The effects of a training program using a phantom to accustom heifers to the automatic milking system.

The introduction of heifers into the automatic milking system (AMS) can be associated with considerable stress for both animals and farm employees, as completely inexperienced heifers initially do not independently enter the unknown milking robot. This study investigated whether training heifers on an AMS phantom provides the possibility of preparing heifers for the following lactation at the AMS. For this purpose, 77 Holstein-Friesian heifers were randomly assigned to one of 2 experimental groups: control (CON) or phantom (PHAN). Four weeks before calving, the PHAN group was given free access to the phantom, which was similar to the actual milking robot, so that they could explore it and be positively conditioned by feeding concentrate in the phantom. The heifers of the CON group had no contact with the phantom or the AMS before the first milking at the AMS. The milking frequency per animal per day was recorded, and the proportion of animals that had to be fetched for milking was determined, to evaluate how the animals accepted the AMS after calving. To assess the stress level of the animals before and after introduction into the AMS, fecal cortisol concentrations and rumination times of the animals were measured. Additionally, lactation performance characteristics (milk yield, milk flow, electrical conductivity of milk, and milk composition) were recorded for 77 animals. The animals trained on the phantom showed a higher milking frequency (DIM 7: 2.70 ± 0.14 visits/d) than the control animals (DIM 7: 2.41 ± 0.14 visits/d) between the 4th and 10th day of lactation. In addition, between d 1 and d 5, the proportion of animals that had to be fetched for milking was lower in PHAN (DIM 1: 35.18 ± 4.16%) than in CON (DIM 1: 48.03 ± 4.46%). The PHAN heifers had unexpectedly high fecal cortisol levels (1 wk prepartum: 43.50 ± 0.93 ng/g of feces), although not considerably elevated compared with CON (1 wk prepartum: 40.76 ± 1.05 ng/g of feces). Training on the phantom had no appreciable influence on rumination time and lactation performance parameters. The increased number of milking visits and the reduced proportion of animals that had to be fetched into the AMS for milking indicate that training on the phantom prepares the animals well for being milked in the AMS. Therefore, training heifers on the phantom offers the possibility to facilitate the start into early lactation for the animals, providing a valuable contribution to improvement of animal welfare.

Seasonal or pathological findings? Morphofunctional characteristics of the equine endometrium during the autumn and spring transition.

The deep anoestrous phase in winter is part of the anovulatory season in mares and is bordered by the autumn and spring transitional periods (ATP/STP). To define an annual time span for effective prognostic biopsy sampling, the aim of this study was to provide a morphofunctional characterization of the endometrium during ATP and STP. To outline both transitional periods, endometrial specimens were taken in September, October and November (n = 76) as well as February, March and April (n = 184) with the requirement of a detailed clinical documentation. Tissue samples were examined histologically with special emphasis on the functional endometrial morphology. Additionally, an immunohistochemical evaluation was performed on selected specimens regarding the expression of oestrogen receptor α, progesterone receptor and Ki67-antigen. An absent to low endometrial activity was ascertained in more than 60% of all specimens from late October onwards, whereas a comparably lacking or low activity in STP was observed until early April. Approximately 30% (ATP) to 22% (STP) of all samples exhibited a predominantly "irregular" endometrial differentiation. During the transitional periods, the clinically evaluated ovarian status (transrectal palpation, transrectal ultrasonography and/or serum progesterone and oestrogen analyses) and the endometrial functional morphology were in accordance with approximately 70% of all cases. The expression of steroid hormone receptors and Ki67-antigen was generally low. Given that endometrial maldifferentiations were frequently found during ATP and STP, its occurence might display a characteristic and physiological feature of the transitional periods. Regarding the functional endometrial morphology, a diagnostic biopsy sampling should therefore be performed between late April and before September.

Glycaemic and insulinaemic responses of adult healthy warm-blooded mares following feeding with Jerusalem artichoke meal.

This study aimed to investigate the impact of the supplementation of a pre-biotic compound [Jerusalem artichoke meal (JAM)] on the glycaemic and insulinaemic response in healthy, non-obese warm-blooded horses. Six adult mares [mean body weight (bwt) 529 ± 38.7 kg; body condition score 5.1 ± 0.49/9] were used. In two equal meals per day, the horses received crushed oat grains (1 g starch/kg bwt per day) and meadow hay (2 kg/100 kg bwt per day) which together were likely to meet the energy recommendation for light work (GfE, ). Additionally, they received either 0.15 g fructo-oligosaccharides and inulin (FOS+INU)/kg bwt per day via commercial JAM or maize cob meal without grains as control (CON) in 2 × 3-week periods according to a crossover design. Blood was collected on d21 of the feeding period at different ante- and postprandial (PP) time points (-60, 0, 30, 60, 90, 120, 180, 240 and 300 min), and the plasma glucose and serum insulin levels were determined. Feeding JAM vs. CON did not change the PP peak of glucose or insulin (glucose: 6.3 ± 0.40 vs. 7.0 ± 0.87 mmol/l; insulin: 0.508 ± 0.087 vs. 0.476 ± 0.082 nmol/l) nor did it cause different AUCs until 120 and 300 min PP for glucose and insulin, respectively (AUC120 , glucose: 997 ± 41.6 vs. 1015 ± 41.63 mmol/l per minute, insulin: 49 ± 6.3 vs. 42 ± 6.3 nmol/l per minute; AUC300 , glucose: 1943 ± 142.3 vs. 2115 ± 142.3 mmol/l per minute, insulin: 94 ± 14.8 vs. 106 ± 14.8 nmol/l per minute; p > 0.05). Following JAM vs. CON feeding, glucose and insulin levels declined more rapidly until 240 min PP and tended to be lower (p = 0.053 and p = 0.056, respectively) at this time point. This result might be promising and should further be studied more detailed.

Effects of isoenergetic quantities of a low-starch muesli feed high in fat and fibre vs. oat grains on the glycemic and insulinemic responses and feed intake patterns in sport ponies.

Aim of this study was to compare glycemic and insulinemic responses and feed intake patterns in sport ponies after feeding isoenergetic quantities of low-starch muesli feed high in fat and fibre (FF) or oat grains (OG). Six sport ponies were randomly assigned to one of these two treatment groups for 2 × 3 weeks according to a crossover-design. Ponies received two equal meals/day of either semi-crushed OG (1 g starch/kg bwt*meal-1 ) or an isoenergetic quantity of FF. Hay was also given in two equal meals/day and provided the remaining metabolisable energy up to 1.3-fold maintenance level. On day 21, blood was sampled 1 h after each pony received 0.5 kg hay (0 min). Then, the concentrate was provided and blood sampled 30, 60, 90, 120, 180, 240 and 300 min thereafter. Plasma glucose and serum insulin were analysed, and the areas under the curve (AUC) was calculated 120 and 300 min postprandial (PP). Feed intake patterns were measured in 4 ponies/group via a modified halter. OG was ingested faster than FF (feed intake time; FITDM in min/kg DM: 8.8 ± 1.6 vs. 15.9 ± 1.62, p < 0.05) combined with a higher chewing frequency (p < 0.05). The AUCsgluc120/300, ins120/300 were statistically higher with OG than FF (mmol/L*min-1 : AUCgluc120 : 776 ± 128 vs. 676 ± 80.4; AUCgluc300 : 1811 ± 295.3 vs. 1569 ± 126.3; nmol/L*min-1 : AUCins120 : 38 ± 18 vs. 22 ± 8.1; AUCins300 : 83 ± 39 vs. 35 ± 12; p < 0.05). Plasma glucose tended to decline following the intake of FF, which might be beneficial for equines with reduced glucose tolerance. This, however, requires further investigation. In this study, the ponies consumed OG unexpectedly rapidly. The rate of feed intake was similar to the results previously reported in the literature for warmblood horses.

[Psychotherapeutic approaches for patients with somatoform disorders]. / Psychotherapeutische Ansätze für Patienten mit somatoformen Störungen.

Patients with somatoform symptoms are considered to be difficult to treat. Clinical studies on treatment of this condition are underrepresented compared to other mental and psychosomatic disorders. Cognitive-behavioral treatment for patients with somatoform symptoms was found to have a significant effect; additionally, some evidence of recently published findings supports psychodynamic therapy. This report provides information on how to effectively deal with those patients. Furthermore it describes transtherapeutic targets and explains three therapeutic phases: establishing a therapeutic relationship, developing a model of the disorder and establishing coping strategies. As a last point a cognitive-behavioral treatment study, a psychodynamically-oriented study, a group intervention study and a new approach, a combination of cognitive-behavioral therapy and emotion regulation training, are presented.

Concentration of platelets and growth factors in canine autologous conditioned plasma.

OBJECTIVES: To report the concentration of blood cells and selected growth factors in canine autologous conditioned plasma (ACP). METHODS: The density of blood cells in whole blood (WB), ACP and standard plasma preparation (SP) of 10 healthy mature dogs was determined. In both ACP and SP, the concentration of insulin-like growth factor-1 (IGF-1), epidermal growth factor, vascular endothelial growth factor, platelet-derived growth factor-AA, platelet-derived growth factor-AB, platelet-derived growth factor-BB, transforming growth factor-ß1 (TGF-ß1), and transforming growth factor-ß2 was measured using the ELISA technique. In another ten dogs, ACP was prepared using an ultra-soft spinning protocol, and again blood cell density was compared to that obtained in WB. RESULTS: The density of platelets in ACP was significantly higher than that in SP (p = 0.0002), but there was not any significant difference between ACP and WB, nor between WB and ACP prepared using softer centrifugations. Interestingly, only for IGF-1, PDGF-BB, and TGF-ß1 could reliable measurements be obtained, showing a significant increase in PDGF-BB and TGF-ß1 concentrations in ACP compared to SP (p = 0.001, p = 0.0028). Regarding IGF-1 content, there was not any significant difference between ACP and SP. CLINICAL SIGNIFICANCE: Canine ACP prepared according to the manufacturer's recommendations, or by using a softer spin does not show the same specifications as human ACP, which shows a doubling in platelet count compared to WB. Even though canine ACP has a similar number of platelets per injected volume and consequently, probably the same amount of injected growth factors than WB, application of canine ACP would not be associated with the proinflammatory potential reported for WB, as it is almost free of erythrocytes and nucleated cells.

[Endocrinologic studies of male psittacine birds for the evaluation of their reproductive status]. / Endokrinologische Untersuchung männlicher Papageienvögel zur Beurteilung ihres Reproduktionsstatus.

OBJECTIVE: Evaluating methods of hormone measurement in different specimens of male parrots in order to assess their reproductive status and stress axis. MATERIAL AND METHODS: Cockatiels and rose-ringed parakeets were chosen as psittaciforme representatives and their endocrine profiles were examined. In various pre-experiments, suitable techniques for the determination of testosterone in plasma, saliva and faeces of male parrots were established. Before analysing the samples by enzyme immunoassay, blood and faeces were extracted using diethyl ether, while saliva could be tested without extraction. Based on the excretion of mainly conjugated testosterone metabolites, parrots' faecal samples were also hydrolysed with ß-glucuronidase/arylsulfatase before extraction. In addition, the levels of the stress hormone corticosterone were determined by radioimmunoassay in order to assess possible relationships between stress and the secretion of testosterone. RESULTS: The examined psittacine species displayed different seasonal secretion patterns for both testosterone and corticosterone. Cockatiels had maximum plasma testosterone levels in February, in contrast, rose-ringed parakeets showed highest concentrations in November. As a consequence of cumulative excretion, both species showed much higher faecal than plasma testosterone concentrations. In rose-ringed parakeets, the levels of corticosterone in plasma were exceptionally high compared to the cockatiels. CONCLUSION: According to this study, we have been able to establish suitable methods for testosterone analysis in blood and faeces of cockatiels and rose-ringed parakeets, supporting the assessment of their reproductive status. At present saliva does not appear to be an ideal medium for reliable hormone level measurement, thus further investigations are required concerning this subject. CLINICAL RELEVANCE: By means of process analysis, it will be possible to detect increasing testosterone levels and/or pathological alterations, which could be considered in breeding programmes.

[Cortisol concentrations in sheep before, during and after sham foot trimming on a tilt table - the suitability of different matrices]. / Kortisolkonzentrationen bei Schafen vor, während und nach der simulierten Klauenbehandlung im Kippstand ­ Eignung verschiedener Matrices.

INTRODUCTION: Matrices that can be collected non-invasively for quantification of a stress response in sheep have received little attention in the veterinary literature. This study examines the suitability of blood, tears and saliva for determining a stress response in sheep undergoing sham foot trimming on a tilt table. The cortisol concentration of blood, tears and saliva and the concentration of cortisol metabolites in faeces were measured in 13 healthy Meat Merino ewes once a day for six days. Sham foot trimming on a tilt table was used as the stressor and was done during a one-hour period on day 4; cortisol concentrations of blood and tears were measured at 0, 10, 20, 30, 40 and 60 minutes. Cortisol concentrations of blood (maximum at 30 minutes) and tears (maximum at 40 minutes) increased during the procedure and then decreased. There were significant correlations between cortisol concentrations of blood and tears (p = 0,04) during sham foot trimming (area under the curve, 0 to 60 minutes). Over the entire 6-day study period, significant correlations were seen between the cortisol concentrations of blood and tears (r = 0,55; p.

INTRODUCTION: Les marqueurs qui peuvent être collectés de manière non invasive pour quantifier une réponse au stress chez le mouton ont fait l'objet de peu d'attention dans la littérature vétérinaire. Cette étude examine la pertinence du sang, des larmes et de la salive pour déterminer une réponse au stress chez des moutons subissant un parage fictif des pieds sur une table basculante. La concentration de cortisol dans le sang, les larmes et la salive ainsi que la concentration de métabolites de cortisol dans les fèces ont été mesurées chez 13 brebis Meat Merino saines une fois par jour pendant six jours. Le parage fictif des pieds sur une table inclinable a été utilisé comme facteur de stress et a été effectué pendant une période d'une heure le jour 4; les concentrations de cortisol dans le sang et les larmes ont été mesurées à 0, 10, 20, 30, 40 et 60 minutes. Les concentrations de cortisol dans le sang (maximum à 30 minutes) et les larmes (maximum à 40 minutes) ont augmenté au cours de la procédure puis ont diminué. Il y avait des corrélations significatives entre les concentrations de cortisol dans le sang et les larmes (p = 0,04) lors du parage fictif des onglons (aire sous la courbe, 0 à 60 minutes). Sur l'ensemble de la période d'étude de 6 jours, des corrélations significatives ont été observées entre les concentrations de cortisol dans le sang et les larmes (r = 0,55 ; p < 0,001), le sang et la salive (r = 0,53 ; p < 0,001) et les larmes et la salive (r = 0,78 ; p < 0,001). La concentration fécale de métabolites de cortisol était significativement augmentée au jour 5 (p 0,05), mais la concentration de cortisol des autres supports n'a pas changé de manière significative au cours de la période d'étude de 6 jours. Le parage fictif des pieds sur une table basculante a été considéré comme un facteur de stress aigu chez les moutons en raison de l'augmentation des concentrations de cortisol dans le sang, des larmes et de l'augmentation des concentrations de métabolites de cortisol dans les selles. La concentration de cortisol dans les larmes était similaire à celle du sang et, par conséquent, la collecte de larmes représente une alternative viable et non invasive au sang pour les tests de cortisol. Le délai des pics entre la concentration maximale de cortisol dans les larmes et le sang doit être pris en compte lors de l'interprétation des résultats.

Investigation of the local expression of the relaxin system in canine mammary tumours.

While mammary tumours are the main reasons of death in bitches, early detection of tumours and metastases is crucial for survival of affected dogs. Invasiveness and angiogenesis, which are important processes of tumour growth and spreading, require connective tissue remodelling. This process is dominantly mediated by matrix metalloproteinases (MMP), which are well known to be positively regulated by relaxin (RLX) in various tissues, including human breast cancer. So far, the presence of RLX and its receptor RXFP-1 as well as their linkage with MMP in canine mammary tumours (CMT) is completely unknown. In the first part of the present study, concentrations of RLX, oestradiol and progesterone from plasma samples of bitches with CMT were compared with clinical and survival data to investigate the predictive value of these hormones. In the second part, the expressions of RLX, RXFP-1 and MMP-2, -9 and -13 were examined by real-time reverse transcriptase polymerase chain reaction (RT-PCR) in 31 CMT samples. Finally, relationships of systemic plasma RLX or locally expressed RLX with expression of MMP in CMT were analyzed for the first time. Comparison of hormone concentrations in 93 bitches in terms of benign or malignant nature of the CMT, lung metastases, recidivation and 12-month survival discovered no significances. The expressions of RLX, RXFP-1 and MMP were independent from plasma RLX, but expressions of local RLX and RXFP-1 showed a strong correlation (p = 0.00004, r = 0.671) as well as RXFP-1 and MMP-2 (p = 0.009, r = 0.463), indicating a possible significant role of the locally produced RLX in CMT pathogenesis as an inducer of connective tissue remodelling.

Effects of dermal dexamethasone application on ACTH and both basal and ACTH-stimulated cortisol concentration in normal horses.

There are no data available regarding the systemic (adverse) effects which might be induced by topical/dermal glucocorticoids (GCs) application in the horse. Besides their widespread use for the treatment of a variety of peripheral inflammatory disorders such as atopic dermatitis, eczemas or arthritis in the horse, their surreptitious application has become a concern in doping cases in competition/performance horses. Assessing both basal and ACTH-stimulated plasma cortisol as well as basal ACTH concentrations following application of dexamethsone-containing dermal ointment is necessary to determine influences on hypothalamus-pituitary-adrenal (HPA) axis. Ten clinically healthy adult standardbred horses (6 mares, 4 geldings) were rubbed twice daily each with 50 g dexamethasone-containing ointment on a defined skin area (30 x 50 cm) for 10 days. RIA and chemiluminescent enzyme immuno-metric assay were used to determine resting and ACTH-stimulated plasma cortisol and basal ACTH concentrations, respectively. HPA feedback sensitivity and adrenal function were measured by a standard ACTH stimulation test. Dermal dexamethasone suppressed significantly the resting plasma cortisol level (to 75-98%) below baseline (P < 0.001) within the first 2 days and decreased further until day 10. ACTH stimulation test showed a markedly reduced rise in plasma cortisol concentrations (P < 0.001 vs. baseline). Plasma ACTH level decreased also during topical dexamethasone application. The number of total lymphocytes and eosinophil granulocytes was reduced, whereas the number of neutrophils increased. No significant change of serum biochemical parameters was noted. Dermal dexamethasone application has the potential to cause an almost complete and transient HPA axis suppression and altered leukocyte distribution in normal horses. The effects on HPA axis function should be considered in relation to the inability of animals to resist stress situations. The data further implicate that percutaneously absorbed dexamethasone (GCs) may cause systemic effects relevant to 'doping'.

Effects of feeding level during lactation on FSH and LH secretion patterns, and follicular development in primiparous sows.

In the primiparous lactating sow undernutrition affects LH, but the effect on FSH is less clear. This study was conducted to investigate the effect of ad libitum (AL; n = 5) and restricted (RE; 70% of the AL feed; n = 5) feeding on the secretion patterns of FSH and LH, and on follicular development in primiparous lactating sows. Body side fat thickness (BSFT) was measured prior to farrowing and at weaning on day 21 postpartum (pp). Sows had an intravenous catheter fitted on day 6 or 7 pp. Blood samples were taken on days 12, 15 and 18 pp, every 15 min starting at 0700 h for 11 h, and plasma analysed for FSH and LH by radioimmunoassay (RIA). Daily transcutaneous ultrasonography of the ovaries was performed between days 10 and 20 pp to monitor follicular growth. Reduction in BSFT was higher in RE than in AL sows (p < 0.05). Mean and basal LH were significantly higher in AL than in RE sows on days 12 and 15 pp. (p < 0.05), and LH pulse frequency tended to be higher (p < 0.1). Mean FSH was higher in AL than in RE sows on days 15 and 18 (p < 0.05), and had a tendency to be higher on day 12 (p = 0.1). Follicle size increased in AL sows between days 10 and 20 of lactation (p < 0.05; r = 0.71), but remained unaffected in RE sows. In conclusion, the study demonstrates that feed restriction of primiparous sows during lactation affects not only LH, but also FSH, as well as lactational follicular growth. An association between those events is suggested.

Radiocarbon constraints on the glacial ocean circulation and its impact on atmospheric CO2.

While the ocean's large-scale overturning circulation is thought to have been significantly different under the climatic conditions of the Last Glacial Maximum (LGM), the exact nature of the glacial circulation and its implications for global carbon cycling continue to be debated. Here we use a global array of ocean-atmosphere radiocarbon disequilibrium estimates to demonstrate a ∼689±53 14C-yr increase in the average residence time of carbon in the deep ocean at the LGM. A predominantly southern-sourced abyssal overturning limb that was more isolated from its shallower northern counterparts is interpreted to have extended from the Southern Ocean, producing a widespread radiocarbon age maximum at mid-depths and depriving the deep ocean of a fast escape route for accumulating respired carbon. While the exact magnitude of the resulting carbon cycle impacts remains to be confirmed, the radiocarbon data suggest an increase in the efficiency of the biological carbon pump that could have accounted for as much as half of the glacial-interglacial CO2 change.

Endocrine pregnancy monitoring in the two-toed sloth (Choloepus didactylus): "Pregnant or not pregnant".

Progesterone (P4), pregnanediol glucuronide (PdG), estradiol-17ß (E2), and estrone sulfate (E1S) were measured in the feces of four female two-toed sloths (Choloepus didactylus) for early pregnancy diagnosis. For individual feces assignment, the examined female sloths were fed with a turquoise food colorant every second day. Fecal samples were collected one to four times per week, depending on the defecation rate throughout the pregnancies and the postpartum periods. The complete course of pregnancy was subdivided into three 16-week intervals (trimester of pregnancy, TP1-3) and a 5-week post-partum period after birth. Progesterone and PdG concentrations started to increase above luteal phase levels 3 weeks after conception (P = 0.028 and 0.005, respectively). At the beginning of TP1, P4 concentrations averaged 345.0 ± 283.0 ng/g and increased approximately 100- to 300-fold to a peak of 7588.0 ± 6717.0 ng/g over the TP3. Progesterone concentrations were considerably lower than PdG concentrations that started with 3206.0 ± 1500.0 ng/g at TP1 and increased up to 12.8556.0 ± 53.744.0 ng/g until birth. In contrast, mean concentrations of E2 (8.2 ± 2.4-11.7 ± 4.2 ng/g) and E1S (12.2 ± 6.7-22.9 ± 13.0 ng/g) elevated insignificantly and were not suitable for pregnancy detection. All hormones analyzed decreased rapidly within the first weeks after birth. Progesterone and PdG, as well as E2 and E1S, highly significantly correlated (r = 0.602, P < 0.001 and r = 0.497, P < 0.001, respectively) at TP1. During the TP2, only P4 and PdG significantly correlated (TP2: r = 0.661, P < 0.001 and postpartum period: r = 0.616, P = 0.009). In summary, only P4 metabolite concentrations were suitable to determine the status of reproduction in the two-toed sloth. Thereby, PdG was ideally suited to diagnose early pregnancy because it was more sensitive and detected pregnancy 2 weeks earlier than P4.

Influence of preoperative dexamethasone therapy on proliferating cell nuclear antigen (PCNA) expression in comparison to other parameters in meningiomas.

We conducted a trial in 42 benign and malignant meningiomas to assess a possible influence of preoperative dexamethasone therapy on mitotic index, labelling indices of proliferating cell nuclear antigen (PCNA), progesterone receptor, epidermal growth factor receptor (EGF-R), c-erbB-2 oncoprotein, cathepsin D, gamma-gamma enolase as well as the mean number of silver-stained nucleolar organizer region-associated proteins (AgNORs). Tumors with preceding dexamethasone therapy for more than 1 day display significantly less immunohistochemical staining for PCNA. A correlation between the labelling index of PCNA and the degree of malignancy could not be identified. There was no significant effect of preoperative dexamethasone therapy on the other parameters. Our data suggest that dexamethasone may selectively inhibit the expression of PCNA in the G1/S-phase of the cell cycle. Thus, we emphasize the necessity to heed factors, e.g. dexamethasone, which may affect the expression of proliferating markers.

Team nursing coverage of prenatal-intrapartum patients at a university hospital. An innovation in obstetric nursing.

A unique approach in utilization of nursing personnel was instituted at the University of Colorado Medical Center to alleviate the following patient care problems: 1) lack of continuity of care of obstetric patients, 2) inadequate prenatal instruction, and 3) the unpredictability of patient census in the labor and delivery area. Half the nurses from the labor and delivery area were organized into four teams of two nurses each who acquired a case load of patients for antepartum care. When a patient was admitted to the hostpial in labor, the nurse from her team who was on call came in and provided all nursing care during the intrapartum period. The problems and benefits of this new approach to nursing care are discussed.

Stereotactic brain biopsy in AIDS.

In the hope of finding a treatable condition, the need for rapid diagnosis in HIV-seropositive patients with brain lesions is apparent. In order to evaluate the efficacy of stereotactic brain biopsy in AIDS patients, we retrospectively studied 25 HIV-infected patients undergoing stereotactic biopsy. Brain lesions were identified with gadolinium-enhanced MRI and/or contrast CT. Brain biopsy was performed using the system of Riechert. From 8 up to 15 small tissue samples from one or two targets were obtained in every patient. The biopsy material was examined cytologically, histologically (including electron microscopy), immunohistochemically and, in part, by animal test and polymerase chain reaction (PCR). A definite diagnosis was achieved in 92%. Diagnosis included primary central nervous system lymphoma (PCNSL) (10), toxoplasmosis (10), progressive multifocal leukoencephalopathy (2) and one case of co-existing toxoplasmosis and cytomegalovirus infection. Two biopsies were non-diagnostic. All PCNSLs showed polymorphic B-cell populations of high malignancy; accurate classification according to the Kiel classification was not possible. In 3 lymphomas Epstein-Barr nuclear antigen (EBNA) 2-mRNA could be detected by PCR and confirmed immunohistochemically by EBNA 2 expression. In 6 cases autopsy confirmed the biopsy diagnosis. Conventional histology was not sufficiently decisive for toxoplasmosis and progressive multifocal leukoencephalopathy, so that immunohistochemistry and animal tests became very important for a final diagnosis. With the help of different morphological and molecular biological techniques stereotactic brain biopsy appears to be an effective method in the diagnosis of HIV-associated brain lesions. In view of the marked radio- and chemosensitivity of PCNSLs it is mandatory to establish an early and accurate histological diagnosis for adequate treatment.

Skin abnormalities in neurofibromatosis 2.

OBJECTIVE: To determine the prevalence, distribution, and histopathological conditions of skin abnormalities in neurofibromatosis 2 (NF2). DESIGN: Case series. SETTING: Hospital neurology department. PATIENTS: Consecutive sample of 88 patients with NF2 referred through workshops and publications, genetic counseling, and referral from neurosurgical departments; 81 patients met the National Institutes of Health, Bethesda, Md, NF2 diagnostic criteria and the diagnosis was established by mutation or segregation analyses in 7 patients. MAIN OUTCOME MEASURES: Prevalence, distribution, and type of skin abnormalities; histopathological features of 29 skin tumors selected primarily for medical indications. RESULTS: Fifty-two patients (59.1%) had 458 skin tumors, which were the first presenting sign in 27.3% of patients and usually appeared as flat dysplastic tumors or subcutaneous spherical nodular tumors of the peripheral nerves, on the limbs and trunk. Although 29 patients (33.0%) had café au lait spots, only 2 patients had as many as 6 spots. compared with patients with milder disease, patients with more severe disease had a significantly greater prevalence of skin tumors (24.0% and 71.0%, P < .001), more than 10 skin tumors (0.0% and 27.4%, P = .004), flat dysplastic skin tumors (8.0% and 54.8%, P < .001), and subcutaneous spherical nodular tumors (24.0% and 58.1%, P = .004). The histologically analyzed tumors were predominantly schwannomas, but 5 were neurofibromas and 2 were mixed tumors. CONCLUSIONS: The prevalence of some skin tumor types in NF2 is high and varies with disease severity, and schwannomas predominate in sampled tumors. The occurrence of neurofibromas is surprising, but could be explained by an interaction between neurofibromin and the NF2 gene product in regulating the ras proto-oncogene.

Epithelial and melanoma antigens in gliosarcoma. An immunohistochemical study.

Gliosarcomas are mixed tumors with malignant glial and mesenchymal elements. The number of GFAP-positive tumor cells decreases with the increase of sarcomatous components, until whole areas may be GFAP negative. These distinct differentiations may, however, lead to false interpretations in small tissue samples. In this connection, it is of interest that, according to other reports, glial tumors may be positive for different anti-keratin antibodies and this prompted us to undertake a systematic investigation of the immunoreactivity of gliosarcomas using a panel of well-characterized monoclonal antibodies against cytokeratins (KL1, AE 1/3, Lu-5, CK-19, CK MNF 116 and Ma-903). These cases were further studied with the anti-epithelial non-cytokeratin antibodies EMA, HEA 125, Ber-EP4, CEA as well as the melanoma-antibody HMB-45, Leu-M1, GFAP and vimentin. As screening study we examined 20 cerebral metastatic carcinomas, 21 malignant gliomas (including 6 gliosarcomas) and 3 metastatic melanomas with the monoclonal antibodies KL1 and HMB-45. All cerebral metastatic carcinomas and 4/6 gliosarcomas were positive for KL1, whereas all melanomas, 2 metastatic carcinomas and 3 gliosarcomas showed an immunostaining with HMB-45. All gliosarcomas were positive with at least one of the tested anti-cytokeratin antibodies. The gliosarcomas did not show an immunoreaction in any of the cases when CEA, HEA 125, Ber-EP4, EMA or Leu M1 were applied. In our opinion, the monoclonal antibodies HEA 125 and Ber-EP4 could obviously be helpful in differentiating gliosarcomas from metastatic carcinomas.

Extracerebral toxoplasmosis in AIDS. Histological and immunohistological findings based on 80 autopsy cases.

Despite the great amount of literature concerning toxoplasmic encephalitis in AIDS patients, little is known about extracerebral toxoplasmosis. Therefore we conducted a study of 80 autopsy cases to estimate the frequency of extracerebral toxoplasmosis. A control group of 50 cases was completely negative for all markers applied. In 35 of the 80 AIDS-cases (43.7%), organisms could be detected. In 13 cases (16.2%) there was an extracerebral toxoplasmosis; 4 cases (5%) showed only extracerebral involvement and in 9 cases (11.2%), extracerebral toxoplasmosis occurred in combination with cerebral manifestations. In 22 cases (27.5%), only cerebral toxoplasmosis was found. The following organs were involved: cardiac muscle (15%), lungs (6.2%), liver (5%), pancreas (5%), gastrointestinal tract (6.2%), adrenal glands (5%), lymph nodes (5%) and testis (3.7%). In individual cases further organs, not mentioned above, were involved. Pseudocysts could be demonstrated within necroses and inflammatory foci by conventional staining, whereas trophozoites became apparent only immunohistologically.