Enhancement of the photodynamic activity of tri-cationic porphyrins towards proliferating keratinocytes by conjugation to poly-S-lysine.

A fast uptake of the tri-cationic 5-(4-carboxyphenyl)-10,15,20-tris(4-methylpyridinium-4-yl)porphyrin tri-iodide (P-H), independent of the presence or absence of proteins in the culture medium, occurs during incubation of NCTC 2544 human keratinocytes with this porphyrin. By contrast, the uptake of the poly-S-lysine conjugate (P-(Lys)(n)) is faster in serum-free medium than in the supplemented medium suggesting that P-(Lys)(n) interacts with serum proteins. The P-(Lys)(n) uptake is almost an order of magnitude greater than that of P-H in serum-free or supplemented culture medium. With histidine as a specific probe of type II photodynamic reactions, the relative photosensitizing effectiveness of the conjugate is only one fourth that of P-H. Nevertheless, the photocytotoxicity of the conjugate is strongly enhanced as compared to that of P-H as a result of its larger uptake. Thus, the doses achieving 50% of photocytotoxicity after incubation with 5 microM of the conjugate and its parent cationic porphyrin are about 20 min and 1 h, respectively. Similarly, the initial rate of the cell lipid peroxidation induced by photosensitization with P-(Lys)(n) is about 8 times higher than that obtained with P-H. Fluorescence microscopy reveals that P-H is more diffusely located in the cytoplasm than P-(Lys)(n) which seems to accumulate in lysosome-like structures. Little if any staining of the nucleus is observed with both photosensitizers.     

Brittle stars (Echinodermata: Ophiuroidea) from La Réunion and the systematic position of Ophiocanops Koehler, 1922

The ophiuroid fauna of La Réunion, Indian Ocean, is still incompletely known. We report 13 different species from coral reefs, including five new records and a probably undescribed species of Amphioplus , which increases the number of ophiuroids known from La Réunion to 25. The most important find consists of three individuals of a previously unknown representative of the genus Ophiocanops . Originally placed in Ophiomyxidae, its systematic position has long been debated, with some authors advocating a placement with the otherwise extinct Palaeozoic Oegophiurida, based on the absence of several skeletal characters. The new species, O. multispina sp. nov. , shows intermediate characters between its previously only known congener, O. fugiens , and the ophiomyxid Renetheo felli . The three species appear to represent a line of taxa with progressively reduced skeleton instead of an ancient condition. We propose to transfer Ophiocanops back to the Ophiomyxidae, to synonymize Renetheo with Ophiocanops , and to invalidate the Ophiocanopidae, for which there is no autapomorphy-based support. © 2008 The Linnean Society of London, Zoological Journal of the Linnean Society , 2008, 153 , 545–560.     

Factors influencing somatic embryogenesis induction in <Emphasis Type="Italic">Eucalyptus globulus</Emphasis> Labill.: basal medium and anti-browning agents

The low induction rates of somatic embryogenesis (SE) in Eucalyptus globulus hamper scaling up the process for commercialization. We analyzed the effectiveness of several media (MS, 1/2MS, B5, WPM, DKW and JADS) during SE induction and expression. MS and B5 were the best media for SE induction and embling regeneration. In general, MS was the best medium for expression, independently of the medium previously used during induction. Several anti-browning compounds (ascorbic acid, charcoal, DTE, DTT, PVP, PVPP and silver nitrate) were added to the expression medium (MS), but all decreased SE potential and only DTE, charcoal and silver nitrate reduced explant browning. When added only during the induction period, anti-browning agents reduced accumulation of phenolics but also severely reduced SE potential. Continuous exposure completely inhibited the SE response. The negative impact of anti-browning agents on SE potential raises a question about the role of production/accumulation of phenolics in the SE process.     

In Vivo Imaging and Tracking of Technetium-99m Labeled Bone Marrow Mesenchymal Stem Cells in Equine Tendinopathy

Recent advances in the application of bone marrow mesenchymal stem cells (BMMSC) for the treatment of tendon and ligament injuries in the horse suggest improved outcome measures in both experimental and clinical studies. Although the BMMSC are implanted into the tendon lesion in large numbers (usually 10 - 20 million cells), only a relatively small number survive (<10%) although these can persist for up to 5 months after implantation. This appears to be a common observation in other species where BMMSC have been implanted into other tissues and it is important to understand when this loss occurs, how many survive the initial implantation process and whether the cells are cleared into other organs. Tracking the fate of the cells can be achieved by radiolabeling the BMMSC prior to implantation which allows non-invasive in vivo imaging of cell location and quantification of cell numbers.This protocol describes a cell labeling procedure that uses Technetium-99m (Tc-99m), and tracking of these cells following implantation into injured flexor tendons in horses. Tc-99m is a short-lived (t_1/2 of 6.01 hr) isotope that emits gamma rays and can be internalized by cells in the presence of the lipophilic compound hexamethylpropyleneamine oxime (HMPAO). These properties make it ideal for use in nuclear medicine clinics for the diagnosis of many different diseases. The fate of the labeled cells can be followed in the short term (up to 36 hr) by gamma scintigraphy to quantify both the number of cells retained in the lesion and distribution of the cells into lungs, thyroid and other organs. This technique is adapted from the labeling of blood leukocytes and could be utilized to image implanted BMMSC in other organs.     

Adipose tissue-derived factors as potential biomarkers in cachectic cancer patients

Cachexia, a paraneoplastic syndrome markedly associated with worsened prognosis in cancer patients, provokes profound wasting of both lean and adipose mass in an association with a state of metabolic “chaos”. The white adipose tissue responds to cachexia with marked local inflammation and may be thus a relevant contributor to systemic inflammation. To address this hypothesis we examined the correlation between tissue expression of adipokines and plasma concentration in cachectic and stable weight patients with or without cancer. Adiponectin and liver-derived CRP concentration were significantly higher in the cachectic groups when compared with stable weight patients (P < 0.01). The concentration of plasma IL-6 was higher (11.4-fold) in the cancer cachectic group when compared with weight-stable controls, and presented a significant correlation with the presence of cancer (P < 0.001). A marked increase (5-fold) in IL-6 as a result of the interaction between the presence of cachexia and the presence of tumour was observed in the subcutaneous tissue of the patients, yet not in the visceral depot. Plasma adiponectin levels were higher in cachectic cancer patients, compared with stable weight cancer patients individually matched by age, sex, and BMI, and the subcutaneous depot was found to be the main contributing tissue, rather than the visceral pad. Based on the results we concluded that the subcutaneous adipose tissue is associated with plasma changes that may function as markers of cachexia.     

In ovo injection with glycerol and insulin-like growth factor (IGF-I): hatchability,intestinal morphometry,performance, and carcass characteristics of broilers

ABSTRACT

The objective of this study was to evaluate the effects of in ovo injection with glycerol (GLY) and insulin-like growth factor (IGF-I) on hatchability, biochemical parameters, intestinal morphometry, performance, and carcass characteristics of broiler chickens. A total of 400 fertilised eggs were distributed into five experimental groups. The treatments were arranged as non-injected (control), saline solution injected (0.9% NaCl solution), GLY solution injected (10 nmol/ml), IGF-I solution injected (100 ng/ml), and GLY + IGF-I solution injected. At 17.5 d of incubation, 0.5 ml of each solution was injected into the amniotic fluid of each egg of the injected groups. The injection of different solutions did not influence the hatchability and incubation time of the eggs. Compared to intact eggs, IGF-I and IGF-I+ GLY increased (p < 0.01) the blood IGF-I at hatching. Higher hepatic glycogen was observed (p < 0.05) with GLY or IGF-I. The tested substances decreased (p = 0.02) the fructose 1,6-biphosfate phosphatase activity but did not affect glycaemia. No difference in performance was observed in the first week. Higher feed intake and weight gain with lower feed conversion ratio was obtained ( p  < 0.05) with IGF-I at 14 d. At 21 d, higher weight gain was obtained (p = 0.05) with IGF-I, GLY, IGF-I, and GLY + IGF-I, resulting (p < 0.01) in birds with greater weight gain at 35 and 42 d of age. GLY provided higher villus height in the ileum at hatching and at 7 d of age. The tested solutions increased the relative weight of the liver at hatching. At 42 d of age, no carcass characteristics were influenced. It is concluded that GLY and IGF-I, together or separately, can be used in the in ovo feeding to improve the post-hatch performance of broilers, without affecting hatchability and carcass composition.     

Ant removal distance,but not seed manipulation and deposition site increases the establishment of a myrmecochorous plant

Oecologia - Myrmecochory (seed dispersal by ants) is a unique seed dispersal syndrome among invertebrates. It comprises three main phases: seed removal, seed manipulation, and seed deposition....