Echocardiography-derived septal curvature correlated with invasive hemodynamics in pediatric pulmonary hypertension.

BACKGROUND: Right ventricular function and afterload are associated with clinical outcomes in pulmonary hypertension (PH). MRI-derived interventricular septal curvature has been associated with invasive hemodynamics in PH patients. This study sought to determine the relationship of echocardiography derived septal curvature with invasive hemodynamics in pediatric PH patients. METHODS: A single center chart review identified 56 pediatric patients with PH and 50 control patients with adequate echocardiography to assess septal curvature within one month of initial cardiac catheterization. Echocardiographic indices of septal flattening including end-systolic eccentricity index (EIs), maximum EI (EImax), minimum septal curvature (SCmin), and average SC (SCavg) were determined. RESULTS: PH patients had a median right ventricular systolic pressure of 64 mmHg (interquartile range (IQR) 48-81), mean pulmonary artery pressure of 44 mmHg (IQR 32-57), pulmonary vascular resistance of 7.9 iWU (IQR 4.8-12.9), and pulmonary capillary wedge pressure of 10 mmHg (IQR 8-12). Patients with PH had higher EIs and EImax and lower SCmin and SCavg compared to control patients. SCavg demonstrated the strongest association with right ventricular systolic pressure (R2 0.73, p < 0.0001), mean pulmonary artery pressure (R2 0.63, p < 0.0001), and pulmonary vascular resistance (R2 0.47, p < 0.0001). All septal curvature indices were associated with the composite adverse outcome, including Potts shunt, lung transplantation, and death. SCmin (HR 0.29; 95%CI 0.07-0.97) and SCavg (HR 0.15; 95%CI 0.03-0.72) were the only septal flattening indices associated with death. CONCLUSIONS: Echocardiography derived septal curvature is a non-invasive marker of ventricular afterload and adverse outcomes.

Cytoarchitectonic and chemoarchitectonic characterization of the prefrontal cortical areas in the mouse.

This study describes cytoarchitectonic criteria to define the prefrontal cortical areas in the mouse brain (C57BL/6 strain). Currently, well-illustrated mouse brain stereotaxic atlases are available, which, however, do not provide a description of the distinctive cytoarchitectonic characteristics of individual prefrontal areas. Such a description is of importance for stereological, neuronal tracing, and physiological, molecular and neuroimaging studies in which a precise parcellation of the prefrontal cortex (PFC) is required. The present study describes and illustrates: the medial prefrontal areas, i.e., the infralimbic, prelimbic, dorsal and ventral anterior cingulate and Fr2 area; areas of the lateral PFC, i.e., the dorsal agranular insular cortical areas and areas of the ventral PFC, i.e., the lateral, ventrolateral, ventral and medial orbital areas. Each cytoarchitectonically defined boundary is corroborated by one or more chemoarchitectonic stainings, i.e., acetylcholine esterase, SMI32, SMI311, dopamine, parvalbumin, calbindin and myelin staining.

Identification and classification of components in flash pyrolysis oil and hydrodeoxygenated oils by two-dimensional gas chromatography and time-of-flight mass spectrometry.

Hydrodeoxygenated pyrolysis oils (HDO) are considered promising renewable liquid energy carriers. To gain insights in the various reaction pathways taking place during the hydrodeoxygenation reaction of pyrolysis oil, two-dimensional gas chromatography with time-of-flight mass spectrometric analyses (2D-GC-TOF-MS) was applied on the feedstock and product oil. Chromatographic parameters like injection temperature and column choice of the 1D-(2)D ensemble are discussed. Fractionation of the oils by hexane extraction was applied to show the distribution of analytes over the phases. Some 1000 and 2000 components in the pyrolysis and HDO oil, respectively could be identified and classified. The TOF-MS detection considerably improved the understanding of the molecular distribution over the 1D-(2)D retention time fields in the contour plot, in order to classify the analytes in functional groups. By group-type classification of the main components (>0.3% relative area), it was possible to characterize the oils by 250 and 350 analytes, respectively pyrolysis oil and HDO oil, describing 75% of the chromatographable fraction. The 2D-GC-TOF-MS method showed to be a useful and fast technique to determine the composition of (upgraded) pyrolysis oil and is potentially a very useful tool for exploratory catalyst research and kinetic studies. The 2D-GC-TOF-MS technique is not only useful for the chemical study as such, but also provides the basic knowledge for method transfer to a 2D-GC-FID (flame ionization detector) application.

The Ruhrverband sewage sludge disposal concept in the conflict between European and German standards and regulations.

The Ruhrverband, acting as a water association responsible for integrated water resources management within the entire natural river basin of the Ruhr, operates a network of 83 wastewater treatment plants (WWTPs) and connected sludge disposal facilities. According to German regulations, the disposal of sewage sludge containing more than 5% of organic dry solids will be prohibited as of 1 June 2005. In Germany, the only future alternative to incineration will be the agricultural utilization of sludge. However, this way of sludge disposal is presently the subject of critical discussions in Germany because of the organic and inorganic toxic substances, which may be contained in sewage sludge, despite the fact that very stringent standards are to be met by agricultural uses. On the other hand, application of sewage sludge to agricultural land is explicitly supported by the European Sewage Sludge Directive 86/278/EEC. In the face of this controversial situation the Ruhrverband has initiated, in 2000, the development of a comprehensive and sustainable sludge and waste disposal concept for all wastewater facilities it operates in the entire Ruhr River Basin. The concept includes de-central sludge digestion and dewatering and subsequent transport to two central sludge incineration plants. It is expected that in future not more than 5% of all sludges produced in Ruhrverband's WWTPs will be used in agriculture. That means, the major part of 95% will have to be incinerated.

Integrated water resources management in the Ruhr River Basin, Germany.

The Ruhr, with an average flow of 80.5 m3/s at its mouth, is a comparatively small tributary to the Rhine River that has to perform an important task: to secure the water supply of more than 5 million people and of the industry in the densely populated region north of the river. The complex water management system and network applied by the Ruhrverband in the natural Ruhr River Basin has been developed step by step, over decades since 1913. And from the beginning, its major goal has been to achieve optimal conditions for the people living in the region. For this purpose, a functional water supply and wastewater disposal infrastructure has been built up. The development of these structures required and still requires multi-dimensional planning and performance. Since the river serves as receiving water and at the same time as a source of drinking water, the above-standard efforts of Ruhrverband for cleaner water also help to conserve nature and wildlife. Ruhrverband has summed up its environmental awareness in the slogan: "For the people and for the environment". This basic water philosophy, successfully applied to the Ruhr for more than 80 years, will be continued in accordance with the new European Water Framework Directive, enacted in 2000, which demands integrated water resources management in natural river basins, by including the good ecological status of surface waterbodies as an additional goal.

Antigen retrieval in formaldehyde-fixed human brain tissue.

Microwave-stimulated antigen retrieval has become a widely accepted method in both pathology and research laboratories. Since the introduction of the method in 1991, many groups have tried to optimize and standardize it. This review describes the present state of the art. A standard method for microwave-stimulated antigen retrieval in formaldehyde-fixed paraffin-embedded and nonembedded tissue is presented that results, in general, in very good staining for antibodies used in neuroscience. However, there are still a few antigens that are retrieved not at all or not in an optimal manner. Factors of importance for microwave antigen retrieval are the pH of the retrieval solution and, related to the pH, the temperature and duration of heating. These factors are discussed.

Clinical and radiographic evaluation of intertrochanteric osteotomy in dogs: a retrospective study of 18 dogs.

OBJECTIVE: To determine whether intertrochanteric osteotomy (ITO) can prevent the progression of degenerative joint disease (DJD) in dysplastic hip joints. STUDY DESIGN: The results of ITO were assessed retrospectively by using owner questionnaires, physical examination, and radiographic evaluation. ANIMALS: Eighteen client-owned dogs (29 coxofemoral joints were evaluated). METHODS: Lameness was scored according to a grading system. A scoring system was also developed to assess radiographically evident osteoarthritis on a ventrodorsal projection of the coxofemoral joints in extension. RESULTS: Twenty-nine ITO were performed in 18 dogs with varying degrees of hip dysplasia. The dogs were lame on 19 of 29 rear limbs on physical examination before surgery. In 22 of the 29 hip joints, palpation caused signs of pain. The median age at the first and second surgical procedure was 14.5 months and 18 months, respectively. Follow-up evaluation was performed on average at 9.77 (FU I), 22.52 (FU II), and 47.50 months (FU III) after surgery. Based on the owners' evaluation, there was a tendency toward improvement after surgery. The results of the physical examination at the preoperative examination and at the three follow-up examinations did not differ significantly. The follow-up radiographic scores showed significantly worse DJD than the preoperative scores. CONCLUSIONS: ITO does not prevent progression of DJD in the dysplastic hip. CLINICAL RELEVANCE: Knowledge of the long-term effects of ITO is essential for surgeons trying to achieve improvement in dogs with hip dysplasia.

An optimal antigen retrieval method suitable for different antibodies on human brain tissue stored for several years in formaldehyde fixative.

In the last 5 years the microwave oven has become a widely accepted apparatus for retrieving all kinds of antigens that are masked by prolonged formaldehyde fixation. However, it has so far not been possible to obtain good results for all the antigens we were interested in with just one method. This study offers a solution for this problem. We tested Tris buffered saline (TBS) pH 8-10 in order to see whether a microwave (MW) pretreatment in a solution with a high pH works on the antigens that have our particular interest: MAP-2, non-phosphorylated part of the neurofilament (SMI-32, SMI-311), phosphorylated part of the neurofilament (SMI-312) and the calcium binding proteins Calbindin D28-K, Parvalbumin and Calretinin. Furthermore we checked whether or not a lower temperature of 90 degrees C (instead of boiling the tissue) could be used to achieve the same good staining results. These tests showed that an MW pretreatment in TBS pH 9.0 at full power (boiling) gave the best results. Pretreatment at a lower temperature gave also good results for most (but not all) antibodies when the irradiation time was prolonged to 30 min. The optimal antigen retrieval method also improved the quality of staining with an antibody against neuropeptide Y (NPY), for which, until now, no antigen retrieval appeared to be necessary, even after a long duration of fixation.

Laminar distribution of neuropeptide Y-immunoreactive neurons in human prefrontal cortex during development.

Neuropeptide Y (NPY) is present in neurons of the adult human cerebral cortex. In view of the reported roles of NPY in the central nervous system in health and during certain disease conditions, we have studied normal development of NPY immunoreactivity (-ir) in the human prefrontal cortex (PFC), Brodmann areas 9 and 46. Twenty-six specimens ranging from the ages of 14 postovulatory weeks to 34 years exhibited patterns that revealed six periods in the development of the laminar distribution and density of NPY-ir neurons. Changes during prenatal and perinatal periods reflect the onset, development, and resolution of the transient fetal telencephalic compartments, including the subplate zone, in which NPY-ir neurons are especially abundant. Before the age of 1 year, the majority of NPY-ir neurons were found in the subplate zone, whereas, after 1 year, the majority were seen in the cortical layers. This is in contrast with the human visual cortex, where the majority of NPY-ir neurons were still located in the white matter. The density of cortical NPY-ir neurons increased in the fifth developmental period (ages 4-7 years), coinciding with the increase of cortical volume and marked progression of cognitive functions. The adult pattern of a relatively low density of cortical NPY-ir neurons was reached in period 6 (from about 8 years), when individual variation also became apparent. Our data point to a protracted maturation of NPY-ir in the human PFC and to different distribution patterns of NPY-ir neurons in different cortical areas.

Long-term results of treatment of traumatic coxofemoral joint dislocation in dogs: 64 cases (1973-1992).

OBJECTIVE: To determine long-term results of various treatments for traumatic coxofemoral joint dislocation in dogs. DESIGN: Retrospective case series. ANIMALS: 64 dogs that underwent closed reduction and bandage stabilization, extracapsular suture stabilization, transacetabular pinning, toggle pinning, DeVita pinning, or femoral head and neck excision. PROCEDURE: Follow-up evaluations included owner evaluation (64 dogs), physical evaluation (23), and radiography (19). Follow-up time ranged from 8 to 156 months. RESULTS: Owner evaluation scores after closed reduction were significantly better than scores after DeVita pinning, extracapsular suture stabilization, and femoral head and neck excision. On physical examination, 6 of 23 dogs were lame on the side of the previous dislocation. Signs of pain and crepitation were evident during palpation of 12 and 8 of 25 joints, respectively. Thirteen of 21 joints had radiographic evidence of degenerative joint disease. There was a greater progression of degenerative joint disease in previously dislocated joints than in unaffected joints. There were not any significant differences between treatments in regard to results of physical and radiographic evaluation. Time between trauma and treatment and existence of concomitant injuries did not influence follow-up results, but there was a significant association between body weight and radiographic evaluation score. CLINICAL IMPLICATIONS: Concomitant injuries do not appear to justify a worse prognosis in dogs with traumatic coxofemoral joint dislocation, nor does a delay in treatment of > 3 days. Gait abnormalities and degenerative joint disease might develop in the long term. Proper body weight should be maintained regardless of treatment.

Intestinal pseudodiverticulosis in a cat.

A two-year-old domestic shorthair presented for estrous behavior four months after ovariohysterectomy. On exploratory laparotomy, ovarian remnant tissue and multiple intestinal diverticula were found. Histopathology identified the diverticula as pseudodiverticula.

Microwave-stimulated antigen retrieval is pH and temperature dependent.

The effects of the pH of solutions used in microwave-stimulated antigen retrieval were investigated for two antibodies commonly used in neurosciences, MAP-2 and SMI-32. A prolonged fixation time (longer than 1 week) prevents good immunohistochemical staining of these and many other antigens in paraffin and vibratome sections. On human brain tissue stored for 17 months in formaldehyde, the effects of different solutions during microwave treatment were compared, i.e., 4% aluminum chloride (pH 2.75), 0.1 N hydrochloric acid (pH 0.98), and citrate buffers (pH 2.5, 4.5, 6.0), together with the influence of temperature and microwave irradiation. We conclude that the pH and temperature of the solutions and the irradiation time are of great importance for the retrieval of antigens, but in a different way for the two antibodies studied. The SMI-32 staining results are optimal with citrate buffer at pH 2.5 and 2-hr irradiation at 90 degrees C, whereas MAP-2 staining results are optimal with citrate buffer at pH 4.5 and 10-min full-power irradiation. In the other conditions the staining is less intensive or less homogeneously distributed over the section.

Effects of microwave pretreatment on immunocytochemical staining of vibratome sections and tissue blocks of human cerebral cortex stored in formaldehyde fixative for long periods.

The effects of the microwave antigen-retrieval method were investigated for five antibodies commonly used in neurosciences, i.e., Parvalbumin, Calbindin D28-K, MAP-2, MAP-5 and SMI-32. Immunocytochemical stainings were performed on free-floating vibratome sections of tissue which had been stored in fixative for as long as 10 months to compare the effects of microwave treatment in one of the different metal solutions or in distilled water. Microwave treatment of free-floating vibratome sections led to a severe wrinkling of the sections. Therefore the possibility of pretreatment of a thick tissue slice before vibratome sectioning was investigated. We conclude that microwave pretreatment of tissue slices in an aluminium chloride solution is the preferable antigen-retrieval procedure for the examined antibody staining when the immunocytochemical staining is lost or markedly reduced after storage in formaldehyde fixative for a long period. In particular SMI-32 and MAP-2, but also Parvalbumin staining, improved enormously after such a pretreatment.

A comparison of 2 modes of administration of recombinant interleukin-2: continuous intravenous infusion alone versus subcutaneous administration plus interferon alpha in patients with advanced renal cell carcinoma.

PURPOSE: To compare 2 treatment modalities with recombinant Interleukin-2 (rIL-2) for patients with advanced Renal Cell carcinoma (RCC): continuous intravenous infusion (CIV) alone versus subcutaneous (s/c) rIL-2 + Interferon-alpha (IFN-alpha). PATIENTS AND METHODS: Data have been collected on 425 patients with RCC, treated CIV rIL-2 alone, (225 patients), or rIL-2 by the s/c route (200 patients). Patients receiving s/c rIL-2 also received s/c IFN-alpha both drugs being administered on an outpatient basis. Patients receiving CIV rIL-2 were treated as inpatients. Patient eligibility criteria were similar on all studies, and included patients with progressive, advanced disease, but with an ambulatory performance status. RESULTS: The overall response rate for the CIV schedules was not significantly different from the s/c regimens: 15% (95% confidence limits (CL) 10-20%) vs 20% (95%CL 14-26%) with 4% CR in both approaches. Durable responses were seen in both CIV and s/c schedules and there was no evidence of a significant difference in survival in multivariate analysis. There was however an important shift in the toxicity profile. The s/c regimens do not induce a clinically detectable capillary leak syndrome, which is the dose limiting toxicity for CIV regimens. CONCLUSION: Although the introduction of CIV regimens of rIL-2 was a major step forward compared to high-dose bolus, because most patients could be treated in a normal oncology ward, the s/c schedule of rIL-2 + IFN-alpha offers the possibility of outpatient (home) therapy, with no evidence of a reduction in efficacy.

Continuous infusion of recombinant interleukin-2 with or without autologous lymphokine activated killer cells for the treatment of advanced renal cell carcinoma.

Data have been analysed for 327 patients with advanced renal cell carcinoma receiving a continuous infusion of recombinant interleukin 2 (rIL-2) alone (225 patients) or rIL-2 plus lymphokine activated killer (LAK) cells (102) on a normal oncology ward. Eligibility criteria were uniform across protocols, all patients having advanced progressive disease, but with an ambulatory performance status. The baseline characteristics of patients receiving rIL-2 alone did not differ significantly from those receiving LAK, with the exception that the LAK treated patients had a better performance status. Despite similar treatment intensity, toxicity was more severe in the patients receiving LAK. The addition of LAK did not lead to higher response rates or to prolonged response duration, progression-free survival or survival. This review confirms the activity of rIL-2 for the treatment of advanced renal cell carcinoma and demonstrates that the addition of LAK cells does not lead to increased efficacy.

Propolis allergy (IV). Studies with further sensitizers from propolis and constituents common to propolis, poplar buds and balsam of Peru.

26 different compounds have been investigated experimentally for their sensitizing capacity in guinea pigs. 19 of these occur in propolis as well as in poplar bud exudates, and 14 of them are also found in balsam of Peru. 4 caffeates and benzyl isoferulate were found to be strong sensitizers. 7 compounds were moderate, and 13 compounds showed only weak sensitizing potency. Methyl cinnamate was negative. Patch tests in 11 propolis-sensitive patients once more revealed 3-methyl-2-butenyl caffeate and phenylethyl caffeate as the major sensitizers. In addition to the 8 compounds already known to occur in propolis as well as in balsam of Peru, we detected 5 further substances that both materials have in common. Among these, benzyl isoferulate is considered a noteworthy sensitizer. Coniferyl benzoate, which was shown to be a moderate sensitizer, is present in fresh samples of balsam of Peru, while in propolis it has been detected only once so far. The flavonoid aglycones occurring in poplar bud exudates, and hence also in propolis, are weak sensitizers which play only a minor role in propolis hypersensitivity.

The development of neutralizing antibodies in a patient receiving subcutaneous recombinant and natural interleukin-2.

Systemic administration of interleukin-2 (IL-2) in humans may induce antibodies specific to IL-2. The case is reported of a patient with metastatic rectal carcinoma who was treated with long-term subcutaneous IL-2 and a combination of subcutaneous IL-2 and interferon-alpha 2b (IFN-alpha 2b). This patient developed nonneutralizing and neutralizing anti-IL-2 antibodies recognizing both the recombinant and natural cytokine. Detectable serum levels of neutralizing antibodies were accompanied by the inhibition of immune responsiveness to systemic IL-2 in vivo.

[The diagnosis of adrenal cortical function in animals using hormone analysis]. / Zur Diagnostik der Nebennierenrindenfunktion bei Tieren aus hormonanalytischer Sicht.

This paper describes the use of hormone analysis in the diagnosis of adrenal cortex dysfunction in the dog, cat and horse. Analytical problems concerning the determination of corticosteroid levels are discussed and the pathology of adrenal dysfunction is briefly presented. The paper focuses on the problems in establishing physiological norms for adrenal function based on the established assays. Own experiences and other reported data are referred to.

Subcutaneous interleukin-2 and interferon-alpha 2b in patients with metastatic renal cell cancer: the German outpatient experience.

A phase II clinical trial was conducted using subcutaneous recombinant human interleukin-2 (rIL-2, EuroCetus) and subcutaneous interferon-alpha 2b (rIFN-alpha 2b, Essex) in patients with advanced cancer. Safety and tolerance of this outpatient regimen were assessed in 17 patients with progressive metastatic renal carcinoma, 14 of whom were evaluable for clinical response to combined rIL-2 and rIFN-alpha 2b. In this study, rIL-2 was administered every 12 hours, at 1.5 million (Cetus) U/m2 on days 1 and 2, followed by 0.3 million U/m2 5 days per week for 6 consecutive weeks. Concomitantly, rIFN-alpha 2b was given as 5 million U/m2 three times weekly for 6 consecutive weeks. Patients presenting with stable or regressive disease after 6 weeks of rIL-2 and rIFN-alpha 2b (11 of 14) were scheduled to repeat combination therapy. After one treatment cycle, five of 14 patients presented with partial remission; two of these patients achieved complete regression of metastatic lesions. After therapy, six patients have been in stable disease for up to 8 months. toxicity of this regimen was moderate, with local inflammation of the injection sites, grade I-II (World Health Organization criteria) fevers, chills, malaise, nausea and/or vomiting, and anorexia in 70% to 100% of patients treated. After 6 weeks of rIL-2 and rIFN-alpha 2b, laboratory evidence of treatment-related hypothyroidism and hyperthyroidism was obtained in one and four patients, respectively. Immunogenicity of sc rIL-2 was mostly limited to the development of nonneutralizing antibodies that occurred in approximately 40% of patients. None of the patients exhibited antibodies specific to rIFN-alpha 2b.