Guidelines for spacing and timing of samples to detect populations of Schistosoma mansoni cercariae in the field

The speed and extent of horizontal dispersion of schistosome cercariae are major factors to be considered in planning the sampling program for a field study on cercariae. Cercariae of Schistosoma mansoni were studied in a circular pool in the laboratory to determine their speed of dispersion from a release point in the centre of the 2 m diameter pool. Within 15 min of release, cercariae were found at the outer margin of the pool. The rate of travel away from the release point was proportional to the initial number of cercariae released.In relating these observations to the known daily emission pattern of cerceriae it was determined that the spacing of samples in flowing water should be proportional to the velocity of the stream and should be conducted after 12 noon.     

Effects of methylazoxymethanol given at different stages of postnatal life on development of the rat brain. Comparison with those of thyroid deficiency

Newborn rats were treated at different stages of their development with low doses of methylazoxymethanol acetate. The postnatal increase of the DNA content of the cerebrum did not differ from that of controls. In the cerebellum, the DNA content was transitorily reduced, but later, the external granular layer became thicker and DNA deposition increased in comparison with controls; finally, the cerebellar DNA returned to a normal value. Morphological abnormalities of the cerebellum, abnormal orientation of migrating cells, scattering of Purkinje cell bodies within the internal granule cells and specially striking abnormalities of the morphology and orientation of Purkinje cell dendrites were noted in rats treated with MAM from birth to day 3. The effects on the Purkinje cell morphogenesis persisted but were much less marked when MAM was given from 4 to 7 or from 8 to 11 days. Neonatal thyroid deficiency, as MAM-treatment between days 0 and 3, leads to an abnormal position of Purkinje cell bodies within the cerebellar cortex; it also leads to morphological abnormalities of their dendritic arborization which closely resemble those observed after MAM-treatment during the second postnatal week. It also alters the cell formation in the cerebellum. Thyroid deficiency probably exerts its effect on cell formation earlier than previous biochemical studies have shown. On another hand, the morphological abnormalities of Purkinje cell arborizations in the thyroid-deficient animals may be partly due to the perturbations of cell formation which persist later in the cerebellum.     

Glucocorticoid hormones increase the activity of γ-glutamyltransferase in a highly differentiated hepatoma cell line

γ-Glutamyltransferase activity was detected in the plasma membrane of the highly differentiated hepatoma cell line Fao, (0.93 mU/mg cell protein). Dexamethasone (1 μM) provoked a 2–3-fold increase in the activity of the enzyme in the presence of fetal calf serum. Maximal induction occurred 48–72 h after addition of the glucocorticoid to the cell culture medium. The hormonal specificity was demonstrated by the relative potencies of several glucocorticoids and sex steroids: hydrocortisone and corticosterone increased γ-glutamyltransferase activity while tetrahydrocorticosterone and all sex steroids tested were ineffective. The effect of dexamethasone on γ-glutamyltransferase activity was specific since the activities of several other plasma membrane enzymes were not modified. The mechanism of the dexamethasone-induced increase in γ-glutamyltransferase activity was neither by modification of the affinity of the enzyme for its substrates nor by alteration of the subcellular distribution of the enzyme. This increase was prevented by cycloheximide and actinomycin D. The data presented are consistent with a specific glucocorticoid receptor-mediated induction of γ-glutamyltransferase activity in Fao cells. The kinetic parameters of the induction process by glucocorticoids are very similar to those found in adult rat liver. These results suggest that the Fao cell line is a very convenient system for the study of the molecular mechanisms of glucocorticoid effects on differentiated cells.     

NARC-1/PCSK9 and its natural mutants: zymogen cleavage and effects on the low density lipoprotein (LDL) receptor and LDL cholesterol

The discovery of autosomal dominant hypercholesterolemic patients with mutations in the PCSK9 gene, encoding the proprotein convertase NARC-1, resulting in the missense mutations suggested a role in low density lipoprotein (LDL) metabolism. We show that the endoplasmic reticulum-localized proNARC-1 to NARC-1 zymogen conversion is Ca2+-independent and that within the zymogen autocatalytic processing site SSVFAQ [downward arrow]SIP Val at P4 and Pro at P3' are critical. The S127R and D374Y mutations result in approximately 50-60% and > or =98% decrease in zymogen processing, respectively. In contrast, the double [D374Y + N157K], F216L, and R218S natural mutants resulted in normal zymogen processing. The cell surface LDL receptor (LDLR) levels are reduced by 35% in lymphoblasts of S127R patients. The LDLR levels are also reduced in stable HepG2 cells overexpressing NARC-1 or its natural mutant S127R, and this reduction is abrogated in the presence of 5 mm ammonium chloride, suggesting that overexpression of NARC-1 increases the turnover rate of the LDLR. Adenoviral expression of wild type human NARC-1 in mice resulted in a maximal approximately 9-fold increase in circulating LDL cholesterol, while in LDLR-/- mice a delayed approximately 2-fold increase in LDL cholesterol was observed. In conclusion, NARC-1 seems to affect both the level of LDLR and that of circulating apoB-containing lipoproteins in an LDLR-dependent and -independent fashion.     

PIAS1-mediated sumoylation of focal adhesion kinase activates its autophosphorylation

Focal adhesion kinase (FAK) is a protein tyrosine kinase enriched in focal adhesions, which plays a critical role in integrin-dependent cell motility and survival. The crucial step in its activation is autophosphorylation on Tyr-397, which promotes the recruitment of several enzymes including Src family kinases and the activation of multiple signaling pathways. We found in a yeast two-hybrid screen that the N-terminal domain of FAK interacted with protein inhibitor of activated STAT1 (PIAS1). This interaction was confirmed and shown to be direct using in vitro assays. PIAS1 was co-immunoprecipitated with FAK from transfected cells and brain extracts. PIAS1 has recently been recognized as a small ubiquitin-like modifier (SUMO) ligase. In the presence of PIAS1 and SUMO-1, FAK was sumoylated in intact cells, whereas PYK2, a closely related enzyme, was not. Sumoylation occurred on Lys-152, a residue conserved in FAK during evolution. Sumoylated FAK, like PIAS1, was recovered predominantly from the nuclear fraction. Sumoylation did not require the catalytic activity or autophosphorylation of FAK. In contrast, sumoylation increased dramatically the ability of FAK to autophosphorylate in intact cells and in immune precipitate kinase assays. Endogenous FAK was sumoylated in the presence of PIAS1 and SUMO-1 independently of cell adhesion, and autophosphorylation of sumoylated FAK was persistently increased in suspended cells. These observations show that sumoylation controls the activity of a protein kinase and suggest that FAK may play a novel role in signaling between the plasma membrane and the nucleus.     

Serum and Lymphocytic Neurotrophins Profiles in Systemic Lupus Erythematosus: a Case-Control Study

Background

Neurotrophins play a central role in the development and maintenance of the nervous system. However, neurotrophins can also modulate B and T cell proliferation and activation, especially via autocrine loops. We hypothesized that both serum and lymphocytic neurotrophin levels may be deregulated in systemic Lupus erythematosus (SLE) and may reflect clinical symptoms of the disease.

Methods

Neurotrophins in the serum (ELISA tests) and lymphocytes (flow cytometry) were measured in 26 SLE patients and 26 control subjects. Th1 (interferon-γ) and Th2 (IL-10) profiles and serum concentration of BAFF were assessed by ELISA in the SLE and control subjects.

Findings

We have demonstrated that both NGF and BDNF serum levels are higher in SLE patients than healthy controls (p=0.003 and p<0.001), independently of Th1 or Th2 profiles. Enhanced serum NT-3 levels (p=0.003) were only found in severe lupus flares (i.e. SLEDAI ≥ 10) and significantly correlated with complement activation (decreased CH 50, Γ=-0.28, p=0.03). Furthermore, there was a negative correlation between serum NGF levels and the number of circulating T regulatory cells (Γ=0.48, p=0.01). In circulating B cells, production of both NGF and BDNF was greater in SLE patients than in healthy controls. In particular, the number of NGF-secreting B cells correlated with decreased complement levels (p=0.05). One month after SLE flare treatment, BDNF levels decreased; in contrast, NGF and NT-3 levels remained unchanged.

Conclusion

This study demonstrates that serum and B cell levels of both NGF and BDNF are increased in SLE, suggesting that the neurotrophin production pathway is deregulated in this disease. These results must be confirmed in a larger study with naive SLE patients, in order to avoid the potential confounding influence of prior immune-modulating treatments on neurotrophin levels.     

Wearing Blue-Blockers in the Morning Could Improve Sleep of Workers on a Permanent Night Schedule: A Pilot Study

Night shiftworkers often complain of disturbed sleep during the day. This could be partly caused by morning sunlight exposure during the commute home, which tends to maintain the circadian clock on a daytime rhythm. The circadian clock is most sensitive to the blue portion of the visible spectrum, so our aim was to determine if blocking short wavelengths of light below 540 nm could improve daytime sleep quality and nighttime vigilance of night shiftworkers. Eight permanent night shiftworkers (32–56 yrs of age) of Quebec City's Canada Post distribution center were evaluated during summertime, and twenty others (24–55 yrs of age) during fall and winter. Timing, efficacy, and fragmentation of daytime sleep were analyzed over four weeks by a wrist activity monitor, and subjective vigilance was additionally assessed at the end of the night shift in the fall–winter group. The first two weeks served as baseline and the remaining two as experimental weeks when workers had to wear blue-blockers glasses, either just before leaving the workplace at the end of their shift (summer group) or 2 h before the end of the night shift (fall–winter group). They all had to wear the glasses when outside during the day until 16:00 h. When wearing the glasses, workers slept, on average ±SD, 32±29 and 34±60 more min/day, increased their sleep efficacy by 1.95±2.17% and 4.56±6.1%, and lowered their sleep fragmentation by 1.74±1.36% and 4.22±9.16% in the summer and fall–winter group, respectively. Subjective vigilance also generally improved on Fridays in the fall–winter group. Blue-blockers seem to improve daytime sleep of permanent night-shift workers.     

Influence of Food Resources on the Ranging Pattern of Northern Pig-tailed Macaques (Macaca leonina)

Food availability may influence primates’ home range size and use. Understanding this relationship may facilitate the design of conservation strategies. We aimed to determine how fruit availability influences the ranging patterns of a group of northern pig-tailed macaques (Macaca leonina) living around the visitor center of Khao Yai National Park, Thailand. We predicted that macaques would increase their range during low fruit abundance periods to gather high-quality food and that they would go where there are more fruits or more fruits of particular species. We also predicted that human food, linked to human presence, would attract the macaques. We followed the macaques and recorded their diet and movements within their home range. We superimposed a grid on kernels defining the monthly home range surface to compare spatially macaques’ travel and the availability of fruits measured on botanical transects. Our results showed that the macaques increased their monthly home range in March, probably to obtain newly available fruits. During high fruit abundance seasons, they spent more time near particular fruit species. In August and September, although fruits became rare again, macaques kept their home range large, perhaps to find enough fruits as supplies dwindled. Finally, from October to February, they decreased their monthly home range size while consuming human food, a high-quality item. In conclusion, the macaques used several ranging strategies according to fruit availability. However, we think that, without human food, macaques would tend to increase their range during low fruit abundance periods, as predicted.