Contrafreeloading and foraging-related behavior in hens differing in laying performance and phylogenetic origin.

Different breeds of domestic and junglefowl differ in foraging strategies indicating that domestication resulted in modified energy saving behavioral strategies. In the present study we investigated foraging strategies and foraging-related behavior in 4 lines of laying hens differing in phylogenetic origin and laying performance to analyze a possible relationship between foraging and the level of egg production. High performing brown and white pure bred lines were compared with their low performing brown and white counterparts. To control possible effects on behavior other than genetic effects, all hens were reared and kept in an identical environment. A total of 72 hens from each line were kept in 6 compartments with 12 hens per compartment, respectively. Observations were done for 3 times during one laying period. Foraging strategy was tested by a contrafreeloading (CFL) paradigm. CFL describes a behavior in which animals prefer food that requires effort to obtain, although at the same time food is freely available. The hens were offered a commercial standard diet in one trough and a mixture of wood shavings and commercial standard diet in another trough. The behavior of hens was video recorded and the activity level of individual hens in the litter area was recorded by an antenna-transponder system. The high performing layers showed less CFL and foraging-related behavior compared with their low performing counterparts in both the white and brown layers. Despite differences in CFL, all hens showed a preference for the commercial standard diet compared to the mixture of wood-shavings. Our results show an association between foraging strategy and level of egg production. This suggests that a high level of egg production is accompanied by behaviors enabling the hens to satisfy their higher energy demand more efficiently. Saving energy by reduced activity probably allows them to reallocate energy into reproduction, that is, laying performance.

Ghrelin plasma concentration does not covary with energy demand in adult laying hens.

The peptide hormone ghrelin is suggested to be involved in food intake regulation in young growing chicken. Whether ghrelin is involved in the regulation of energetic balance associated with laying performance in adult laying hens was studied by use of 4 chicken lines that differ in laying performance and phylogeny (4 lines; 16 hens per line). As housing conditions are also known to affect energy demand, half of the hens per line were housed in single cages and the other half of hens were maintained in a floor housing system. Plasma samples were collected at 17 to 19, 33 to 35, 49 to 51, and 72 wk of age and analyzed with a chicken ghrelin ELISA Kit. From caged hens, individual food consumption and laying performance additionally was recorded. Due to its function in growth and its relationship with ghrelin, also GH plasma concentrations were analyzed. Ghrelin concentrations did not differ between the 4 lines at any of the test periods (all P > 0.05). Ghrelin was negatively related to food consumption only in the growing period of the high-performing lines (both P < 0.0001). During this phase, floor-housed hens showed greater ghrelin concentrations compared with caged hens (P < 0.0001). Our results suggest that in adult layers ghrelin is not involved in regulating energy intake related to laying performance but rather seems to be related to body growth and housing condition before start of lay, the latter possibly due to differences in hens' behavioral activity.

Effects of nemorosone, isolated from the plant Clusia rosea, on the cell cycle and gene expression in MCF-7 BUS breast cancer cell lines.

BACKGROUND: Breast cancer is the cause of considerable morbidity and mortality in women. While estrogen receptor antagonists have been widely used in breast cancer treatment, patients have increasingly shown resistance to these agents and the identification of novel targeted therapies is therefore required. Nemorosone is the major constituent of the floral resin from Clusia rosea and belongs to the class of polycyclic polyisoprenylated benzophenones of the acylphloroglucinol group. The cytotoxicity of nemorosone in human cancer cell lines has been reported in recent years and has been related to estrogen receptors in breast cancer cells. METHODS: Changes induced by nemorosone in the cell cycle and gene expression of the MCF-7 BUS (estrogen-dependent) breast cancer cell line were analyzed using flow cytometry and the RT(2) Profiler PCR array, respectively. RESULTS: In comparison to breast cancer cells without treatment, nemorosone induced discrete cell cycle arrest in the G1 phase and significant depletion in the G2 phase. Moreover, the compound altered the expression of 19 genes related to different pathways, especially the cell cycle, apoptosis and hormone receptors. CONCLUSION: These promising results justify further studies to clarify mechanisms of action of nemorosone, in view of evaluate the possible use of this benzophenone as adjuvant in the treatment of breast cancer.

Carbopol improves the early cellular immune responses induced by the modified-life vaccine Ingelvac PRRS® MLV.

Adjuvants enhance both the magnitude and duration of immune responses, therefore representing a central component of vaccines. The nature of the adjuvant can determine the particular type of immune response, which may be skewed toward cytotoxic T cell (CTL) responses, antibody responses, or particular classes of T helper (Th) responses and antibody isotypes. Traditionally, adjuvants have been added to intrinsically poor immunogenic vaccines, such as those using whole killed organisms or subunit vaccines. Here, we have compared cellular immune responses induced by the immunogenic modified life-attenuated vaccine Ingelvac PRRS® MLV when administered alone or in combination with carbopol, a widely used adjuvant in veterinary medicine. Using functional readouts (IFN-γ ELISpot and cell proliferation) and analyzing phenotypical hallmarks of CD4T cell differentiation, we show that carbopol improves cellular immunity by inducing early IFN-γ-producing cells and by preferentially driving T cell differentiation to effector phenotypes. Our data suggest that adjuvants may enhance and modulate life-attenuated--not only subunit/inactivated--vaccines.

Effect of rocker shoe design features on forefoot plantar pressures in people with and without diabetes.

BACKGROUND: There is no consensus on the precise rocker shoe outsole design that will optimally reduce plantar pressure in people with diabetes. This study aimed to understand how peak plantar pressure is influenced by systematically varying three design features which characterise a curved rocker shoe: apex angle, apex position and rocker angle. METHODS: A total of 12 different rocker shoe designs, spanning a range of each of the three design features, were tested in 24 people with diabetes and 24 healthy participants. Each subject also wore a flexible control shoe. Peak plantar pressure, in four anatomical regions, was recorded for each of the 13 shoes during walking at a controlled speed. FINDINGS: There were a number of significant main effects for each of the three design features, however, the precise effect of each feature varied between the different regions. The results demonstrated maximum pressure reduction in the 2nd-4th metatarsal regions (39%) but that lower rocker angles (<20°) and anterior apex positions (>60% shoe length) should be avoided for this region. The effect of apex angle was most pronounced in the 1st metatarsophalangeal region with a clear decrease in pressure as the apex angle was increased to 100°. INTERPRETATION: We suggest that an outsole design with a 95° apex angle, apex position at 60% of shoe length and 20° rocker angle may achieve an optimal balance for offloading different regions of the forefoot. However, future studies incorporating additional design feature combinations, on high risk patients, are required to make definitive recommendations.

[General practitioner-centred health-care in Germany. The general practitioner as gatekeeper]. / Hausarztzentrierte Versorgung in Deutschland. Der Hausarzt als Lotse?

In many countries, the gatekeeper system in health care has been existent for several years. The health reform 2004 has strengthened the position of the general practitioner (GP) in German health care. The introduction of the so-called general practitioner programmes (Hausarztprogramme) represents a new form of health care. The GP is credited with a gate-keeping function. The aim of GP-centred care is to avoid expensive double check-ups and specialist consultations which are not necessary from a medical point of view. Optimisation and co-operation of people and institutions within the health care system are also intended. The GP programme offers advantages for both parties. The position of the GPs is being strengthened. For example, the insured have fewer expenses, shorter waiting times at the GP practice and benefit of the co-ordination of more complex treatments by the GP. Regarding this topic, a postal survey with members of a compulsory health insurance fund interviewed 962 participants of a GP programme and 644 non-participants (control group). The study's results show that the GP's gatekeeper function is accepted by most of the insured, but that there is still a need to optimise the daily implementation of GP-centred care.

[Company centredness in the rehabilitation system of the German Democratic Republic, GDR]. / Die Betriebszentrierung im Rehabilitationssystem der DDR.

This article describes major aspects of the rehabilitation system of the former German Democratic Republic, GDR. Based on presentation of the actors in the medical and occupational rehabilitation system, differences are pointed out in comparison to the notion of rehabilitation in the former Federal Republic of Germany, FRG. The article focuses on the system of rehabilitation after stroke, which is outlined with relevant scientific literature. The health and rehabilitation system of the GDR was company-centred. Patients in rehabilitation received comprehensive care and guidance, and especially the companies played an integrative part and adjusted the occupational activity of the patients in rehabilitation to their physical and mental abilities in order to avoid early retirement. On account of the political and social transformation process as well as transfer of the health and rehabilitation legislation of the old FRG, the company-centred rehabilitation system of the GDR was replaced, implementing West German rehabilitation structures in East Germany as well.

An easy-to-use semiquantitative food record validated for energy intake by using doubly labelled water technique.

BACKGROUND: Estimating dietary intake is important for both epidemiological and clinical studies, but often lacks accuracy. OBJECTIVE: To investigate the accuracy and validity of energy intake estimated by an easy-to-use semiquantitative food record (EI(SQFR)) compared to total energy expenditure (TEE) estimated by doubly labelled water technique (EE(DLW)). DESIGN: TEE was measured in 29 nonobese subjects using the doubly labelled water method over a period of 14 days. Within this period, subjects reported their food consumption by a newly developed semiquantitative food record for 4 consecutive days. Energy intake was calculated using the German Food Code and Nutrition Data Base BLS II.3. RESULTS: A good correlation was observed between EI(SQFR) and EE(DLW) (r = 0.685, P < 0.001). The mean difference between EI(SQFR) and EE(DLW) was -1.7+/-2.6 MJ/day (-14+/-21%, P = 0.002). An underestimation of EI(SQFR) <10% was observed in nine subjects (31%), of 10-20% in six subjects (21%), and of >20% in nine subjects (31%). In five subjects (17%), an overestimation of EI(SQFR) was observed. CONCLUSIONS: The easy-to-use semiquantitative food record provided good estimates of EI in free-living and nonobese adults without prior detailed verbal instructions. The presented food record has limitations regarding accuracy at the individual level.

Expression and function of the peptide transporters in escape variants of human renal cell carcinomas.

The transporter associated with antigen processing (TAP) complex is a heterodimeric transmembrane pump consisting of the TAP-1 and TAP-2 subunits; these subunits translocate peptides from the cytoplasm into the lumen of the endoplasmic reticulum, where they bind nascent major histocompatibility complex (MHC) class I molecules. Loss or reduced expression of the TAP genes results in the synthesis of unstable peptide free MHC class I molecules that are only weakly expressed on the cell surface. In a number of human tumor cell lines, downregulation of MHC class I expression has been found to be associated with reduced or absent TAP expression. To investigate whether alterations in MHC class I expression occur during transformation and subsequent progression and whether MHC class I suppression is caused by impaired TAP function, we analyzed the protein expression of MHC class I heavy and light chain and TAP-1 in three renal cell carcinoma (RCC) cell lines and short-term cultures from corresponding normal kidney tissue. In one case a cell line established from a metastatic lesion was also available. Compared with normal epithelial cells, suppression of TAP-1 and MHC class I molecules was detected in all three primary RCC cells and was even more pronounced in the metastatic cell line. In contrast to normal epithelial cells, MHC class I membrane expression of two RCC lines was enhanced by culture in the presence of MHC class I binding peptides or at low temperature (26 degrees C) instead of 37 degrees C. Unstable MHC class I surface expression is caused by dissociation of the MHC class I heavy and light chain molecules as a result of functional defects in the antigen processing machinery, e.g., impaired peptide transport. Attempts to counteract the reduced immunogenicity by transferring the TAP genes into these cells demonstrated that TAP-1-modified RCC cells expressed higher levels of MHC class I molecules. These data indicate that downregulation and instability of MHC class I surface expression in RCC cells is at least partially caused by deficient loading with endogenous peptides and can be restored by TAP gene transfer.

IFN-gamma-mediated coordinated transcriptional regulation of the human TAP-1 and LMP-2 genes in human renal cell carcinoma.

Some human tumor cells exhibit deficient expression of the peptide transporters TAP1 and TAP2 and of the proteasome subunits low molecular weight protein (LMP)-2 and LMP-7, which could be partially restored by cytokine treatment. Here, we show that IFN-gamma stimulation of human renal cell carcinoma lines increased the MHC class I, transporter associated with antigen processing (TAP), and LMP transcript and protein levels, but TAP and LMP expression are more rapidly induced by IFN-gamma than MHC class I molecules. No correlation between the level of induction of the MHC class I antigen presentation genes and IFN sensitivity/resistance was detected. The IFN-gamma-mediated increase of MHC class I, TAP-1, and LMP-2 expression was independent of de novo protein synthesis. Analysis of the dual TAP-1/LMP-2 promoter activity revealed that TAP-1 and LMP-2 expression are controlled by IFN-gamma at the transcriptional level. Site-specific mutations in the IFN-gamma-responsive element of the TAP-1/LMP-2 promoter blocked induction by IFN-gamma. Thus, the IFN-gamma-mediated coordinated transcriptional up-regulation of TAP-1 and LMP-2 expression occurs through the use of a common regulatory element, which might result in enhanced recognition of renal cell carcinoma cells by the immune system.

Impaired HLA-class-I stability in a sarcoma cell line which stimulates exclusively HLA-class-II-restricted autologous T cells.

Defects in the generation and transport of antigenic peptides within tumor cells will lead to the expression of unstable HLA-class-I molecules on the cell surface. These defects will allow tumor cells to escape an MHC-class-I-restricted T-cell response. Recently, we described an exclusively HLA-class-II-restricted autologous T-cell response against a human sarcoma cell line MZ-MES-1 in vitro. Here, we show that surface HLA-class-I molecules of MZ-MES-1 cells are unstable at physiological temperature. HLA-class-I surface expression of MZ-MES-1 cells could be strongly enhanced by culture at low temperature in contrast to various other cell lines analyzed in parallel. Furthermore, culture at low temperature decreased shedding of HLA-class-I molecules by MZ-MES-1 cells. Incubation with allele-specific HLA-class-I-binding peptides strongly increased HLA-class-I surface expression of MZ-MES-1 sarcoma cells and TAP-deficient T2 cells in contrast to other tumor and B-cell lines tested in parallel. IFN-gamma enhanced the expression of TAP, LMP and HLA-class-I proteins in MZ-MES-1 cells. However, the impaired stability of HLA-class-I surface molecules of MZ-MES-1 could not be reversed by IFN-gamma. These results show a new example of impaired HLA-class-I stability of a human tumor which coincides with lack of HLA-class-I-restricted autologous T cells recognizing this tumor. It underlines the importance of MHC-class-II-restricted T-cell responses against tumors with these defects.

Reduced membrane major histocompatibility complex class I density and stability in a subset of human renal cell carcinomas with low TAP and LMP expression.

Suppression of MHC class I expression is thought to allow tumor cells to escape immune surveillance mediated by CD8(+) CTLs. For stable MHC class I surface expression, multiple protein interactions are required for efficient assembly of MHC class I heavy chain and beta 2-microglobulin with endogenous peptides. Peptide processing and transport into the endoplasmic reticulum involves the genes of the transporters associated with antigen processing, TAP-1 and TAP-2, and the two components of the proteasome complex, the low molecular weight proteins LMP-2 and LMP-7. We selected human renal cell carcinoma (RCC) cells derived from a tumor that is thought to be controlled by host immunity to study the MHC class I antigen presentation machinery. Eleven RCC lines established from primary tumors were investigated for the mRNA and protein expression of MHC class I, TAP, and LMP genes. In addition, membrane stability of MHC class I was determined by incubation of the RCC cell lines at low temperature and in the presence of exogenous HLA-binding peptides. Our results revealed the existence of two different phenotypes of RCC cell lines. Group I displayed temperature-stable MHC class I surface expression associated with high, and in most cases coordinated, expression of MHC class I heavy and light chain, TAP and LMP transcripts, and proteins. Group II demonstrated a marked MHC class I instability at 37 degreesC associated with low but coordinated expression of the respective transcripts and proteins. MHC class I membrane expression of group II, but not of group I RCC cells, could be stabilized by incubation with specific MHC class I binding peptides. These results suggest an important role of the genes of the antigen presentation machinery in stable and efficient MHC class I surface expression of RCC cells. However, it has still to be defined whether deficient antigen processing is one of the mechanisms of RCC cells to escape the surveillance of the immune system.

Analysis of the major histocompatibility complex class I antigen presentation machinery in normal and malignant renal cells: evidence for deficiencies associated with transformation and progression.

In some human tumors, reduced or defective MHC class I surface expression has been attributed to functional deficiencies of the genes of the antigen-processing machinery, the proteasome subunits low molecular weight (LMP)-2 and LMP-7, as well as the peptide transporters associated with antigen processing (TAP)-1 and TAP-2. Using normal epithelial kidney cells (MZ1851NN) and renal cell carcinoma cell lines established from the primary tumor (MZ1851RC) and a lymph node metastasis (MZ1851LN) of the same patient, we investigated whether the modulation of MHC class I antigens, TAP and LMP molecules, occurs during transformation and subsequent progression. The mRNA and protein expression of MHC class I heavy and light chain TAP and LMP was strongly reduced in MZ1851RC when compared to the corresponding normal kidney cells MZ1851NN, and this suppression was even more pronounced in the metastatic cell line MZ1851LN. In addition, the activity of the TAP molecules, as measured by peptide translocation assays, was also markedly diminished in MZ1851RC compared to MZ1851NN cells and was further down-regulated in cells of the metastatic lesion. MHC class I surface expression was enhanced by either culturing MZ1851RC and MZ1851LN cells at 26 degrees C instead of 37 degrees C or by incubation of both cell lines with class I-specific binding peptides, whereas MHC class I surface expression of MZ1851NN cells was not affected under these culture conditions. IFN-alpha and in particular IFN-gamma treatment enhances the steady-state mRNA and/or protein levels of TAP, LMP, and MHC class I genes of MZ1851 cell lines but had no additional effect on the stability of MCH class I surface expression. These data indicate that malignant transformation and subsequent in vivo selection of renal tubular cells can lead to the recovery of carcinoma cells that show stable expression of an immune escape phenotype. Deficiencies associated with this phenotype involve all levels of the MHC class I-restricted antigen presentation machinery, are at least partially reversible by IFN treatment, and are even more pronounced in cells that had acquired metastatic potential.

c-erbA and v-erbA modulate growth and gene expression of a mouse glial precursor cell line.

The c-erbA alpha protooncogene coding for the thyroid hormone (T3) receptor (TR alpha 1) and the viral, mutated v-erbA oncogene were expressed in an immortal mouse glial cell line (B3.1) using retroviral vectors. c-erbA alpha expression led to a decrease in cell proliferation in high and low serum conditions, both in the presence and in the absence of T3. In serum-free medium, c-erbA-expressing cells (B3.1 + TR alpha 1) were completely arrested, whereas cells expressing v-erbA (B3.1 + v-erbA) showed a higher DNA synthesis rate than normal B3.1 cells. Although proliferation of all three cell types was stimulated by platelet-derived growth factor and basic fibroblast growth factor, differences were also observed in the response to these agents. B3.1 + TR alpha 1 cells were more sensitive to platelet-derived growth factor than B3.1 and B3.1 + v-erbA cells. In contrast, B3.1 cells responded to basic fibroblast growth factor better than B3.1 + TR alpha 1 or B3.1 + v-erbA cells. Insulin-like growth factor I potentiated the action of platelet-derived growth factor and basic fibroblast growth factor. Again, different responses to treatment with insulin-like growth factor I alone were observed; B3.1 + TR alpha 1 cells did not respond to it, whereas B3.1 + v-erbA cells showed a dramatic stimulation by this agent. Interestingly, in the presence of T3, the blockade in B3.1 + TR alpha 1 cell proliferation was accompanied by the down-regulation of the typical astrocytic genes, glial fibrillary acidic protein and vimentin. These hormone effects were not found in v-erbA-expressing cells. In addition, v-erbA inhibited the basal expression of the cyclic nucleotide phosphodiesterase gene, an oligodendrocytic marker.(ABSTRACT TRUNCATED AT 250 WORDS)

[Left ventricular hemodynamics at rest and during exercise in patients with idiopathic mitral valve prolapse syndrome (author's transl)]. / Ruhe- und Belastungshämodynamik bei Patienten mit idiopathischem Mitralsegelprolaps.

Idiopathic Mitral Valve Prolapse has been regarded by some authors as a primary cardiomyopathy. To determine, if left ventricular hemodynamics at rest and during exercise are impaired, 20 young symptomatic patients with idiopathic mitral valve prolapse, proven by echo- and phonocardiography, without coronary artery disease or severe mitral regurgitation, underwent supine bicycle exercise stress testing. Pulmonary artery pressure, central venous oxygen saturation and cardiac index as well as heart rate were measured with a Swan-Ganz thermodilution catheter and the results compared with those of 10 voluntary normal subjects. Variance analysis showed no significant differences in global left ventricular hemodynamics between patients with idiopathic mitral valve prolapse and normal subjects at rest as well as during exercise. In addition no significant differences were found between patients with echocardiographical holosystolic or mesosystolic mitral valve prolapse. In patients with idiopathic mitral valve prolapse, global left ventricular hemodynamics are not impaired neither at rest nor during exercise.