A quantitative case study assessment of changes to hepatic metabolism from nonlactating grazing dairy cows consuming a large proportion of their diet as fodder beet.

Because of its high yield and the ability of cows to graze it in situ, fodder beet (FB) has become a popular crop in grazing systems, particularly for nonlactating cows. Due to its high sugar content, however, the transition to FB must be managed carefully to avoid rumen acidosis and associated metabolic dysfunction. The initial consumption of FB reduces ruminal pH; however, it is unclear whether this affects liver metabolism and results in systemic inflammation, as has been reported during subacute ruminal acidosis from high-grain diets. We used a quantitative case study approach to undertake additional measurements on a project demonstrating the effects of FB on urinary nitrogen excretion. The objective of our component, therefore, was to determine whether the inclusion of high rates of FB in the diet of nonlactating cows changed indicators of hepatic metabolism relative to a standard diet for nonlactating grazing cows. During the nonlactating period, multiparous, pregnant Holstein-Friesian cows were randomly assigned (n = 15 per treatment) to either pasture (8 kg of DM/cow per day) with corn silage (4 kg of DM/cow per day; PA) or transitioning onto an FB diet (8 kg of DM/cow per day) with pasture silage (4 kg of DM/cow per day; BT) over 14 d. Blood was sampled and the liver was biopsied during the adaptation period and after 7 d of full diet allocation. The hepatic expression of genes involved in peroxisomal oxidation was increased in cows adapting to FB, whereas the expression of genes involved in mitochondrial oxidation was increased when cows were on their full allocation of FB. These results indicate changes to fatty acid metabolism with FB consumption. Expression of 2 genes involved in the unfolded protein response was greater during the adaptation period in cows consuming FB, potentially reflecting negative effects of transitioning onto the FB diet on hepatic metabolism. Interestingly, expression of genes involved in the methionine cycle was increased in the BT cows. We hypothesize that this is a result of FB betaine absorption, although it is unclear to what extent betaine escapes ruminal degradation. While on the full diet allocation, there were lower serum concentrations of markers of hepatic stress in BT cows and no difference in expression of genes involved in oxidative stress compared with pasture-fed cows. However, there was an increase in plasma haptoglobin concentrations, indicative of an acute inflammatory response in BT cows. From this case study, we conclude that the results indicate no negative effects of the FB diet on liver metabolism and, possibly, positive effects on hepatic function. It appears, therefore, that the transition of nonlactating cows onto an FB diet can be managed to minimize the negative effects of the high sugar intake. Further research on the amount of betaine that escapes ruminal degradation in cows consuming FB would be of value to better understand whether betaine reduces liver damage in dairy cows consuming FB.

Feeding fodder beet (Beta vulgaris L.) with either barley straw or pasture silage to non-lactating dairy cows.

AIMS: To determine the suitability of diets containing either approximately 85% fodder beet (Beta vulgaris L.) with barley straw or 65% fodder beet with pasture silage when fed to non-lactating dairy cows, by measuring intakes, digestibility, rumen function including microbial growth, and N excretion. METHODS: Holstein-Friesian cows fitted with permanent rumen fistulae were fed either 65% fodder beet with pasture silage (Silage; n=8) or 85% fodder beet with barley (Hordeum vulgare L.) straw (Straw; n=8) in an indoor facility over a 9-day period, for measurement of intakes, digestibility, rumen function and urine production. The cows were adapted to the diets over 2 weeks before the indoor measurements. Feed was available for about 6 hours/day, as practiced commercially for wintering non-lactating cows. RESULTS: Five cows fed the Straw diet had to be removed from the trial because of acute acidosis; four on Day 1 of the measurement period and one on Day 7. One cow allocated to the Silage diet refused to eat fodder beet bulbs and was also removed from the trial. Two cows fed the Silage diet were also treated for acidosis. DM intakes were lower with the Straw than Silage diets (6.4 (SE 0.4) vs. 8.3 (SE 0.5) kg/day) and organic matter (OM) digestibility was lower with the Straw than Silage diets (77 (SE 1) vs. 83 (SE 1) g/100g). The N content of the two diets was 1.14 and 1.75 g/100 g DM and there was a net loss of N by cows fed the Straw diet (-22.7 (SE 7) g/day). Rumen microbial N production was much lower in cows fed the Straw than the Silage diet (6.6 (SE 1.3) vs. 15.8 (SE 0.7) g microbial N/kg digestible OM intake). Concentrations of ammonia in rumen liquid collected on Days 5-6 were below detection limits (<0.1 mmol/L) in 36/48 (75%) samples collected from cows fed the Straw diet and in 27/48 (56%) cows fed the Silage diet. Mean urinary N excretion was lower in cows fed the Straw than the Silage diet (52.0 (SE 5.8) vs. 87.7 (SE 5.9) g/day). CONCLUSION AND CLINICAL RELEVENCE: An over-wintering diet for dry cows comprising about 65% fodder beet with 35% pasture silage provided adequate nutrition, although there was some risk of acidosis. In contrast, the diet containing about 85% fodder beet with barley straw resulted in lower DM intakes, poor rumen function, negative N balance so that both nutrition and welfare were compromised.

Fear of cancer recurrence in young women with a history of early-stage breast cancer: a cross-sectional study of prevalence and association with health behaviours.

PURPOSE: Fear of cancer recurrence (FCR) is common and associated with younger age. This study aimed to explore the prevalence and correlates of FCR amongst younger survivors of early breast cancer. SUBJECTS: A total of 218 women aged 18-45 were diagnosed with stage 0-2 breast cancer at least 1 year earlier. METHODS: The participants completed a web-based survey including a validated measure of FCR and items exploring medical surveillance practices and health care use. RESULTS: A total of 70% of participants reported clinical levels of FCR. Higher FCR was associated with higher frequency of unscheduled visits to the GP, higher frequency of breast self-examination and other forms of self-examination for cancer, not having mammograms or ultrasounds or other forms of cancer screening in the past year, more complementary therapy use and the use of counselling and support groups. CONCLUSIONS: Young women with breast cancer are particularly vulnerable to FCR. The present study provides preliminary evidence that FCR is associated with higher health costs and lower surveillance rates which may compromise health outcomes. Routine screening for FCR in follow-up care is recommended.

Outcomes and predicting response in anaemic chemotherapy patients treated with epoetin alfa. A multicentre, 4-month, open-label study in Australia and New Zealand.

BACKGROUND: The aim of the study was to evaluate the effectiveness, safety, and clinical outcomes of erythropoietin therapy in the treatment of anaemic cancer subjects receiving chemotherapy and to examine hypochromic red blood cell measurement as an indicator of functional iron sufficiency and as a predictor of responsiveness or non-responsiveness to erythropoietin therapy. METHODS: Patients who had a non-myeloid malignancy, had Hb < or = 11.0 g/dL, had a life expectancy of more than 6 months, were 18 years or older, were receiving chemotherapy and would continue to be treated for at least 2 months were given s.c. epoetin alfa three times a week. RESULTS: Haemoglobin levels increased significantly at all time periods compared with baseline and the number of transfusions received decreased significantly at all time periods compared with baseline. Quality of life as measured by Functional Assessment of Cancer Therapy-Anaemia showed significant increases at months 2 and 4 and there were significant improvements in the fatigue subscale at both time points (P < 0.05). Significant improvements at end-point were observed for the physical, emotional and functional well-being, and additional concern subscales (all P < 0.05). Haematocrit and reticulocytes increased significantly at end-point compared with at baseline (haematocrit 33.4 vs 28.3%, P < 0.001; reticulocytes 105.8 vs 78.6 x 10(9)/dL, P = 0.005). The percentage of hypochromic red blood cells did not show predictive value for response to treatment status. CONCLUSION: Epoetin alfa improved haemoglobin levels and quality of life in anaemic cancer patients receiving chemotherapy.

Development and evaluation of a test for tuberculosis in live European badgers (Meles meles) based on measurement of gamma interferon mRNA by real-time PCR.

A real-time PCR assay for the measurement of gamma interferon (IFN-gamma) mRNA in European badger (Meles meles) blood cultures was developed. The levels of IFN-gamma mRNA in blood cultures stimulated with either bovine or avian tuberculin or specific mycobacterial antigens were compared with those in a nonstimulated control blood culture as the basis for determining the tuberculosis (TB) status of live badgers. The assay was validated by testing 247 animals for which there were matching data from postmortem examination and culture of tissues. Relative changes in the levels of IFN-gamma mRNA in response to bovine tuberculin and specific antigens were found to be greater among badgers with tissues positive for TB on culture. The test was at its most accurate (87% of test results were correct) by using blood cultures containing bovine tuberculin as the antigen and when the response to avian tuberculin was taken into account by subtracting the avian tuberculin response from the bovine tuberculin response. At a specificity of 90.7%, the test was 70.6% sensitive. At the same specificity, the current serological enzyme-linked immunosorbent assay for TB in badgers was only 53% sensitive. This work demonstrates that measurement of IFN-gamma mRNA by real-time PCR is a valid method for the detection of TB in live badgers and may provide an alternative to the current serological methods of diagnosis, the Brock test. The testing procedure can be completed within 5 h of receipt of the blood culture samples. In addition, the use of a molecular biology-based test offers the potential to fully automate the testing procedure through the use of robotics.

Milking cows once daily influences behavior and udder firmness at peak and mid lactation.

Our aim was to evaluate the effects of once-daily milking on the welfare of dairy cows in a pastoral-based farming system. There are concerns that cows milked only once daily may experience discomfort associated with udder distension. We evaluated the behavior, including lying time and postures, grazing patterns, and kicking behavior in the parlor, and measures of udder firmness in cows milked once daily in 2 experiments: 1) at peak lactation, comparing cows milked once (1x) or twice-daily (2x) from the time of calving (n = 4 groups; 1x and 2x treatments); and 2) during the transition from 2x to 1x milking at mid lactation (n = 12 groups; 1x, 2x, and 2x to 1x treatments or TRANS). At peak lactation, cows milked 1x were more likely to lie with their hind legs touching the body than cows milked 2x. There were no differences in udder firmness (1x: 10.0 vs. 2x: 9.9 +/- 0.13 g of force) or kicking (1x: 25 vs. 2x: 24 +/- 7.8%) behavior in the parlor. In the second experiment, milking frequency was reduced from 2x to 1x and this change resulted in an increase in udder firmness (1 x: 9.1, 2x: 9.3, and TRANS: 9.5 +/- 0.14 g of force). Despite these differences in udder firmness, lying time (1x: 10.1, 2x: 9.5, and TRANS: 9.6 +/- 0.38 h/24 h), postures, and concentrations of fecal glucocorticoid metabolites (1x: 10.0, 2x: 10.5, and TRANS: 10.4 +/- 0.49 ng/g of feces in the 3 d after transition) in cows milked 1x or 2x were similar. Although milking frequency did not influence total time spent grazing, the diurnal grazing pattern of cows milked 2x was disrupted by afternoon milking at both peak and mid lactation. Cows milked 2x were able to compensate for this by grazing more after the return to pasture (DIM 153 to 155, 1x: 52 vs. 2x: 66 +/- 6.2 min/90 min of grazing). Cows milked 1x had slightly longer strides than 2x cows at mid lactation (1x: 0.73, 2x: 0.70, and TRANS: 0.70 +/- 0.014 strides/m). In conclusion, under our experimental conditions, we found no overt changes in behavior or physiology to indicate that the welfare of cows milked 1x is impaired.

Yields and persistency of lactation in Friesian and Jersey cows milked once daily.

Effects of milking cows once daily throughout lactation at high stocking rates (17% more cows/ha than for those milked twice daily) were studied in 2 Friesian and 2 Jersey herds during 3 lactations. Cows were allocated to 2 herds within breed and were milked either once or twice daily, based on age, genetic merit, and previous performance. Cows remained in their original herd and were milked at the same milking frequency during all lactations. Culled cows (20% per year) were replaced by 2-yr-old heifers. Yields of milk, lactose, protein, and fat were measured every 2 wk by commercial herd test. Cubic splines (5 knots) were used to approximate the lactation curve for each cow-yr to provide estimates of performance for each day of lactation. Yields of milk were greater for Friesian and Jersey cows milked twice daily (4,751 +/- 89 and 3,067 +/- 81 kg/cow) than for cows milked once daily (3,329 +/- 80 and 2,431 +/- 75 kg/cow), respectively. Cows milked once daily had lesser total and peak yields of milk, lactose, protein, and fat than cows milked twice daily. Friesians had greater total and peak yields than Jerseys. Peak production for all milk components occurred earlier in lactation for cows milked once daily than twice daily (d 24 to 39 vs. 32 to 44). Three measures of persistency of lactation were considered for each cow with 2 measures (Pers1 and Pers2) indicating that cows milked twice daily had better persistency than those milked once daily. Ranking of herds in persistency tended to match the ranking based on total yields. Measures of persistency (Pers1 and Pers2) were positively related to total yield in the Jerseys milked once daily and negatively related to peak yield in the Friesians. The third persistency measure (Pers3) ranked once-daily Jerseys first and twice-daily Friesians last, and was negatively correlated with total yield in the Friesian herds and negatively correlated with peak yield in all herds. For most performance measures, cows milked twice daily had better total yields and persistency than cows milked once daily.

The safety and immunogenicity of Bacillus Calmette-Guérin (BCG) vaccine in European badgers (Meles meles).

European badgers (Meles meles) are a wildlife reservoir for Mycobacterium bovis (M. bovis) in Great Britain (GB) and the Republic of Ireland and therefore constitute a potential source of infection for cattle. Reduction of badger densities in the Republic of Ireland has resulted in an associated reduction in the risk of a herd break-down with bovine tuberculosis and a study to determine whether this is also the case in GB has been running since 1997. If badgers are a significant source of M. bovis infection for cattle, vaccinating badgers with Bacillus Calmette-Guérin (BCG) might prove to be a long term, cost-effective strategy for controlling bovine tuberculosis whilst preserving badger populations. As a first step towards BCG vaccination of wild badgers, it was necessary to demonstrate safety of the vaccine in captive badgers. Therefore, captive badgers were vaccinated with a commercial source of BCG that is already licensed for administration to humans in GB-BCG Danish SSI. Using a protocol prescribed by the Veterinary Medicines Directorate (VMD) of GB, badgers were vaccinated with two consecutive doses of BCG via either the subcutaneous (s.c.) or intra-muscular (i.m.) routes. The first dose was high, ranging from 16 to 22 x 10(7) colony-forming units (CFU), and was followed 15 weeks later by a lower dose in the range of 4-7 x 10(5)CFU. Local reaction at the site of injection and general responses (body temperature, haematology and blood serum chemistry), behaviour and excretion of BCG were monitored for 28 weeks from the time of the first vaccination. The only side-effect observed was the occurrence of localised swelling at the site of BCG injection that disappeared 48 days after i.m. vaccination but persisted longer in the group vaccinated by the s.c. route. Immunological responses were measured at regular intervals. Strong cellular responses were observed 13 days after the first vaccination, which persisted for 76 days. The lower dose induced a weaker and shorter-lived response.

A systems comparison of once- versus twice-daily milking of pastured dairy cows.

The objective of this study was to compare the effect of milking frequency (once vs. twice-daily milking) and breed (Holstein-Friesians vs. Jerseys) on milk and milk solids (MS; milk fat + milk protein), yield per cow, milk composition, somatic cell count and lactation length; cow body weight, body condition score, and reproductive performance over a 4-yr period. Total cow numbers in each herd were 30, 35, 36, and 42 for Holstein-Friesians milked once or twice daily, and Jerseys milked once or twice daily, respectively. Forty hectares of pasture were subdivided into 4 smaller pastures of 10 ha each. Stocking rates for the once-daily herds were 16.7% greater than the twice-daily herd in their respective breed. An increased stocking rate was chosen to achieve equal milk and MS per ha from the 2 milking frequencies. Annual milk, fat, protein, and lactose yields per cow were less for once-daily than for twice-daily milking. Interactions were detected between milking frequency and breed for annual milk, fat, protein, and lactose yields per cow, because Jerseys were relatively less affected by once-daily than by twice-daily milking than Holstein-Friesians. Holstein-Friesian cows milked once daily produced 31.2% less milk and 29.4% less MS per cow than their twice-daily counterparts. In contrast, Jersey cows milked once daily produced 22.1% less milk and 19.9% less MS per cow than their twice-daily counterparts. Milk per ha was 17.7 and 9% less for the once-daily Holstein-Friesians and once-daily Jersey herds, respectively, compared with their twice-daily counterparts, because the greater stocking rate for the once-daily herds did not fully compensate for the milk loss per cow. Milking once daily increased somatic cell count throughout the year in both breeds. Cows milked once daily conceived 3 d earlier, took 5 d less from calving to conception, and needed 11% fewer controlled internal drug release devices than those milked twice daily. Milking once daily is a viable milking option for New Zealand farmers who are prepared to trade-off loss of MS income for increased time to accomplish other non-milking activities.

Fertility- and menopause-related information needs of younger women with a diagnosis of early breast cancer.

PURPOSE: The use of chemotherapy and endocrine therapies in the treatment of premenopausal women carries with it reproductive and gynecologic implications that young women may find distressing and discordant with plans for childbearing. This multicenter study aimed to investigate fertility- and menopause-related information needs among young women with a diagnosis of early-stage breast cancer. PATIENTS AND METHODS: Two hundred twenty-eight women with a diagnosis of early-stage breast cancer who were aged 40 years or younger at diagnosis and who were 6 to 60 months after diagnosis were entered onto the trial. Participants completed a mailed self-report questionnaire that included a purposely designed fertility- and menopause-related information needs survey and standardized measures of distress, anxiety, quality of life, menopausal symptoms, and information-seeking style. RESULTS: Seventy-one percent of participants discussed fertility-related issues with a health professional as part of their breast cancer treatment, and 86% discussed menopause-related issues. Consultation with a fertility or menopause specialist was the most preferred method of obtaining this information. Receiving fertility-related information was rated as being significantly more important than receiving menopause-related information at time of diagnosis (P < .001) and at treatment decision making (P = .058). Receiving menopause-related information was rated as being significantly more important than receiving fertility-related information during adjuvant treatment (P < .05), at completion of adjuvant treatment (P < .001), and during follow-up (P < .001). Common questions, sources of information, and correlates of perceived importance were identified. CONCLUSION: The results of this study suggest that younger women have unmet needs for fertility- and menopause-related information and provide preliminary empirical data to guide the development of better fertility- and menopause-related patient education materials for younger women with a diagnosis of early breast cancer.

Cloning and sequencing of badger (Meles meles) interferon gamma and its detection in badger lymphocytes.

The European badger (Meles meles) has been identified as a reservoir for Mycobacterium bovis and is implicated in the maintenance and transmission of tuberculosis in cattle. There is a need for a sensitive test of M. bovis infection in badgers and the current serodiagnostic test used for this purpose has low sensitivity. As observed for other species, assay of interferon-gamma (IFNgamma) produced in response to M. bovis antigens is a more sensitive test of tuberculosis. With this objective in sight, we report the first step in the development of an ELISA for badger IFNgamma. The badger IFNgamma gene was cloned and sequenced and used to generate a specific polyclonal antibody to the cytokine. The gene sequence demonstrated regions that were conserved within the IFNgamma genes of other mammals. The badger sequence was most similar to the canine, showing similar structural organisation of the gene and 88% amino acid identity. Rabbits were immunised with DNA encoding badger IFNgamma and the resulting polyclonal antiserum demonstrated specificity for canine IFNgamma by immunoblot of a commercial recombinant canine IFNgamma. The antiserum was used to detect intracellular badger IFNgamma by flow cytometry analysis of badger lymphocytes stimulated with mitogen.

A low dietary cation-anion difference precalving and calcium supplementation postcalving increase plasma calcium but not milk production in a pasture-based system.

It was hypothesized that a reduction in the dietary cation-anion difference (DCAD) before calving, combined with an increase in Ca intake after calving, would reduce the incidence of periparturient hypocalcaemia and increase milk production in pasture-based dairy cows. Cows (n = 40) were assigned to one of two DCAD levels before calving (i.e., +7 and +50 mEq/100 g). Each group was then assigned to one of two dietary Ca concentrations after calving (i.e., 1.0 and 0.7%) in a 2 x 2 factorial design. The lower DCAD resulted in a nonrespiratory reduction in systemic pH as indicated by a lower urine pH. This acidosis resulted in an increased concentration of Ca in urine before calving. The lower precalving DCAD helped prevent the decline in blood Ca caused by the onset of lactation, even though blood Ca concentration was lower before calving compared with cows receiving a high DCAD. Supplementation of cows with Ca after calving increased plasma Ca concentration on the day of calving and during the subsequent 14 d. Milk production was not affected by pre- or postcalving treatments.

Dietary cation-anion difference and the health and production of pasture-fed dairy cows. 1. Dairy cows in early lactation.

Diets offered to lactating dairy cows in the pasture-based dairy systems in southeastern Australia can vary in their dietary cation-anion difference (DCAD) from 0 to +76 mEq/100 g. The effects of such a range of DCAD on the health and production of cows, on a predominantly pasture-based diet, were examined in an indoor feeding experiment. Four groups of five cows were offered a diet of 5 kg of barley and ad libitum pasture, which is a diet representative of what is offered to cows in early lactation in the region. The cows were supplemented twice daily, with varying levels of salt combinations to alter the DCAD, which ranged from +21 to +127 mEq/100 g. Although a reduction in DCAD to +21 mEq/100 g caused a nonrespiratory systemic acidosis, there was a threshold value, above which blood and urine pH did not appear affected, although the strong ion difference of blood and urine and the blood bicarbonate concentration increased linearly (P < 0.05, 0.001, and 0.01, respectively). A DCAD above +21 mEq/100 g linearly reduced dry matter intake (P < 0.1), average daily bodyweight gain (P < 0.05), and milk protein yield (P < 0.05) but did not have a significant effect on the concentration of fat, protein, or lactose in milk. Although data were consistent with a tendency for milk yield to decrease as dietary cation-anion differences increased, this trend was not statistically significant. Urine hydroxyproline to creatinine ratio increased (P < 0.001) as dietary cation-anion difference increased, possibly suggesting an increased rate of uterine involution. It is concluded that a range in the dietary cation-anion difference, above +52 mEq/100 g, may have deleterious effects on dry matter intake and milk production.

Dietary cation-anion difference and the health and production of pasture-fed dairy cows 2. Nonlactating periparturient cows.

Anecdotal observations of reduced hypocalcemia due to small reductions in the precalving dietary cation-anion difference (DCAD) are widely reported in Australia and New Zealand. Diets offered to nonlactating, periparturient dairy cows in pasture-based dairy systems in southeastern Australia can vary in their cation-anion difference from 0 to +76 mEq/100 g. The effects of such a range in the DCAD on the health and production of cows, on a pasture-based diet, were examined in an indoor feeding experiment. Four groups of four cows were offered pasture-hay and freshly cut pasture, a periparturient diet typical of that associated with the grazing system in Australia and New Zealand. Varying levels of salt supplementation were used to alter the dietary cation-anion difference, which ranged from -12 to +69 mEq/100 g. Blood and urine pH and mineral concentrations and urine hydroxyproline were measured. The addition of anions to the diet, to produce a negative DCAD, resulted in a nonrespiratory systemic acidosis. With decreasing DCAD, the pH of blood and urine and the strong ion difference of urine decreased curvilinearly, blood bicarbonate decreased linearly and the urinary ratio of Ca to creatinine increased curvilinearly. Although systemic pH was not reduced at a DCAD of +16 mEq/100 g, urine Ca-to-creatinine ratio had begun to rise, probably indicating increased calcium absorption. The absorption and renal excretion of Mg increased with decreasing DCAD. No differences were observed in urine hydroxyproline concentrations and no significant differences in milk production were measured.

Progesterone clearance rate in lactating dairy cows with two levels of dry matter and metabolisable energy intakes.

The aim of these studies was to determine the effect of levels of dry matter (DM) and metabolisable energy (ME) intakes on clearance rate of progesterone (P4) in dairy cows. Thirty-two lactating Holstein-Friesian cows were selected for the study and were fed indoors in individual stalls for a period of 5 weeks. They were individually offered a diet of combinations of pasture, hay and pelleted cereal grain to achieve two different levels of DM and ME. In the first trial, 16 cows were allocated to two groups: (i) high DM (HDM), and (ii) low DM (LDM) intakes, while the amount of ME intake was constant. In the second trial, 16 cows were allocated to two groups: (i) high ME, and (ii) low ME intakes with similar amount of DM intake. A GnRH-agonist (deslorelin) was initially implanted in the ear of each cow to block endogenous P4 secretion. Then 3 weeks later, a CIDR device was inserted into the vagina of each cow and left in place for 11 days. Chromic oxide (Cr(2)O(3)) capsules were administered to allow daily faecal output (FO) to be estimated. Daily blood, faecal and milk samples were taken during the period of the experiment for P4 and faecal P4 metabolites analyses. Trial 1: The average milk yield was similar among cows in high and LDM intake groups (26.7 versus 25.0 l per day, P = 0.2). The average daily FO was 7.8 kg DM in the HDM and 5.7 in the LDM cows (P < 0.0001). Average daily DM intakes were 17.3 kg and 15.4 kg in the HDM and LDM groups, respectively (P < 0.0001). The average plasma P4 concentrations were similar between the two groups (1.56 versus 1.60 ng/ml, P = 0.7) but milk P4 concentrations were higher in LDM cows (4.6 versus 3.6 ng/ml, P = 0.02). The average daily excretion rate of P4 into the milk was higher in LDM cows (122.3 versus 88.5 microg, P = 0.002). The concentrations of faecal P4 metabolites (FP4M) were not influenced by the level of daily DM intake (2.85 versus 2.90 microg/g, P = 0.6). The average daily yields of FP4M were higher among cows in the HDM group (23.2 versus 16.3mg, P = 0.01). Trial 2: The average milk yield was 31.2l per day in HME cows compared to 25.0l per day in LME cows (P < 0.0001). The average daily FO was 7.8 kg DM in LME and 5.8 kg DM in HME cows (P < 0.0001), and the average DM content of faeces was higher in LME cows (15.8 versus 12.7%, P = 0.01). The average daily ME intake was 213MJ per day in HME group compared to 183MJ per day in LME group (P<0.0001). The average plasma and milk P4 concentrations were similar between the two groups (plasma P4 = 1.54 versus 1.56 ng/ml, P = 0.4; milk P4: 3.7 versus 3.6 ng/ml, P = 0.6). The average daily excretion rate of P4 into the milk was higher in HME cows (114 versus 88.5 microg, P = 0.03). Concentrations of FP4M were not influenced by the level of daily ME intake (2.5 versus 2.85 micro g/g, P = 0.08). However, daily yields of FP4M were greater in the LME group (23.2 versus 14.4 mg, P = 0.01). In conclusion, this study was unable to establish a relationship between the level of DM and ME in the diet with the excretion rates of FP4M metabolites and plasma P4 concentrations.

Immunological responses of Eurasian badgers (Meles meles) vaccinated with Mycobacterium bovis BCG (bacillus calmette guerin).

Wildlife species, such as the badger (Meles meles), may act as maintenance hosts for Mycobacterium bovis and contribute to the spread and persistence of tuberculosis in associated cattle populations. Targeted vaccination of badgers against tuberculosis is an option that, if successfully employed, could directly facilitate the advancement of bovine tuberculosis eradication in affected areas. In this study, the immunological responses of a group of badgers vaccinated subcutaneously with low doses of Mycobacterium bovis bacillus calmette guerin (BCG) were measured in vitro and compared with non-vaccinated control animals over a period of 42 weeks. Peripheral blood mononuclear cells (PBMC) from badgers which had received repeated booster injections of BCG proliferated in response to culture with PPD-bovine (purified protein derivative of tuberculin). The proliferation was significantly greater than that seen in the non-vaccinated control group. In contrast, the proliferative response of PBMC from vaccinated badgers to PPD-avian declined relative to the control group. These results demonstrate that repeated vaccination of badgers with M. bovis BCG induced a population of T-lymphocytes responsive to specific antigens in PPD-bovine. Throughout the course of the study, the sera from all animals were tested (BrockTest) by an enzyme-linked immunosorbent assay (ELISA) system for the presence of antibodies to MPB83, a serodominant antigen whose expression is high in M. bovis, but very low in BCG (Pasteur). No animals at any stage showed seroconversion to the antigen, consistent with the tuberculosis-free status of the badgers under study.

Open label, multi-centre phase II study of raltitrexed ('Tomudex') in patients with inoperable squamous-cell carcinoma of head and neck.

BACKGROUND: Raltitrexed ('Tomudex') is a folate based inhibitor of thymidylate synthase which has been registered in Europe and Australia for the treatment of advanced colorectal cancer. In a European phase I trial of raltitrexed anti-tumour activity was seen in two patients with head and neck cancer, prompting the current study. PATIENTS AND METHODS: From November 1996 to December 1998, 24 patients with metastatic or recurrent squamous-cell carcinoma of the head and neck from 7 Australian centres received raltitrexed, 3 mg/m2 given intravenously over 15 minutes every 3 weeks, for a maximum of 6 cycles. Patients were required to be chemotherapy naïve and have measurable disease, age > 18 years, WHO performance status initially < or = 2, no significant intercurrent illness or organ dysfunction and a life expectancy > 12 weeks. RESULTS: Twenty-two men and two women, median age 65 years, median performance status 1 were enrolled. Fifteen patients (63%) had received both prior surgery and radiotherapy. In 15 patients (63%) there was recurrent locoregional disease only. Twelve patients (50%) received one cycle of treatment with only four patients (17%) receiving four or more cycles of treatment. No patient achieved a complete or partial response, although 5 patients experienced stable disease which lasted a median of 188 days (range 61-436). The median survival for the whole group was 101 days (range 20436). Raltitrexed was generally well tolerated with minimal antiproliferative toxicity. CONCLUSIONS: Single-agent raltitrexed does not demonstrate significant anti-tumour response rates in patients with predominantly locally recurrent head and neck cancer.

Capecitabine, an oral fluoropyrimidine carbamate with substantial activity in advanced colorectal cancer: results of a randomized phase II study.

PURPOSE: To evaluate in patients with advanced colorectal cancer (CRC) three treatment regimens of oral capecitabine in order to select the most appropriate regimen for testing in phase III. PATIENTS AND METHODS: Three capecitabine schedules were evaluated in a randomized phase II design: arm A, 1,331 mg/m(2)/d bid continuously; arm B, 2,510 mg/m(2)/d bid intermittently (2 weeks on/1 week off); and arm C, 1,657 mg/m(2)/d plus oral leucovorin 60 mg/d bid intermittently (2 weeks on/1 week off). RESULTS: One hundred nine patients were randomized; 39 patients were assessable for efficacy in arm A, 34 in arm B, and 35 in arm C. Patient characteristics were balanced in the arms. Confirmed tumor responses (partial response [PR] + complete response [CR]) were reported for eight patients with two CRs (21%; 95% confidence interval [CI], 9% to 36%) in arm A, eight patients with one CR (24%; 95% CI, 11% to 41%) in arm B, and eight patients with two CRs (23%; 95% CI, 10% to 40%) in arm C. Median times to progression (TTP) in arms A, B, and C were 127, 230, and 165 days, respectively. Overall, more toxicity was seen with capecitabine plus leucovorin, particularly diarrhea and hand-foot syndrome. There was no grade 3 or 4 marrow toxicity. CONCLUSION: Capecitabine offers a new, effective treatment option as an oral single agent in advanced CRC. Promising overall response rates were reported for all three regimens. The addition of leucovorin to the intermittent regimen had no marked effect on tumor response or median TTP. The intermittent single-agent capecitabine schedule is proposed for phase III evaluation, based on considerations of toxicity, dose-intensity, response rate, and TTP.

Phase I clinical trial of the chimeric monoclonal antibody (c30.6) in patients with metastatic colorectal cancer.

The murine antibody 30.6 recognizes an antigen that is expressed on a high proportion of colorectal carcinomas and their metastases. We report the results of single-dose escalation studies of the chimeric 30.6 (c30.6) monoclonal antibody in metastatic colorectal cancer, to evaluate its safety, pharmacokinetics, and biodistribution. Recombinant c30.6 (IgG1kappa) antibody was secreted from Chinese hamster ovary cells and purified by a multistep chromatography process. Seventeen patients with metastatic colorectal cancer were enrolled in this dose escalation study. The first four patients were treated with 3 mg of 123I-labeled c30.6, whereas the next 13 received a single dose of unlabeled antibody (maximum dose, 50 mg/m2). The most frequent side effect was a novel syndrome of severe burning and erythema of the face, chest, neck, ears, palms, soles, and genitalia. The frequency of this syndrome was markedly reduced in those patients premedicated with high doses of histamine receptor 1 and histamine receptor 2 blockers. Other side effects were mild and predictable. Biodistribution studies showed a rapid and intensive hepatic uptake. At the 50 mg/m2 level the half-life and maximum serum concentration were 81 +/- 15 h and 7.9 microg/ml, respectively. One patient developed a low-level human anti-c30.6 response. Tumor response was assessed by computed tomography, positron emission tomography scanning, and serial carcinoembryonic antigen measurements. There were no partial responses, although positron emission tomography scanning demonstrated some reduction in tumor activity in three individuals. The chimerized c30.6 antibody is not immunogenic in humans and appears worthy of further study. It does, however, produce a unique profile of side effects that can be well controlled with premedication.