[Water requirements, water supply and thermoregulation in small ruminants in pasture-based husbandry systems]. / Wasserbedarf, Wasserversorgung und Thermoregulation kleiner Wiederkäuer bei Weidehaltung.

Water is an essential source of life and is available to animals as free water, water content of feed, film water (e. g. dew) and metabolic water. The water requirements of small ruminants are influenced by the type of feed, climate, stage of production, type and length of the fleece or hair coat, husbandry factors and the general health of the animal. Differences in water metabolism, drinking behaviour and the efficiency of temperature regulation are further influenced by species, breed, production type, husbandry system, acclimatisation and adaptation. Small ruminants have been, and are still predominantly kept in extensive husbandry systems. They are therefore genetically and phenotypically well adapted to these conditions and possess a range of physiological and behavioural mechanisms to deal with adverse and suboptimal weather conditions. Regarding animal welfare, there is considerable debate in the discussion and assessment of what constitutes a sufficient water supply for small ruminants under different husbandry conditions, often involving differences between theoretical demands and practical experience. This publication reviews and summarises the current literature regarding water requirements, water metabolism and thermoregulatory mechanisms of small ruminants to provide the basis for an informed assessment of extensive husbandry systems in terms of compliance with animal-welfare requirements.

[Influence of lameness caused by hoof disease on welfare in sheep and goats. A comment on the guidelines of the German Small Ruminant Veterinary Association]. / Der Einfluss von Klauenerkrankungen auf das Tierwohl von Schaf und Ziege. Ein Kommentar zu den Haltungsempfehlungen der DVG.

The publication of the "Recommendations of husbandry and welfare of sheep and goats" by the German Small Ruminant Veterinary Association provides guidelines that are also used for assessing animal welfare. The included statements concerning lameness are reviewed according to the available literature and commented on with the help of practical examples. Diagnostic tools, criteria, appropriate measures and target values are discussed to facilitate the application of the guidelines.

The future is now: early life events preset adult behaviour.

To consider the evidence that human and animal behaviours are epigenetically programmed by lifetime experiences. Extensive PubMed searches were carried out to gain a broad view of the topic, in particular from the perspective of human psychopathologies such as mood and anxiety disorders. The selected literature cited is complemented by previously unpublished data from the authors' laboratories. Evidence that physiological and behavioural functions are particularly sensitive to the programming effects of environmental factors such as stress and nutrition during early life, and perhaps at later stages of life, is reviewed and extended. Definition of stimulus- and function-specific critical periods of programmability together with deeper understanding of the molecular basis of epigenetic regulation will deliver greater appreciation of the full potential of the brain's plasticity while providing evidence-based social, psychological and pharmacological interventions to promote lifetime well-being.

DNA memories of early social life.

The foundations of brain architecture are established early in life through a continuous series of dynamic interactions in which environmental conditions and personal experiences have a significant impact on how genetic predispositions are expressed. New scientific research shows that early social experiences can actually influence how genes are expressed. Thus, the old-school concepts that genes are "chiseled in stone" or that they alone determine development have been disproven. The discovery of the epigenome provides an explanation, at the molecular level, for why and how early positive and negative social experiences give rise to a biological memory that can have lifelong impacts. Signatures associated with the epigenome can be temporary or permanent, affect multiple organ systems, and increase the risk not only for poor physical and mental health outcomes but also for impairments in future learning capacity and behavior. Here, we focus on recent evidence for a role of epigenetic DNA modifications as a potential mechanism that explains how early social life experiences become embedded in the circuitry of the developing brain and are associated with lifelong consequences.

[Recommendations for the husbandry and welfare of sheep and goats by the German Small Ruminant Veterinary Association. Part 2]. / Empfehlung für die Haltung von Schafen und Ziegen der Deutschen Gesellschaft für die Krankheiten der kleinen Wiederkäuer, Fachgruppe der DVG. Teil 2.

The second part of the recommendations deals with the healthcare and the regulatory framework for the husbandry of sheep and goats. The suggested concept for healthcare aims to develop an individual health plan for every flock. This health plan focuses not only on the prevention of notifiable diseases, but also on chronic and slow infections as well as on parasite monitoring. The emphasis is on early detection of diseases and prophylaxis. In conjunction with this, the handling of lameness, shearing, animal trade and quarantine as well as cleaning and disinfection in sheep and goat flocks are intensively discussed. There are detailed federal and European legal regulations concerning the transport and the physical well-being of animals. These laws are clearly presented and advice for their practical implementation is provided.

[Infectious causes of lameness in sheep - treatment options in veterinary practice]. / Infektiöse Lahmheiten beim Schaf - Therapiemöglichkeiten in der tierärztlichen Praxis.

Lameness in sheep induced by infectious agents can cause problems regarding animal welfare, diagnosis and treatment. Individual lame animals and lameness caused by systemic disease, but especially flock problems due to infectious causes of lameness, such as foot rot, can for various reasons pose a problem for veterinary treatment. The causes of lameness in sheep are described with a special focus on infectious flock problems, and the different treatment options such as foot trimming, foot baths and local and systemic antibiotic therapy as well as vaccination are discussed based on a review of recent international research. The limited choice of drugs licensed for use in sheep in Germany is highlighted. Treatment, therefore, often requires off-label use or the import of footbathing agents licensed in other European countries. The legal consequences of this lack of nationally licensed veterinary products in dealing with a "minor species" are discussed, with a final call for political solutions that will help improve this unsatisfactory situation.

[Recommendations for the husbandry and welfare of sheep and goats by the German Small Ruminant Veterinary Association. Part 1]. / Empfehlung für die Haltung von Schafen und Ziegen der Deutschen Gesellschaft für die Krankheiten der kleinen Wiederkäuer, Fachgruppe der DVG. Teil 1.

Recommendations for the different forms of sheep and goat husbandry based on the legal regulations are summarized. These are given in particular respect to transhumance, tending, alpine farming, and indoor housing. The requirements for pasture, housing, supply of water and food, lambing, rearing of lambs, and health management are intensively discussed. The general requirements of the extensive as well as of the intensive husbandry of sheep and goats are defined. Examples of species-specific capabilities for adaption, the limits of adaption, and signs of decompensation are provided. Compliance with these recommendations should accord the animals entrusted to our care the "five freedoms (13)": 1. Freedom from hunger and thirst, 2. freedom from discomfort, 3. freedom from pain, injury, or disease, 4. freedom to express normal behaviour, and 5. freedom from fear and distress.

In vivo macrophage recruitment by murine intervertebral disc cells.

SUMMARY: An in vivo murine experiment was conducted to measure the capacities of viable intervertebral disc cells to recruit inflammatory cells. The objective was to determine whether compounds secreted from viable cells induce inflammation or whether inflammation in disc herniation simply requires exposure to structural cell or matrix components. Three tissue preparations were inserted into the right lower peritoneal cavity of male mice: tissue with viable annulus fibrosus and nucleus pulposus cells, tissue with viable annulus fibrosus cells, or devitalized annulus fibrosus and nucleus pulposus tissue. Controls included sham-operated and nonoperated groups. Mice were killed 1, 2, or 7 days after surgery. Macrophage recruitment occurred after exposure to viable disc tissue but not after exposure to devitalized disc components; recruitment increased over time. Viable disc cells play a role in the etiology of inflammation in disc herniation.

Chondrocyte differentiation is modulated by frequency and duration of cyclic compressive loading.

As part of a program of research aimed at determining the role of mechanical forces in connective tissue differentiation, we have developed a model for investigating the effects of dynamic compressive loading on chondrocyte differentiation in vitro. In the current study, we examined the influence of cyclic compressive loading of chick limb bud mesenchymal cells to a constant peak stress of 9.25 kPa during each of the first 3 days in culture. Cells embedded in agarose gel were subjected to uniaxial, cyclic compression at 0.03, 0.15, or 0.33 Hz for 2 h. In addition, load durations of 12, 54, or 120 min were evaluated while holding frequency constant at 0.33 Hz. For a 2 h duration, there was no response to loading at 0.03 Hz. A significant increase in chondrocyte differentiation was associated with loading at 0.15 Hz, and an even greater increase with loading at 0.33 Hz. Holding frequency constant at 0.33 Hz, a loading duration of 12 min elicited no response, whereas chondrocyte differentiation was enhanced by loading for either 54 or 120 min. Although not statistically significant from the 120 min response, average cartilage nodule density and glycosaminoglycan synthesis rate were highest in the 54 min duration group. This result suggests that cells may be sensitive to the level of cumulative (nonrecoverable) compressive strain, as well as to the dynamic strain history.

Alternative splicing of the imprinted candidate tumor suppressor gene ZAC regulates its antiproliferative and DNA binding activities.

ZAC encodes a zinc finger protein with antiproliferative activity, is maternally imprinted and is a candidate for the tumor suppressor gene on 6q24. ZAC expression is frequently lost in breast and ovary tumor-derived cell lines and down-regulated in breast primary tumors. In this report, we describe ZACDelta2, an alternatively spliced variant of ZAC lacking the sequence encoding the two N-terminal zinc fingers. Messenger RNAs encoding ZAC or ZACDelta2 were equally abundant and both proteins were nuclear. ZACDelta2 displayed an improved transactivation activity and an enhanced affinity for a ZAC binding site, suggesting that the two N-terminal zinc fingers negatively regulated ZAC binding to its target DNA sequences. Both proteins were equally efficient in preventing colony formation, indicating similar overall antiproliferative activities. However, these activities resulted from a differential regulation of apoptosis vs cell cycle progression since ZACDelta2 was more efficient at induction of cell cycle arrest than ZAC, whereas it was the reverse for apoptosis induction. Hence, these data further underline that ZAC gene is critically controlled, both at the transcriptional level through imprinting and at the functional level through alternative splicing.

Beta(2)-adrenergic receptors potentiate glucocorticoid receptor transactivation via G protein beta gamma-subunits and the phosphoinositide 3-kinase pathway.

Glucocorticoid hormones influence manifold neuronal processes including learning, memory, and emotion via the glucocorticoid receptor (GR). Catecholamines further modulate these functions, although the underlying molecular mechanisms are poorly understood. Here, we show that epinephrine and norepinephrine potentiate ligand-dependent GR transactivation in a hippocampal cell line (HT22) via beta(2)-adrenergic receptors. This enhancement was strongest at low concentrations of glucocorticoids and was accompanied by increased GR binding to a glucocorticoid-responsive element (GRE). beta(2)-Adrenergic receptor-mediated GR enhancement was relayed via G protein beta gamma-subunits, insensitive to pertussis toxin and independent of protein kinase A (PKA). In contrast, the catecholamine-evoked GR enhancement was strongly reduced by wortmannin, suggesting a critical role for phosphoinositide 3-kinase (PI3-K). In agreement, epinephrine directly activated PI3-K in vivo. Similarly, stimulation of tyrosine kinase receptors coupled to PI3-K activation, e.g. receptors for insulin-like growth factor I (IGF-I) or fibroblast growth factor (FGF), increased GR transactivation. Further analysis indicated that G protein-coupled receptor (GPCR) and tyrosine kinase receptor signals converge on PI3-K through separate mechanisms. Blockade of GR enhancement by wortmannin was partially overcome by expression of the downstream-acting protein kinase B (PKB/Akt). Collectively, our findings demonstrate that GPCRs can regulate GR transactivation by stimulating PI3-K. This novel cross-talk may provide new insights into the molecular processes of learning and memory and the treatment of stress-related disorders.

Matrix metalloproteinase-3-dependent generation of a macrophage chemoattractant in a model of herniated disc resorption.

Herniated disc (HD) is a common health problem that is resolved by surgery unless spontaneous resorption occurs. HD tissue contains abundant macrophage infiltration and high levels of matrix metalloproteinases (MMPs) MMP-3 and MMP-7. We developed a model system in which disc tissue or isolated chondrocytes from wild-type or MMP-null mice were cocultured with peritoneal macrophages and used this system to investigate the role of MMPs and chondrocyte/macrophage interactions in disc resorption. We observed a marked enhancement of MMP-3 protein and mRNA in chondrocytes after exposure to macrophages. Chondrocytic MMP-3, but not MMP-7, was required for disc resorption, as determined by assaying for a reduction in wet weight and proteoglycan content after 3 days of coculture. Surprisingly, chondrocyte MMP-3 was required for the generation of a macrophage chemoattractant and the subsequent infiltration of the disc tissue by proteolytically active macrophages. We conclude that macrophage induction of chondrocyte MMP-3 plays a major role in disc resorption by mechanisms that include the generation of a bioactive macrophage chemoattractant.

Matrix metalloproteinase-7-dependent release of tumor necrosis factor-alpha in a model of herniated disc resorption.

Herniated disc (HD), one of the major causes of low back pain, is often resolved spontaneously without surgical intervention. Resorption is associated with a marked increase in infiltrating macrophages, and the matrix metalloproteinases (MMP) MMP-3 and MMP-7 have been implicated in this phenomenon. We developed a murine organ culture model in which intact intervertebral discs were cocultured with peritoneal macrophages to investigate the role of MMPs in HD resorption. Using macrophages isolated from MMP-null mice, we report that macrophage-produced MMP-7 was required for proteoglycan degradation, loss of wet weight, and macrophage infiltration of cocultured discs. The inability of MMP-7-deficient macrophages to infiltrate discs could not be attributed to a defect in macrophage migration. MMP-7 was required for the release of the cytokine TNF-alpha from peritoneal macrophages. The generation of soluble TNF-alpha was essential for the induction of MMP-3 in disc cocultures, which in turn is required for the generation of a macrophage chemoattractant and subsequent macrophage infiltration. TNF-alpha release from macrophages was necessary but insufficient for disc resorption, which required macrophage infiltration. We conclude that there is extensive communication between macrophages and chondrocytes in HD resorption and that an essential component of this communication is the requirement for MMPs to release soluble bioactive factors.

The expression of the antiproliferative gene ZAC is lost or highly reduced in nonfunctioning pituitary adenomas.

The ZAC gene encodes a new zinc-finger protein that concomitantly induces apoptosis and cell cycle arrest and localizes to chromosome 6q24-q25, a well-known hot spot related to cancer. ZAC is highly expressed in the anterior pituitary gland, and its ablation by antisense targeting promotes pituitary cell proliferation. Here we investigate ZAC status in pituitary tumors to evaluate its role in pituitary tumorigenesis. Interest ingly, a strong reduction or absence of ZAC mRNA and protein expres sion was detected in nonfunctioning pituitary adenomas, whereas in clin ically active pituitary neoplasias, the decrease in ZAC expression was variable. Loss of expression was not associated with a mutation of the ZAC gene. Our observations suggest that alternative mechanisms of gene inactivation and/or altered regulation of the ZAC gene occur in nonfunctioning pituitary adenomas.

Loss of expression of the candidate tumor suppressor gene ZAC in breast cancer cell lines and primary tumors.

Loss of chromosome 6q21-qter is the second most frequent loss of chromosomal material in sporadic breast neoplasms suggesting the presence of at least one tumor suppressor gene on 6q. We recently isolated a cDNA encoding a new zinc finger protein which we named ZAC according to its functional properties, namely induction of apoptosis and control of cell cycle progression. ZAC is expressed in normal mammary gland and maps to 6q24-q25, a recognized breast cancer hot spot on 6q. In the present report, we investigated the possible inactivation of ZAC in breast cancer cell lines and primary tumors. We detected no mutation in ZAC coding region in a panel of 45 breast tumors with allelic imbalance of 6q24-q25. However, a survey of eight breast cancer cell lines showed a deeply reduced (three cell lines) or complete loss of (five cell lines) ZAC expression. Treatment of three of these cell lines with the methylation-interfering agent 5-azacytidine induced ZAC re-expression. In addition, Northern blot and RNase protection assay analysis of ZAC expression in 23 unselected primary breast tumors showed a reduced expression in several samples. Together with its functional properties and chromosomal localization, these findings substantiate ZAC as a good candidate for the tumor suppressor gene on 6q24-q25.

Risk factors associated with methicillin-resistant staphylococcal wound infection after spinal surgery.

We used the data from a retrospective case controlled study to identify risk factors for methicillin-resistant staphylococcal wound infection after spinal surgery. Thirty-five cases and 35 uninfected control patients were matched for indication for initial surgery and approximate operative date. Preoperative, intraoperative, and postoperative risk factors were examined. At our institution between 1989 and 1995, 35 adult patients developed spinal wound infection requiring operative debridement; 16 infections were caused by methicillin-resistant staphylococci (MRS). Significant risk factors for MRS infection were lymphopenia, history of chronic infections, alcohol abuse, recent hospitalization, and prolonged postoperative wound drainage. Patients with MRS infections were also somewhat less likely to have received vancomycin prophylaxis. In contrast, the only factor associated with infection caused by other pathogens was alcohol abuse. A number of preoperative risk factors were significantly associated with subsequent MRS spinal wound infection. Chemoprophylaxis with vancomycin should be targeted to patients at increased risk, because overuse may promote the emergence of vancomycin-resistant pathogens.

Up-regulated expression of matrilysin and neutrophil collagenase in human herniated discs.

Spontaneous resorption of herniated nucleus pulposus (HNP) is commonly observed when there is substantial contact of the disc with the spinal canal. We already demonstrated the expression of matrix metalloproteinase (MMP)-3 (stromelysin-1) in the granulation tissues of HNP, suggesting its role in the resorption process of HNP. Recent studies of osteoarthritic cartilages reported an up-regulated expression of metalloproteinases including MMP-7 (matrilysin) and MMP-8 (neutrophil collagenase), suggesting their roles in the matrix degradation. To clarify the expression of MMP-7 and MMP-8 in HNP, immunohistological analysis of various types of HNP was performed. We found MMP-7 was expressed in infiltrated mononuclear cells and chondrocytes, whereas MMP-8 was specifically expressed in chondrocytes. The positive rate for both MMP-7 and MMP-8 significantly increased when HNP was exposed to the epidural space (p < 0.01). Our data suggest that not only MMP-3 but also MMP-7 and MMP-8 may play a role in the resorption process of HNP.

Induction of the PAC1-R (PACAP-type I receptor) gene by p53 and Zac.

Pituitary adenylate cyclase-activating polypeptides and PAC1-R are expressed during early embryogenesis and PACAP's neurotrophic action supports a role in neuronal development. In the adult brain PACAP functions as a neuroprotective factor that attenuates the neuronal damage resulting from various insults. The tumor suppressor gene p53 and the new zinc finger protein Zac regulate apoptosis and cell cycle arrest through unrelated pathways and both genes are up-regulated under cerebral ischemia. We report here that p53 and Zac induce expression of the PAC1-R gene. By this mechanism p53 and Zac could fine-tune the balance between death promoting and protective signals and may thus fulfil a dual role in ischemia.

Inhibition of Zac1, a new gene differentially expressed in the anterior pituitary, increases cell proliferation.

Zac1 is a new zinc finger protein that concomitantly controls apoptosis and cell cycle arrest through separate pathways. The mouse Zac1 gene is mainly expressed in the pituitary gland and in different brain areas. In this study regional and cellular expression of Zac1 in the pituitary gland was determined by in situ hybridization. Zac1 messenger RNA was abundantly expressed in the anterior pituitary lobe compared with that in the intermediate and posterior lobes. Zac1 transcripts were found in all hormone-secreting cell types, with the highest levels in GH- and PRL-producing cells. To investigate the impact of Zac1 in pituitary cell proliferation, we ablated the endogenous Zac1 gene by antisense treatment in two murine cell types, AtT-20 and TtT/GF, that are representative of granular and agranular cell lineages, respectively. The decline in Zac1 protein levels under antisense treatment was accompanied by increased DNA synthesis in clonal corticotroph and folliculo-stellate cells, as demonstrated by enhanced [3H]thymidine incorporation (36% and 50%, respectively). Antisense oligonucleotides against Zac1 controlled cell proliferation in a dose-dependent way, and mutagenized antisense oligonucleotides were inert. Conclusively, our data provide the first evidence of a role for Zac1 in pituitary growth control.

Results of elective lumbar discectomy for patients involved in the workers' compensation system.

We compared the outcomes from lumbar discectomy for patients who were workers' compensation claimants and/or who were involved in active litigation with patients who underwent elective lumbar discectomy, but who were not involved with either compensation or litigation. Eighty-two consecutive patients who underwent elective lumbar discectomy by the senior author were identified from 1989 through 1994. Those patients who underwent a primary discectomy with a minimum of 6 months' follow-up were studied. Patients were excluded if a spinal fusion was performed or if a multilevel laminectomy procedure was required. Patients were classified as compensation patients if they were involved in either worker's compensation claims or active litigation at the time of the lumbar discectomy. The compensation group was further divided into three subsets of patients: those involved in active litigation without compensation, those involved in both compensation and litigation, and those pursuing workers' compensation claims without litigation. The control group was comprised of patients who were not in any way involved with compensation or litigation. Outcome assessment and ratings were determined independently by the coauthors, not the primary surgeon. Outcome was based on pain, employment status, analgesic use, and level of activity. Fifty-four patients met the inclusion criteria. Average follow-up for the compensation patients was 40 weeks. Follow-up for the noncompensation patients averaged 51 weeks. Eighty-one percent of our patients in the noncompensation group achieved a good result. Only 1 of 27 patients was categorized as having a poor outcome. Conversely, patients who were actively involved in the compensation and/or litigation process had significantly poorer outcomes, with only 29% of the patients receiving a good outcome evaluation (p = < 0.0002). Legal involvement was associated with poorer outcome in compensation patients (p = < 0.001).