Characterization by human autoantibody of a nuclear antigen related to the cell cycle

Using a serum from a patient with an autoimmune disease, we have recently described a novel 55 000-dalton antigen (p55) in the nucleus of several animal cells including human ones. This antigen, designated PSL, was not related to the previously defined antigens recognized by sera from patients with systemic rheumatic diseases (Sm, n-RNP, SS-B, Scl-70). We have now found that p55 is associated with chromatin structures as it is released from the nucleus of mink cell fibroblasts by saline + DNase treatments. Analysis by sucrose gradient centrifugation of the nuclear material released in these conditions indicated that p55 co-migrated with core histones. Meanwhile, p55 was absent from the residual nuclear matrices (achromatinic nuclei). Localization of p55 in synchronized cells was performed by indirect immunofluorescence and immunoprecipitation. P55 appeared to accumulate in the nucleus during the S phase. Finally, it was not recognized by an anti-SV40 tumor serum that specifically precipitated the protein p53, which has been recently related to cell proliferation. Thus, PSL an p53, although apparently not antigenically related, appear to be implicated in the same step of the cell cycle.     

Avian Use of Wetlands in Reclaimed Minelands in Southwestern Indiana

We studied the use of mineland wetlands by birds and the relationship between avian communities and wetland characteristics. Data were collected from 20 wetlands in Pike County, Indiana, and included wetland size, depth, water conductivity and salinity, aquatic macroinvertebrate abundance, vegetation, and bird use. Principal component analysis showed that physical variables could be explained by two principal component scores and that wetlands could be grouped on the basis of size and conductivity. Principal component analysis could not reduce vegetation variables to fewer principal component scores, meaning that wetland vegetation characteristics were independent of one another and did not show any trend. Most wetlands had low invertebrate density, and wetlands with higher invertebrate density had low invertebrate diversity. Wetlands with similar habitat characteristics (physical, vegetative, and invertebrate) did not necessarily show similarities in bird assemblages. Bird similarity index values ranged from 0 to 59%, implying that each wetland has its own bird community. Stepwise multiple regression analysis (α= 0.05) relating bird use and habitat characteristics showed that bird species richness increased with the species richness of submergent vegetation and was correlated negatively with the species richness of emergent vegetation. There was no significant relationship between bird species richness or bird species diversity and wetland size. The number of species within different avian guilds correlated with different habitat characteristics. The species richness of submergent plants was a factor that correlated positively with the number of species of several guilds (dabblers, wading birds, and plunge divers). Wetland age was not a factor that determined bird use.     

Is Clinical Practice Concordant with the Changes in Guidelines for Antiretroviral Therapy Initiation during Primary and Chronic HIV-1 Infection? The ANRS PRIMO and COPANA Cohorts

Objective

Guidelines for initiating HIV treatment are regularly revised. We explored how physicians in France have applied these evolving guidelines for ART initiation over the last decade in two different situations: chronic (CHI) and primary HIV-1 infection (PHI), since specific recommendations for PHI are also provided in France.

Methods

Data came from the ANRS PRIMO (1267 patients enrolled during PHI in 1996–2010) and COPANA (800 subjects enrolled at HIV diagnosis in 2004–2008) cohorts. We defined as guidelines-inconsistent during PHI and CHI, patients meeting criteria for ART initiation and not treated in the following month and during the next 6 months, respectively.

Results

ART initiation during PHI dramatically decreased from 91% of patients in 1996–99 to 22% in 2007 and increased to 60% in 2010, following changes in recommendations. In 2007, however, after the CD4 count threshold was raised to 350 cells/mm³ in 2006, only 55% of the patients with CD4≤350 were treated and 66% in 2008. During CHI, ART was more frequently initiated in patients who met the criteria at entry (96%) than during follow-up: 83% when recommendation to treat was 200 and 73% when it was 350 cells/mm³. Independent risk factors for not being treated during CHI despite meeting the criteria were lower viral load, lower educational level, and poorer living conditions.

Conclusion

HIV ART initiation guidelines are largely followed by practitioners in France. What can still be improved, however, is time to treat when CD4 cell counts reach the threshold to treat. Risk factors for lack of timely treatment highlight the need to understand better how patients’ living conditions and physicians’ perceptions influence the decision to initiate treatment.     

G Protein betagamma subunits augment UVB-induced apoptosis by stimulating the release of soluble heparin-binding epidermal growth factor from human keratinocytes

UV radiation induces various cellular responses by regulating the activity of many UV-responsive enzymes, including MAPKs. The betagamma subunit of the heterotrimeric GTP-binding protein (Gbetagamma) was found to mediate UV-induced p38 activation via epidermal growth factor receptor (EGFR). However, it is not known how Gbetagamma mediates the UVB-induced activation of EGFR, and thus we undertook this study to elucidate the mechanism. Treatment of HaCaT-immortalized human keratinocytes with conditioned medium obtained from UVB-irradiated cells induced the phosphorylations of EGFR, p38, and ERK but not that of JNK. Blockade of heparin-binding EGF-like growth factor (HB-EGF) by neutralizing antibody or CRM197 toxin inhibited the UVB-induced activations of EGFR, p38, and ERK in normal human epidermal keratinocytes and in HaCaT cells. Treatment with HB-EGF also activated EGFR, p38, and ERK. UVB radiation stimulated the processing of pro-HB-EGF and increased the secretion of soluble HB-EGF in medium, which was quantified by immunoblotting and protein staining. In addition, treatment with CRM179 toxin blocked UV-induced apoptosis, but HB-EGF augmented this apoptosis. Moreover, UVB-induced apoptosis was reduced by inhibiting EGFR or p38. The overexpression of Gbeta(1)gamma(2) increased EGFR-activating activity and soluble HB-EGF content in conditioned medium, but the sequestration of Gbetagamma by the carboxyl terminus of G protein-coupled receptor kinase 2 (GRK2ct) produced the opposite effect. The activation of Src increased UVB-induced, Gbetagamma-mediated HB-EGF secretion, but the inhibition of Src blocked that. Overexpression of Gbetagamma increased UVB-induced apoptosis, and the overexpression of GRK2ct decreased this apoptosis. We conclude that Gbetagamma mediates UVB-induced human keratinocyte apoptosis by augmenting the ectodomain shedding of HB-EGF, which sequentially activates EGFR and p38.     

Évaluation de méthodes automatiques de segmentation des volumes tumoraux en tomographie par émission de positons par comparaison avec des contours manuels réalisés par un groupe d’experts

Objective

Set up a framework for evaluating automatic segmentation methods of tumour volumes on PET images.

Patient and methods

This study was performed with PET images of 18 patients with non-Hodgkin's lymphoma. One target lesion per patient was pointed out. Each lesion was then three times manually delineated by five experts. Four automatic methods (the application of a threshold of 42% of the maximum SUV, the MIP-based method, the Daisne et al. method and the Nestle et al. method) were evaluated by comparison with the set of manual delineations.

Results

From the manual delineations, we have concluded to a moderate intra-operator variability and to a reduced interoperator reproducibility. From statistical tests performed on various quantitative criteria, there was no significant difference between the MIP-based method, the Daisne et al. method and the Nestle et al. one. The application of a threshold of 42% of the maximum SUV appears to be less efficient.

Conclusion

This work proposes a comparison and an evaluation protocol for segmentation methods. The generated data set will be distributed online for the community to simplify the evaluation of any new method of segmentation.     

The effect of cysteine and glutathione on sperm and oxidative stress parameters of post-thawed bull semen

The aim of this study was to determine the effects of antioxidants such as reduced glutathione (GSH) and cysteine in Laiciphose® extender on semen parameters, fertilizing ability, lipid peroxidation (LPO) level and glutathione peroxidise (GPx) activity of post-thawed bull semen. Totally 54 ejaculates of three bulls were used in the study. Five groups, namely; GSH (0.5 and 2 mM), cysteine (5 and 10 mM) and control group, were conducted to test the antioxidants in Laiciphose®. Insemination doses were processed that each 0.25-mL straw contained 15 × 10⁶ sperm. The addition of antioxidants did not present any significant effect on the percentages of post-thaw sperm morphology (acrosome and total abnormalities), subjective, CASA and progressive motilities, as well as sperm motility characteristics (VAP, VSL, VCL, LIN and ALH), compared to the control groups (P > 0.05). GSH 0.5 mM (55.5 ± 7.38%) and cysteine 10 mM (48 ± 5.65%) led to lower rates of DNA damage, compared to control (P < 0.05). As regards to MDA level, cysteine at 10 mM dose gave the highest level (4.99 ± 0.44 nmol/L) (P < 0.001). GPx activity was demonstrated to be higher level upon the addition of 5 mM cysteine when compared to the other groups (P < 0.05). With respect to fertility results based on 60-day non-returns, the supplementation of antioxidants did not present significant differences (P > 0.05). The results of this study may provide an useful information for the future studies in this area. So, further studies could be suggested to achieve better information in terms of the DNA damage and fertilizing capacity of bull sperm frozen with effective antioxidants.     

Nuclear ribonucleoproteins recognized by human antinuclear antibodies in retrovirus-infected cells

Antibodies present in sera of patients with auto immune diseases (systemic Lupus erythematosus, mixed connective tissue disease) were used to react with nuclear ribonucleoproteins (HnRNPs) from normal cells and cells infected with retroviruses. Only antibodies directed against Sm and RNP antigens precipitated particles with definite spectra of small nuclear RNAs (SnRNA) and proteins. No difference could be found between infected and uninfected cells, suggesting that virus replication is dependent on normal cellular fonctions.