Toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in the developing male Wistar(Han) rat II: chronic dosing causes developmental delay

We have investigated whether fetal exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) causes defects in the male reproductive system of the rat, using chronically exposed rats to ensure continuous exposure of the fetus. 5-6 week old rats were exposed to control diet, or diet containing TCDD, to attain an average dose of 2.4, 8 and 46 ng TCDD kg-1 day-1 for twelve weeks, whereupon the rats were mated, and allowed to litter; rats were switched to control diet after parturition. Male offspring were allowed to develop until kills on PND70 (25 per group), or PND120 (all remaining animals). Offspring from the high dose group showed an increase in total litter loss, and the number of animals alive on post-natal day (PND) 4 in the high dose group was ~26% less than control. The high and medium dose offspring showed decreased weights at various ages. Balano-preputial separation was significantly delayed in all three dose groups, compared to control. There were no significant effects of maternal treatment when the offspring were subjected to a functional observational battery, or learning tests, with the exception that the high dose group showed a deficit in motor activity. 20 rats per group were mated to females, and there were no significant effects of maternal treatment on the fertility of these rats, nor on the F1 or F2 sex ratio. Sperm parameters at PND70 and 120 showed no significant effect of maternal treatment, with the exception that there was an increase in the proportion of abnormal sperm in the high dose group at PND70; this is associated with the developmental delay in puberty in this dose group. There were no remarkable findings of maternal treatment on organ weights, with the exception that testis weights were reduced by ~10% at PND70 (but not PND120), and although the experiment was sufficiently powered to detect small changes, ventral prostate weight was not reduced. There were no significant effects of maternal treatment upon histopathological comparison of high dose and control group organs. These data confirm that developmental exposure to TCDD shows no potent effect on adult sperm parameters or accessory sexual organs, but show that delay in BPS occurs after exposure to low doses of TCDD, and this is dependent upon whether TCDD is administered acutely or chronically.

Characterization of Copper Covetic Bulk and Films: Copper with High Carbon Content

Incorporation of carbon nanostructures in metals is desirable to combine the strongly bonded electrons in the metal and the free electrons in carbon nanostructures that give rise to high ampacity and high conductivity, respectively. Carbon in copper has the potential to impact industries such as: building construction, power generation and transmission, and microelectronics. This thesis focuses on the structure and properties of bulk and thin films of a new material, Cu covetic, that contains carbon in concentrations up to 16 at.%. X-ray photoelectron spectroscopy (XPS) shows C 1s peak with both sp2 and sp3 bonded C measuring up to 3.5 wt.% (16 at.%). High resolution transmission electron microscopy and electron diffraction of bulk covetic samples show a modulated structure of ≈ 1.6 nm along several crystallographic directions in regions that have high C content suggesting that the carbon incorporates into the copper lattice forming a network. Electron energy loss spectra (EELS) from covetics reveal that the level of graphitization from the source material, activated carbon, is maintained in the covetic structure. Bulk Cu covetics have a slight increase in the lattice constant, as well as <111> texturing, or possibly a different structure, compared to pure Cu. Density functional theory calculations predict bonding between C and Cu at the edges and defects of graphene sheets. The electrical resistivity of bulk covetics first increases and then decreases with increasing C content. Cu covetic films were deposited using e-beam and pulsed laser deposition (PLD) at different temperatures. No copper oxide or any allotropes of carbon are present in the films. The e-beam films show enhanced electrical and optical properties when compared to pure Cu films of the same thickness even though no carbon was detected by XPS or EELS. They also have slightly higher ampacity than Cu metal films. EELS analysis of the C-K-edge in the PLD films indicate that graphitic carbon is transferred from the bulk into the films with uniform carbon distribution. PLD films exhibit flatter and higher transmittance curves and sheet resistance two orders of magnitude lower than e-beam films leading to a high figure of merit as transparent conductors.

A contribution towards the performance and structural understanding of copper and nickel salts as ammonia stores

The ongoing depletion of fossil fuels and the severe consequences of the greenhouse effect make the development of alternative energy systems crucially important. While hydrogen is, in principle, a promising alternative, releasing nothing but energy and pure water. Hydrogen storage is complicated and no completely viable technique has been proposed so far. This work is concerned with the study of one potential alternative to pure hydrogen: ammonia, and more specifically its storage in solids. Ammonia, NH3, can be regarded as a chemical hydrogen carrier with the advantages of strongly reduced flammability and explosiveness as compared to hydrogen. Furthermore, ammine metal salts presented here as promising ammonia stores easily store up to 50 wt.-% ammonia, giving them a volumetric energy density comparable to natural gas. The model system NiX2–NH3 ( X = Cl, Br, I) is studied thoroughly with respect to ammine salt formation, thermal decomposition, air stability and structural effects. The system CuX2–NH3 ( X = Cl, Br) has an adverse thermal decomposition behaviour, making it impractical for use as an ammonia store. This system is, however, most interesting from a structural point of view and some work concerning the study of the structural behaviour of this system is presented. Finally, close chemical relatives to the metal ammine halides, the metal ammine nitrates are studied. They exhibit interesting anion arrangements, which is an impressive showcase for the combination of diffraction and spectroscopic information. The characterisation techniques in this thesis range from powder diffraction over single crystal diffraction, spectroscopy, computational modelling, thermal analyses to gravimetric uptake experiments. Further highlights are the structure solutions and refinements from powder data of (NH4)2[NiCl4(H2O)(NH3)] and Ni(NH3)2(NO3)2, the combination of crystallographic and chemical information for the elucidation of the (NH4)2[NiCl4(H2O)(NH3)] formation reaction and the growth of single crystals under ammonia flow, a technique allowing the first documented successful growth and single crystal diffraction measurement for [Cu(NH3)6]Cl2.

Molecular and biochemical mechanisms involved in the toxicity of a marine cyanobacteria compound on cancer cell lines

In recent years marine biotechnology has revealed a crucial role in the future of bioindustry. Among the many marine resources, cyanobacteria have shown great potential in the production of bioactive compounds with diverse applicability. The pharmacological potential of these organisms has been one of the most explored areas in particular its antibacterial, antifungal and anticancer potential. This work was based on the assessment of potential anticancer compound E13010 F 5.4 isolated from marine cyanobacteria strain Synechocystis salina LEGE 06099. Thus the aim of this work was to explore molecular and biochemical mechanisms underlying the bioactivity detected in human cancer cells, specifically in lines RKO colon carcinoma and HT-29. The isolation of the compound was performed from biomass obtained by large-scale culture. To obtain the compound fractionation was carried and confirmation and isolation performed by Nuclear Magnetic Resonance (NMR), Thin Layer Chromatography (TLC) and High-Performance Liquid Chromatography (HPLC). Cell viability assays were performed based on reduction of 3- (4,5-dimetiltiaziol-2-yl) -2,5-diphenyltetrazolium bromide (MTT) to assess the cytotoxic potential of the compound. From the battery of cell lines RKO (colon carcinoma), HT-29 (colorectal adenocarcinoma), MG-63 (osteosarcoma) and T47D (breast carcinoma) the cell lines RKO and HT-29 were selected for elucidation of mechanisms of cytotoxicity. For the elucidation of the mechanisms involved in cytotoxicity the cell lines RKO and HT29 were exposed to the compound. A genomic approach based in the mRNA expression of genes involved in apoptosis and cell cycle by Real-Time PCR and a proteomic approach based on the separation of proteins by two-dimensional electrophoresis (2DGE) was performed. For mRNA expression were selected the genes RPL8, HPRT1, VDAC, SHMT2, CCNE, CCNB1, P21CIP, BCL-2 and BAD and for proteomics isoelectric focussing between 3 – 10 and molecular weight of 19 – 117 kDa separated by polyacrylamide gels (2DGE). The MTT results confirmed the reduction of the cell viability. The RT-PCR results for the expression of genes studied were not yet fully elucidative. For the cell line RKO there was a significant reduction in the expression of the gene P21CIP, and a tendency for reduction in the BAD gene expression and for increased expression of gene CCNB1, pointing to an effort for cell proliferation. In HT-29 cell line, there was a tendency for increase in the expression of P21CIP and BAD, which may explain the reduction in cell viability. The 2DGE results indicate proteomic patterns with differentially altered spots in the treated and control cells with both qualitative and quantitative differences, and differences in response between the RKO and HT-29 cell lines.

Acute burns of the hands – physiotherapy perspective.

Background: Acute burns of the hands are complex and may impact on various aspects of a person’s life. Physiotherapy rehabilitation and restoration of hand function is critical for the patient’s independence and re-integration into society. Purpose: This study aimed to explore the perceptions and experiences of physiotherapists in the management of patients with their hand burn injuries. Method: Five focus groups consisting of physiotherapists and physiotherapy assistants working with burn injured patients from each of the five selected public hospitals in KwaZulu-Natal were recruited. An explorative qualitative approach was adopted. Results: Physiotherapists emphasised that the acute management of the hand was trivialised due to a primary focus on the survival of the burn sufferer. Therapists identified several factors that determined the patients’ level of participation and motivation in therapy one of which was the procedural pain experienced. The role of the therapists’ within the rehabilitation framework was found to be critical to their recovery however there appeared to be a breakdown in the collaboration and communication among health care professionals to the detriment of effective intervention. Conclusion: A multidisciplinary team approach is the foundation in the management of acute burn injuries and during the trajectory of the trauma care continuum.

The effects of acute administration of Chinese aphrodisiacs sold in Blantyre City on sperm characteristics and fertility profile in guinea pigs

Background Chinese aphrodisiacs have become popular remedy for sexual dysfunction and improvement of libido in men in Malawi. However, selling of these drugs seems not to be well regulated. Probably the aphrodisiacs that are currently on the market have unknown efficacy, potency and safety profiles. The aim of this study was to assess the efficacy of imported Chinese aphrodisiacs using guinea pigs as a model. Materials and Methods Two types of drugs were purchased from vendors in Blantyre City. Tonic tea, which was purported to improve erectile function and libido, and sperm multiplier tablets which were claimed to increase the sperm count. The tonic tea was prepared by soaking one tea bag in 100ml boiling water. The tea was cooled and administered to eight male experimental animals in varying doses. Each animal was introduced into a separate cage with a female guinea pig. Sexual behaviour such as mounting, sniffing behind the female were observed and recorded. Each sperm multiplier tablet was dissolved in distilled water and administered to the experimental animals in the morning and evening for seven days. At the end of the treatment, the experimental and control animals were sacrificed, their semen collected and analysed sperm motility, concentration and morphology. Results For the tonic tea, there were no statistical differences between the experimental and the control animals in terms of the number of mountings and sniffing behind the female. The sperm multiplier drug showed statistically significant differences between the experimental and the control animals in terms of the sperm motility (78.24 ± 1.35 vs. 86.54 ± 1.88, p< 0.05), and concentration (54.28 ± 1.24 vs. 67.59 ± 2.12, p<0.05). Conclusion The tonic tea did not show any efficacy in improving erection and libido. The sperm multiplier tablets, purported to increase sperm production, significantly increased the sperm motility, sperm concentration in the treated animals.

A truncation in the Aryl Hydrocarbon Receptor of the CRL:WI(Han) rat does not affect the developmental toxicity of TCDD

The Aryl Hydrocarbon Receptor (AhR) is required for the toxicity of TCDD, and so the AhR of CRL:WI and CRL:WI(Han) rats was characterised. Western blot showed AhR proteins of ~110 and ~97 kDa in individual rats from both strains. The AhR cDNA from a CRL:WI(Han) rat with the ~110kDa protein revealed a sequence that was identical to that of the CRL:WI and SD rat. However, cloning of the AhR from a rat with the ~97kDa protein revealed a point mutation, and five variants encoding two C-terminally truncated variants of the AhR protein, arising from a point mutation in the intron/exon junction and consequent differential splicing. These C-terminally truncated variants were expressed and shown to give rise to a protein of ~97kDa; the recombinant AhR bound TCDD with an affinity that was not statistically different from the full-length protein. A single-nucleotide polymorphism (SNP) assay was developed, and showed that both alleles were represented in a Hardy-Weinberg equilibrium in samples of CRL:WI and CRL:WI(Han) populations; both alleles are abundant. Rats from two studies of TCDD developmental toxicity were genotyped, and the association with toxicity investigated using statistical analysis. There was no plausible evidence that the AhR allele had a significant effect on the toxic endpoints examined. These data show that the two AhR alleles are common in two strains of Wistar rat, and that the AhR alleles had no effect on TCDD-induced developmental toxicity in two independent studies.

Comparative toxicity of antifouling compounds on the development of sea urchin

In the present study, embryotoxicity experiments using the sea urchin Lytechinus variegatus were carried out to better clarify the ecotoxicological effects of tributyltin (TBT) and triphenyltin (TPT) (the recently banned antifouling agents), and Irgarol and Diuron (two of the new commonly used booster biocides). Organisms were individually examined to evaluate the intensity and type of effects on embryo-larval development, this procedure has not been commonly used, however it showed to be a potentially suitable approach for toxicity assessment. NOEC and LOEC were similar for compounds of same chemical class, and IC10 values were very close and showed overlapping of confidence intervals between TBT and TPT, and between Diuron and Irgarol. In addition, IC10 were similar to NOEC values. Regardless of this, the observed effects were different. Embryo development was interrupted at the gastrula and blastula stages at 1.25 and 2.5 mu g l(-1) of TBT, respectively, whereas pluteus stage was reached with the corresponding concentrations of TPT. Furthermore, embryos reached the prism and morula stages at 5 mu g l(-1) of TPT and TBT, respectively. The effects induced by Irgarol were also more pronounced than those caused by Diuron. Pluteus stage was always reached at any tested Diuron concentration, while embryogenesis was interrupted at blastula/gastrula stages at the highest concentrations of Irgarol. Therefore, this study proposes a complementary approach for interpreting embryo-larval responses that may be employed together with the traditional way of analysis. Consequently, this application leads to a more powerful ecotoxicological assessment tool focused on embryotoxicity.

Effect of copper on the activation of the acid phosphatase from the green algae Pseudokirchneriella subcapitata.

The in vitro activation effect of copper on the acid phosphatase of the green algae Pseudokirch- neriella subcapitata (formely Selenastrum capricor- nutum) under preincubation condition. Apparent Michaelis constant values of 1.21 and 0.37 mM, and activation energy values of 26.8 and 13.6 kJ mol -1 were determined in the absence and in the presence of 0.2 mM Cu2?, respectively. The dissociation constant value for Cu2? binding to the enzyme was determined to be 22.04 lM. The decrease of the apparent Michaelis constant (Km) and activation energy values in the presence of Cu2? correlates well with its activating effect on the acid phosphatase activity. This propriety could be used as a sensitive bioindicator for copper in environmental samples.

Release of copper complexes from a nanostructured sol–gel titania for cancer treatment

Copper complexes containing inorganic ligands were loaded on a functionalized titania (F-TiO2) to obtain drug delivery systems. The as-received copper complexes and those released from titania were tested as toxic agents on different cancer cell lines. The sol–gel method was used for the synthesis and surface functionalization of the titania, as well as for loading the copper complexes, all in a single step. The resultant Cu/F-TiO2 materials were characterized by several techniques. An “in vitro” releasing test was developed using an aqueous medium. Different concentrations (15.6–1000 µg mL−1) of each copper complex, those loaded on titania (Cu/F-TiO2), functionalized titania, and cis-Pt as a reference material, were incubated on RG2, C6, U373, and B16 cancer cell lines for 24 h. The morphology of functionalized titania and the different Cu/F-TiO2 materials obtained consists of aggregated nanoparticles, which generate mesopores. The amorphous phase (in dominant proportion) and the anatase phase were the structures identified through the X-ray diffraction profiles. These results agree with high-resolution transmission electron microscopy. Theoretical studies indicate that the copper compounds were released by a Fickian diffusion mechanism. It was found that independently of the copper complex and also the cell line used, low concentrations of each copper compound were sufficient to kill almost 100 % of cancer cells. When the cancer cells were treated with increasing concentrations of the Cu/F-TiO2 materials the number of survival cells decreased. Both copper complexes alone as well as those loaded on TiO2 had higher toxic effect than cis-Pt.

Recrystallization kinetic study of copper-bearing strip cast steels

The effects of copper on as-cast structure, recrystallization and precipitation kinetics of strip cast low carbon steel were investigated. As-cast microstructure mainly consists of polygonal ferrite and Widmanstatten ferrite. Recrystallization responses were strongly dependent on initial microstructure and Cu content. Precipitation strengthening was observed in high copper content alloys.

Thermal behavior of copper processed by ECAP with and without back pressure

Samples of electrolytic tough pitch (ETP) pure copper were subjected to 12 passes of Equal-Channel Angular Pressing (ECAP) at room temperature with and without back pressure. Subsequent annealing was performed to evaluate the influence of back pressure during ECAP on the thermal behavior of ultrafine-grained copper. The microstructural and hardness changes caused by annealing were characterized by orientation imaging microscopy (OIM) and microhardness measurements. The application of back pressure resulted in an earlier drop in hardness upon annealing, which is believed to be associated with a lower critical temperature for the initiation of recrystallization and a rapid coarsening of microstructure. Regardless of whether back pressure was applied or not, structure coarsening during short-time annealing of ECAP-processed copper was governed by discontinuous static recrystallization. This is seen as a result of microstructure heterogeneity. Analysis of recrystallization kinetics was carried out based on observations of the microstructure after annealing in terms of the Avrami equation. The magnitude of the apparent activation energies for recrystallization in the absence of back pressure and in the case of back pressure of 100 MPa was estimated to be ~99 kJ/mol and ~91 kJ/mol, respectively. The reasons for reduced activation energy in the case of processing with back pressure are discussed.

Corticomotor excitability is increased following an acute bout of blood flow restriction resistance exercise

We used transcranial magnetic stimulation (TMS) to investigate whether an acute bout of resistance exercise with blood flow restriction (BFR) stimulated changes in corticomotor excitability (motor evoked potential, MEP) and short-interval intracortical inhibition (SICI), and compared the responses to two traditional resistance exercise methods. Ten males completed four unilateral elbow flexion exercise trials in a balanced, randomized crossover design: (1) heavy-load (HL: 80% one-repetition maximum [1-RM]); (2) light-load (LL; 20% 1-RM) and two other light-load trials with BFR applied; (3) continuously at 80% resting systolic blood pressure (BFR-C); or (4) intermittently at 130% resting systolic blood pressure (BFR-I). MEP amplitude and SICI were measured using TMS at baseline, and at four time-points over a 60 min post-exercise period. MEP amplitude increased rapidly (within 5 min post-exercise) for BFR-C and remained elevated for 60 min post-exercise compared with all other trials. MEP amplitudes increased for up to 20 and 40 min for LL and BFR-I, respectively. These findings provide evidence that BFR resistance exercise can modulate corticomotor excitability, possibly due to altered sensory feedback via group III and IV afferents. This response may be an acute indication of neuromuscular adaptations that underpin changes in muscle strength following a BFR resistance training programme.

The acute effects of exercise on cigarette cravings, withdrawal symptoms, affect, and smoking behaviour: systematic review update and meta-analysis

RATIONALE: Smoking cessation is associated with cigarette cravings and tobacco withdrawal symptoms (TWS), and exercise appears to ameliorate many of these negative effects. A number of studies have examined the relationships between exercise, cigarette cravings, and TWS. OBJECTIVES: The objectives of this study were (a) to review and update the literature examining the effects of short bouts of exercise on cigarette cravings, TWS, affect, and smoking behaviour and (b) to conduct meta-analyses of the effect of exercise on cigarette cravings. METHODS: A systematic review of all studies published between January 2006 and June 2011 was conducted. RESULTS: Fifteen new studies were identified, 12 of which found a positive effect of exercise on cigarette cravings. The magnitude of statistically significant effect sizes for 'desire to smoke' and 'strength of desire to smoke' ranged from 0.4 to 1.98 in favour of exercise compared to passive control conditions, and peaked either during or soon after treatment. Effects were found up to 30 min post-exercise. Cigarette cravings were reduced following exercise with a wide range of intensities from isometric exercise and yoga to activity as high as 80-85 % heart rate reserve. Meta-analyses revealed weighted mean differences of -1.90 and -2.41 in 'desire to smoke' and 'strength of desire to smoke' outcomes, respectively. Measures of TWS and negative affect were reduced following light-moderate intensity exercise, but increased during vigorous exercise. CONCLUSIONS: Exercise can have a positive effect on cigarette cravings and TWS. However, the most effective exercise intensity to reduce cravings and the underlying mechanisms associated with this effect remain unclear.

Effect of an acute bout of plyometric exercise on neuromuscular fatigue and recovery in recreational athletes

Although plyometric training is widely used by sports coaches as a method of improving explosive power in athletes, many prescribe volumes in excess of the National Strength and Conditioning Association recommendations. The purpose of this study was to assess voluntary and evoked muscle characteristics to assess the neuromuscular impact of a high-volume bout of plyometric exercise that was non-exhaustive. Ten athletes who did not have plyometric training experience and were in their competitive season for club-level sport volunteered for the study. After at least 2 days without high-intensity activity, subjects were assessed on maximal twitch torque, time to peak torque, rate of twitch torque development, twitch half-relaxation time, rate of twitch relaxation, and voluntary activation by the interpolated twitch technique before, immediately after, and 2 hours after a high-volume plyometric training program (212 ground contacts). Data were analyzed by repeated-measures analysis of variance and described as mean +/- SD and Cohen d. Statistically significant decrements appeared immediately after the training protocol in the total torque generated by maximal voluntary contractions (p < 0.05, d = -0.51) and twitch (p < 0.01, d = -0.92), rate of twitch torque development (p < 0.01, d = -0.77), and rate of relaxation (p < 0.01, d = -0.73). However, we did not observe any differences that remained statistically different after 2 hours. There were no significant differences observed at any time point in time to peak twitch, half-relaxation time, or voluntary activation. We conclude that high-volume plyometric training results primarily in peripheral fatigue that substantially impairs force and rate of force development. We recommend that coaches carefully monitor the volume of plyometric training sessions to avoid neuromuscular impairments that can result in suboptimal training.