Prochloraz causes irreversible masculinization of zebrafish (Danio rerio).

The aim of the present study was to investigate the persistence of endocrine effects by prochloraz, a fungicide known to have multiple effects on the endocrine system of vertebrates. Since discontinuous exposure is particularly relevant in aquatic ecosystems, an exposure scenario with an exposure phase and a subsequent recovery period was chosen to assess the potential for reversibility of effects by prochloraz on the sexual development of zebrafish (Danio rerio). Zebrafish were exposed to different concentrations of prochloraz (10-300 µg/L) until 60 days post hatch (dph), which includes the period of sexual differentiation. For the subsequent 40 days, fish were either held in clean water for depuration or under further continuous exposure. Histological investigations of the gonads revealed persistent effects on sexual differentiation. The sex ratio was skewed towards males and significantly more intersex individuals were found after exposure to prochloraz at 60 dph. No intersex fish, but masculinized sex ratios were still present after the depuration period, documenting that prochloraz irreversibly affects the sexual development of zebrafish.

Combining displacement field and grip force information to determine mechanical properties of planar tissue with complicated geometry.

Performing planar biaxial testing and using nominal stress-strain curves for soft-tissue characterization is most suitable when (1) the test produces homogeneous strain fields, (2) fibers are aligned with the coordinate axes, and (3) strains are measured far from boundaries. Some tissue types [such as lamellae of the annulus fibrosus (AF)] may not allow for these conditions to be met due to their natural geometry and constitution. The objective of this work was to develop and test a method utilizing a surface displacement field, grip force-stretch data, and finite-element (FE) modeling to facilitate analysis of such complex samples. We evaluated the method by regressing a simple structural model to simulated and experimental data. Three different tissues with different characteristics were used: Superficial pectoralis major (SPM) (anisotropic, aligned with axes), facet capsular ligament (FCL) (anisotropic, aligned with axes, bone attached), and a lamella from the AF (anisotropic, aligned off-axis, bone attached). We found that the surface displacement field or the grip force-stretch data information alone is insufficient to determine a unique parameter set. Utilizing both data types provided tight confidence regions (CRs) of the regressed parameters and low parameter sensitivity to initial guess. This combined fitting approach provided robust characterization of tissues with varying fiber orientations and boundaries and is applicable to tissues that are poorly suited to standard biaxial testing. The structural model, a set of C++ finite-element routines, and a Matlab routine to do the fitting based on a set of force/displacement data is provided in the on-line supplementary material.

Persistence of endocrine disruption in zebrafish (Danio rerio) after discontinued exposure to the androgen 17ß-trenbolone.

The aim of the present study was to investigate the effects of the androgenic endocrine disruptor 17ß-trenbolone on the sexual development of zebrafish (Danio rerio) with special emphasis on the question of whether adverse outcomes of developmental exposure are reversible or persistent. An exposure scenario including a recovery phase was chosen to assess the potential reversibility of androgenic effects. Zebrafish were exposed to environmentally relevant concentrations of 17ß-trenbolone (1 ng/L-30 ng/L) from fertilization until completion of gonad sexual differentiation (60 d posthatch). Thereafter, exposure was either followed by 40 d of recovery in clean water or continued until 100 d posthatch, the age when zebrafish start being able to reproduce. Fish exposed for 100 d to 10 ng/L or 30 ng/L 17ß-trenbolone were masculinized at different biological effect levels, as evidenced from a concentration-dependent shift of the sex ratio toward males as well as a significantly increased maturity of testes. Gonad morphological masculinization occurred in parallel with decreased vitellogenin concentrations in both sexes. Changes of brain aromatase (cyp19b) mRNA expression showed no consistent trend with respect to either exposure duration or concentration. Gonad morphological masculinization as well as the decrease of vitellogenin persisted after depuration over 40 d in clean water. This lack of recovery suggests that androgenic effects on sexual development of zebrafish are irreversible.

Quantitative T2* (T2 star) relaxation times predict site specific proteoglycan content and residual mechanics of the intervertebral disc throughout degeneration.

Degeneration alters the biochemical composition of the disc, affecting the mechanical integrity leading to spinal instability. Quantitative T2* MRI probes water mobility within the macromolecular network, a potentially more sensitive assessment of disc health. We determined the relationship between T2* relaxation time and proteoglycan content, collagen content, and compressive mechanics throughout the degenerative spectrum. Eighteen human cadaveric lumbar (L4-L5) discs were imaged using T2* MRI. The T2* relaxation time at five locations (nucleous pulposus or NP, anterior annulus fibrosis or AF, posterior AF, inner AF, and outer AF) was correlated with sulfated-glycosaminoglycan (s-GAG) content, hydroxyproline content, and residual stress and strain at each location. T2* relaxation times were significantly correlated with s-GAG contents in all test locations and were particularly strong in the NP (r = 0.944; p < 0.001) and inner AF (r = 0.782; p < 0.001). T2* relaxation times were also significantly correlated with both residual stresses and excised strains in the NP (r = 0.857; p < 0.001: r = 0.816; p < 0.001), inner AF (r = 0.535; p = 0.022: r = 0.516; p = 0.028), and outer AF (r = 0.668; p = 0.002: r = 0.458; p = 0.041). These strong correlations highlight T2* MRI's ability to predict the biochemical and mechanical health of the disc. T2* MRI assessment of disc health is a clinically viable tool showing promise as a biomarker for distinguishing degenerative changes.

Quantification of continuous in vivo flexion-extension kinematics and intervertebral strains.

PURPOSE: Healthy subjects performed lumbar flexion and were assessed by video fluoroscopy to measure the in vivo kinematics of the lower lumbar motion segments. METHODS: Fifteen healthy subjects (8 male, 7 female, 28 ± 10 years) performed lumbar flexion and extension back to neutral while their vertebrae were imaged. The sagittal plane vertebral margins of L3-S1 were identified. Lumbar angle, segmental margin strains, axial displacements, anterior-posterior (A-P) translations, and segmental rotations over the course of flexion were measured. RESULTS: L4-L5 had the largest posterior margin Green strain (65%). Each segment displayed more axial displacement than A-P translation. Peak vertebral angulation occurred at approximately 75% of peak flexion during the extension phase. CONCLUSION: L4-L5 exhibited the largest anterior and posterior margin strains (29 and 65%, respectively). Strains in the disc during in vivo lumbar flexion are due to both angular rotation and linear translation.

The maturity index as a tool to facilitate the interpretation of changes in vitellogenin production and sex ratio in the Fish Sexual Development Test.

In July 2011, the Fish Sexual Development Test (FSDT) has officially been adopted as OECD test guideline 234 for the detection of endocrine disrupting chemicals (EDCs). Sex ratio and vitellogenin (VTG) induction are the mandatory endocrine endpoints within this test, whereas gonad staging is only included as an option. In the present study, five FSDTs with zebrafish (Danio rerio) were conducted with EDCs with different modes of action (17α-ethinylestradiol, dihydrotestosterone, 17ß-trenbolone, prochloraz and 4-tert-pentylphenol). Results document that not only sex ratio and VTG production of the exposed fish were massively affected, but also gonad maturation. As a novel approach for the quantification of gonad maturation in zebrafish, the maturity index was developed to allow not only an improved assessment of dose-dependent EDC-related effects on gonad maturation, but also statistical analysis of histological data. VTG induction and maturity index showed an excellent correlation for all five EDCs tested. Most importantly, the maturity index often helped to find appropriate interpretations for results that seemed contradictory at first sight. Results show that histological analyses and their predictive power for population fitness are currently underestimated and should become a standard component in the evaluation of potential EDCs.