Tumour, but not Age-associated, Increase of Senescence Markers γH2AX and p21 in the Canine Eye.

Senescent cells display an irreversible cell cycle arrest with resistance to apoptosis. They are known to accumulate with age in mice, monkeys and man, and are suspected to drive the development and progression of neoplasia. Eyes develop age-associated changes, most commonly in the retina, cornea and lens. The aim of this study was to test whether senescent cells increase with age in the canine eye in general and in the microenvironment of ocular tumours in particular. The senescence markers γH2AX and p21 were tested in young (n = 10, age ≤2 years) versus old (n = 9, age range 9.5-12.4 years) canine eyes, as well as in the microenvironment of intraocular tumours, namely uveal melanocytomas (n = 13) and ciliary body adenomas (n = 9). To consider a potential association of senescence with biological behaviour, we compared the expression of both markers in tumour cells of benign uveal melanocytomas (n = 13) versus malignant conjunctival melanomas (n = 7). Canine eyes showed no age-dependent changes in senescent cells. However, a significant increase of the percentage of γH2AX- or p21-labelled cells was found in the retina, uvea and lens of tumour-bearing eyes. Tumour cells in conjunctival melanomas had a significantly increased percentage of p21-expressing cells compared with uveal melanocytomas. We conclude, that senescent cells do not accumulate with age in otherwise normal canine eyes and that a senescent microenvironment of intraocular tumours is unlikely to be age driven. In addition, as in man, the percentage of p21-positive cells was increased in melanomas, supporting the theory that malignant tumours may override the senescence-associated cell cycle arrest.

An innovative multistage treatment system for sanitary landfill leachate depuration: Studies at pilot-scale.

In this work, an innovative methodology for the treatment of landfill leachates, after aerobic lagooning, is proposed and adjusted at pilot-scale. This methodology involves an aerobic activated sludge biological pre-oxidation (ASBO), a coagulation/sedimentation step (240mgFe3+/L, at pH4.2) and a photo-oxidation through a photo-Fenton (PF) reaction (60mg Fe2+, at pH2.8) combining solar and artificial light. The ASBO process applied to a leachate after aerobic lagooning, with high organic and nitrogen content (1.1-1.5gC/L; 0.8-3.0gN/L) and low biodegradability (BOD5/COD =0.07-0.13), is capable to oxidise 62-99% of the ammonium nitrogen, consuming only the affluent alkalinity (70-100%). The coagulation/sedimentation stage led to the humic acids precipitation, promoting a marked change in leachate colour, from dark-brown to yellowish-brown (related to fulvic acids), accompanied by a reduction of 60%, 58% and 88% on DOC, COD and TSS, respectively. The PF system promoted the degradation of the recalcitrant organic molecules into more easily biodegradable ones. According to Zahn-Wellens biodegradability test, a leachate with 419mg DOC/L after coagulation, would have to be photo-oxidized until DOC <256mg/L, consuming 117mM of H2O2 and 10.4kJ/L of accumulated UV energy, to achieve an effluent that can be biologically treated in compliance with the COD discharge limit (150mg O2/L) into water bodies. The biological process downstream from the photocatalytic system would promote a mineralization >60%. The PF step cost to treat 100m3/day of leachate was 6.41€/m3, combining 1339m2 of CPCs with 31 lamps.

Insights into solar photo-Fenton reaction parameters in the oxidation of a sanitary landfill leachate at lab-scale.

This work evaluates the effect of the main photo-Fenton (PF) reaction variables on the treatment of a sanitary landfill leachate collected at the outlet of a leachate treatment plant, which includes aerated lagooning followed by aerated activated sludge and a final coagulation-flocculation step. The PF experiments were performed in a lab-scale compound parabolic collector (CPC) photoreactor using artificial solar radiation. The photocatalytic reaction rate was determined while varying the total dissolved iron concentration (20-100 mg Fe(2+)/L), solution pH (2.0-3.6), operating temperature (10-50 °C), type of acid used for acidification (H2SO4, HCl and H2SO4 + HCl) and UV irradiance (22-68 W/m(2)). This work also tries to elucidate the role of ferric hydroxides, ferric sulphate and ferric chloride species, by taking advantage of ferric speciation diagrams, in the efficiency of the PF reaction when applied to leachate oxidation. The molar fraction of the most photoactive ferric species, FeOH(2+), was linearly correlated with the PF pseudo-first order kinetic constants obtained at different solution pH and temperature values. Ferric ion speciation diagrams also showed that the presence of high amounts of chloride ions negatively affected the PF reaction, due to the decrease of ferric ions solubility and scavenging of hydroxyl radicals for chlorine radical formation. The increment of the PF reaction rates with temperature was mainly associated with the increase of the molar fraction of FeOH(2+). The optimal parameters for the photo-Fenton reaction were: pH = 2.8 (acidification agent: H2SO4); T = 30 °C; [Fe(2+)] = 60 mg/L and UV irradiance = 44 WUV/m(2), achieving 72% mineralization after 25 kJUV/L of accumulated UV energy and 149 mM of H2O2 consumed.

Enhancement of a solar photo-Fenton reaction with ferric-organic ligands for the treatment of acrylic-textile dyeing wastewater.

Literature describes a kinetic mineralization profile for most of acrylic-textile dyeing wastewaters using a photo-Fenton reaction characterized by a slow degradation process and high reactants consumption. This work tries to elucidate that the slow decay on DOC concentration is associated with the formation of stable complexes between Fe(3+) and textile auxiliary products, limiting the photoreduction of Fe(3+). This work also evaluates the enhancement of a solar photo-Fenton reaction through the use of different ferric-organic ligands applied to the treatment of a simulated acrylic-textile dyeing wastewater, as a pre-oxidation step to enhance its biodegradability. The photo-Fenton reaction was negatively affected by two dyeing auxiliary products: i) Sera(®) Tard A-AS, a surfactant mainly composed of alkyl dimethyl benzyl ammonium chloride and ii) Sera(®) Sperse M-IW, a dispersing agent composed of polyglycol solvents. The catalytic activity of the organic ligands toward the ferrous-catalysed system followed this order: Fe(III)-Oxalate > Fe(III)-Citrate > Fe(III)-EDDS, and all were better than the traditional photo-Fenton reaction. Different design parameters such as iron concentration, pH, temperature, flow conditions, UV irradiance and H2O2 addition strategy and dose were evaluated. The ferrioxalate induced photo-Fenton process presented the best results, achieving 87% mineralization after 9.3 kJUV L(-1) and allowing to work until near neutral pH values. As expected, the biodegradability of the textile wastewater was significantly enhanced during the photo-Fenton treatment, achieving a value of 73%, consuming 32.4 mM of H2O2 and 5.7 kJUV L(-1).

Performance evaluation of different solar advanced oxidation processes applied to the treatment of a real textile dyeing wastewater.

The performance of different solar-driven advanced oxidation processes (AOPs), such as TiO2/UV, TiO2/H2O2/UV, and Fe(2+)/H2O2/UV-visible in the treatment of a real textile effluent using a pilot plant with compound parabolic collectors (CPCs), was investigated. The influence of the main photo-Fenton reaction variables such as iron concentration (20-100 mg Fe(2+) L(-1)), pH (2.4-4.5), temperature (10-50 °C), and irradiance (22-68 WUV m(-2)) was evaluated in a lab-scale prototype using artificial solar radiation. The real textile wastewater presented a beige color, with a maximum absorbance peak at 641 nm, alkaline pH (8.1), moderate organic content (dissolved organic carbon (DOC) = 129 mg C L(-1) and chemical oxygen demand (COD) = 496 mg O2 L(-1)), and high conductivity mainly associated to the high concentration of chloride (1.1 g Cl(-) L(-1)), sulfate (0.4 g SO 4 (2 -) L(- 1)), and sodium (1.2 g Na(+) L(-1)) ions. Although all the processes tested contributed to complete decolorization and effective mineralization, the most efficient process was the solar photo-Fenton with an optimum catalyst concentration of 60 mg Fe(2+) L(-1), leading to 70 % mineralization (DOCfinal = 41 mg C L(-1); CODfinal < 150 mg O2 L(-1)) at pH 3.6, requiring a UV energy dose of 3.5 kJUV L(-1) (t 30 W = 22.4 min; [Formula: see text]; [Formula: see text]) and consuming 18.5 mM of H2O2.

Biodegradability and toxicity assessment of a real textile wastewater effluent treated by an optimized electrocoagulation process.

In this work, the application of an iron electrode-based electrocoagulation (EC) process on the treatment of a real textile wastewater (RTW) was investigated. In order to perform an efficient integration of the EC process with a biological oxidation one, an enhancement in the biodegradability and low toxicity of final compounds was sought. Optimal values of EC reactor operation parameters (pH, current density and electrolysis time) were achieved by applying a full factorial 3(3) experimental design. Biodegradability and toxicity assays were performed on treated RTW samples obtained at the optimal values of: pH of the solution (7.0), current density (142.9 A m(-2)) and different electrolysis times. As response variables for the biodegradability and toxicity assessment, the Zahn-Wellens test (Dt), the ratio values of dissolved organic carbon (DOC) relative to low-molecular-weight carboxylates anions (LMCA) and lethal concentration 50 (LC50) were used. According to the Dt, the DOC/LMCA ratio and LC50, an electrolysis time of 15 min along with the optimal values of pH and current density were suggested as suitable for a next stage of treatment based on a biological oxidation process.

Insights into real cotton-textile dyeing wastewater treatment using solar advanced oxidation processes.

Different advanced oxidation processes (AOPs) were applied to the treatment of a real cotton-textile dyeing wastewater as a pre-oxidation step to enhance the biodegradability of the recalcitrant compounds, which can be further oxidized using a biological process. Tests were conducted on a lab-scale prototype using artificial solar radiation and at pilot scale with compound parabolic collectors using natural solar radiation. The cotton-textile dyeing wastewater presents a lilac color, with a maximum absorbance peak at 641 nm, alkaline pH (pH = 8.2), moderate organic content (DOC = 152 mg C L(-1), COD = 684 mg O2 L(-1)) and low-moderate biodegradability (40 % after 28 days in Zahn-Wellens test). All the tested processes contributed to an effective decolorization and mineralization, but the most efficient process was the solar-photo-Fenton with an optimum catalyst concentration of 60 mg Fe(2+) L(-1), leading to 98.5% decolorization and 85.5% mineralization after less than 0.1 and 5.8 kJUV L(-1), respectively. In order to achieve a final wastewater with a COD below 250 mg O2 L(-1) (discharge limit into water bodies imposed by the Portuguese Legislation-Portaria no. 423/97 of 25 June 1997), considering the combination of a solar-photo-Fenton reaction with a biological process, the phototreatment energy required is 0.5 kJUV L(-1), consuming 7.5 mM hydrogen peroxide, resulting in 58.4% of mineralization [Formula: see text].

Gene expression profiling in subcutaneous, visceral and epigastric adipose tissues of patients with extreme obesity.

OBJECTIVE: The goal of the present study was to identify differences in gene expression between SAT, VAT and EAT depots in Class III severely obese individuals. DESIGN: Human subcutaneous (SAT) and visceral (VAT) adipose tissues exhibit differential gene expression profiles. There is little information, however, about the other proximal white adipose tissue, epigastric (EAT), in terms of its function and contribution to metabolism. SUBJECTS AND METHODS: Using RNA from adipose biospecimens obtained from Class III severely obese patients undergoing open Roux-en-Y gastric bypass surgery, we compared gene expression profiles between SAT, VAT and EAT, using microarrays validated by real-time quantitative PCR. RESULTS: The three depots were found to share 1907 genes. VAT had the greatest number of genes (66) expressed exclusively in this depot, followed by SAT (23), and then EAT (14). Moreover, VAT shared more genes with EAT (65) than with SAT (38). Further analyses using ratios of SAT/EAT, VAT/EAT and SAT/VAT identified specific as well as overlapping networks and pathways of genes representing dermatological diseases, inflammation, cell cycle and growth, cancer and development. Targeted analysis of genes, having a role in adipose tissue development and function, revealed that Peroxisome proliferator-activated receptor Gamma Coactivator 1-alpha (PGC1-α) that regulates the precursor of the hormone Irisin (FNCD5) were abundantly expressed in all three fat depots, along with fibroblast growth factors (FGF) FGF1, FGF7 and FGF10, whereas, FGF19 and FGF21 were undetectable. CONCLUSIONS: These data indicate that EAT has more in common with VAT, suggesting similar metabolic potential. The human epigastric adipose depot could have a significant functional role in metabolic diseases and should be further investigated.

Tension pneumothorax precluding laparoscopic repair of diaphragmatic hernia.

Pneumothorax can result from laparoscopic procedures in the abdomen. Usually, pneumothoraxes are mild and asymptomatic and do not require conversion to an open procedure. We report a case of tension pneumothorax that developed during the course of a laparoscopic repair of a diaphragmatic hernia. In this patient, the tension pneumothorax did not respond to conventional means of therapy and required conversion to a laparotomy. A large diaphragmatic hernia with communication between the peritoneal and pleural cavities may be a contraindication to minimally invasive laparoscopic procedures.

Modification of antibody isoelectric point affects biodistribution of 111-indium-labeled antibody.

We evaluated the influence of changes in charge on the biodistribution of 111In-labeled purified rabbit antihuman serum albumin (R-HSA) IgG conjugated to diethylenetriaminepentaacetic dianhydride (DTPA). Optimization of isoelectric point (pI) may influence the biodistribution profile, especially retention in vital organs, which ultimately affects radioimmunoimaging. Experiments were designed to modify the pI of R-HSA by conjugating various molar ratios of DTPA (DTPA:R-HSA ratios 5:1 to 100:1). The pI of the conjugates was determined by isoelectricfocusing (IEF). 111In-labeled DTPA:R-HSA with known pI range was injected intraperitoneally into BALB/c mice to evaluate biodistribution. There was a proportional relationship between the molar ratio of DTPA to R-HSA IgG and the number of DTPA substituted. Molar ratios of 5, 10, 20, 50 and 100 gave, on average, 2.0, 3.6, 5.1, 9.5 and 16.0 DTPA per R-HSA IgG, respectively. An anodal shift in the pI of 111In-labeled R-HSA IgG was noted with increased number of DTPA conjugation. Biodistribution studies at both 4 and 24 h showed sequential increase in the liver activity with increasing number of DTPA per antibody, whereas colon and small intestine showed a decrease in the activity at 4 h. The organ-specific increase (e.g., liver) or decrease (e.g., colon and small intestine) in the activity may depend on a critical balance of charge of a particular organ and its interaction with the amount of negative charge carried by the antibody conjugate. The results suggest that pI optimized 111In-labeled antibody could be used to increase or decrease colon and hepatic retention for more efficient radioimmunoimaging of colon tumors and their hepatic metastasis.

Desialylation of metastatic human colorectal carcinoma cells facilitates binding to Kupffer cells.

Cell surface hypersialylation of human colorectal carcinoma (HCRC) cells correlates with increased metastatic potential after intrasplenic injection, while desialylation with various agents has been shown to inhibit hepatic metastases. In this study we examined the effects of desialylation of HCRC cell lines with a novel intracellular inhibitor of the CMP-sialic acid transport protein (KI-8110). HCRC cells, which are poorly differentiated and poorly metastatic in nude mice (Clone A and MIP-101) were compared to well-differentiated, highly metastatic cells (CX-1 and CCL-235). KI-8110 treatment has previously been shown to reduce sialic acid levels in each of these cell lines and to reduce hepatic metastases in CX-1 and CCL-235 cell lines. This study attempts to identify a mechanism by which desialylation inhibits hepatic metastases. After KI-8110 treatment, in vitro adhesion assays were performed with each cell line to examine binding to Kupffer cells and the extracellular matrix protein fibronectin. Binding of Clone A, CX-1, and CCL-235 to Kupffer cells was significantly increased after KI-8110 treatment. Desialylation had no significant effect on binding of HCRC cell lines to fibronectin. While the metastatic cascade involves many complex interactions, the cytotoxic effects of Kupffer cells in the hepatic sinusoid are known to be an important mechanism of host defense against tumor cells. Cell surface sialic acids may well mask Kupffer cell binding to HCRC cells, preventing their cytotoxic effects and enhancing the metastatic potential of circulating tumor cells.

Carcinoembryonic antigen: enhancement of liver colonisation through retention of human colorectal carcinoma cells.

Carcinoembryonic antigen (CEA) is an oncofetal antigen whose function in the progression of colorectal carcinoma remains unclear although recent studies suggest it participates in homotypic cellular adhesion. We have previously shown that 40 micrograms of CEA injected intravenously into athymic nude mice enhances experimental metastasis in liver and lung by two human colorectal carcinoma cell lines that are injected intrasplenically 30 min later. The metastatic potential of another three moderately to highly metastatic colorectal carcinoma cell lines and of one weakly metastatic line has now been analysed in this model. CEA pretreatment only enhanced colony formation by cell lines that were weakly metastatic in untreated nude mice; it did not affect experimental metastasis by highly metastatic lines. CEA pretreatment enhanced the retention of 125I Idudr-labelled weakly metastatic tumour cells within the liver and lungs 4 h after intrasplenic injection but not the retention of highly metastatic tumour cells or inert latex beads. A significant correlation existed between the formation of experimental metastases and the early retention of tumour cells within the liver after intrasplenic injection. Aggregation did not appear to be important for retention in liver because CEA did not aggregate colorectal carcinoma cells in vitro. Also CEA did not alter natural host effector cell function in a cytolysis assay in vitro. We suggest that CEA facilitates liver colonisation by three of eight human colorectal carcinomas in athymic nude mice by increasing the hepatic retention of tumour cells. The potential mechanisms by which CEA may increase the retention of tumour cells in the liver are discussed.

A peptide sequence on carcinoembryonic antigen binds to a 80kD protein on Kupffer cells.

Clearance of carcinoembryonic antigen (CEA) from the circulation is by binding to Kupffer cells in the liver. We have shown that CEA binding to Kupffer cells occurs via a peptide sequence YPELPK representing amino acids 107-112 of the CEA sequence. This peptide sequence is located in the region between the N-terminal and the first immunoglobulin like loop domain. Using native CEA and peptides containing this sequence complexed with a heterobifunctional crosslinking agent and ligand blotting with biotinylated CEA and NCA we have shown binding to an 80kD protein on the Kupffer cell surface. This binding protein may be important in the development of hepatic metastases.