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Omega-1, a glycosylated T2 ribonuclease (RNase) secreted by Schistosoma mansoni eggs and abundantly present in soluble egg antigen, has recently been shown to condition dendritic cells (DCs) to prime Th2 responses. However, the molecular mechanisms underlying this effect remain unknown. We show in this study by site-directed mutagenesis of omega-1 that both the glycosylation and the RNase activity are essential to condition DCs for Th2 polarization. Mechanistically, we demonstrate that omega-1 is bound and internalized via its glycans by the mannose receptor (MR) and subsequently impairs protein synthesis by degrading both ribosomal and messenger RNA. These experiments reveal an unrecognized pathway involving MR and interference with protein synthesis that conditions DCs for Th2 priming.

Für viele Wissenschaftler sind Membranproteine ein faszinierendes Forschungsobjekt, für viele aber auch der schlimmste Alptraum, dem sie je begegnet sind. Letztere kämpfen mit intrinsischen Problemen: Expression, Reinigung, Stabilität, biochemische und biophysikalische Charakterisierung; die anderen begeistert die Funktion von Membranproteinen — Informationsaustausch über biologische Membranen, ohne die wahrscheinlich das Leben, wie wir es heute kennen, nicht existieren würde.

Quantitative utilization of L-cysteate (2-amino-3-sulphopropionate) as the sole source of carbon and energy for growth of the aerobic, marine bacterium Silicibacter pomeroyi DSS-3T was observed. The sulphonate moiety was recovered in the medium largely as sulphite, and the appropriate amount of the ammonium ion was also observed. Genes [suyAB (3-sulpholactate sulpho-lyase)] encoding the known desulphonation reaction in cysteate degradation were absent from the genome, but a homologue of a putative sulphate exporter gene (suyZ) was found, and its neighbour, annotated as a D-cysteine desulphhydrase, was postulated to encode pyridoxal 5′-phosphate-coupled L-cysteate sulpho-lyase (CuyA), a novel enzyme. Inducible CuyA was detected in cysteate-grown cells. The enzyme released equimolar pyruvate, sulphite and the ammonium ion from L-cysteate and was purified to homogeneity by anion-exchange, hydrophobic-interaction and gel-filtration chromatography. The N-terminal amino acid sequence of this 39-kDa subunit confirmed the identification of the cuyA gene. The native enzyme was soluble and homomultimeric. The Km-value for L-cysteate was high (11.7 mM) and the enzyme also catalysed the D-cysteine desulphhydrase reaction. The gene cuyZ, encoding the putative sulphite exporter, was co-transcribed with cuyA. Sulphite was exported despite the presence of a ferricyanide-coupled sulphite dehydrogenase. CuyA was found in many bacteria that utilize cysteate.

ABSTRACTInfection with human immunodeficiency virus (HIV) and the resulting immunosuppression are associated with an increased risk for human papillomavirus (HPV) persistence and related malignancies. In the present study we investigated the prevalence of HPV in urine samples from 104 HIV-infected men with low CD4+cell counts (<100 per mm3) and 115 urine samples from HIV-negative men. A high prevalence of HPV DNA (39.4%) was found in the HIV patients. Most of the HPV types were high risk (81.4%), with HPV 52 as the most prevalent type (12.5%), followed by HPV 18 (6.7%), HPV 35 (5.8%), and HPV 70 (4.8%). Multiple HPV genotypes were observed in 17 (41%) of the 41 HPV- and HIV-positive men. In contrast, only 11 (9.6%) HPV DNA-positive cases were observed among the 115 HIV-uninfected men, and 3 (27.3%) contained multiple genotypes. Quantitative analyses indicated that the HPV viral load, as measured in urine samples, is significantly higher in HIV-positive men compared to HIV-negative men. In the present study we show that urine samples are useful for detecting HPV DNA, there is a high prevalence of HPV in HIV-positive men, and the HPV viral load is substantially higher in HIV-positive than in HIV-negative men. More studies are needed to evaluate the risk and natural development of HPV-related malignancies in HIV-positive men.

The objective of the present experiment was to investigate the effects of transportation, lairage, and preslaughter stressor treatment on glycolytic potential and pork quality of the glycolytic longissimus and the oxidative supraspinatus (SSP) or serratus ventralis (SV) muscles. In a 2 × 2 × 2 factorial design, 384 pigs were assigned randomly either to short (50 min) and smooth or long (3 h) and rough transport, long (3 h) or short (< 45 min) lairage, and minimal or high preslaughter stress. Muscle samples were taken from the LM at 135 min and from the SSP at 160 min postmortem for determination of the glycolytic potential and rate of glycolysis. At 23 h postmortem, pork quality was assessed in the LM and the SV. Effects of transport and lairage conditions were similar in both muscle types. Long transport increased (P< 0.01) the glycolytic potential and muscle lactate concentrations compared with short transport. Both long transportation and short lairage decreased (P< 0.01) redness (a* values) and yellowness (b* values) of the LM and SV. In combination with short lairage, long transport decreased (P< 0.05) pork lightness (lower L* values), and electrical conductivity was increased (P< 0.05) after long transport. Several interactions between stress level and muscle type (P< 0.001) were observed. High preslaughter stress decreased (P< 0.001) muscle glycogen in both the LM and SSP, but this decrease was greater in the LM. Lactate concentrations were increased (P< 0.001) only in the LM by high preslaughter stress. Increases in ultimate pH (P< 0.001) and decreases in a* values (P< 0.01) were greatest in the SV, whereas increases in electrical conductivity (P< 0.001) were greatest in the LM. The lack of interactions among transportation, lairage, and muscle type was attributed to the relatively minor differences in stress among treatments. It was concluded that, in glycolytic muscle types such as the LM, the high physical and psychological stress levels associated with stress in the immediate preslaughter period have a greater effect on the water-holding capacity of the meat and may promote PSE development. Conversely, oxidative muscle types tend to have higher ultimate pH values and produce DFD pork in response to intense physical activity and/or high psychological stress levels preslaughter.

The objectives of the present experiment were 1) to study the effects of transport conditions and lairage duration on stress level, muscle glycolytic potential, and pork quality; and 2) to investigate whether the negative effects of high stress immediately preslaughter are affected by preceding handling factors (transport and lairage). In a 2 × 2 × 2 factorial design, halothane-free pigs (n = 384) were assigned to either short (50 min) and smooth or long (3 h) and rough transport; long (3 h) or short (<45 min) lairage; and minimal or high preslaughter stress. Pigs were processed in eight groups (48 pigs per group) on various days at a commercial plant. Blood samples were taken at exsanguination to measure plasma cortisol and lactate concentrations. Muscle pH and temperature were measured at 30 and 40 min, respectively, and both were measured at 3 h, postmortem. A LM sample was taken 135 min postmortem to estimate glycogen content and rate of glycolysis. Pork quality attributes were assessed 23 h postmortem. Short transport increased cortisol when followed by short lairage (transport × lairage; P< 0.01). Long transport, but not lairage (P> 0.30), tended to increase (P= 0.06) muscle glycolytic potential. Long transport tended to increase (P= 0.08) electrical conductivity, and decreased a* (P< 0.01) and b* (P< 0.02) values. Decreasing lairage from 3 h to <45 min decreased (P< 0.05) the L* value, but it did not (P> 0.10) affect other pork quality traits. High stress decreased (P< 0.001) muscle glycolytic potential, and increased (P< 0.001) plasma lactate, cortisol, muscle temperature, rate of pH decline, and ultimate pH. Except for decreased (P< 0.001) b* values, pork color was not (P> 0.40) affected by high stress, but water-holding properties (measured by electrical conductivity, filter paper moisture, and drip loss) were impaired (P< 0.001) by high stress. Fiber optic-measured light scattering and Warner-Bratzler shear force were not (P> 0.12) affected by any treatment. Comparisons with the “optimal” handling (short transport, long lairage, and minimal stress) revealed that, with regard to water-holding properties, the negative effects of high stress were aggravated by suboptimal transport and lairage conditions. High stress alone increased electrical conductivity by 56%, whereas high stress in combination with short lairage led to an 88% increase. However, high preslaughter stress contributed most and was the major factor responsible for reductions in pork quality.

The human gastric pathogen Helicobacter pylori spontaneously switches lipopolysaccharide (LPS) Lewis (Le) antigens on and off (phase-variable expression), but the biological significance of this is unclear. Here, we report that Le+ H. pylori variants are able to bind to the C-type lectin DC-SIGN and present on gastric dendritic cells (DCs), and demonstrate that this interaction blocks T helper cell (Th)1 development. In contrast, Le− variants escape binding to DCs and induce a strong Th1 cell response. In addition, in gastric biopsies challenged ex vivo with Le+ variants that bind DC-SIGN, interleukin 6 production is decreased, indicative of increased immune suppression. Our data indicate a role for LPS phase variation and Le antigen expression by H. pylori in suppressing immune responses through DC-SIGN.

The objective of the present experiment was to study physiological changes elicited in response to stress in the immediate preslaughter period and to link them to pork quality characteristics. Crossbred, halothane-free pigs (n = 192) were processed in eight groups (24 pigs per group) on various days at one of two commercial processing plants operating different stunning systems (electrical and CO2stunning in Plants A and B, respectively). In each group, half the pigs were exposed to either minimal or high preslaughter stress. Blood samples were taken at exsanguination, and lactate, cortisol, and catecholamines, as well as blood pH and temperature, were assessed and linked to various longissimus muscle quality attributes. Additionally, muscle pH and temperature were measured 30 min postmortem, and muscle glycolytic potential was determined 22 h postmortem. At both processing plants, high preslaughter stress resulted in higher (P< 0.05) blood cortisol and lactate; however, the effects of preslaughter stress on catecholamines and blood pH were believed to be biased by the different stunning methods employed at the plants. High preslaughter stress increased (P< 0.05) blood temperature at Plant A but not at Plant B. At both plants, high stress increased (P< 0.05) 30-min muscle temperature and decreased (P< 0.05) 30-min muscle pH. Ultimate pH was increased (P< 0.05) and muscle glycolytic potential was decreased (P< 0.05) by high preslaughter stress. At both plants, high stress resulted in inferior pork quality attributes (P< 0.05), including reflectance, electrical conductivity, filter paper moisture, drip loss, and L* value. The effect of stress was greater on water-holding capacity than on pork color, with drip losses increased by 56%. Of all stress indicators measured at exsanguination, only blood lactate was strongly correlated with pork quality attributes. Regression analyses revealed that blood lactate and glycolytic potential accounted for 52 and 48% of the variation in drip loss and L* value, respectively. In combination with high preslaughter stress, high glycolytic potentials were related to increased drip losses. We conclude that high preslaughter stress leads to impaired pork quality, with high muscle energy levels aggravating the negative effects of preslaughter stress. Monitoring stress level by blood lactate measurement in combination with strategies to control muscle energy present at slaughter may help to improve meat quality.

The present experiment investigated whether increasing chilling rate could improve meat quality in pigs exposed to either minimal or high stress immediately preslaughter. Pigs (n = 192) were offspring of halothane-free lines. On various days, four groups of 48 pigs were processed at a commercial plant. Within each group, half the pigs were exposed to either minimal or high preslaughter stress. Before entering the cooler at 45 min postmortem, carcasses of both minimal and high preslaughter stress treatments were allocated randomly to either conventional (+4°C for 22 h) or rapid (three-phase chilling tunnel: −15, −10, and −1°C for 15, 38, and 38 min, respectively, followed by storage at 4°C until 22 h postmortem) chilling. Temperature and pH were measured in the blood at exsanguination and in the longissimus lumborum (LL) and semimembranosus (SM) muscle at 0.5, 2.5, 4.5, 6.5, and 22 h postmortem. Meat quality attributes (water-holding capacity and objective color measurements) were assessed on the LL. Preslaughter stress level affected pH and temperature in both blood and muscle, with lower (P< 0.001) pH values and higher (P< 0.001) temperatures for pigs exposed to high vs. minimal stress. Rapid chilling led to a faster (P< 0.001) temperature decline regardless of preslaughter stress level. Rapid chilling did not (P> 0.05) influence the rate of pH decline in the LL muscle, but reduced (P= 0.061) pH decline in the SM. Rapid chilling, as opposed to conventional chilling, decreased (P< 0.05) electrical conductivity in the LL, regardless of preslaughter stress; however, it could not compensate for the detrimental effect (P< 0.05) of stress on drip loss, filter paper moisture absorption, and meat color (L* value). Results from the present study indicated that increasing chilling rate is not a suitable method to resolve pork quality problems caused by inadequate preslaughter handling.

Percutaneous thrombolysis has become an accepted treatment of thrombosed haemodialysis grafts. Several devices have been developed for mechanical thrombolysis, which macerate the clot using different mechanisms such as aspiration and fragmentation. The aim of our study was to compare the efficacy of three devices for mechanical thrombolysis in removing the thrombus from thrombosed haemodialysis access grafts and to determine the initial technical and clinical success, complication rates of each device, and graft patency after the procedure.

Interleukin 12 (IL-12) is a potent enhancer of interferon γ production by activated T cells. The high-affinity IL-12 receptor (IL-12R) is a heterodimer of a β1 and a β2 subunit. Expression of the signaling IL-12Rβ2 chain is usually low, as compared with the more abundant β1 chain, and may be rate-limiting for IL-12 sensitivity. Little is known about the mechanisms controllingIL-12Rβ2gene expression. Reporter gene assays in IL-12Rβ2-expressing Jurkat cells showed that truncation of the region from −151 to −61 abrogated promoter activity. The proximal promoter region does not contain a typical TATA box, suggesting a role for SP-1. Indeed, mutagenesis of the −63 SP-1 consensus site decreased transcription by 50%. Electrophoretic mobility shift experiments confirmed the binding of SP-1 and SP-3 at this site. In contrast, truncation of −252 to −192 increased promoter activity. Likewise, mutagenesis of the consensus nuclear factor of activated T cells site at −206 increased promoter activity by 70%, suggesting silencer activity of this element. Electrophoretic mobility shift experiments with primary Th (T helper) cells showed the formation of a specific, T-cell receptor-inducible complex at this site that is sensitive to cyclosporin A and supershifted with anti-NFATc2 in both Th1 and Th2 cells. Accordingly, cyclosporin A dose-dependently increased IL-12Rβ2 mRNA expression. These first data onIL-12Rβ2gene regulation indicate a TATA-less promoter, depending on SP-1/SP-3 transcription factors, and a negative regulatory NFAT element at −206. This element may contribute to the overall low level of IL-12Rβ2 expression on Th cells.

Human papillomavirus (HPV) can be detected by DNA amplification from clinical samples. The aim of the present study was to compare the HPV status in both cervical scrape and biopsy specimens obtained from 174 patients, using the recently developed broad spectrum SPF10PCR‐LiPA method. The detection rate of HPV in these materials was determined and the spectrum of HPV genotypes was compared. Cervical scrapes and biopsy specimens were obtained, either on the same day (group I), or with an interval of up to almost 2 years (group II, mean interval 97 days, range 1–469 days). HPV DNA was amplified by SPF10PCR and detected in a microtitre plate hybridization assay. Of the HPV‐positive cases, the genotype was determined by reverse hybridization of the same SPF10amplimer on a line probe assay (LiPA), discriminating between HPV genotypes 6, 11, 16, 18, 31, 33–35, 39, 40, 42–45, 51–54, 56, 58, 59, 66, 68, 70, and 74. The results showed that the detection rate and the spectrum of HPV genotypes in cervical scrapes and the corresponding biopsy specimens were highly comparable in both patient groups, even when multiple genotypes were present. In both groups, multiple HPV genotypes were more frequently detected in cervical scrapes than in the corresponding biopsy specimens. In conclusion, HPV infection can be diagnosed in cervical scrapes and biopsy specimens using the SPF10PCR‐LiPA system. Analysis of cervical scrapes accurately reflects the spectrum of HPV genotypes in the patient's cervical region, even with a sampling interval between the cervical scrape and the biopsy specimen. Copyright © 2001 John Wiley & Sons, Ltd.

IL-12 is a potent inducer of IFN-γ production and drives the development of Th1 cells. Human polarized Th2 cells do not express the signaling β2-subunit of the IL-12R and, therefore, do not signal in response to IL-12. The question was raised as to what extent the loss of the IL-12Rβ2 chain in Th2 cells has bearing on the stability of the human Th2 phenotype. In the present report, we show that restimulation of human fully polarized Th2 cells in the presence of IL-12 primes for a shift towards Th0/Th1 phenotypes, accompanied by suppression of GATA-3 expression and induction of T-bet expression. These reversed cells are further characterized by a marked IL-12Rβ2 chain expression and fully restored IL-12-inducible STAT4 activation. The IL-12-induced phenotypic shift proved to be stable as a subsequent restimulation in the presence of IL-4 and in the absence of IL-12 could not undo the accomplished changes. Identical results were obtained with cells from atopic patients, both with polyclonal Th2 cell lines and allergen-specific Th2 cell clones. These findings suggest the possibility of restoring IL-12 responsiveness in established Th2 cells of atopic patients by stimulation in the presence of IL-12, and that IL-12-promoting immunotherapy can be beneficial for Th2-mediated immune disorders, targeting both naive and memory effector T cells.

The host‐searching behaviour of Heterorhabditis megidisstrain NLH‐E 87.3 in the presence of insect hosts and plant roots, offered individually and in combination, was studied using a newly developed Y‐tube olfactometer filled with sand. Within a period of 24 hours infective juveniles (IJs) were significantly attracted to living G. mellonellalarvae and caused 100% larval mortality. Otiorhynchus sulcatuslarvae, however, did not elicit host‐oriented movement of IJs and no larval mortality was observed. Roots of strawberry plants induced a negative response in IJs. The combination of strawberry roots and O. sulcatuslarvae, however, strongly attracted IJs leading to 37% host mortality. It was shown that this type of Y‐tube choice arena is a useful tool in studying the searching behaviour of entomopathogenic nematodes in a semi‐natural habitat.

Genetic lack of interleukin 12 receptor β1 (IL-12Rβ1) surface expression predisposes to severe infections by poorly pathogenic mycobacteria or Salmonella and causes strongly decreased, but not completely abrogated, interferon (IFN)-γ production. To study IL-12Rβ1–independent residual IFN-γ production, we have generated mycobacterium–specific T cell clones (TCCs) from IL-12Rβ1–deficient individuals. All TCCs displayed a T helper type 1 phenotype and the majority responded to IL-12 by increased IFN-γ production and proliferative responses upon activation. This response to IL-12 could be further augmented by exogenous IL-18. IL-12Rβ2 was found to be normally expressed in the absence of IL-12Rβ1, and could be upregulated by IFN-α. Expression of IL-12Rβ2 alone, however, was insufficient to induce signal transducer and activator of transcription (Stat)4 activation in response to IL-12, whereas IFN-α/IFN-αR ligation resulted in Stat4 activation in both control and IL-12Rβ1–deficient cells. IL-12 failed to upregulate cell surface expression of IL-18R, integrin α6, and IL-12Rβ2 on IL-12Rβ1–deficient cells, whereas this was normal on control cells. IL-12–induced IFN-γ production in IL-12Rβ1–deficient T cells could be inhibited by the p38 mitogen-activated protein kinase (MAP) kinase inhibitor SB203580 and the MAP kinase kinase (MEK) 1/2 inhibitor U0126, suggesting involvement of MAP kinases in this alternative, Stat4-independent, IL-12 signaling pathway. Collectively, these results indicate that IL-12 acts as a partial agonist in the absence of IL-12Rβ1. Moreover, the results reveal the presence of a novel IL-12Rβ1/Stat4–independent pathway of IL-12 responsiveness in activated human T cells involving MAP kinases. This pathway is likely to play a role in the residual type 1 immunity in IL-12Rβ1 deficiency.

Virulence and speed of action, as related to dose, are important effectiveness-determining properties of insect-pathogenic biocontrol agents. We used the droplet-feeding bioassay to compare dose responses between two wild-type baculoviruses, Autographa californicamulticapsid nucleopolyhedrovirus (AcMNPV) and Spodoptera exiguaMNPV (SeMNPV), and three deletion mutants of AcMNPV in S. exigualarvae. In each mutant one gene was deleted by genetic engineering: pp34,coding for the polyhedral membrane; egt,coding for ecdysteroid UDP–glucosyltransferase; or p10,coding for fibrillar structures in infected insect cells. SeMNPV had the lowest median lethal dose (LD50) as well as the highest speed of action (LT50) of all viruses investigated. In our comparative bioassays the only significant effect of gene deletions in AcMNPV was a slightly lower speed of action for the p10deletion mutant. Otherwise, wild-type and recombinant AcMNPVs had similar biological activities. Our results suggest, in contrast to what is generally assumed, that gene deletions in AcMNPV for improved insecticidal activity should be critically assessed in each host system prior to further implementation as a control agent. Insertion of foreign genes coding for entomotoxins is less questionable and more promising in this respect.

Photorhabdus luminescensisolates were exchanged between four entomopathogenic nematode strains; two isolates ofHeterorhabditis megidisfrom The Netherlands and two isolates ofH. bacteriophora, one from Australia and one from Moldavia. When cultured onH. megidissymbionts only fewH. bacteriophorainfective juveniles retained the symbiotic bacterium. These infective juveniles, without symbiotic bacteria, were not pathogenic to insects. WhenP. luminescenswas injected into insect larvae it could killGalleria mellonella(Lepidoptera: Galleridae) larvae but it was not pathogenic toTipula oleracea(Diptera: Tipulidae). The combination of nematode and bacterium killedT. oleracea, showing that the nematode is more than just a vector for the bacterium. Small differences in pathogenicity between the combinations could only be observed inT. oleracea, not in the highly susceptibleG. mellonella. The pathogenicity of a combination depends on the pathogenicity of the bacterium, the pathogenicity of the nematode and the interaction between them. The pathogenicity ofH. bacteriophorastrains againstT. oleraceawas low, partly because of the low penetration rate of these strains.

The nematode Steinernema feltiae(Nematoda: Steinemematidae) was tested for its ability to control two main mushroom pests i.e. the sciarid Lycoriella auripila(Diptera: Sciaridae) and the phorid Megaselia halterata(Diptera: Phoridae) in growing-rooms filled with spawned compost. A clear difference between female and male sciarid control was observed. A nematode application 1 day after casing preceded by an application 1 day before casing on the compost caused an almost complete control (97) of the F1-generation of female sciarids. The F2-generation of females was similarly controlled (95) by an application 7 days after casing. A dosage of 1 × 106nematodes m-2was found to be equally effective as higher dosages. Diflubenzuron remained active throughout entire the cropping period with high sciarid mortality rates varying from 72 to 99.

Experiments were conducted to examine the location of oviposition by the phorid fly Megaselia halterata(Wood) (Diptera: Phoridae) in uncased and cased compost. Clearly, a majority of the gravid females choose oviposition sites directly after entering the top layer of the compost. In uncased compost, 60% of all adults emerged from the top of four compost layers of equal thickness. When the compost was covered by a casing layer which was still uncolonized by Agaricus bisporus, oviposition was further concentrated in the top compost layer. In this situation, 91% of all adults emerged from the top compost layer whereas only 1.5% emerged from the casing. When the casing layer was colonized by mushroom mycelium, 45% of all adults emerged from the casing layer and 53% emerged from the top compost layer. Further concentration in the top compost layer and the casing layer occurred as a result of upward migration of larvae. When compost was cased after oviposition, up to 43% of all adults emerged from the casing layer. We concluded that in the control of phorid infestations with insect pathogenic nematodes, applications in uncased compost can be restricted to the upper compost layer. When compost and casing are filled simultaneously, nematode applications in the casing layer only could be considered.

In a comparative investigation of Critical Fusion Frequency (C. F. F.) values of 10 squirrels and 12 hedgehogs, it was shown that the C. F. F. values of a diurnal retina (squirrel) lie between about 30 and higher than 103 per sec., whereas these for a nocturnal retina (hedgehog) are about 30 per sec. or lower.Electroretinographic Critical Fusion Frequency determinations (C. F. F.) can be used to distinguish between « rod » and « cone » systems.

The existence of a fibre connection from the deep layers of the dorsal lateral geniculate body to the lateral posterior thalamic nucleus has been confirmed with histological methods in the cat. A fibre tract from the lateral posterior thalamic nucles to the ventral anterior thalamic nucleus and to the central lateral thalamic neucleus has been demonstrated.Electrical stimulation of the lateral posterior thalamic nucleus resulted in ipsilateral head movements, loss of posture, jerking and fully developed seizures with loss of bladder control, and with spikewave activity in the EEG in the freely moving cat.Electrical stimulation of anterior thalamic regions at the ventral anterior thalamic nucleus and at the central lateral thalamic nucleus produced only some jerking of the legs, which stopped on termination of the stimulus and was never combined with loss of bladder control.Electrical stimulation of the dorsal lateral geniculate body and of the rostral colliculus and of the nucleus cuneiformus confirmed earlier results from the literature.The lateral posterior thalamic nucleus is thought to be a major part of the centrencephalic epileptic system with a direct connection to the occipital cortex and with an indirect connection, through the anterior thalamic nuclei, to the frontal cortex.

Early Xenopusdevelopment is characterized by a poor correlation between global mRNA and protein abundances due to maternal mRNA and protein loading. Therefore, proteome profiling is necessary to study gene expression dynamics during early Xenopusdevelopment. In contrast to mammals, single Xenopuseggs and embryos contain enough protein to allow identification and quantification of thousands of proteins using mass spectrometry-based proteomics. In addition to investigating developmental processes, single egg or blastomere proteomes can be used to study cell-to-cell variability at an unprecedented depth. In this protocol, we describe a mass spectrometry-based proteomics approach for the identification and absolute quantification of Xenopus laevisegg or embryo proteomes, including sample preparation, peptide fractionation and separation, and data analysis.

Objective:This study explores older adults’ perspectives on healthy living, and their interactions with professionals regarding healthy living. This perspective is necessary for health professionals when they engage in tailored health promotion in their daily work routines. Method:In a qualitative study, 18 semi-structured interviews were carried out with older adults (aged 55-98) living in the Netherlands. The framework analysis method was used to analyze the transcripts. Results:Three themes emerged from the data—(a) healthy living: daily routines and staying active, (b) enacting healthy living: accepting and adapting, (c) interaction with health professionals with regard to healthy living: autonomy and reciprocity. Discussion:Older adults experience healthy living in a holistic way in which they prefer to live active and independent lives. Health professionals should focus on building an equal relationship of trust and focus on positive health outcomes, such as autonomy and self-sufficiency when communicating about healthy living.