Prognostic significance of aromatase and estrone sulfatase enzymes in human breast cancer

The aromatase and estrone sulfatase enzymes are important sources of local synthesis of biologically active estrogens in human breast cancer. Significant intratumoral aromatase activity was detected in 91/145 (63%) of tumors and estrone sulfatase was detected in 93/104 (89%) of tumors. There was no relationship between aromatase activity and tumor size, site, nodal status, menopousal status or estrogen receptor status. There was a significant correlation between the aromatase activity and histological grade, with an excess of aromatase-positive in the high grade tumors (P = 0.03). There was a marginally inverse correlation between the aromatase activity and time to relapse (P < 0.1), a significant correlation between aromatase activity and survival after relapse (P < 0.05) but not with overall survival (P < 0.1). Intratumoral estrone sulfatase activity was not significantly correlated to any putative prognostic factors, nor with time to relapse nor overall survival time.     

Surgical castration of pigs affects the behavioural response to a low-dose lipopolysaccharide (LPS) challenge after weaning

The objective of the present study was to evaluate the effect of an acute stressor in the early postnatal life of pigs, surgical castration, on post-weaning behaviour, and on the behavioural, endocrine and immune responses elicited by a low-dose lipopolysaccharide (LPS) challenge after weaning. At 5-days-of-age, 64 male piglets were randomly assigned to undergo surgical castration or were left untreated (treatment). Pigs were weaned at 28 days-of-age. Behaviour post-weaning and mixing was assessed during a 1-h period, during which agonistic interactions were recorded. One day post-weaning, pigs were injected intraperitoneally with a single dose of 0 or 5 μg/kg of BW of LPS from Escherichia coli (challenge). Sickness behaviour was studied by scan sampling every 5 min for 45 min at 0, 1, 2, 3, 4, 6 and 8 h after the challenge. Blood samples were taken at 0, 2, 12 or 24 h after injection and were analysed for plasma concentrations of tumor necrosis factor-alpha (TNF-), interleukin-1beta (IL-1β), C-reactive protein (CRP), serum amyloid A (SAA) and cortisol. Results showed that non-castrated pigs were more aggressive than castrated pigs immediately after weaning (P < 0.05). Administration of LPS provoked behaviours characteristic of sickness including a reduction in general activity, as well as decreased eating and exploratory behaviours (P < 0.05). These altered behaviours occurred predominantly 3-h post injection (P < 0.05). Significant treatment by challenge interactions showed that castration reduced the occurrence of sickness behaviours induced by LPS, such as depressed general activity (P < 0.01), anorexia (P < 0.01) and reduced exploratory behaviours (P < 0.05). LPS administration increased TNF- levels (P < 0.05), with peak concentrations 2 h after injection (P < 0.01). CRP levels of LPS-treated pigs were higher than saline-treated animals at 12 h (P < 0.05). LPS administration tended to increase plasma SAA levels (P < 0.1), but did not increase cortisol levels (P > 0.1). However, castration did not affect the response of pro-inflammatory cytokines, acute phase proteins and cortisol to the challenge. These results show that surgical castration reduces aggressiveness at weaning and affects specific sickness behaviours but not the endocrine and immune responses elicited by low-dose endotoxin challenge in weaned pigs.     

Sequential endocrine changes and behaviour during oestrus and metoestrus in repeat breeder and virgin heifers

The aim of the experiment was to study the oestrous behaviour and the peripheral blood plasma profiles of luteinizing hormone (LH), progesterone and the prostaglandin metabolite, 15-keto-13,14-dihydro-PGF₂, during oestrus and metoestrus in repeat breeder (RBH) and virgin heifers (VH). Ten animals of each category were utilized. The RBH had a history of at least three inseminations without conception, and the VH were sexually mature animals not previously inseminated or mated. Oestrous symptoms were recorded and blood collected from the onset of prooestrus to 7 days after oestrus. The animals were inseminated during oestrus and their embryos were collected by a nonsurgical technique 7 days after insemination. The morphology of the embryos was evaluated.

The duration of oestrus was longer (P < 0.05) in the RBH (31.5 ± 3.6 h) than in the VH (23.8 ± 2.0 h). No differences in duration of prooestrus or in the interval from the end of oestrus to postoestrous bleeding were found between the heifer categories. The interval from the onset of oestrus to the preovulatory LH peak was longer (P < 0.05) in the RBH (12.2 ± 2.8 h) than in the VH (4.8 ± 1.5 h). There was a lower LH release in the RBH than in the VH, measured as the magnitude of the preovulatory LH peak (P < 0.05; 28.0 ± 4.0 vs. 40.7 ± 3.6 μg/l) or as the area under the curve of the LH peak (P < 0.01; 1141 ± 164 vs. 1765 ± 144 mm²). The progesterone levels were higher (P < 0.05) in the RBH than in the VH during the interval 5–48 h and lower (P < 0.05) during the interval 121–168 h after the LH peak. Peaks of the prostaglandin metabolite were seen during oestrus in both heifer groups. There were more prostaglandin metabolite peaks in the RBH than in the VH during the interval 13–24 h after the LH peak. Fewer normal embryos (P < 0.05) and more degenerated embryos (P < 0.01) were found in the RBH than in the VH group 7 days after insemination. No apparent relation was found between the morphology of the embryos and the hormonal changes.

The result of the study indicates a hormonal imbalance in the RBH. The hormonal asynchronism starts before or early in oestrus, which presumably leads to a sequence of improper hormonal changes responsible for the elevated embryonic loss in repeat breeder animals.     

The effect of two teeth resection procedures on the welfare of piglets in farrowing crates. Part 1

Teeth resection is a method of controlling the injurious effects of the aggression displayed when newborn piglets fight to establish a teat order. Recent European legislation discourages the practice. The objective of this study was to assess the effect of clipping and grinding piglets’ needle teeth, compared to leaving them intact, on the welfare of piglets in farrowing crates.

Six days pre-partum, 60 sows were assigned to one of three treatments. Litters had their teeth clipped (C), ground (G) or left intact (I) at birth. The time taken to carry out each procedure was recorded. Piglet weights and facial lesions, which were scored according to severity, were recorded on days 1, 4, 11, 18 and 27. Piglet weights were also recorded at birth. Mouth lesions were recorded on days 1, 4 and 27. Instantaneous scan samples of piglet behaviour were carried out for 30 min post-teeth resection procedure (1 min intervals), and for 6 h on days 1, 4, 8, 14, 21 and 26 (5 min intervals). One male and one female piglet per litter were chosen as focal animals and observed for 5 min each post-procedure and for 10 min each twice per day on days 1, 5, 12, 20 and 26. Mortalities were recorded throughout lactation.

Grinding took significantly longer than clipping the teeth or leaving them intact (F = 638.87, P < 0.001). I piglets had higher facial lesion scores than C and G piglets (F = 10.58, P < 0.001). A smaller proportion of piglets in I litters than C and G litters and a smaller proportion of piglets in G litters than C litters had at least one mouth lesion (F = 4.74, P < 0.001). During 30 min post-procedure, I piglets were active on the heatpad in more observations than C and G piglets (F = 3.49, P < 0.05). During 5 min post-procedure C piglets spent longer chomping than I piglets (F = 5.92, P = 0.05). On day 21, I piglets were active in more observations than G piglets (F = 2.11, P < 0.05). On day 26, G piglets were inactive in more observations than C and I piglets (F = 5.02, P < 0.05). On days 14 and 26, C piglets were sleeping in more observations than G piglets (F = 2.87, P = 0.05). There was a tendency for a larger proportion of I than C piglets to die due to overlying (F = 2.68, P = 0.08).

In conclusion, although all three options were associated with welfare problems, grinding can be recommended in preference to clipping or leaving the teeth intact.     

P450-dependent enzymes as targets for prostate cancer therapy

Metastatic prostate adenocarcinoma is a leading cause of cancer-related deaths among men. First line treatment is primarily aimed at blocking the synthesis and action of androgens. As primary endocrine treatment, androgen deprivation is usually achieved by orchidectomy or LHRH analogues, frequently combined with androgen receptor antagonists in order to block the residual adrenal androgens. However, nearly all the patients will eventually relapse. Available or potential second line therapies include, among others, alternative endocrine manipulations and chemotherapy.

Cytochrome P450-dependent enzymes are involved in the synthesis and/or degradation of many endogenous compounds, such as steroids and retinoic acid. Some of these enzymes represent suitable targets for the treatment of prostate cancer.

In first line therapy, inhibitors of the P450-dependent 17,20-lyase may achieve a maximal androgen ablation with a single drug treatment. Ketoconazole at high dose blocks both testicular and adrenal androgen biosynthesis but its side-effects, mainly gastric discomfort, limit its widespread use. A series of newly synthesized, more selective, steroidal 17,20-lyase inhibitors related to 17-(3-pyridyl)androsta-5,16-dien-3β-ol, may open new perspectives in this field.

In prostate cancer patients who relapse after surgical or medical castration, therapies aiming at suppressing the remaining adrenal androgen biosynthesis (ketoconazole) or producing a medical adrenalectomy (aminoglutethimide + hydrocortisone) have been used, but are becoming obsolete with the generalization of maximal androgen blockade in first line treatment. The role of inhibition of aromatase in prostate cancer therapy, which was postulated for aminoglutethimide, could not be confirmed by the use of more selective aromatase inhibitors, such as formestane.

An alternative approach is represented by liarozole fumarate (LIA), a compound that blocks the P450-dependent catabolism of retinoic acid (RA). In vitro, it enhances the antiproliferative and differentiation effects of RA in cell lines that express RA metabolism, such as F9 teratocarcinoma and MCF-7 breast carcinoma cells. In vivo, monotherapy with LIA increases RA plasma levels and, to a greater extent, endogenous tissue RA levels leading to retinoid-mimetic effects. In the rat Dunning prostate cancer models, it inhibits the growth of androgen-independent as well as androgen-dependent carcinomas relapsing after castration. Concurrently, changes in the pattern of cytokeratins characteristic of increased differentiation were observed. Early clinical trials show that LIA, in second or third line therapy in metastatic prostate cancer, induces PSA responses in about 30% of unselected patients. In some patients regression of soft tissue metastasis has been observed. In a subgroup of patients, an important relief of metastatic bone pain was also noted.     

Efficiency of asbestos shading for growth of Barki rams during hot summer

Twenty-four growing Barki rams, 5–6 months old with an average body weight of 22 kg, were divided into asbestos shaded and unshaded groups during summer. Ad libitum feeding on roughage with 0.5 kg of concentrates per head per day was practiced. Drinking water was available twice daily in the early morning and afternoon. Biweekly observations included rectal temperature (RT; °C), and respiration rate (RR; breaths per minute) at 06:00 h and 14:00 h were recorded. Meteorological data, plasma glucose, calcium, inorganic phosphorus and packed cell volume were also measured. Results indicated that experimental animals developed hyperthermia during June to September, as evidenced by higher (P < 0.01) RT and RR than normal (40 ± 0.05 and 103.9 ± 3.87 vs. 38.9 ± 0.10 and 40 ± 6.56, respectively). Diurnal variations in these physiological phenomena were closely associated (P < 0.05) with ambient temperature changes. As compared with the unshaded group, providing an asbestos shed reduced (P < 0.05) RT and RR in the hyperthermic animals during the day (39.9 ± 0.07 and 94.7 ± 3.75 vs. 40.1 ± 0.08 and 113.1 ± 4.74, respectively), but it increased (P < 0.05) these parameters during summer nights (39.5 ± 0.05 and 82.4 ± 0.95 vs. 39.2 ± 0.07 and 71.6 ± 2.41, respectively). It also increased (P < 0.05) packed cell volume 1 h after morning drinking (35 ± 0.97 vs. 33.2 ± 0.60); reduced (P < 0.05) plasma glucose (43.3 ± 5.88 vs. 53.2 ± 6.31); caused hypocalcemia (10.9 ± 0.35 vs. 11.5 +- 0.43; P < 0.05) and hypophosphatemia (3.9 ±0.35 vs. 4.6 ± 0.34; P < 0.05) as a result of hyperthermia. Monthly variations in all parameters studied were higher (P < 0.01) during early summer. It is concluded that providing an asbestos shed for growing Barki rams under Egyptian summer conditions will not protect the animals from hyperthermia by day and night, as it interferes with extra body heat dissipation to the surroundings during summer nights. Although the unshaded animals were more hyperthermic during summer days, they tended to be normal during the night.     

Effects of low testosterone levels and of adrenal androgens on growth of prostate tumor models in nude mice

Two transplantable, androgen dependent prostate tumor models of human origin, PC-82 and PC-EW, were used to study the effect of low androgen levels and adrenal androgens on prostate tumor cell proliferation. Tumor load of the PC-82 and PC-EW tumors could be maintained constant when plasma testosterone levels were 0.8 and 0.9 nmol/l, respectively, corresponding with an intratissue 5-dihydrotestosterone level of 3–4 pmol/g tissue. This critical androgen level for prostate tumor growth stimulation amounted to 2–3 times the castration level and proved to be similar for both tumor models. Relatively high levels of androstenedione resulted in physiological levels of plasma testosterone causing androgen concentrations in PC-82 tumor tissue exceeding the critical level for tumor growth. These results indicate that submaximal suppression of androgens can stop tumor growth in these prostate tumor models.     

Testosterone increases acetylcholine receptor number in the “levator ani” muscle of the rat

The “levator ani” muscle of male rats provides a neuromuscular system in which both the muscle and its motoneurons have high levels of androgen receptors. Two weeks of castration caused a 48% loss of acetylcholine receptors in this muscle. One week of testosterone propionate injections initiated one week after castration increased receptor number by 27% over untreated castrate levels. These changes paralleled changes in muscle protein content. In contrast, castration and testosterone treatments of castrates had no effect on total, Triton X-100-extractable acetylcholinesterase activity. This system may provide a useful model of synaptic plasticity.     

Prognostic value of pS2 protein and receptors for epidermal growth factor (EGF-R), Insulin-like growth factor-1 (IGF-1-R) and somatostatin (SS-R) in patients with breast and ovarian cancer

The prognostic value of EGF-R, IGF-1-R and SS-R, and of cytosolic estrogen-regulated pS2 protein, was studied in patients (pts) with primary breast and advanced ovarian cancer. Ovarian cancer tissues were negative for pS2 (by immunoradiometric assay) IGF-1-R and EGF-R contents (by ligand binding assay, LBA) were of no or moderate prognostic value for breast cancer pts (n = 214). For advanced ovarian cancer pts, EGF-R content determined by LBA (n = 55) showed no prognostic value, whereas EGF-R status (n = 55) determined by immunohistochemistry (MoAb 2E9) signiificantly correlated with progression of disease (P < 0.05). In breast cancer pts, both SS-R and pS2 showed no association with tumor size, nodal status and grade. For pS2 the best cut-off level with respect to relapse-free (RFS) and overall survival (OS) was found to be 11 ng/mg protein. Both SS-R (1 g% SS-R+, n = 135; P < 0.04) and pS2 (27% pS2+, n = 197; P < 0.001), which were mainly positive in ER+ tumors, were of prognostic value, especially within the subgroups with ER+/PgR+ tumors. Also within N+ and No pts the 5-yr RFS and OS showed a difference between pS2+ and pS2- (33 and 54% for N+, and 31 and 13% difference for No pts). In summary, SS-R and pS2 are valuable pronosticators in breast cancer pts, and prognostic significance of EGF-R in ovarian cancer pts needs further study.     

The effect of two piglet teeth resection procedures on the welfare of sows in farrowing crates. Part 2

Recent EU legislation discourages the practice of resecting piglets’ needle teeth. However, the effect of leaving piglets’ teeth intact on the welfare of sows in farrowing crates is poorly understood. Therefore, the aim of the study was to compare the effects of clipping and grinding piglets’ needle teeth, compared to leaving them intact, on the welfare of sows in farrowing crates.

Six days pre-partum 60 multiparous sows were assigned to one of three treatments. Litters had their teeth clipped (C), ground (G) or left intact (I) at birth. Sows’ teats were inspected for lesions pre-partum (day −3) and on days 1, 4, 11, 18 and 27 post-partum. Instantaneous scan samples (5 min intervals) of sow behaviour were carried out during three 2 h periods on days 1, 4, 8, 14, 21 and 26. On days 1, 4 and 11 all piglets were removed from the crate for 60 min. On re-introduction of the piglets, sow maternal behaviour was recorded continuously for 20 min.

The number of sows with teat lesions tended to differ between treatments on days 11 (P = 0.06) and 18 (P = 0.10). There was an interactive effect between treatment and day on sow dog-sitting behaviour throughout lactation (P < 0.001) and a tendency for an interactive effect on posture-changing behaviour (P = 0.08). On day 21, I sows were dog-sitting in more observations than C sows (P < 0.05) and on day 26 in more observations than C and G sows (P < 0.001). There was an interaction between treatment and day in the latency of sows to suckle their piglets following 60 min separation (P < 0.05). On day 4, I sows had a shorter latency to suckle than C and G sows (P < 0.05). There was an effect of treatment on the number of sows that terminated bouts of post-suckling udder massage (P < 0.05). On day 4, more I than C sows terminated post-suckling udder massage (P = 0.01). Finally, there was an effect of treatment on the time spent lying in the ventral posture in the observations of maternal behaviour (P < 0.05). C sows spent less time lying in the ventral posture than G and I sows (P < 0.05).

There were indications that leaving the teeth intact and to a lesser extent grinding caused injury and disturbance to sows. In farrowing crates, leaving piglets’ teeth intact cannot be recommended.     

Treatment of newly diagnosed state D2 prostate cancer with leuprolide and flutamide or leuprolide alone, phase III, intergroup study 0036

In order to test the hypothesis of complete androgen blockade for advanced prostate cancer (D₂CaP), an intergroup trial was instituted in 1985 comparing leuprolide (L) alone to the combination of L with flutamide (F). Eligibility requirements included previously untreated histologically confirmed stage D₂CaP, measurable bone or soft tissue metastases, performance status (PS) of 3 or better, acceptable renal and hepatic function, no severe cardiac disease, and no prior or concomitant endocrine therapy. Stratification at entry was on the basis of PS and none or minimal disease (MD) versus severe degree (SD) of bone metastases. Six hundred and seventeen patients were entered into this study between March 1985 and April 1986. At the present time, there is a 3-month difference in the median progression-free survival (13.9 vs 16.9 months; P = 0.039) and a 7.1-month difference in survival (27.9 vs 35.01 months; P = 0.035) favoring L + F. In L + F-treated patients with good PS-MD, the median survival recently has been reached and is 51.9 months vs 39.6 months for L + P patients. The 107 black patients in the study had median survival of 26.4 months vs 33.3 months for whites. Discussions of racial differences in survival as well as other prognostic factors will be presented. The combination of L + F is superior to treatment with L alone. The benefits appear greatest in patients with minimal disease.     

CELL POPULATION GROWTH IN THE CASTRATE MOUSE PROSTATE COMPLEX: EXPERIMENTAL VERIFICATION OF COMPUTER SIMULATION

The stimulatory action of androgen on cell proliferation in the castrate mouse seminal vesicle and coagulating gland has been studied by DNA measurements in mice 14 days after castration. 100 hr after continuous androgen treatment the level of DNA had increased 2.5-fold in the seminal vesicle and coagulating gland compared with 14 days castrated controls. A mathematical model predicted this new experimental data when the parameters employed in the simulation were constrained by the results of previous experiments.     

The effects of farrowing nest size and width on sow and piglet behaviour and piglet survival

The ‘Werribee farrowing pen' (WFP) was developed as a loose housing alternative to the farrowing crate. The WFP occupies about twice the space of a crate and comprises two compartments, a ‘nest' and a ‘non-nest' area. In this experiment, we investigated the effects of reducing total pen space by modifying the dimensions of the ‘nest'. The hypothesis was that modifying ‘nest' size and width would not negatively affect piglet survival. A reduction in total floor space in the WFP may increase attractiveness for adoption of the system by pig producers. The experiment had a 2×2 factorial design with nine replicates and a total of 72 primiparous sows (Large White×Landrace) and their litters. All subjects were included to day 4 of lactation, but production data to weaning (day 23) was restricted to 36 litters. We examined the effects of ‘nest' size (large: L vs. small: S) and width (wide: W vs. narrow: N) on sow and piglet behaviours and piglet survival. The W, as compared to N treatment sows, had longer mean bouts of standing in the ‘nest' during 16–8 h pre-farrowing (3.7 vs. 2.0 min, P<0.01), suggesting that pre-farrowing (‘nesting') behaviour may have been adversely affected by narrow nest width. On the first 2 days of lactation, sow suckling grunts occurred more in the W than N treatment (14.1 and 8.0% of observations, P<0.01). However, this was due to an approximately 50% lower incidence in the SN treatment, as compared to the other treatments combined (5.9 and 12.7%). Piglets spent more (P<0.01) time at the udder in L vs. S (41 and 30%) and W vs. N nests (40 and 31%) and performed more (P<0.05) sucking behaviour (i.e., drinking milk from a teat) in L vs. S nests (2.5 and 1.2%). These differences were due to SN treatment litters sucking less than other treatments combined (0.9 vs. 2.2%). Of the 680 piglets born (n=72 litters), 3.8% were stillborn and 6.8% died between birth and day 4 of lactation. From day 5 to weaning (n=36 litters), 3.5% of piglets died, all due to Escherichia coli infection. While there were no effects of treatment on piglet survival, liveborn piglets were less likely (P<0.05) to die from E. coli in the L vs. S treatments (9/24 vs. 23/33 piglets). In conclusion, our results provide evidence that the size and width of the farrowing ‘nest' in a loose farrowing system may effect sow and piglet behaviours that may be relevant to piglet survival.     

Impact of circulating cholesterol levels on growth and intratumoral androgen concentration of prostate tumors

Prostate cancer (PCa) is the second most common cancer in men. Androgen deprivation therapy (ADT) leads to tumor involution and reduction of tumor burden. However, tumors eventually reemerge that have overcome the absence of gonadal androgens, termed castration resistant PCa (CRPC). Theories underlying the development of CRPC include androgen receptor (AR) mutation allowing for promiscuous activation by non-androgens, AR amplification and overexpression leading to hypersensitivity to low androgen levels, and/or tumoral uptake and conversion of adrenally derived androgens. More recently it has been proposed that prostate tumor cells synthesize their own androgens through de novo steroidogenesis, which involves the step-wise synthesis of androgens from cholesterol. Using the in vivo LNCaP PCa xenograft model, previous data from our group demonstrated that a hypercholesterolemia diet potentiates prostatic tumor growth via induction of angiogenesis. Using this same model we now demonstrate that circulating cholesterol levels are significantly associated with tumor size (R = 0.3957, p = 0.0049) and intratumoral levels of testosterone (R = 0.41, p = 0.0023) in LNCaP tumors grown in hormonally intact mice. We demonstrate tumoral expression of cholesterol uptake genes as well as the spectrum of steroidogenic enzymes necessary for androgen biosynthesis from cholesterol. Moreover, we show that circulating cholesterol levels are directly correlated with tumoral expression of CYP17A, the critical enzyme required for de novo synthesis of androgens from cholesterol (R = 0.4073, p = 0.025) Since hypercholesterolemia does not raise circulating androgen levels and the adrenal gland of the mouse synthesizes minimal androgens, this study provides evidence that hypercholesterolemia increases intratumoral de novo steroidogenesis. Our results are consistent with the hypothesis that cholesterol-fueled intratumoral androgen synthesis may accelerate the growth of prostate tumors, and suggest that treatment of CRPC may be optimized by inclusion of cholesterol reduction therapies in conjunction with therapies targeting androgen synthesis and the AR.     

A scanning electron-microscopic study of femoral bone surfaces from castrate rats treated with dichloromethylene biphosphonate

This study compared SEM appearances of endosteal and periosteal surfaces of anorganic femoral diaphyses from three groups of adult male rats: (1) control; (2) castrate (osteoporotic), and (3) castrate + injections of dichloromethylene biphosphonate (Cl2 entrances; (2) area of vascular canal entrances; (3) number of osteoblast lacunae, and (4) percent area of bone surface types. Endosteal surfaces from untreated osteoporotic rats had significantly more osteoblast lacunae, larger vascular canal entrances, and less resting surface than those of the other groups. There were no differences in endosteal surfaces between controls and Cl2MBP-treated castrates. Periosteal surfaces demonstrated no significant differences in percent area of bone surface types between groups, untreated castrates had more osteoblast lacunae than other groups, and vascular canal entrances were smaller in castrate groups than in controls. Castrates treated with Cl2MBP had significantly fewer periosteal vascular canal entrances, and many canal entrances and osteoblast lacunae appeared to be plugged with mineral deposits when compared with the other groups. The results indicate that Cl2MBP treatment prevented endosteal bone surface changes that occurred with castration osteoporosis, and may have disrupted normal vascularization on periosteal surfaces.     

Preferences of sheep for different types of pen flooring

In countries where the climate makes it practical with indoor housing during cold periods of the year and access to straw is limited, e.g. Iceland and Norway, housing of sheep on slats or expanded metal floors is common practice. However, European regulations for organic farming require that all animals should have access to a lying area with solid floor. The objective of this experiment was to investigate sheep preferences for different types of pen flooring.

In Experiment 1, a total number of 16 ewes, divided into four groups, were subjected to four different treatments. In each treatment, the ewes could choose between two lying areas with one of the following flooring materials: (1) solid wood versus rubber mats, (2) expanded metal versus solid wood, (3) solid wood versus straw, (4) expanded metal versus straw. In Experiment 2, a total number of eight ewes were individually subjected to the same treatments, first fully coated and then sheared. The ewes were video taped for 48 h in each treatment period.

In Experiment 1, the first animal that lay down after feeding preferred to lie down on straw or wood to expanded metal and straw to wooden floor (P < 0.05), but this first choice did not reflect the overall flooring preference for unsheared ewes. The groups of unsheared ewes showed no significant preferences for lying area.

In Experiment 2, single housed, unsheared ewes preferred wooden floor to rubber mats (P < 0.05), and tended to prefer expanded metal floor to straw (P = 0.08). There were no significant preferences in the two other treatments. After shearing, the ewes’ preferred wooden floor to expanded metal (P < 0.05), straw to wooden floor (P < 0.05), and straw to expanded metal floor (P< 0.0001). There were no significant preferences between rubber mats and wooden floor. Mean lying time (% of observations) for all treatments was 64.7% for unsheared ewes, and there were no significant differences between treatments. Mean lying time (% of observations) for all treatments the first 2–3 days after shearing was 43%. Significant differences in pre-shearing versus post-shearing lying times (% of observations) existed when the ewes were housed in pens with no straw (P < 0.05), this was not the case when the ewes had access to straw.

In conclusion, sheared but not unsheared ewes, preferred softer floors with low thermal conductivity (straw and wood). The less dramatically reduction in lying time (% of observations) after shearing when the ewes had access to straw, suggest that access to straw the first weeks after shearing may improve animal welfare.     

Effects of promethazine HCl on osteoporotic femora of adult castrated male rats

Promethazine HCl (Phenergan) was added to the drinking water (2.4-9.6 mg/kg/day) of castrated adult male Holtzman rats. One group of castrates and a group of normal rats received no drug and served as controls. Treatment was started at castration and continued for 4 months. After controlling for body weight, analyses of covariance and Student-Newman-Keuls tests revealed that promethazine did not prevent the development of femoral osteoporosis in the castrate rats. However, control rats (normal and untreated castrates) had significantly wider femora and thicker cortices at midshaft than did promethazine-treated animals. The results indicated that promethazine HCl had no effect on the development of femoral osteoporosis and retarded normal femoral expansion in the adult castrate male rats.     

Extraadrenal expression of steroid 21-hydroxylase and 11β-hydroxylase by a benign testicular leydig cell tumor

We present an unusual case with bilateral testicular Leydig cell tumors displaying extraadrenal expression of steroid 21-hydroxylase and 11β-hydroxylase. Histological examination of a 38-yr-old man infertile due to azoospermia showed him to have bilateral testicular Leydig cell tumors. The in vitro steroidogenic potential of the tumors and their adjacent testicular tissue was evaluated using organ culture. Tumor tissue was found to secrete deoxycorticosterone (DOC), corticosterone (B) and cortisol, which are not produced in normal adult testis, into the medium, while testicular tissue adjacent to the tumors secreted a small amount of DOC and B. Northern blot analysis with cytochrome P-450_C21 complementary DNA (cDNA) and P-450_11β cDNA as probes revealed that the tumor contained a considerable amount of mRNA for P-450_C21 and P-450_11β, while the mRNAs were not detected in the testicular tissues adjacent to the tumors. It is suggested that the high local levels of estrogen and/or progesterone within the Leydig cell tumors and their adjacent testicular tissues induced extraadrenal expression of steroid 21-hydroxylase and 11β-hydroxylase by the tumors and their adjacent testicular tissues.