[Studies on Factors Influencing Influenza Vaccination Rates in Patients with Chronic Obstructive Pulmonary Disease]. / Untersuchungen zu Einflussfaktoren auf die Influenza-Impfraten bei Patienten mit chronisch obstruktiver Lungenerkrankung.

BACKGROUND : With low influenza vaccination rates among the chronically ill, approaches to increase these rates among risk patients with chronic obstructive pulmonary disease (COPD) are to be uncovered. METHODS : 120 COPD patients from Magdeburg filled out a questionnaire and were analyzed regarding the influenza vaccination status 2015/2016 or 2016/2017. Vaccinated and unvaccinated were compared in socio-epidemiological factors, the health belief model (HBM), self-efficacy (GESIS-ASKU), anxiety/depression (HADS-D) and disease processing (FKV-LIS). RESULTS : 62.5 % (n = 75) were vaccinated, 31.7 % (n = 38) unvaccinated, 5.8 % (n = 7) made no statement. In over or equal to 60-year-olds 76 % were vaccinated, in under 60-year-olds 42 % were vaccinated. 60 % (n = 72) knew to belong to a risk group. Unvaccinated indicated greater concern about side effects of the vaccination (p = .004) and drew a worse benefit-expense balance (p = .001). Unvaccinated were more often uncertain about the vaccination protection and the severity of influenza (p ≤ .001). Vaccinated were highly motivated to think about vaccination themselves and more often had a positive vaccination history (p = .001). COPD patients showed a lower self-efficacy than the reference group of the German general population (p = .000), vaccinated and unvaccinated did not differ (p = .418). No difference between vaccinated and unvaccinated was found in the processing of the disease and in depression and anxiety, but unvaccinated tended to give higher anxiety values. CONCLUSION : Measures should particularly target COPD patients under 60 years of age with a negative vaccination history and sensitize them as risk patients. Widespread uncertainties about the severity of influenza and vaccination protection should be addressed. It should be communicated that influenza vaccination does not lead to exacerbation. The vaccination recommendation should increasingly be made by pulmonologists.

The influence of cartilage thickness at the sigmoid notch on inclination at the distal radioulnar joint.

AIMS: The aims of this study were: firstly, to investigate the influence of the thickness of cartilage at the sigmoid notch on the inclination of the distal radioulnar joint (DRUJ), and secondly, to compare the sensitivity and specificity of MRI with plain radiographs for the assessment of the inclination of the articular surface of the DRUJ in the coronal plane. PATIENTS AND METHODS: Contemporaneous MRI images and radiographs of 100 wrists from 98 asymptomatic patients (mean age 43 years, (16 to 67); 52 male, 53%) with no history of a fracture involving the wrist or surgery to the wrist, were reviewed. The thickness of the cartilage at the sigmoid notch, inclination of the DRUJ and Tolat Type of each DRUJ were determined. RESULTS: The assessment using MRI scans and cortical bone correlated well with radiographs, with a kappa value of 0.83. The mean difference between the inclination using the cortex and cartilage on MRI scans was 12°, leading to a change of Tolat type of inclination in 66% of wrists. No reverse oblique (Type 3) inclinations were found when using the cartilage to assess inclination. CONCLUSION: These data revealed that when measuring the inclination of the DRUJ using cartilage, reverse oblique inclinations might not exist. The data suggest that performing an ulna shortening osteotomy might be reasonable even in distal radioulnar joints where the plain radiographic appearance suggests an unfavourable reverse oblique inclination in the coronal plane. We recommend using MRI to validate radiographs in those that appear to be reverse oblique (Tolat Type 3), as the true inclination might be different, thereby removing one possible contraindication to ulnar shortening. Cite this article: Bone Joint J 2017;99-B:369-75.

Interaction of rat alveolar macrophages with dental composite dust.

BACKGROUND: Dental composites have become the standard filling material to restore teeth, but during the placement of these restorations, high amounts of respirable composite dust (<5 µm) including many nano-sized particles may be released in the breathing zone of the patient and dental operator. Here we tested the respirable fraction of several composite particles for their cytotoxic effect using an alveolar macrophage model system. ​METHODS: Composite dust was generated following a clinical protocol, and the dust particles were collected under sterile circumstances. Dust was dispersed in fluid, and 5-µm-filtered to enrich the respirable fractions. Quartz DQ12 and corundum were used as positive and negative control, respectively. Four concentrations (22.5 µg/ml, 45 µg/ml, 90 µg/ml and 180 µg/ml) were applied to NR8383 alveolar macrophages. Light and electron microscopy were used for subcellular localization of particles. Culture supernatants were tested for release of lactate dehydrogenase, glucuronidase, TNF-α, and H2O2. RESULTS: Characterization of the suspended particles revealed numerous nano-sized particles but also many high volume particles, most of which could be removed by filtering. Even at the highest concentration (180 µg/ml), cells completely cleared settled particles from the bottom of the culture vessel. Accordingly, a mixture of nano- and micron-scaled particles was observed inside cells where they were confined to phagolysosomes. The filtered particle fractions elicited largely uniform dose-dependent responses, which were elevated compared to the control only at the highest concentration, which equaled a mean cellular dose of 120 pg/cell. A low inflammatory potential was identified due to dose-dependent release of H2O2 and TNF-α. However, compared to the positive control, the released levels of H2O2 and TNF-α were still moderate, but their release profiles depended on the type of composite. CONCLUSIONS: Alveolar macrophages are able to phagocytize respirable composite dust particle inclusive nanoparticles. Since NR8383 cells tolerate a comparatively high cell burden (60 pg/cell) of each of the five materials with minimal signs of cytotoxicity or inflammation, the toxic potential of respirable composite dust seems to be low. These results are reassuring for dental personnel, but more research is needed to characterize the actual exposure and uptake especially of the pure nano fraction.

Nanoparticle release from dental composites.

Dental composites typically contain high amounts (up to 60 vol.%) of nanosized filler particles. There is a current concern that dental personnel (and patients) may inhale nanosized dust particles (<100 nm) during abrasive procedures to shape, finish or remove restorations but, so far, whether airborne nanoparticles are released has never been investigated. In this study, composite dust was analyzed in real work conditions. Exposure measurements of dust in a dental clinic revealed high peak concentrations of nanoparticles in the breathing zone of both dentist and patient, especially during aesthetic treatments or treatments of worn teeth with composite build-ups. Further laboratory assessment confirmed that all tested composites released very high concentrations of airborne particles in the nanorange (>10(6)cm(-3)). The median diameter of airborne composite dust varied between 38 and 70 nm. Electron microscopic and energy dispersive X-ray analysis confirmed that the airborne particles originated from the composite, and revealed that the dust particles consisted of filler particles or resin or both. Though composite dust exhibited no significant oxidative reactivity, more toxicological research is needed. To conclude, on manipulation with the bur, dental composites release high concentrations of nanoparticles that may enter deeply into the lungs.

[Neuropsychopharmaca influence the intracellular pH value of central neurons]. / Neuropsychopharmaka verändern den intrazellulären pH-Wert von zentralen Neuronen.

The intracellular pH (pHi) of neurons is tightly regulated, mainly by membrane-bound transporters acting as acid extruders or acid loaders. Regulation of pHi helps to control neuronal excitability, as increased bioelectric activity moderately lowers pHi and, in the sense of a negative feedback loop, intracellular acidosis mostly reduces neuronal excitability. Moreover, a change of pHi widely influences complex cellular functions. With respect to neuropsychopharmaca, little is known about whether or not they may affect neuronal H ( + )-homeostasis. To this aim, we tested several antipsychotics, antidepressants, anticonvulsants, and lithium for effects on neuronal pHi, using guinea pig hippocampal slice preparations in which CA 3 pyramidal neurons were loaded with the pHi-sensitive dye BCECF-AM. All antipsychotics, most antidepressants and about half of the anticonvulsants tested so far elicited reversible changes of neuronal pHi when applied at therapeutic and supratherapeutic concentrations. Although these results await confirmatory in vivo experiments, we believe that the pHi activity of neuropsychopharmaca needs further attention, especially when therapeutic mechanisms or even harmful side effects are discussed.

Effects of essential oils on milk production, milk composition, and rumen microbiota in Chios dairy ewes.

The effect of the addition of an essential oil (EO) preparation (containing a mixture of natural and nature-identical EO) on the performance of dairy ewes of the Chios breed was investigated. Eighty lactating ewes were allocated into 4 equal groups in a randomized block design, each with 4 replicates of 5 ewes housed in the same pen. The 4 groups were fed the same total mixed ration allowance, the roughage being a mixture of corn silage, lucerne hay, and wheat straw, and the concentrate based on cereals and oil cakes. Control ewes were fed their daily allowance of total mixed ration without any EO. The other 3 groups were supplemented with EO at levels of 50, 100, and 150 mg/kg of the concentrated feed, respectively. Individual milk yield was recorded daily and feed refusals were recorded on a pen basis weekly during the first 5 mo of lactation. Milk samples were analyzed for chemical composition, somatic cell count, and urea content. Rumen samples were analyzed for pH, NH(3)-N content, and protozoa, cellulolytic, hyper-ammonia-producing, and total viable bacteria counts. Results showed that inclusion of EO increased milk production per ewe, the effect being dose dependent [1.565, 1.681, 1.876, and 2.119 L/d (standard error of the difference ± 0.176) for the control, 50, 100, and 150 mg of EO/kg of concentrate diets, respectively], and thus improved feed utilization. Although the inclusion of EO did not affect milk composition, it lowered urea concentration and somatic cell count in milk samples at the highest supplementation level compared with the control. Total counts of viable and cellulolytic bacteria and protozoa were not influenced by EO supplementation; however, counts of hyper-ammonia-producing bacteria were decreased at the 2 highest supplementation levels compared with the control group. Rumen pH was not affected by EO supplementation, but rumen NH(3)-N was reduced at the highest EO supplementation level, and acetate rumen concentrations tended to decrease and propionate to increase in a dose-dependent manner. In conclusion, EO supplementation may improve feed utilization and performance of the high-yielding dairy Chios ewes; however, the underlying mechanisms leading to this improvement merit further investigation.

The NHE3 inhibitor AVE1599 stimulates phrenic nerve activity in the rat.

The effect of AVE1599, an inhibitor of the sodium/proton exchanger type 3 (NHE3), on phrenic nerve (PN) activity was investigated using the working heart brainstem preparation (WHBP). Hypercapnia (Delta pH: -0.1) applied for 10 min reversibly increased PN frequency (f) by 66.0 +/- 19.5% and decreased burst duration by 23.3 +/- 3% (mean +/- SE, n = 21). Similarly, AVE1599 (0.3 microM) increased f after 10 and 30 min by 75.1 +/- 13.2 and 176 +/- 36.2% (n = 10), respectively, and reduced duration of PN bursts by 24.9 +/- 10.8%. Hypercapnia-induced increases of f were attenuated by AVE1599. An elevated concentration of AVE1599 (0.9 microM) had no significant effect on PN. As AVE1599 accumulates in brain tissue and might interfere with the less affine NHE1, we furthermore tested the NHE1-inhibitor HOE642. In fact, HOE642 (0.9 microM) diminished f by 88.5 +/- 9.2 and 58.6 +/- 10.0% after 10 and 30 min (n = 6), respectively, but did not abolish hypercapnic responses. We conclude that AVE1599 augments central respiratory drive in the WHBP via NHE3 but not NHE1 inhibition.

Pharmacological impact on loop gain properties to prevent irregular breathing.

Theory predicts respiratory instabilities at elevated system loop gain (G), determined by such factors as ventilatory CO(2) sensitivity, set-point PCO(2), and metabolic rate. In anesthetized rabbits, the effects on G of carbonic anhydrase (CA) inhibitors and of different sodium/proton exchanger type 3 (NHE3) inhibitors were studied. Acetazolamide significantly reduced G by 42.0 +/- 9.3% and methazolamide by 35.0 +/- 9.5% (each n = 7, P<0.01). Irrespective of the substance, NHE3 inhibition reduced G by 33.0 +/- 7.8% (n = 10, P<0.01) at 35.5 +/- 1.6 mmHg PaCO(2) (mean +/-SE), but not at lower arterial CO(2) levels (n=5). Since high baseline PCO(2) coincides with elevated brainstem NHE3 mRNA expression, this may also account for a higher risk of sleep apnea (or even occurrence of sudden infant death). Therefore, NHE3 inhibitors may gain similar therapeutic importance in the treatment of irregular breathing as CA inhibitors. Generally, effective treatment should aim at a low system loop gain, by reducing respiratory chemosensitivity, improving blood gases and preventing low metabolic rates.

Expression of anion exchanger 3 influences respiratory rate in awake and isoflurane anesthetized mice.

The anion exchanger 3 (AE3) is involved in neuronal pH regulation of which may include chemosensitive neurons. Here we examined the effect of AE3 expression on respiratory rate (RR) in vivo. AE3 knockout (KO, n=5) and wild type (WT, n=6) mice were subjected to body plethysmography, both while awake and during isoflurane anesthesia. RR was significantly lower in awake AE3 KO (162+/-7SE min(-1)) than in WT mice (212+/-20 min(-1), P=0.036). The same was found during isoflurane anesthesia at 0.5 MAC (KO: 123+/-9 min(-1), WT: 168+/-15 min(-1), P=0.026) and 1.0 MAC (KO: 51+/-6 min(-1), WT: 94+/-6 min(-1), P=0.001). Hypercapnia (5% CO2) increased RR in awake and decreased RR in nesthetized (1.0 MAC) mice, whereby relative changes were larger in AE3 KO mice. Recovery from isoflurane anesthesia in respect to RR regaining baseline values was more pronounced in AE3 KO. Results show that AE3 expression profoundly influences control of breathing in mice.

[Biomaterial studies in cultures of human stapedial bone-like cells]. / Untersuchung unterschiedlicher Biomaterialien in einer Knochenzellkultur des menschlichen Steigbügels.

BACKGROUND: Cell culture studies may provide information on the behavior of biomaterials in the intended implant environment. Cell cultures from such an environment could be used for the development of middle ear implants. MATERIAL AND METHODS: Secondary bone-like cell cultures derived from human stapes were exposed to different materials [Al(2)O(3) ceramic, glass ceramic (Ceravital), gold and titanium]. Proliferation was studied for up to 40 days. RESULTS: The proliferation of cultured stapes bone-like cells did not differ significantly between the four tested biomaterials. The well known cytotoxic effect of copper, which was used as a control, was evident. CONCLUSIONS: Four biomaterials [Al(2)O(3) ceramic, glass ceramic (Ceravital), gold and titanium] have similar biocompatibility and no toxicity when tested in human stapes cell cultures. This in vitro model may be of considerable value for the further development of middle ear implants, e.g., when coated with bone morphogenetic proteins.

Inhuman shields--children caught in the crossfire of domestic violence.

BACKGROUND: Child abuse is a worldwide scourge. One of its most devastating manifestations is non-accidental head injury (NAHI). METHODS: This is a retrospective chart review of children presenting to the Red Cross Children's Hospital trauma unit with a diagnosis of NAHI over a 3-year period. RESULTS: Sixty-eight children were included in the study and 2 different groups were identified. Fifty-three per cent of the children were deliberately injured (median age 2 years), while 47% were allegedly not the intended target of the assailant (median age 9 months). The assailant was male in 65% of the intentional assaults and male in 100% of the unintentional assaults, with the intended adult victim female in 85% of the latter cases. Overall, 85% of the assaults were committed in the child's own home. CONCLUSIONS: The high proportion of cases in which a young child was injured unintentionally suggests that these infants effectively become shields in assaults committed by adults. In this context any attempts to deal with child abuse must also address the concurrent intimate partner violence.

HSP27 in patients with ovarian carcinoma: still an independent prognostic indicator at 60 months follow-up.

OBJECTIVE: Heat shock protein 27 (HSP27) is produced in response to pathophysiologic stress in animal cells. The authors have previously shown that HSP27 is an independent prognostic indicator in patients with ovarian carcinoma. The present study was performed to see whether HSP27 remained an independent prognostic indicator with longer follow-up. METHODS: One hundred and three consecutive patients with epithelial ovarian carcinoma were studied. Slides were prepared from fresh tissue. HPS27 staining was performed as previously described. Patient records were examined for FIGO stage, grade, histology, level of cytoreduction and survival. RESULTS: One hundred and three patients were followed for a mean of 60 months. Twenty patients had FIGO Stage I disease, four Stage II, 59 Stage III, and 20 Stage IV. Immunohistochemical (IHC) staining for HSP27 was not related to histologic grade, level of cytoreduction or histologic subtype. A statistically significant decrease in HSP27 staining was found to correlate with increased FIGO stage (p = 0.008). Using cox-regression analysis, HSP27 staining (p = 0.025), stage (p = 0.0012), and level of cytoreduction (p < 0.0001) were independent predictors of survival in these patients. CONCLUSION: Cox-regression analysis found HSP27 to be an independent indicator of prognosis and survival in patients with ovarian carcinoma who had longer follow-up. Decreased HSP27 staining was related to decreased survival. This study confirms the authors' earlier report on the importance of HSP27 as a prognostic indicator in ovarian carcinoma.

Heat shock protein 27 in the placentas of women with and without severe preeclampsia.

BACKGROUND: Although not fully understood, heat shock proteins (HSP) are well known stress response proteins. The purpose of this analysis was to determine whether staining for HSP27 was different between placentas from pregnancies complicated by severe pre-eclampsia with intrauterine growth restriction (IUGR) as compared to controls. METHODS: Sterile placental tissue was collected from ten women whose pregnancies were complicated by severe preeclampsia with IUGR and from ten women with uncomplicated by severe pre-eclampsia with IUGR and from ten women with uncomplicated term pregnancies. The tissue was then stained for HSP27. RESULTS: The median age of the patients was 27 years (mean 27, range 17-37). The median estimated gestational age at delivery was 38 weeks (mean 37, range 29-41). Overall 12 of 20 placentas stained positively for HSP27 (nuclear and/or cytoplasmic). Eight of ten placentas from women with pre-eclampsia and IUGR stained positively for HSP27 (p = 0.046). CONCLUSION: HSP27 staining of the placenta is twice as common in patients with severe preeclampsia as compared to patients with normal term gestations. These preliminary results warrant the inauguration of a similar but larger study to examine the significance of these findings.

Nuclear and cytoplasmic c-myc staining in endometrial carcinoma and their relationship to survival.

OBJECTIVE: The role of the c-myc proto-oncogene in genomic instability is just becoming more fully understood. However, its role in endometrial cancer is essentially unknown. The objective of this study was to determine the relationship between cytoplasmic and nuclear c-myc staining, DNA index, and survival in patients with endometrial carcinoma. METHODS: One hundred and twenty-one patients with endometrial carcinoma were studied. Image analysis was used to determine DNA index. In addition to cytoplasmic and nuclear c-myc staining and DNA index, histologic type, stage, grade, depth of invasion, lymphvascular space invasion, and peritoneal cytology were evaluated as prognostic indicators. Univariate and multivariate analyses were performed. RESULTS: One hundred and twenty-one patients were followed for over 5 years. c-myc cytoplasmic staining was present in 75.2% of the patients' tumors, and nuclear staining was present in 66.9% (P = 0.99). DNA index was significantly higher in patients with nuclear c-myc staining and no cytoplasmic staining (DNA index 1.38) as compared to those patients whose tumors displayed cytoplasmic c-myc staining but no nuclear c-myc staining (1.18) (P = 0.016). Patients whose tumors stained positively for nuclear c-myc and negatively for cytoplasmic c-myc had significantly worse survival by Kaplan-Meier analysis (P < 0.0001). Seventeen patients died during the follow-up period of this study. By multivariate analysis, positive cytoplasmic c-myc staining with negative nuclear staining (P = 0.0076), negative cytoplasmic c-myc staining with positive nuclear staining (P = 0.011) and FIGO stage (P < 0.0001) were shown to be independent prognostic indicators predictive of survival. CONCLUSION: Nuclear and cytoplasmic c-myc staining, as well as FIGO stage, when assessed by multivariate analysis, were demonstrated to be important factors in predicting survival in the 121 patients in this study. While increasing FIGO stage was prognostic of decreased survival, the specific location of c-myc staining was also associated with prognosis. The expression of the c-myc protein is related to survival in patients with adenocarcinoma of the endometrium.

Nuclear size, shape, and density in endometrial carcinoma: relationship to survival at over 5 years of follow-up. Does analyzing only cells occupying the G0-G1 peak add useful information?

OBJECTIVE: The authors, using image analysis, previously demonstrated nuclear size and summed optical density to be independent prognostic indicators of recurrence in patients with endometrial carcinoma. The same tumors were analyzed by studying the optical features in the G0-G1 peak to see if this changed the values found as well as their importance as prognostic features at greater than 5 years of follow-up. METHODS: Tumors from 74 consecutive patients, surgically treated, with endometrial cancer, were evaluated. Survival, depth of invasion, lymphvascular space invasion, FIGO stage, grade, histology were analyzed. DNA index, progesterone receptor status, as well as nuclear size (NUSZ), shape (NUSH), and summed optical density (NUSD) were evaluated. NUSZ, NUSH, and NUSD were quantified using image analysis. RESULTS: Fifteen patients died from disease during the observation period of the study. Mean follow-up was 82 months with a median of 84 months. Forty-nine patients had stage I cancers, five stage II, 17 stage III, and three stage IV. NUSZ and NUSD were all significantly different between the original (entire cell cycle) and the re-measured (G0G1 only) values (both P < 0.001). Multivariate analysis showed both the original (P = 0.0001) and G0G1-only (P = 0.046) NUSZ and the original (P = 0.0002) and G0G1-only (P = 0.018) NUSD to be independent prognosticators of survival. CONCLUSION: Image analysis is able to quantify cellular and nuclear parameters not otherwise quantifiable. NUSD and NUSZ correlated with traditional prognostic indicators, were demonstrated independent predictors of survival at over 5 years of follow-up. Although the re-measured NUSZ and NUSD from only the G0-G1 peak were significantly different from the original NUSZ and NUSD, they were not as valuable as prognostic factors. Nuclear size and summed optical density measured from the entire cell cycle are independent prognostic indicators of survival at greater than 5 years of follow-up. Measuring nuclear morphometric features in the G0-G1 peak only does not add any new prognostic information.

Chemotherapy for ovarian cancer: an evidence-based approach.

Although cytoreductive surgery is the most influential factor in treatment for ovarian cancer, chemotherapy is needed for almost all patients diagnosed with this disease. The mainstay of chemotherapy is platinum. Different platinum compounds are used for different histologies, and different combinations are used for different histologies also. We will present the data so that each reader can understand the knowledge behind chemotherapy decisions.

Interferon-gamma or granulocyte-macrophage colony-stimulating factor administered as adjuvants with a vaccine of irradiated autologous tumor cells from short-term cell line cultures: a randomized phase 2 trial of the cancer biotherapy research group.

The objective was to study the effects of patient-specific vaccine immunotherapy administered with either interferon-gamma (IFNgamma) or granulocyte-macrophage colony stimulating factor (GM-CSF) in patients with metastatic cancer. Short-term cell lines were established from cancer tissue resected from patients with metastatic cancer for use as autologous tumor cell vaccines. Successful cultures were expanded to 1 to 2 x 108 cells, irradiated, and cryopreserved in aliquots of 106 cells for intradermal testing of delayed tumor hypersensitivity and 107 cells for subcutaneous vaccinations. The study design was that of a randomized phase 2 trial. Patients were stratified by tumor type and by whether they had measurable disease at the time vaccination was to commence, and then randomized to receive either 100 MIU IFNgamma subcutaneously or 500 microg GM-CSF subcutaneously at the time of each tumor cell vaccination. Following a baseline test of delayed-type hypersensitivity (DTH) to an intradermal injection of 106 irradiated autologous tumor cells, patients received 3 weekly subcutaneous injections of 107 cells, had a repeat DTH test at week 4, then received monthly vaccinations for 5 months. A positive DTH test was defined as at least 10 mm of induration; survival was determined from the first DTH test. There were 98 patients enrolled with a median follow-up of over 4 years. The most prevalent diagnoses were melanoma (51), renal cell carcinoma (18), and soft-tissue sarcoma (14). There were 49 patients (26 men, 23 women, average age 50.4 years) randomized to IFNgamma and 49 (28 men, 21 women, average age 54.1 years) to GM-CSF. The average numbers of vaccine and adjuvant injections were 6.3 and 5.9, respectively. For the patients who received IFNgamma, the objective response rate was 0 of 21; for patients who received GM-CSF the response rate was 1 of 26. Only eight patients (four from each arm) had a positive baseline DTH reaction to autologous tumor. The tumor DTH test converted from negative to positive in 13 of 45 of the IFNgamma group and 11 of 43 of the GM-CSF group. With 29 patients deceased in the IFNgamma arm and 31 in the GM-CSF arm, the 2-year and 5-year survival rates were 45% and 29% for the IFNgamma arm and 41% and 23% for the GM-CSF arm (NSD). Both adjuvants were well tolerated and results were similar in both arms of the study. Both adjuvants were associated with a 25% to 30% rate of DTH conversion and a 25% 5-year survival rate. Immune recognition of autologous tumor can be induced with this approach.

[Optimizing histological image data for 3-D reconstruction using an image equalizer]. / Optimierung histologischer Bilddaten für die 3D-Rekonstruktion mit Hilfe eines Image-Equalizers.

Bone cells form a wired network within the extracellular bone matrix. To analyse this complex 3D structure, we employed a confocal fluorescence imaging procedure to visualize live bone cells within their native surrounding. By means of newly developed image processing software, the "Image-Equalizer", we aimed to enhanced the contrast and eliminize artefacts in such a way that cell bodies as well as fine interconnecting processes were visible.

A reporter-cell assay for the detection of BMP-2 immobilized on porous and nonporous materials.

Human recombinant bone morphogenetic protein-2 (rhBMP-2) immobilized on the surface of metal implants can facilitate osseointegration. Here, we describe a cell reporter assay useful for quantifying small amounts of immobilized rhBMP-2 on various materials. The peptide was dotted and heat-fixed on titanium, 316L stainless steel, nitrocellulose, or glass, and its distribution was monitored by in situ biotinylation followed by detection with the avidin-biotin method. Bioactivity of rhBMP-2 was demonstrated by means of a confluent layer of osteoblastic MC3T3-E1 cells that evenly covered rhBMP-2-free and rhBMP-2-loaded surface areas, as shown with epifluorescence microscopy of calcein acetoxymethyl (AM)-loaded cells. Expression of osteocalcin, fibronectin, actin, and vimentin increased where cells were located on rhBMP-2 dotted areas, but the signal:noise ratio was too low to bioassay the peptide. However, local pronounced expression of alkaline phosphatase was used to quantify BMP-2 in the range of 5-80 ng/dot by means of a cytochemical color reaction for alkaline phosphatase and image analysis of resulting dots. The lower detection limit was in the order nitrocellulose > glass > titanium > 316L steel. We conclude that the cell reporter assay is useful to assess biological activity of rhBMP-2 even after immobilization on three-dimensional implant materials.

Ventrolateral neurons of medullary organotypic cultures: intracellular pH regulation and bioelectric activity.

The hypothesized role of the intracellular pH (pH(i)) as a proximate stimulus for central chemosensitive neurons is reviewed on the basis of data obtained from organotypic cultures of the medulla oblongata (obex level) of new born rats (OMC). Within OMC a subset of neurons responds to hypercapnia as do neurons in the same (or similar) brain areas in vivo. Maneuvers altering intra- and/or extracellular pH (pH(o)) such as hypercapnia, bicarbonate-withdrawal, or ammonium pre-pulses, evoked well defined changes of the neuronal pH(i). During hypercapnia (pH(o) 7.0) or bicarbonate-withdrawal (pH(o) 7.4) most ventrolateral neurons adopted a pH(i) which was < or = 0.2 pH units below the steady state pH(i), while signs of pH(i)-regulation occurred only in a small fraction of neurons. During all treatments leading to intracellular acidosis, bioelectric activity of chemosensitive neurons increased and was often indistinguishable from the response to hypercapnia, regardless of whether pH(o) was unchanged, decreased or increased during the treatment. These data strongly suggest that the pH(i) acts as proximate stimulus. The mode of acid extrusion of chemosensitive neurons is, therefore, of major importance for the control of central chemosensitivity. Immunocytochemical data, pH(i) measurements and neuropharmacological studies with novel drugs pointed to the Na(+)/H(+) exchanger subtype 3 (NHE3) as a main acid extruder in ventrolateral chemosensitive neurons. Possible functions and neuropharmacological strategies arising from this very local NHE3 expression are discussed.