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Biomedical subjects

L Duchateau

Publications and source records attributed to L Duchateau.

At least 37 records · Page 2Linked to original sources

Inhibition of bovine sperm-zona binding by bovine herpesvirus-1.

The purpose of the present study was to identify a potential interference of bovine herpesvirus-1 (BoHV-1) with sperm-oocyte interactions during bovine in vitro fertilization. An inhibition of almost 70% of sperm-zona binding was observed when bovine cumulus-denuded oocytes were inseminated in the presence of 10(7) 50% tissue culture infective dose/ml BoHV-1. The inhibitory effect of BoHV-1 on sperm-zona binding was mediated by an interaction of the virus with spermatozoa, but not with oocytes. Treatment of spermatozoa with BoHV-1, however, did not affect sperm motility and acrosomal status. Antiserum against BoHV-1 prevented the virus-induced inhibition of sperm-zona binding, indicating that BoHV-1 itself affects the fertilization process. In order to investigate which BoHV-1 glycoprotein(s) are responsible for the virus-sperm interaction, BoHV-1 was treated with monoclonal antibodies against the viral glycoproteins gB, gC, gD and gH prior to insemination. Anti-gC completely prevented the inhibitory effect of BoHV-1 on sperm-zona binding, while anti-gD caused a reduction of this inhibition. Further evidence for the involvement of gC and gD in the virus-sperm interaction was provided by the fact that purified gC and gD decreased sperm-zona binding in a dose-dependent way with gC being more effective than gD. These results indicated that BoHV-1 inhibits bovine sperm-zona binding by interacting with spermatozoa. The binding of BoHV-1 to a spermatozoon is mediated by the viral glycoproteins gC and gD, and therefore seems to be comparable with the mechanisms of BoHV-1 attachment to its natural host cell.

Acrosome Reaction↗

Association between somatic cell count in early lactation and culling of dairy heifers using cox frailty models.

The association between somatic cell count (SCC) of dairy heifers in early lactation [SCCel; measured between 5 and 14 d in milk (DIM)] and the culling hazard during the first lactation was studied using Cox frailty models. Udder health problems were the culling reason for 10% of the culled heifers in this study. For each unit increase in the log-transformed SCCel (LnSCCel), the culling hazard increased by 11% [Hazard ratio (HR) = 1.11]. The strength of the association depended on 5 factors. Firstly, the association was stronger when SCCel was recorded after 10 DIM than at an earlier DIM. Secondly, the association was stronger if only culling events for udder disorders were considered (HR = 1.32) instead of all culling events (HR = 1.11). Furthermore, for each unit increase of test-day LnSCC after 14 DIM, modeled as a time-varying covariate, the culling hazard in the first lactation increased by 26% (HR = 1.26). Including LnSCC in the model already containing LnSCCel, reduced the estimate of LnSCCel slightly. Fourth, a higher test-day milk yield, modeled as a time-varying covariate, protected against culling and reduced the magnitude of the effect of LnSCCel as well when taken into account. Finally, the association between LnSCCel and culling was still present, although smaller, in the group of heifers with a second test-day SCC </=50,000 cells/mL.

Animals↗

Effect of carprofen treatment following experimentally induced Escherichia coli mastitis in primiparous cows.

Acute Escherichia coli mastitis is one of the major sources of economic loss in the dairy industry due to reduced milk production, treatment costs, discarded milk, and occasional fatal disease. Nonsteroidal anti-inflammatory drugs (NSAIDs) are frequently used as adjunctive therapy to antibiotics. The objective of the current study was to evaluate the effect of carprofen treatment following infusion of Escherichia coli into the mammary glands of primiparous cows during the periparturient period. Severity of mastitis was scored based on the average milk production in the uninfected quarters on d +2 postinoculation and a clinical severity score. Carprofen was administered intravenously at 9 h postchallenge, when clinical signs of mastitis appeared. In previous work, efficacy of NSAIDs was mainly evaluated using clinical symptoms. In the present study, the effect of carprofen on innate immune response was also assessed by quantification of inflammatory mediators. All primiparous cows reacted as moderate responders throughout the experimental period. Primiparous cows were intramammarily inoculated with 1 x 10(4) cfu of E. coli P4:O32 in 2 left quarters. Analysis of blood and milk parameters, including IL-8, complement component C5a, lipopolysaccharide-binding protein (LBP), soluble CD14, prostaglandin E2, and thromboxane B2 was performed from d 0 to d +6 relative to intramammary inoculation. Rectal temperature in carprofen-treated animals was lower than in control animals at 3 and 6 h posttreatment. Treatment also restored the decreased reticulorumen motility that occurs during E. coli mastitis to preinfection levels faster than in control animals. Carprofen treatment resulted in an earlier normalization of the clinical severity score. Eicosanoid (prostaglandin E2 and thromboxane B2) production in milk tended to be inhibited by carprofen. No significant differences in the kinetic patterns of somatic cell count, IL-8, complement component C5a, LBP, and soluble CD14 were observed. In conclusion, carprofen treatment improved general clinical condition by effective antipyrexia and restoration of reticulorumen motility but did not significantly inhibit eicosanoid production. Carprofen treatment did not result in a significant decrease of chemotactic inflammatory mediators, IL-8 and C5a, and early innate immune molecules, sCD14 and LBP. Therefore, major modulatory effects from NSAID administration were not observed in this mastitis model, although a larger study might confirm some apparent trends obtained in the present results.

Animals↗

Risk factors for the herd-level bacteriologic prevalence of Salmonella in Belgian slaughter pigs.

Herd-level risk factors for salmonellosis in pigs were investigated in a cross-sectional study on 62 Belgian farrow-to-finish pig herds belonging to one slaughterhouse cooperative. Data concerning housing and ventilation, management, hygiene, biosecurity, production parameters, feeding, disease control and transport to the slaughterhouse were collected during a herd visit by means of a questionnaire. The percentage of positive animals in a slaughterhouse delivery, as determined by qualitative Salmonella isolation in the mesenteric lymph nodes taken from 30 slaughter pigs, was the outcome variable. All samples were taken in 4 different slaughterhouses. Variables first were submitted to a univariable analysis using a logistic mixed regression model, with herd as random effect. Variables which were related to the Salmonella prevalence (P < 0.05) were analysed further in a multivariable model. The clustering of Salmonella infection within a pen also was studied in a generalised mixed model with pen as random effect. Salmonella isolates were identified by serotype. In 57 (92%) of the herds, at least one sample was found positive for Salmonella. The median percentage of positive Salmonella samples per delivery was 64% (range: 0-100%). In the multivariable model, only type of floor was related significantly to the prevalence: 100% (95% CI 88-100) for herds with <50% slatted floors to 54% (36-70) for herds with fully slatted floors. The results from the analysis should be interpreted with care because only 62 herds were included in the study. Clustering between pigs from the same pen could not be demonstrated (variance +/- S.D.: 0.11 +/- 0.16). S. typhimurium (30%) and S. derby (20%) were most common among the 23 different serotypes that were found.

Abattoirs↗

Variations in the immune response to natural Schistosoma mattheei infections in calves born to infected mothers.

During previous work Schistosoma antibodies and circulating antigens were detected at birth in the serum from some calves born to Schistosoma mattheei infected mothers. The objectives of the present survey were: (1) to investigate the proportion of calves, born to cows infected with S. mattheei, which have specific antibodies and circulating schistosome antigens present in their serum at birth and (2) to investigate whether the presence or absence of these specific antibodies and/or circulating antigens at birth may affect the pattern of a natural S. mattheei infection in calves from 4 to 5 months of age, when the colostral antibodies are thought to be of negligible importance. A total of 28 calves born to infected mothers were randomly selected. Faeces, serum and colostrum samples were collected from the cows at calving, serum samples were collected from the calves at birth (day 0), after intake of colostrum (day 1) and monthly thereafter up to the age of 10 months. Both serum and colostrum samples were analysed for IgG(H+L) against SWAP mattheei and schistosome circulating anodic antigen (CAA) levels. The calves were exposed to a natural challenge from the age of 4-5 months. Faecal samples were collected from the calves monthly, starting at an age of 5 months up to 10 months, and were examined for faecal egg counts. Nine (group 1) out of the 28 calves were found to have specific antibodies in their serum at birth, in 5 of them CAA levels were also detected. In the other 19 calves (group 2) no IgG(H+L) or CAA were detected. At the end of the study faecal egg counts and CAA levels were significantly lower in calves from group 1 compared to group 2. Results confirm earlier work that specific antibodies and circulating antigens may be present in serum from calves at birth, and show that these calves have lower faecal egg counts and CAA levels after exposure to a natural challenge.

Animals↗

Inhibition of bovine sperm-oocyte fusion by the p-aminophenyl derivative of D-mannose.

Several steps in the process of mammalian fertilization are mediated by carbohydrates. This study investigated the role of the p-aminophenyl derivative of d-mannose (APMP) during bovine fertilization. Inseminating cumulus-oocyte complexes (COCs) in the presence of increasing APMP concentrations resulted in a significant dose-dependent decrease of the fertilization rate (P < 0.05). No negative effect of 50 mM APMP on total sperm motility and progressive motility was found. Subsequently, the fertilization steps at which this blocking effect could be exerted were investigated, i.e., sperm penetration of the cumulus oophorus, sperm-zona binding, acrosome reaction, sperm-oolemma binding, and/or sperm-oocyte fusion. Inseminating cumulus-enclosed and cumulus-denuded oocytes in the presence of 50 mM APMP significantly decreased the fertilization rate to a comparable minimum level (P < 0.05). There was no significant relationship between the number of spermatozoa bound to the zona pellucida and the APMP concentration, and APMP nor d-mannosylated bovine serum albumin (BSA) suppressed or stimulated sperm acrosomal status. Inseminating zona-free oocytes in the presence of 50 mM APMP did not influence sperm-oolemma binding, but significantly inhibited sperm-oocyte fusion (P < 0.05). Preincubating zona-free oocytes with 200 microg/ml Con A but not with 50 mM APMP inhibited the sperm-oocyte fusion rate to the same extent as when the gametes were simultaneously exposed to 50 mM APMP. These data indicate that the blocking effect of APMP on bovine fertilization is mainly due to an inhibition of sperm-oocyte fusion, probably by specific obstruction of the sperm receptor sites that are responsible for the fusion process.

Acrosome Reaction↗

Carbohydrates and glycoproteins involved in bovine fertilization in vitro.

In the present study, efforts were made towards identifying carbohydrates and glycoproteins involved in bovine in vitro fertilization (IVF). In vitro matured cumulus-oocyte complexes (COCs) were inseminated in the presence of a variety of carbohydrates and glycoproteins to determine which glycoconjugates act as competitive inhibitors of oocyte penetration. Among the carbohydrates and glycoproteins tested, D-mannose, fucoidan, dextran sulfate, and fibronectin were the most potent inhibitors of oocyte penetration (90% or more inhibition), while L-fucose and vitronectin inhibited the penetration rate to a lesser extent (around 50% inhibition). Other carbohydrates caused less than 40% inhibition (i.e., D-galactose, N-acetyl-D-galactosamine, D-fucose, and sialic acid) or were not effective as inhibitors of oocyte penetration (i.e., mannan, N-acetyl-D-glucosamine, dextran, and heparan sulfate). Heparin was the only carbohydrate that significantly increased the penetration rate. To exclude a possible toxic effect on spermatozoa, sperm motility was evaluated over time by means of computer-assisted sperm analysis in the presence of carbohydrates and/or glycoproteins that inhibited the penetration rate with 40% or more. L-fucose, dextran sulfate, and vitronectin did not significantly influence total and progressive sperm motility, whereas D-mannose, fucoidan, and fibronectin caused a significant, but slight reduction in both motility parameters. These results are indicative for the involvement of D-mannose, L-fucose, fucoidan, dextran sulfate, fibronectin, and vitronectin in bovine IVF.

Animals↗

Risk factors for mortality in grow-finishing pigs in Belgium.

The present study investigated risk factors for mortality in grow-finishing pigs of 137 pig herds belonging to one integration company during a period of 2.5 years. Mortality data, expressed as the number of dead pigs divided by the number of pigs placed in the fattening unit were investigated retrospectively. The following potential risk factors were evaluated: type of pig herd, season and year of placement in the fattening unit, pig density in the municipality, management practices (density of the pigs in the barn, origin of the pigs), housing conditions and feeding practices. The overall average mortality percentage was 4.70%. Three variables in a multivariable regression model were significantly associated with mortality: season of placement in the fattening unit, origin of the piglets and duration of the fattening period. Pigs placed in October, November and December, were at higher risk than pigs placed in other months. Herds that purchased pigs from a merchant, used pigs from other herds with an excess of piglets for filling their fattening units or herds that purchased pigs from more than five origin herds also suffered higher mortality. The mortality also increased in case of longer duration of the fattening period. This study documented for the first time that in addition to seasonal effects and a longer duration of the fattening period, purchasing feeder pigs from one or a limited number of herds is of crucial importance to achieve low mortality in the grow-finishing pigs.

Animal Husbandry↗

Moderate inflammatory reaction during experimental Escherichia coli mastitis in primiparous cows.

Nineteen primiparous cows were experimentally infected in 2 quarters with 1 x 10(4) (group A) or 1 x 10(6) (group B) cfu of Escherichia coli P4:O32 per quarter within 2 to 4 wk after parturition. Blood and milk samples were collected from all primiparous cows at regular time intervals from d -4 to d +3 relative to inoculation. Milk production rapidly decreased in both groups during E. coli mastitis, but recovery appeared to be faster in group B at d + 1 postinfusion (p.i.). The milk production losses in the noninfected quarters were substantial on the day of inoculation, which is probably due to pronounced systemic effects. However, on d + 2 p.i. milk production in the noninfected quarters nearly reached preinfection levels, indicating a moderate clinical severity following intramammary inoculation. None of the other severity criteria evolved towards a severe response pattern. Reticulorumen motility was inhibited in both groups during E. coli mastitis. The clinical episode was short lasting in both groups. Rectal temperature, heart rate, blood leukocyte count, number of colony-forming units, milk somatic cell count and several indicators for the disintegration of the blood-milk barrier returned to normal values within 24 to 72 h p.i. Primiparous cows reacted with a moderate inflammatory response following intramammary infusion with a relatively high dose of E. coli. Despite the use of a high inoculum dose, primiparous cows in both groups showed pronounced resistance against severe intramammary E. coli infection. A possible effect of the inoculum dose could be present, however, further research into the effect of the inoculum dose on the inflammatory response should be performed.

Animals↗

All-trans retinoic acid is increased in the acute phase-related hyporetinemia during Escherichia coli mastitis.

Blood vitamin A profiles, including concentrations of retinol and its active metabolite retinoic acid, were assessed during the peripartum period and during experimentally induced Escherichia coli mastitis in heifers. Serum retinol decreased in all animals in the immediate postpartum period and normalized within 1 wk after parturition. No significant changes were detectable in the concentrations of retinoic acid isomers during puerperium. Following intramammary E. coli infusion, all cows showed moderate symptoms of systemic disease besides the local signs of inflammation. The presence of a systemic acute-phase reaction was documented by fever, increase in serum amyloid A, and decrease in serum albumin. Retinol concentration in serum also decreased spectacularly during coliform mastitis, and the decline was clearly related to the timing of the acute-phase response. Moreover, a significant increase of all-trans retinoic acid, mirrored by a lowering of 13-cis retinoic acid, was detected during the same time period. The 9-cis isomer of retinoic acid was present in all samples, but it remained below the quantification limit. Results confirmed the decrease in serum retinol during the peripartum period of dairy cows. Furthermore, the study established that profound changes in vitamin A metabolism occur during the acute-phase reaction of coliform mastitis in heifers. The bovine infection model reproduced the acute phase-related hyporetinemia, as previously observed in humans and rats. In addition, all-trans retinoic acid was found to be the most abundant circulating acid isomer during mastitis, providing an indication for a possible key role of all-trans retinoic acid in the modulation of the immune response.

Acute-Phase Reaction↗

Differential effects of steroids and retinoids on bovine myelopoiesis in vitro.

Pregnancy and parturition are associated with physiological changes caused by steroid hormones. Alterations in number, maturity, and function of polymorphonuclear leukocytes observed in dairy cows at parturition suggest a common causative relationship with steroid hormones. This study was designed to investigate the effects of progesterone, 17-beta-estradiol, and hydrocortisone on the proliferation of bovine progenitor cells. An in vitro culturing system was used, and colonies were scored after 7 d of incubation. At low concentrations, 17-beta-estradiol inhibited proliferation of granulocyte progenitor cells. Hydrocortisone reduced growth of granulocyte and monocyte colonies, whereas myelopoiesis was not altered by progesterone. Furthermore, we studied the effect of retinoids on colony formation of bovine bone marrow cells. All-trans- and 9-cis-retinoic acid stimulated growth of granulocyte colonies and inhibited proliferation of the monocyte lineage. The addition of the 13-cis-isomer also increased numbers of granulocyte colony-forming units. This study indicates that steroid hormones may be responsible for alterations in the bovine hematopoietic profiles observed in circulation during the postpartum period. White blood cells, especially polymorphonuclear leukocytes, which are derived from bone marrow, are an important first line defense against mastitis. Therefore, these effects of steroids might contribute to the increased susceptibility of dairy cows to Escherichia coli mastitis. We furthermore hypothesize that an important role might be attributed to retinoic acid in its regulation of bovine myelopoiesis. Modulation of myelopoiesis in favor of the granulocyte lineage during the acute-phase reaction may be an adaptive mechanism designed to increase the capacity of first-line defense to intramammary infections.

Animals↗

Proteolysis in milk during experimental Escherichia coli mastitis.

This work consisted of the intramammary infections (IMI) of 8 heifers by high doses of Escherichia coli to study both the proteolytic activity in milk and the resulting peptides. Therefore, a milking kinetic has been followed, and several parameters have been studied, such as proteose peptones (PP) fraction (quantitative and qualitative changes), plasmin activity (PA), milk somatic cell count (SCC), and bacterial count. A qualitative study of milk proteins and PP was performed by sodium dodecyl sulfate-PAGE, and the peptides recovered in PP during the acute phase of inflammation were amino-terminal micro-sequenced. A BSA increase in milk over time supported the hypothesis of an increase in the permeability of the epithelial barrier. A significant increase in PP content, considered to be an indicator of proteolysis, was observed from postinfusion hours (PIH) 12 to 48. Both the E. coli bacterial count and the SCC increased from PIH 3 to 216. Plasmin activity was increased noticeably from PIH 15 to 24. The respective increases in SCC, bacterial count, and PA suggest their involvement in a global mechanism responsible for the increase in proteolysis in milk after E. coli challenge. Somatic cell count and E. coli may be involved from PIH 3 to 216, and PA involvement might be highlighted during the maximum proteolysis, from PIH 15 to 24. A qualitative study of PP fraction by electrophoresis revealed the apparition of 5 peptide bands: P1 and P2 previously recovered during the lipopolysaccharide challenge, and E1 (27.0 kDa), E2 (15.5 kDa), and E3 (9.0 kDa) were specific to E. coli challenge; E1, E2, and E3 contained casein fragments. The roles played by leukocytes and E. coli are discussed.

Animals↗

Influence of 17beta-estradiol, progesterone, and dexamethasone on diapedesis and viability of bovine blood polymorphonuclear leukocytes.

The aim of the current study was to investigate whether polymorphonuclear leukocyte (PMN) diapedesis and viability are influenced by steroid hormones. Using an in vitro model with different types of cell layers (bovine mammary epithelial cells and fibroblasts), we investigate whether steroid hormone treatments (17beta-estradiol, progesterone, and dexamethasone) have an influence on the diapedesis capacity and viability of PMN. In addition, we studied apoptosis of PMN in the in vitro model and evaluated the influence of different types of cell layers and steroid hormone treatments on this process. A significant decrease in the number of viable PMN in the lower compartment of the in vitro model (i.e., number of migrated PMN x viability after migration) was found after 17beta-estradiol treatment, whereas no influence was detected after progesterone or dexamethasone treatment. The effect of 17beta-estradiol was not due to a lower viability before migration as none of the treatments caused a significant effect on the viability before diapedesis. This treatment effect was not influenced by endogenous 17beta-estradiol or progesterone levels before isolation because there was no correlation between these plasma levels and PMN diapedesis capacity or viability. Furthermore, migration through epithelial cells caused a significant decrease in viability of PMN due to increased apoptosis but not necrosis.

Animals↗

L-selectin and chemotaxis throughout bone marrow granulocyte maturation in the bovine.

Polymorphonuclear neutrophilic leukocytes (PMNL) play a pivotal role during inflammation. Bone marrow (BM) reserves are depleted as cells are released into circulation for recruitment to infection sites. Expression of L-selectin on the cell membrane allows neutrophils to roll along the activated endothelium. Whereas mechanisms leading to recruitment to infection sites are well established, expression of BM adhesion molecules in cows is limited. In this study, we assessed L-selectin expression and chemotactic response to zymosan-activated serum (ZAS) in bovine BM cells and in circulating neutrophils. Isolated blood PMNL and BM cells were used from 9 dairy cows, for quantifying L-selectin expression using flow cytometry, and from 12 dairy cows for chemotaxis studies. All granulocytic maturation stages expressed L-selectin. The percentage of cells fluorescing increased significantly in BM band and mature granulocytes and reached maximal expression on circulating neutrophils. Bone marrow band and segmented cells showed the highest L-selectin density. Chemotaxis through micropore filters in response to zymosan-activated fetal bovine serum was first observed in the myelocytic and metamyelocytic stages, and it increased with maturation and release into the blood stream. From these results, we conclude that L-selectin expression varies among stages of granulocytic maturation within the BM and differs from circulating PMNL. Further, BM cells are capable of migration starting at the metamyelocytic stage, and compared with BM cells, circulating neutrophils are more chemotactively active.

Animals↗

Increase of Escherichia coli inoculum doses induces faster innate immune response in primiparous cows.

The objective of the current study was to evaluate the dynamics of infection and the immunological response to varying numbers of Escherichia coli injected into the mammary glands of primiparous cows during the periparturient period. Primiparous cows have been shown to be more resistant to intramammary E. coli challenge, and an increase of the inoculum dose by 2 log10 units induced a more rapid clinical response and clearance of the organisms. Recognition of lipopolysaccharide (LPS) is a key event in the innate immunity response to gram-negative infection and is mediated by the accessory molecules CD14 and LPS-binding protein (LBP). Primiparous cows were inoculated with 1 x 10(4) (Group A; n=8) or 1 x 10(6) (Group B; n=8) cfu E. coli P4:O32 in their 2 left quarters during the periparturient period. Clinical examination and analysis of blood and milk parameters, including IL-8, complement fragment 5a (C5a), LBP, and soluble CD14 (sCD14), were performed from d -4 to d +3 relative to infection. Primiparous cows in Group B initiated a more rapid clinical response following intramammary infection (IMI), resulting in typical clinical signs and changes in blood and milk parameters approximately 3 h earlier compared with primiparous cows in Group A. Based on average milk production in the noninfected quarters on d +2 postinoculation, all heifers reacted as moderate responders. Distinct differences in the kinetic patterns of rectal temperature, somatic cell count (SCC), IL-8, C5a, LBP, and sCD14 were observed between both groups during the early phase of inflammation. Both C5a and IL-8 increased before cellular influx into the infected glands, followed by increases in sCD14 and LBP. In conclusion, primiparous cows are able to clear an intramammary E. coli infection efficiently. Moreover, increasing the inoculum dose induces a more rapid inflammatory reaction, mainly because of early activation of the innate host immune response.

Animals↗

Viability of milk neutrophils and severity of bovine coliform mastitis.

To study the host-pathogen interactions during Escherichia coli mastitis, we first determined whether E. coli infection would change blood and milk polymorphonuclear neutrophil (PMN) chemiluminescence (CL) and viability. We then hypothesized that when E. coli invade the mammary gland, the viable PMN in milk would efficiently phagocytose and destroy E. coli before establishment of infection. We observed that the phagocytosis-dependent and independent CL were closely linked to PMN viability and were crucial to the outcome of mastitis. Maximal PMN influx and colony-forming units in infected quarters appeared at postinfection hours (PIH) 6 to 24. This further boosted PMN recruitment through bone marrow-blood barrier as well as blood-milk barrier. The survival of recruited PMN in the E. coli-infected quarters was much higher than that of noninfected quarters. Chemiluminescence activity of PMN from the infected quarters significantly increased following E. coli infection, even exceeding that of blood at PIH 6, 12, and 18 to 24; no such increase was observed in noninfected quarters, suggesting that the various responses of milk PMN to stimuli resulted largely from PMN viability. The highest CL intensity and durability was observed in milk PMN from infected quarters at PIH 12. Whereas an increased viability of PMN in the noninfected quarters was only significant at PIH 6, the viability of PMN in infected quarters was long lasting and significantly higher at PIH 6 to 72. Importantly, higher preinfection milk PMN viability correlated with bacterial clearance, which was accompanied by faster recovery. Our study strongly supports the hypothesis that boosting milk PMN viability could be a strategy with which to prevent or reduce the severity of coliform mastitis in dairy cows. This strategy might be achieved through strengthening bone marrow functionality.

Animals↗

Apoptosis in the canine endometrium during the estrous cycle.

Apoptotic cell death in the endometria of 58 female dogs in different stages of the estrous cycle was assessed (in formalin-fixed, paraffin-embedded sections) with both the terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate nick end labeling (TUNEL) assay and immunohistochemical detection of caspase-3 activity. For both techniques, the apoptotic index was determined in the surface epithelium, stroma, crypts, and basal glands by counting the percentage of stained cells in a total of 500 cells in each category. In the surface epithelium and stroma, TUNEL- and caspase-3-positive cells were rare (apoptotic index<1) throughout the estrous cycle. However, caspase-3 detection showed a significant increase in the apoptotic index in the stroma during anestrus as well as an increase in the index in both the stroma and surface epithelium in late metestrus. The apoptotic index increased during late metestrus and anestrus in the crypts and basal glands; in the crypts, this increase was significant only when caspase-3 detection was used, whereas in basal glands, significant differences were found for both techniques. In conclusion, apoptosis was present in canine endometrial cells during the estrous cycle, but caspase-3 detection showed more significant differences than the TUNEL assay. Furthermore, a high apoptotic index (suggestive of endometrial desquamation) was not detected in the surface epithelium and there was no significant correlation between the apoptotic index in any cell group and serum progesterone concentrations.

Animals↗

Continuing chemotherapy or not after the induction treatment in advanced breast cancer patients. clinical outcomes and oncologists' preferences.

The optimal duration of cytostatic treatment for metastatic breast cancer is still a matter of debate. Possible gain in the duration of remission has to be weighed against the side-effects of treatment. Our aim was to define the optimal duration of cyclophosphamide, methotrexate, 5-fluorouracil (CMF) treatment by studying the time to treatment failure, overall survival and using a Q-TWiST analysis. The treating physician's opinion was asked. The European Organization for Research and Treatment of Cancer (EORTC) Breast Cancer Group conducted a randomised trial in 204 non-progressing metastatic breast cancer patients after induction chemotherapy (CMF) to stop or continue treatment. Progression-free (PFS) and overall survival (OS) were studied. To gain more insight into the burden of treatment-related side-effects, Q-TWiST was analysed. In addition, we asked for oncologists' preferences as patients are likely to be influenced by their physicians' opinion. Continuation of CMF had a significantly longer time to treatment failure (TTF) 5.2 versus 3.5 months (P=0.011). There was no overall survival (OS) difference 14.0 versus 14.4 months (P=0.77). Mean quality-adjusted survival time was equal to 8.4 months for no further treatment and decreased to 7.9 months for continuation of CMF (95% Confidence Interval (CI) of difference equals 0.5+/-2.5 months). Almost half of the oncologists said they would favour continuous treatment for a 3-month gain in time to progression-a difference which was not found in this study. Based on these data, an interruption of chemotherapy (CMF), if this is the wish of the patient, is justified.

Adult↗