status, histologic grade, uni-multifocality and PGF/ER status, significant associations were, for ER status: PGR status (OR = 34.01, 95% CI:20.08-57.80), histology (OR = 3.24, 95% CI:1.85-5.67), histologic grade (OR = 2.18, 95% CI:1.38-3.42), menopausal status (OR = 2.17, 95% CI:1.26-3.74), age (OR = 34.01, 95% CI:20.08-57.80), menopausal status (OR = 5.27, 95% CI:1.43-3.33), age (OR = 1.71, 95% CI:1.13-2.59). The finding that estrogen receptor positivity was more prevalent among tumors with lobular histology seems to suggest the possibility of fundamental differences in tumor biology ductal and lobular cancers.

Does a relationship exist between trends in estrogen receptor levels and breast cancer incidence and mortality?

Estrogen receptor data from 4,049 patients with primary breast cancer treated at the National Cancer Institute of Naples between 1984 and 1996, have been evaluated to analyze temporal trend of this tumor marker. The prevalence rate of estrogen receptor levels falls from >75% in women older than 60 years to <70% in younger patients. The analysis of these data by birth cohort shows a trend very similar to that of breast cancer incidence in Italy, suggesting that the breast cancer appearance could be modulated in different period of life.

M-phase promoting factor (MPF) and mitogen activated protein kinases (MAPK) activities of domestic cat oocytes matured in vitro and in vivo.

This work was undertaken in order to examine M-phase promoting factor (MPF) and mitogen-activated protein kinases (MAPK) activities during meiotic progression of cat oocytes cultured in two different media for two different incubation times and preovulatory cat oocytes that reached MII in vivo. Oocytes recovered from ovaries of ovariectomized cats were cultured either in TCM 199 or SOF for 24 h and 40 h. In vivo matured oocytes were recovered by follicular aspiration from ovaries of domestic cats ovariectomized 24 h to 26 h after hormonal treatment. Results showed that the kinetic of MPF and MAPK activity was similar during meiotic progression of cat oocytes matured in TCM 199 and SOF. After 24 h of incubation, MII oocytes had significantly (p < 0.001) higher MPF and MAPK levels than MII oocytes cultured for 40 h in both culture media. MPF and MAPK activity was significantly (p < 0.01) lower in the oocytes matured in vitro than in those matured in vivo. This study provides evidence that the two different maturation media did not determine differences in MPF and MAPK fluctuations and levels during meiotic progression of cat oocytes and that the time of maturation influenced the level of the two kinases. Moreover, it shows that MPF and MPK activity is higher in in vivo matured oocytes than in in vitro matured oocytes, suggesting a possible incomplete cytoplasmic maturation after culture.

Pattern and concentration of free and acetylated polyamines in urine of cirrhotic patients.

The pattern and concentration of urinary, free, monoacetylated and total polyamines were determined in 31 cirrhotic patients, divided into three classes according to Child's classification, and in 28 healthy subjects. Cirrhotic patients had increased levels of free, monoacetylated and total polyamines. They also showed a significant increase in N1-acetylspermidine to N8-acetylspermidine molar ratio. Urinary polyamine excretion was not related to the severity of liver disease nor to the values of laboratory liver function tests. Furthermore, polyamine excretion was not significantly different in cirrhotics with or without diabetes or IGT, while plasma insulin and glucagon levels were increased in all cirrhotic patients. The results suggest that enhanced polyamine biosynthesis and catabolism, particularly N1-acetylation, occur in cirrhotic patients, probably due to hepatic regeneration and/or increased levels of insulin and glucagon.

Differences in polyamine metabolism between carcinomatous and uninvolved human breast tissues.

The polyamine biosynthetic enzymes ODC and SAMDC show higher activity in carcinomatous human breast tissue than in uninvolved tissue of the same breast; the interconversion enzyme PAO shows significantly lower activity in carcinomatous than uninvolved tissue. The polyamine metabolism in carcinomatous human breast tissue thus appears to differ from that in uninvolved tissue. Intracellular polyamine concentrations, particularly spermine, are high in carcinomatous tissue. This increase and that of the biosynthetic enzymes suggest that a higher polyamine concentration is needed for carcinomatous cell growth. If this is the case, the lower PAO activity in carcinomatous tissue may be explained as a mechanism that conserves the high intracellular polyamine concentration.

Epidermal growth factor receptor in human breast cancer comparison with steroid receptors and other prognostic factors.

The relationship between epidermal growth factor receptor (EGFR) status and various prognostic factors was investigated in 70 human breast cancer specimens. Epidermal growth factor receptor was determined by radioligand binding assay, standardized by the EORTC Receptor Study Group, using hydroxyapatite to separate receptor-bound and free ligand. The percentage of EGFR positivity was 80% when the cutoff was set at 5 fmol/mg of membrane protein; this percentage was among the highest hitherto reported. Regression analysis of EGFR versus ER and PR levels confirmed an inverse relationship between EGFR and ER (p = 0.022) as well as between EGFR and PR (p = 0.024). Univariate analysis of the EGFR data stratified according to steroid hormone receptor status showed EGFR to be negatively associated with ER and PR. No association was found between EGFR and menopausal status, axillary lymph node involvement, tumor size, and differentiation grade. A direct association between EGFR status and Ki-67 positive cell rate could be demonstrated.

Arundo donax cane as a precursor for activated carbons preparation by phosphoric acid activation.

Canes from Arundo donax, a herbaceous rapid-growing plant, were used as precursor for activated carbon preparation by phosphoric acid activation under a self-generated atmosphere. The influence of the carbonization temperature in the range 400-550 degrees C and of the weight ratio phosphoric acid to precursor (R = 1.5-2.5) on the developed porous structure of the resulting carbons was studied for 1 h of carbonization time. Surface properties of the activated carbons were dependent on a combined effect of the conditions employed. Carbons developed either with R = 1.5 over the range 400-500 degrees C, or with R = 2 at 500 degrees C exhibited surface areas of around 1100 m2/g, the latter conditions promoting a larger pore volume and enhanced mesoporous character. For both ratios, temperature above 500 degrees C led to reduction in porosity development. A similar effect was found for the highest ratio (R = 2.5) and 500 degrees C. The influence of carrying out the carbonization either for times shorter than 1 h or under flowing N2 was also examined at selected conditions (R = 2, 500 degrees C). Shorter times induced increase in the surface area (approximately 1300 m2/g), yielding carbons with smaller mean pore radius. Activated carbons obtained under flowing N2 possessed predominant microporous structures and larger ash contents than the samples derived in the self-generated atmosphere.

Effect of pyrolysis temperature on composition, surface properties and thermal degradation rates of Brazil Nut shells.

Changes in chemical and surface characteristics of Brazil Nut shells (Bertholletia excelsa) due to pyrolysis at different temperatures (350 degrees C, 600 degrees C, 850 degrees C) were examined. For this purpose, proximate and ultimate analyses, physical adsorption measurements of N2 (-196 degrees C) and CO, (25 degrees C) as well as samples visualisation by scanning electronic microscopy (SEM) were performed. Appreciable differences in the residue characteristics, depending markedly on the pyrolysis temperature, were observed. Release of volatile matter led to the development of pores of different sizes. Progressive increases in micropore development with increasing pyrolysis temperature took place, whereas a maximum development of larger pores occurred at 600 degrees C. Furthermore, kinetics measurements of Brazil Nut shells pyrolysis from ambient temperature up to 900 degrees C were performed by non-isothermal thermogravimetric analysis. A model taking into account the significant changes in the residue during pyrolysis, through an increase in the activation energy with temperature and solid conversion, were found to properly fit the kinetics data over the wide range of degradation investigated.

Polyamine oxidase activity in carcinoma-bearing human breast: significant decreased activity in carcinomatous tissue.

The paper reports a comparative study of the activity and distribution of polyamine oxidase (PAO) in carcinomatous human breast tissue and in the healthy uninvolved tissue of the same breast. The results indicated that in the carcinomatous tissue the PAO activity is lower than in the healthy uninvolved tissue.