Anabolic androgenic steroids reverse the beneficial effect of exercise on tendon biomechanics: An experimental study

Background The effect of anabolic androgenic steroids on tendons has not yet been fully elucidated. Aim of the present study was the evaluation of the impact of anabolic androgenic steroids on the biomechanical and histological characteristics of Achilles tendons. Methods Twenty-four male Wistar rats were randomized into four groups with exercise and anabolic steroids (nandrolone decanoate) serving as variables. Protocol duration was 12 weeks. Following euthanasia, tendons’ biomechanical properties were tested with the use of a modified clamping configuration. Histological examination with light and electron microscopy were also performed. Results In the group of anabolic steroids and exercise the lowest fracture stress values were observed, while in the exercise group the highest ones. Histological examination by light and electron microscopy revealed areas of collagen dysplasia and an increased epitendon in the groups receiving anabolic steroids and exercise. Conclusions These findings suggest that anabolic androgenic steroids reverse the beneficial effect of exercise, thus resulting in inferior maximal stress values

Assessing the effect of oxidative enzymes and stem anatomy on adventitious rooting of Olea europaea (L.) leafy cuttings

Aim of study: To assess the role of polyphenol oxidase (PPO), peroxidase (POD) and indole-3-acetic acid oxidase (IAAox) during adventitious rooting (Ar) in semi-hardwood cuttings of the easy-to-root olive cv. ‘Arbequina’ and the difficult-to-root cv. ‘Kalamata’. Simultaneously, a histological study was carried out in both cultivars to investigate the tissue related with Ar development.Area of study: The rooting experiments were carried out in ‘Kostelenos’ nurseries (Troizinia, Greece) and in Agricultural University of Athens.Material and methods: Plant material to set up the experiment was collected from current year shoots from 15-year-old mother plants of ‘Arbequina’ and ‘Kalamata’ at three different seasons (summer, autumn and spring). The auxin indole-3-butyric acid (IBA) at 2000 mg L-1 was used as rooting inducer.Main results: Analysis revealed that ‘Kalamata’ had significantly higher enzymatic activities before experiment onset and during Ar compared to ‘Arbequina’. Control cuttings of both cultivars exhibited increased enzymatic activities compared to IBA treated ones. IAAox was on average three times higher in ‘Kalamata’ than in ‘Arbequina’ and exhibited significant peaks during Ar. Similar peaks of POD and PPO activities were also detected. Histological analyses in ‘Kalamata’ revealed a continuous sheath of sclerenchyma ring and increased cortex thickness. Significant cell proliferation occurred in the phloem region in ‘Arbequina’ 15 days after planting and afterwards the root initials started developing in the secondary phloem from cambial cells.Research highlights: The differences in enzymatic activities as well as in stem anatomy could partly justify the different rooting ability of both cultivars

Molecular and biochemical characterization of the carbonic anhydrase isotypes of Caenorhabditis elegans

The genome of C. elegans was queried for the identification of genes, possibly encoding members of the carbonic anhydrase (CA) family. Eight genes were found encoding proteins with homology to characterized CAs. Six belonging to the a class (cah-1, cah-2, cah-3, cah-4, cah-5, and cah-6) and two belonging to the β class (bca-1 and y116a8c.28). Cah-4 has two products a and b. Cah-4a has a predicted leader peptide for localization in the mitochondria while the rest are predicted to be cytoplasmic. Proteins Cah-3, Cah-4, Cah-5, Bca-1 and Y116a8c.28 contain conserved aminoacids that are considered important for the CA activity of their corresponding class. Proteins Cah-1, Cah-2 and Cah-6 are missing all or several of these aminoacids. Genes cah-3, cah-4, bca-1 and y116a8c.28 are expressed throughout the development of the worm with the highest levels at the stages L1-3, while cah-1, cah-2, cah-5, and cah-6, are expressed at much lower levels. The yeast complementation experiment and the in vitro measurements performed using purified enzymes after expression in E. coli, showed that Cah-3, Cah-4 and Y116a8c.28 are active enzymes. The rest are either inactive, or their activity cannot be detected by the methods used. The expression of the genes was measured under 0% CO₂ or 0% O₂ and 0% CO₂. Expression levels of genes implicated in various metabolic pathways were also measured. The results showed that the expression of cah-3, cah-4, bca-1 and y116a8c.28 are affected by these stress conditions and that each gene is redulated differently. Silencing the genes by RNAi did not reveal any visual phenotypes both under normal or stress conditions involving 0% CO₂, 0% O₂ and 0% CO₂, or acid or basic pH. The only phenotype scored was a decrease in the maximum life span of worms fed with dsRNA for cah-3 and cah-4. Finally, the genome of C. elegans was found to contain genes encoding two a- CAs, four encoding proteins that could be referred to as CA related proteins (CA-RPs) and two encoding ß-CAs. None of these genes are required individually for the viability of the worm. The four CAs, both a and β, are expressed and functional under different conditions and probably serve different functions in the worms physiology.Το γονιδίωμα του C. elegans μελετήθηκε με σκοπό την αναζήτηση γονιδίων που πιθανά κωδικοποιούν ισοτύπους της καρβονικής ανυδράσης (CA). Οκτώ γονίδια βρέθηκαν με προϊόντα που εμφανίζουν ομολογία με χαρακτηρισμένες CA. Τα έξι είναι α τύπου (cah-1, cah-2, cah-3, cah-4, cah-5, και cah-6), ενώ τα δύο είναι β (bca-1 και y116a8c.28). Το γονίδιο cah-4 έχει δύο προϊόντα, το a και το b. To Cah-4a, πιθανότατα, φέρει πεπτίδιο οδηγό για εντοπισμό στα μιτοχόνδρια, ενώ οι υπόλοιποι ισότυποι είναι κυτταροπλασματικοί. Οι πρωτεΐνες Cah-3, Cah-4, Cah-5, Bca-1 και Y116a8c.28 φέρουν τα συντηρημένα αμινοξέα, στα ενεργά τους κέντρα, τα οποία θεωρούνται απαραίτητα για την καταλυτική δράση των CA του αντίστοιχου τύπου. Τα γονίδια cah-3, cah-4, bca-1 και y116a8c.28 εκφράζονται σε όλα τα στάδια ανάπτυξης του σκουληκιού και κυρίως στα στάδια L1-3, ενώ τα γονίδια cah-1, cah-2, cah-5, και cah-6, εκφράζονται σε πολύ χαμηλότερα επίπεδα. Από πείραμα συμπληρωματικότητας φαινοτύπου σε ζύμη και από in vitro μετρήσεις ενεργότητας ενζύμων, μετά από έκφραση σε E. coli και καθαρισμό, προέκυψε ότι τα Cah-3, Cah-4 και Y116a8c.28 είναι ενεργά ένζυμα. Τα υπόλοιπα, είτε δεν έχουν δράση CA, είτε δεν εμφανίζουν ενεργότητα κάτω από τις συνθήκες αυτές. Η έκφραση των γονιδίων μετρήθηκε και σε συνθήκες με 0% CO₂ ή 0% Ο₂ και 20% CO₂. Ταυτόχρονα, μετρήθηκε και η έκφραση γονιδίων που συμμετέχουν στον ενεργειακό μεταβολισμό του σκουληκιού. Τα αποτελέσματα έδειξαν ότι η έκφραση ων γονιδίων cah-3, cah-4, bca-1 και y116a8c.28 επηρεάζεται από τις συνθήκες αυτές και μάλιστα το κάθε γονίδιο συμπεριφέρεται διαφορετικά. Η σίγαση των γονιδίων, με RNAi, δεν εμφάνισε κάποιο ορατό φαινότυπο κάτω από φυσιολογικές συνθήκες, αλλά ούτε και υπό συνθήκες stress με ακραίες συγκεντρώσεις Ο₂ και CΟ₂ ή σε όξινο και βασικό περιβάλλον. Το μόνο που παρατηρήθηκε ήταν μια μείωση στον μέγιστο χρόνο ζωής στα σκουλήκια που μεγάλωσαν σε τρυβλία με dsRNA για το cah-3 και το cah-4. Συμπερασματικά, ο C. elegans, φαίνεται πως φέρει δύο α-CA, τέσσερις πρωτεΐνες που θα μπορούσαν να χαρακτηριστούν ως συγγενικές με τις CA (CA-RPs) και δύο β-CA. Κανένα από αυτά τα γονίδια αυτά δεν είναι απαραίτητο για την βιωσιμότητα του C. elegans. Οι τέσσερις CA, a και β τύπου, εκφράζονται και λειτουργούν κάτω από διαφορετικές συνθήκες πιθανότατα εξυπηρετώντας διαφορετικούς σκοπούς

Additional file 2: Figure S1. of Epidermal growth factor receptor status and Notch inhibition in non-small cell lung cancer cells

Representative plots for apoptosis and necrosis detection in H23, A549, H661 and HCC827 cells as described in “Materials and Methods” section. Control: untreated cells and DAPT: cells treated with DAPT. Four populations are distinguished: the viable (An-/PI-) [region 3], the early apoptotic (An+/PI-) [region 4], the late apoptotic (An+/PI+) [region 2] and the necrotic (An-/PI+) [region 1] cells. (PDF 399 kb

Additional file 3: Figure S2. of Epidermal growth factor receptor status and Notch inhibition in non-small cell lung cancer cells

Representative plots for cell cycle analysis in H23, A549, H661 and HCC827 cells as described in “Materials and Methods” section. Control: untreated cells, DAPT: cells treated with DAPT. The G0/G1, S and G2/M phases are indicated with blue, red and green colour, respectively. (PDF 113 kb