The influence of age and sex on phagocyte chemiluminescence

The process of ageing is associated with increased susceptibility to infection. Phagocytes form the primary defence mechanism against infecting microorganisms, but the influence of ageing on phagocyte function remains controversial. In this study we have applied a microtitre plate phagocyte chemiluminescence (CL) assay suitable for clinical use to compare phagocyte oxidative metabolism in younger healthy subjects (age 20–60 years) and healthy older (60–70 years) subjects. Polymorphonuclear leukocytes (PMNL) and monocytes were stimulated using phorbol myristate acetate (PMA), serum opsonized zymosan (SOZ), and non-opsonized zymosan (ZYM) in the presence of both lucigenin and luminol. Monocytes showed a higher luminolenhanced CL response to PMA in males compared with females in the younger age group. No PMNL differences were observed between the sexes. Although no difference were found in relation to age when cells were stimulated with PMA and SOZ, significantly lower background (unstimulated) CL was obtained from PMNL with luminol. PMNL luminol-enhanced CL responses were also lower in response to ZYM. The findings suggest a reduced response of PMNL from older subjects to minimal stimulation. This could be related to abnormalities in the triggering of the respiratory burst or myeloperoxidase release due to ageing. The influence of age and sex should be taken into account in clinical studies of phagocyte CL.     

Hot-water treatment before tissue culture reduces initial contamination in Lilium and Acer

Plant Cell, Tissue and Organ Culture (PCTOC) -     

Is the Sacred ibis a real threat to biodiversity? Long-term study of its diet in non-native areas compared to native areas

This paper presents the results of a 14-year study about the diet of the Sacred ibis in its main introduction area in France and its impact on native bird species, and compares the data to literature from its native area. During an initial period (1993–2004), the diet was essentially composed of invertebrates such as common aquatic insects (correlated with flooding) or Eristalis larvae picked from the mud (a vacant food niche in France), while scraps of meat taken from rubbish dumps were minor. These traditional preys taken from the same ecosystems as in its native area did not result in an exponential increase of the number of breeding Sacred ibises. Invasive Red swamp crayfish recently replaced other foods in its diet with a resulting sharp increase in breeding pair numbers (R² = 0.48). As in other parts of the world, vertebrates constituted very accidental preys, and no bird species were really threatened by such predation. Conversely, the Sacred ibis can have a positive effect as a predator of invasive crayfish. Adding the species to the DAISIE list of the 100 most invasive alien species in Europe therefore appears debatable.     

The adult fast isozyme of myosin is present in a nerve-muscle tissue culture system

Abstract. Organotypic nerve-muscle cultures were prepared from foetal mouse spinal cord and adult mouse muscle fibres. In this system, the adult fibres degenerate and new myotubes form. The regenerated muscle fibres become innervated, develop cross-striations, and survive for several months. We have investigated the isozymes of myosin present in these muscle fibres using histochemistry and immunocytochemistry with antibodies to rat embryonic, neonatal, and adult fast myosins. We demonstrate that some of the regenerated fibres contain adult fast but not embryonic or neonatal myosin. This is the first demonstration of the production of adult myosin heavy chain in tissue culture. This system therefore offers the possibility for further study of the development of adult myosin isoforms in vitro.     

5S ribosomal gene clusters in wheat: pulsed field gel electrophoresis reveals a high degree of polymorphism

Summary The long-range structure of 5S rRNA gene clusters has been investigated in wheat (Triticum aestivum L.) by means of pulsed field gel electrophoresis. Using aneuploid stocks, 5S rRNA gene clusters were assigned to sites on chromosomes 1B, 1D, 513 and 5D. Cluster sizes were evaluated and the copy number of 5S DNA repeats was estimated at 4700-5200 copies for the short repeating unit (410 bp) and about 3100 copies for the long repeat (500 bp) per haploid genome. A comparison of wheat cultivars revealed extremely high levels of polymorphism in the 5S rRNA gene clusters. With one restriction enzyme digest all varieties tested gave unique banding patterns and, on a per fragment basis, 21-fold more polymorphism was detected among cultivars for 5S DNA compared to standard restriction fragment length polymorphisms (RFLPs) detected with single copy clones. Experiments with aneuploid stocks suggest that the 5S rRNA gene clusters at several chromosomal sites contribute to this polymorphism. A number of previous reports have shown that wheat cultivars are not easily distinguished by isozymes or RFLPs. The high level of variation detected in 5S rRNA gene clusters therefore offers the possibility of a sensitive fingerprinting method for wheat. 5S DNA and other macro-satellite sequences may also serve as hypervariable Mendelian markers for genetic and breeding experiments in wheat.     

Enigma Prevents Cbl-c-Mediated Ubiquitination and Degradation of RETMEN2A

The Cbl proteins (Cbl, Cbl-b, and Cbl-c) are a highly conserved family of RING finger ubiquitin ligases (E3s) that function as negative regulators of tyrosine kinases in a wide variety of signal transduction pathways. In this study, we identify a new Cbl-c interacting protein, Enigma (PDLIM7). This interaction is specific to Cbl-c as Enigma fails to bind either of its closely related homologues, Cbl and Cbl-b. The binding between Enigma and Cbl-c is mediated through the LIM domains of Enigma as removal of all three LIM domains abrogates this interaction, while only LIM1 is sufficient for binding. Here we show that Cbl-c binds wild-type and MEN2A isoforms of the receptor tyrosine kinase, RET, and that Cbl-c enhances ubiquitination and degradation of activated RET. Enigma blocks Cbl-c-mediated RETMEN2A ubiquitination and degradation. Cbl-c decreased downstream ERK activation by RETMEN2A and co-expression of Enigma blocked the Cbl-c-mediated decrease in ERK activation. Enigma showed no detectable effect on Cbl-c-mediated ubiquitination of activated EGFR suggesting that this effect is specific to RET. Through mapping studies, we show that Cbl-c and Enigma bind RETMEN2A at different residues. However, binding of Enigma to RETMENA prevents Cbl-c recruitment to RETMEN2A. Consistent with these biochemical data, exploratory analyses of breast cancer patients with high expression of RET suggest that high expression of Cbl-c correlates with a good outcome, and high expression of Enigma correlates with a poor outcome. Together, these data demonstrate that Cbl-c can ubiquitinate and downregulate RETMEN2A and implicate Enigma as a positive regulator of RETMEN2A through blocking of Cbl-mediated ubiquitination and degradation.     

Interspike interval patterns of taste neurons in the hamster solitary nucleus

The central nervous system first processes taste informationin the solitary nucleus, which has been almost exclusively studiedin terms of average firing rate. We analyzed interspike intervals(ISI's) of 25 taste-responsive single units in the hamster (Mesocricetusauratus) solitary nucleus. ISI's were measured during spontaneousactivity and during stimulation with NaCl, KCl, sucrose, ora mixture of the three, and graphed on semi-logarithmic plots.Two different ISI patterns were evident: simple (13 units) andcomplex (12 units). Simple ISI patterns had a single broad peakat 284.7 ± 70.4 ms spontaneous and 78.8 ± 12.8ms stimulated. All complex ISI patterns had one distinct, sharppeak for an interval about 10 ms (11.3 ± 0.4 ms: spontaneous,9.3 ± 0.5 ms: stimulated), and a second, broader peakat 273.9 ± 45.9 ms spontaneous and 71.5 ± 14.6ms stimulated. As rate of firing increased peaks in ISI patternspredictably moved towards lower intervals, but ISI pattern-typedid not change. This constancy of ISI pattern held for responsesof a unit across all stimuli and did not depend upon the stimulusspecificity or location of the unit within the rostral poleof the solitary nucleus. Apparently, the pattern that a tasteneuron generates is intrinsic to the neuron and may relate tothe way it processes tast information.     

Health-Related Quality of Life,Self-Efficacy and Enjoyment Keep the Socially Vulnerable Physically Active in Community-Based Physical Activity Programs: A Sequential Cohort Study

Physical inactivity is most commonly found in socially vulnerable groups. Dutch policies target these groups through community-based health-enhancing physical activity (CBHEPA) programs. As robust evidence on the effectiveness of this approach is limited, this study investigated whether CBHEPA programs contribute to an increase in and the maintenance of physical activity in socially vulnerable groups. In four successive cohorts, starting at a six-month interval, 268 participants from 19 groups were monitored for twelve months in seven CBHEPA programs. Data collection was based on repeated questionnaires. Socio-economic indicators, program participation and coping ability were measured at baseline. Physical activity, health-related quality of life and on-going program participation were measured three times. Self-efficacy and enjoyment were measured at baseline and at twelve months. Statistical analyses were based on a quasi-RCT design (independent t-tests), a comparison of participants and dropouts (Mann-Whitney test), and multilevel modelling to assess change in individual physical activity, including group level characteristics. Participants of CBHEPA programs are socially vulnerable in terms of low education (48.6%), low income (52.4%), non-Dutch origin (64.6%) and health-related quality of life outcomes. Physical activity levels were not below the Dutch average. No increase in physical activity levels over time was observed. The multilevel models showed significant positive associations between health-related quality of life, self-efficacy and enjoyment, and leisure-time physical activity over time. Short CBHEPA programs (10–13 weeks) with multiple trainers and gender-homogeneous groups were associated with lower physical activity levels over time. At twelve months, dropouts'' leisure-time physical activity levels were significantly lower compared to continuing participants, as were health-related quality of life, self-efficacy and enjoyment outcomes. BMI and care consumption scored significantly higher among dropouts. In conclusion, Dutch CBHEPA programs reach socially vulnerable, but not necessarily inactive, groups in terms of socio-economic and health-related quality of life outcomes. Our findings suggest that CBHEPA programs particularly contribute to physical activity maintenance in socially vulnerable groups, rather than to an increase in physical activity behaviour over time.