Is tree growth reduction related to direct foliar injuries or soil chemistry modifications?

In the context of intense emissions causing atmospheric pollution, tree growth reductions could be related to soil chemistry modifications or direct foliar injuries. To verify these hypotheses, mineral soils were sampled in an area (Murdochville, Canada) where previous studies had demonstrated that tree growth was impacted by smelter emissions and that forest floor lead concentrations could be used as a proxy for atmospheric pollutant depositions. Samples were analysed for Al, Pb (concentrations and isotope ratios), basic cations (Ca, K, P, and Mg) and Zr. Mass balance calculations were performed on soil profiles to assess vertical migration of elements. Pb concentrations in litter diminished gradually with distance from the smelter. The Pb isotope ratios in these organic soil layers were close to those measured in the Murdochville ores. These patterns were not encountered in mineral soil layers. Pb isotope ratios in these layers were close to those measured in uncontaminated geological materials, and Pb concentrations and basic cation depletions were not related to the proximity of the smelter. Growth reduction was closely associated with litter Pb concentrations, which were used as a proxy for atmospheric deposition, but was not correlated with any elemental concentration or cation depletion measured in mineral soil layers. Our overall results suggest that trees responded mainly to direct atmospheric emissions, which caused foliar damage, rather than to soil chemistry modifications.

Spatiotemporal reconstruction of lead contamination using tree rings and organic soil layers.

An atmospheric dispersion model predicting ground-level concentrations from a point source of metal emissions (Murdochville smelter) was calibrated on tree rings in black spruce (Picea mariana Mill. B.S.P.) in order to reconstruct the spatial and temporal Pb-contamination pattern in the Gaspé Peninsula (Canada). Model predictions were validated with forest-floor Pb concentrations that resulted from accumulation of this element over the years and that provide a robust spatial-deposition pattern. Atmospheric emission records were also used to verify the good agreement between the smelter emissions and the temporal-information pattern present in tree rings. Tree rings that formed during the period of smelter emissions exhibited Pb concentrations that correlated with those measured in humus. Temporal variability in tree-ring concentrations was closely associated with the smelter emissions, suggesting that black spruce trees were able to record Pb pollution from a point source. However, a time gap of at least 15years must be considered between the emissions and the actual uptake and incorporation of Pb in the tree rings. A decrease in the level of contamination in the area was associated with the decrease in smelting activities, suggesting a natural resilience of the forest ecosystems to the contamination. This study highlights the strong potential for combining dendrochemical, soil, and modeling approaches in environmental research.

Metal contamination in the lichen Alectoria sarmentosa near the copper smelter of Murdochville, Québec.

Lichens were used to evaluate the metal pollution in a forest ecosystem around the smelter of Murdochville, Canada. As reached values 5.8 times higher in the smelter vicinity than in the 'background' sites. This enrichment was 2 times higher for 3 metals (Cu, Cd and Pb). The highest As, Ba, Cd, Cu, and Pb concentrations in lichens were 4, 112, 1, 23, 50 and 952 mg/kg respectively. Contamination declined exponentially with increasing distance from the smelter and was related to elevation and slope exposition to the smelter flux. (206)Pb/(207)Pb and (206)Pb/(204)Pb ratios were low close to the smelter (1.16 and 18), but increased with distance to constant values (1.19 and 18.7) and showed an inverse correlation with lead concentrations. Forest contamination was detectable up to 30 km from the smelter.

Heat shock protein expression and drug resistance in breast cancer patients treated with induction chemotherapy.

Heat shock proteins (Hsps) are induced in vitro by several cytotoxic drugs; in human breast cancer cells these proteins appear to be involved in anti-cancer drug resistance. The present report was designed to analyze whether chemotherapy affects in vivo the expression of Hsp27, Hsp70, Hsc70 and Hsp90 in breast cancer patients treated with induction chemotherapy and whether these proteins may be determinants of tumor resistance to drug administration. We have analyzed 35 biopsies from breast cancer patients treated with induction chemotherapy. Expression of the Hsps in the tumors was compared with (i) histological and clinical responses to chemotherapy, (ii) tumor cell proliferation measured by proliferating cell nuclear antigen (PCNA) immunostaining and nucleolar organizer regions (AgNORs) staining and (iii) the expression of estrogen and progesterone receptors. We also compared disease-free survival (DFS) and overall survival (OS) with the expression of the Hsps studied. After chemotherapy, nuclear Hsp27 and Hsp70 expression was increased and Hsp70 and Hsc70 cytoplasmic expression was decreased. A high nuclear proportion of Hsp70 in tumor cells (>10%) correlated significantly with drug resistance. We also observed that patients whose tumors expressed nuclear or a high cytoplasmic proportion (>66%) of Hsp27 had shorter DFS. The combination of Hsp27 and Hsp70 levels showed a strong correlation with DFS. Neither the cellular proliferation nor the levels of steroid receptors showed any significant difference before or after drug administration or during follow-up of patients. Our results suggest that Hsp27 and Hsp70 are involved in drug resistance in breast cancer patients treated with combination chemotherapies.

Heat shock proteins and cell proliferation in human breast cancer biopsy samples.

Human breast cancers may overexpress certain heat shock protein (hsp) family members, proteins which are involved with cell proliferation and differentiation as well as with disease prognosis and drug resistance. Here, we have studied the relationship between the expression of two hsps (hsp27 and hsp70) and the proliferative activity of tumor cells in 40 biopsies from breast cancer patients. Twenty of these tumors were selected for a detailed colocalization study. Immunocytochemistry was done using specific antibodies against hsp27 and hsp70. Cell proliferation was studied analyzing the expression of proliferating cell nuclear antigen (PCNA) (late G1, S, and G2 phases of the cell cycle) and the number of silver-staining nucleolar organizer regions (AgNORs) (G1 phase). The colocalization study revealed a statistically significant inverse correlation between hsp27 expression and cell proliferation in 16/19 (84%) of the cases evaluated by PCNA immunostaining, and in 11/16 (69%) of the cases evaluated by AgNORs. In contrast, a statistically significant positive correlation between hsp70 expression and elevated cell proliferation was seen in almost 85% of the cases evaluated by PCNA staining, and in almost 50% of the cases evaluated by AgNORs. Moreover, in 22% (9/40) of the breast cancer samples examined, hsp70 was clearly associated with the mitotic spindle. A Western blot analysis revealed that hsp70 was coprecipitated with taxol-polymerized tubulin. The association of hsp70 with the mitotic spindle was not clearly noted in lung carcinoma samples (N = 20) or in normal cells displaying elevated mitotic activity. These studies thus demonstrate that in a significant percentage of clinical breast cancers hsp27 overexpression is inversely correlated with cell proliferation, while hsp70 is clearly associated with the mitotic spindle and cell proliferation. These results add evidence to the concept that in human breast cancers hsp27 may be involved in cell growth arrest and increased differentiation while, in contrast, hsp70 may be involved in cell proliferation; further studies will be necessary to elucidate these possible cause-and-effect relationships.

Human papillomavirus infection and transitional cell carcinoma of the bladder. Immunohistochemistry and in situ hybridization.

The aim of the present research was to investigate human papillomavirus (HPV) infection by means of immunohistochemistry and in situ hybridization in 76 bladder cancer specimens. A biotinylated DNA probe that recognizes HPV 6/11, HPV 16/18 and HPV 31/33/35 was used for in situ hybridization. A polyclonal antibody recognizing HPV capsid antigen (HPVcAg) was used for immunohistochemistry. In situ hybridization and immunohistochemistry were developed by alkaline phosphatase and immunogold-silver techniques respectively. Our results showed that 25 (32.8%) out of 76 bladder carcinoma specimens reacted with HPVcAg. Twelve (15.7%) out of 76 cases were positive for HPV 16/18-DNA using non-isotopic in situ hybridization. Sixteen cases had koilocytosis. No positive signals were found for HPV 6/11 or 31/33/35-DNA probes.

Corticotropin-releasing hormone, luteinizing hormone-releasing hormone, growth hormone-releasing hormone, and somatostatin-like immunoreactivities in biopsies from breast cancer patients.

The presence of immunoreactive adrenocorticotropin-releasing hormone (CRH), luteinizing hormone-releasing hormone (LHRH), growth hormone-releasing hormone (GHRH), and somatostatin has been investigated by immunohistochemistry in forty biopsies from breast cancer patients. All of these hypothalamic hormones were found in about 30% of the samples, seen in the cytoplasm or in the nuclei of the tumor cells. Positive immunostaining for the hypothalamic hormones was present in colloid, lobular, and infiltrating ductal carcinomas. There was not a clear relationship between occurrence of staining for the hypothalamic hormones and the histologic grade of tumors or the clinical stage of the disease. Immunoreactive LHRH was more frequently found in breast tumors with estrogen and progesterone receptors. On the other hand, preneoplastic breast lesions expressed mainly somatostatin, while immunoreactivity was absent in normal mammary tissue.