Sexing accuracy and indicators of maturation status in captive Murray cod Maccullochella peelii peelii using non-invasive ultrasonic imagery

Macroscopic- and histological-based assessments of gonad condition were compared with ultrasound images to determine the feasibility of this technology as a non-invasive diagnostic tool for identifying sex and assessing maturation status of Murray cod. Four age-classes (1⁺, 2⁺, 3⁺ and 6⁺ years), were sub-sampled at monthly intervals throughout their annual reproductive cycle and scanned with a 5 MHz linear transducer. An interpretation of sex was made from the resulting images and maximum cross-sectional gonad diameter and area were recorded. Fish were subsequently dissected to confirm gender, and the weights and maturation status of gonads determined and then compared with their respective image profile. Ovaries of females were usually a distinctive feature in ultrasound images, being particularly obvious in older and/or more developed fish. In contrast, the identification of male testis was more problematic. Nonetheless, identifying sex from ultrasound images was consistently achieved by recording the presence/absence of a female ovary (96% total sexing accuracy). Maximum cross-sectional ovary diameter and area were highly correlated with gonad weight (r² = 0.90 and 0.89, respectively) suggesting that indices of maturation status, comparable to the gonadosomatic index (GSI), can be obtained non-destructively from ultrasound scans of females. A less distinct relationship occurred between these dimensions and weight of testes (r² = 0.41). Significant increases (P < 0.05) in mean gonad index (GI, calculated from gonad diameter) occurred for most gonad development stages. However, differences in mean GI between maturation stages were confounded by phenotypic variability, indicating that GI may be limited to population level studies. Nevertheless, ultrasound images of ovaries at each development stage were visually distinctive and enabled qualitative evaluations of maturity, thereby complementing quantitative GI assessments. Repeated serial-monitoring of the same population using ultrasound appears to have great potential for tracking maturation-induced changes in broodfish.

Epidermal growth factor-induced ovarian carcinoma cell migration is associated with JAK2/STAT3 signals and changes in the abundance and localization of α6β1 integrin

Peritoneal dissemination of ovarian carcinoma is mediated by epithelial–mesenchymal interconversions leading to the disruption of cell–cell contact and modulation of cell–extracellular matrix (ECM) interactions. The present study was designed to evaluate the effects of epidermal growth factor (EGF) as a modulator of Janus kinase 2 (JAK2) and signal transducer and activator of transcription 3 (STAT3) signalling and changes in integrin expression during the process similar to EMT. A fibroblastic morphology with reduced intercellular cell contacts and increased cell motility was observed in ovarian cancer cell lines in response to EGF and was concomitant with the up regulation of EMT-associated N-cadherin and vimentin expression. These changes were accompanied by an increase in α2, α6 and β1 integrin subunits and activation of JAK2 and STAT3 signalling which was suppressed by a specific JAK2 inhibitor. Consistent with the suppression of STAT3 activity, N-cadherin and vimentin expression were abrogated and was coherent with the loss of cell motility and the expression of α6 and β1 integrin subunits. Neutralizing antibodies against α6 and β1 subunits inhibited cancer cell migration. A strong correlation between the expression of N-cadherin, vimentin and JAK2/STAT3 levels were detected in high-grade ovarian tumors and was consistent with the previously reported enhanced expression of α6 integrin subunit in advanced tumors [Ahmed N, Riley C, Oliva K, Rice G, Quinn M. Ascites induces modulation of α6β1 integrin and urokinase plasminogen activator receptor expression and associated functions in ovarian carcinoma. British Journal of Cancer 2005;92:1475–85]. Our data incorporating the clinical samples and the cancer cell lines is the first to demonstrate that JAK2/STAT3 pathway may be one of the downstream events in EMT-like process and α6β1 integrin-mediated signalling in ovarian carcinomas.

Elevated intracardiac angiotensin II leads to cardiac hypertrophy and mechanical dysfunction in normotensive mice

Introduction
Angiotensin II (Ang II) is known to induce cardiac growth and modulate myocardial contractility. It has been reported that elevated levels of endogenous Ang II contribute to the development of cardiac hypertrophy in hypertensives. However, the long-term functional effects of cardiac exposure to Ang II in normotensives is unclear.

A recently developed transgenic mouse (TG1306/1R), in which cardiac-specific overproduction of Ang II produces primary hypertrophy, provides a new experimental model for investigation of this phenotype. The aim of the present study was to use this model to investigate whether there is a functional deficit in primary hypertrophy that may predispose to cardiac failure and sudden death. We hypothesised that primary cardiac hypertrophy is associated with mechanical dysfunction in the basal state.

Methods
Normotensive heterozygous TG1306/1R mice harbouring multiple copies of a cardiac-specific rat angiotensinogen gene were studied at age 30—40 weeks and compared with age-matched wild-type littermates. Left ventricular function was measured ex vivo in bicarbonate buffer-perfused, Langendorffmounted hearts ( at a perfusion pressure of 80 mmHg, 37°C) using a fluid-filled PVC balloon interfaced to a pressure transducer and digital data acquisition system.

Results
There was no difference in the mean (±SEM) intrinsic heart rate of TG1306/1R and wild-type control mice (357.4±11.8 vs. 367.5±20.9 bpm, n=9 & 7). Under standardised end-diastolic pressure conditions, TG1306/1R hearts exhibited a significant reduction in peak developed pressure (132.2±9.4 vs. 161.5±3.1 mmHg, n=9 & 7, p<0.05) and maximum rate of pressure development (3566.7±323.7 vs. 4486.3±109.4 mmHg, n=9 & 7, p<0.05). TG1306/1R mice show a significant correlation between incidence of arrhythmia and increasing heart size (Spearman's correlation coefficient 0.61).

Conclusion
These data demonstrate that chronic in vivo exposure to elevated levels of intra-cardiac Ang II is associated with significant contractile abnormalities evident in the ex vivo intact heart. Our findings suggest that endogenous overproduction of cardiac Ang II, independent of changes in blood pressure, is sufficient to induce ventricular remodelling that culminates in impaired cardiac function which may precede failure.

A potential application of a piezoelectric atomiser for ophthalmic drug delivery

A liquid atomiser composed of a piezoelectric transducer and a metal plate with numerous micro-apertures is studied to identify the most influential factors on its atomising performance. The Taguchi method is employed in the experiment design and analysis of the study on how each factor acts in the atomising process. An optimal condition is determined for producing a stream of droplets. The study shows that the droplet size and the spraying velocity are suitable for ophthalmic drug delivery application, with an even distribution of the drug over most of the eyeball surface area due to the controllable cross-sectional area of the droplet stream. This greatly improves the treatment effectiveness and efficiency of eye therapy. Finally, a structure of the ophthalmic drug delivery system is proposed.

Development of a contact probe incorporating a bragg grating strain sensor for nano coordinate measuring machines

This paper presents a novel optical fibre based micro contact probe system with high sensitivity and repeatability. In this optical fibre probe with a fused spherical tip, a fibre Bragg grating has been utilized as a strain sensor in the probe stem. When the probe tip contacts the surface of the part, a strain will be induced along the probe stem and will produce a Bragg wavelength shift. The contact signal can be issued once the wavelength shift signal is produced and demodulated. With the fibre grating sensor element integrated into the probe directly, the probe system shows a high sensitivity. In this work, the strain distributions along the probe stem with the probe under axial and lateral load are analysed. A simulation of the strain distribution was performed using the finite element package ANSYS 11. Performance tests using a piezoelectric transducer stage with a displacement resolution of 1.5 nm yielded a measurement resolution of 60 nm under axial loading.

Comparison of corticomotor excitability during visuomotor dynamic and static tasks

The human central nervous system (CNS) has the ability to modulate its activity during the performance of different movements. Recent evidence, however, suggests that the CNS can also modulate its activity in the same movement but with increased precision during a visuomotor static task. This study aimed to extend on these findings by using transcranial magnetic stimulation (TMS) to measure the CNS during the performance of two visuomotor dynamic tasks. Twelve volunteers participated in this study, performing two separate motor tasks. Study I (“Position Tracking”) involved participants to perform a visuomotor tracking task using a dial potentiometer and matching their response icon to the computer generated tracking icon whilst holding a pincer grip. Study II (“Force Tracking”) involved participants to perform a similar visuomotor tracking task by applying or releasing pressure against a fixed force transducer. Tasks were conducted at two speeds (“slow” being one tracking cycle in 10 s; and “fast” being two tracking cycles in 10 s) and compared to a visuomotor static task at a similar muscle contraction level. Results showed corticospinal changes with significant increases (p = 0.002) in excitability demonstrated during Study I (42.3 ± 16.8%) and Study II (56.3 ± 34.2%) slow speed tasks. Moreover, significant reduction in corticospinal inhibition was also observed during both tracking tasks at slow (59.3 ± 13.7%; p = 0.001) and fast speeds (31.9 ± 12.3%; p = 0.001). The findings may provide information on the underlying physiology during the early stages of motor skill acquisition.

Conjugated polymer−titania nanoparticle hybrid films : random lasing action and ultrasensitive detection of explosive vapors

We have first demonstrated that a random laser action generated by a hybrid film composed of a semiconducting organic polymer (SOP) and TiO₂ nanoparticles can be used to detect 2,4,6-trinitrotoluene (TNT) vapors. The hybrid film was fabricated by spin-casting SOP solution dispersed with nanosized TiO₂ particles on quartz glass. The SOP in the hybrid film functioned as both the gain medium and the sensory transducer. A random lasing action was observed with a certain pump power when the size (diameter of 50 nm) and concentration (8.9 - 10¹²/cm³) of TiO₂ nanoparticles were optimized. Measurements of fluorescence quenching behavior of the hybrid film in TNT vapor atmosphere (10 ppb) showed that attenuated lasing in optically pumped hybrid film displayed a sensitivity to vapors of explosives more than 20 times higher than was observed from spontaneous emission. This phenomenon has been explained with the four-level laser model. Since the sensory transducer used in the hybrid polymer/nanoparticles system could be replaced by other functional materials, the concept developed could be extended to more general domains of chemical or environment detection.

Evolution of JAK-STAT pathway components : mechanisms and role in immune system development

Background
Lying downstream of a myriad of cytokine receptors, the Janus kinase (JAK) – Signal transducer and activator of transcription (STAT) pathway is pivotal for the development and function of the immune system, with additional important roles in other biological systems. To gain further insight into immune system evolution, we have performed a comprehensive bioinformatic analysis of the JAK-STAT pathway components, including the key negative regulators of this pathway, the SH2-domain containing tyrosine phosphatase (SHP), Protein inhibitors against Stats (PIAS), and Suppressor of cytokine signaling (SOCS) proteins across a diverse range of organisms.

Results
Our analysis has demonstrated significant expansion of JAK-STAT pathway components co-incident with the emergence of adaptive immunity, with whole genome duplication being the principal mechanism for generating this additional diversity. In contrast, expansion of upstream cytokine receptors appears to be a pivotal driver for the differential diversification of specific pathway components.

Conclusion
Diversification of JAK-STAT pathway components during early vertebrate development occurred concurrently with a major expansion of upstream cytokine receptors and two rounds of whole genome duplications. This produced an intricate cell-cell communication system that has made a significant contribution to the evolution of the immune system, particularly the emergence of adaptive immunity.

Alternative TEL-JAK2 fusions associated with T-cell acute lymphoblastic leukemia and atypical chronic myelogenous leukemia dissected in zebrafish

Background Chromosomal translocations resulting in alternative fusions of the human TEL (ETV6) and JAK2 genes have been observed in cases of acute lymphoblastic leukemia and chronic myelogenous leukemia, but a full understanding of their role in disease etiology has remained elusive. In this study potential differences between these alternative TEL-JAK2 fusions, including their lineage specificity, were investigated.

Design and Methods TEL-JAK2 fusion types derived from both T-cell acute lymphoblastic leukemia and atypical chronic myelogenous leukemia were generated using the corresponding zebrafish tel and jak2a genes and placed under the control of either the white blood cell-specific spi1 promoter or the ubiquitously-expressed cytomegalovirus promoter. These constructs were injected into zebrafish embryos and their effects on hematopoiesis examined using a range of molecular approaches. In addition, the functional properties of the alternative fusions were investigated in vitro.

Results Injection of the T-cell acute lymphoblastic leukemia-derived tel-jak2a significantly perturbed lymphopoiesis with a lesser effect on myelopoiesis in zebrafish embryos. In contrast, injection of the atypical chronic myelogenous leukemia-derived tel-jak2a resulted in significant perturbation of the myeloid compartment. These phenotypes were observed regardless of whether expressed in a white blood cell-specific or ubiquitous manner, with no overt cellular proliferation outside of the hematopoietic cells. Functional studies revealed subtle differences between the alternative forms, with the acute lymphoblastic leukemia variant showing higher activity, but reduced downstream signal transducer and activator of transcription activation and decreased sensitivity to JAK2 inhibition. JAK2 activity was required to mediate the effects of both variants on zebrafish hematopoiesis.

Conclusions This study indicates that the molecular structure of alternative TEL-JAK2 fusions likely contributes to the etiology of disease. The data further suggest that this class of oncogene exerts its effects in a cell lineage-specific manner, which may be due to differences in downstream signaling.

Bacterial biosensors for measuring availability of environmental pollutants

Biosensors are projected to find many applications due to their high selectivity and sensitivity, rapid reaction, economy and ease of handling in field measurements. Even though biosensors for a wide range of environmental pollutants have been extensively reported in the literature, the decision to develop a suitable biosensing system that can be approved by a regulatory perspective for environmental applications is fraught with technical issues. These issues mainly concern the biological recognition element, the physico-chemical transducer and the interfaces between the biological and the physical components, but also aspects of fluidics, electronics, and software for data processing. This article focused on the development of microbial-based biosensors for environmental applications especially metal contaminants such as arsenic.

The small molecule, genistein, increases hepcidin expression in human hepatocytes

Hepcidin, a peptide hormone that decreases intestinal iron absorption and macrophage iron release, is a potential drug target for patients with iron overload syndromes because its levels are inappropriately low in these individuals. Endogenous stimulants of Hepcidin transcription include bone morphogenic protein 6 (BMP6) and interleukin-6 (IL-6) by effects on mothers against decapentaplegic homolog (Smad)4 or signal transducer and activator of transcription (Stat)3, respectively. We conducted a small-scale chemical screen in zebrafish embryos to identify small molecules that modulate hepcidin expression. We found that treatment with the isoflavone, genistein, from 28-52 hours postfertilization in zebrafish embryos enhanced Hepcidin transcript levels, as assessed by whole-mount in situ hybridization and quantitative real-time reverse-transcriptase polymerase chain reaction. Genistein's stimulatory effect was conserved in human hepatocytes: Genistein treatment of HepG2 cells increased both Hepcidin transcript levels and promoter activity. We found that genistein's effect on Hepcidin expression did not depend on estrogen receptor signaling or increased cellular iron uptake, but was impaired by mutation of either BMP response elements or the Stat3-binding site in the Hepcidin promoter. RNA sequencing of transcripts from genistein-treated hepatocytes indicated that genistein up-regulated 68% of the transcripts that were up-regulated by BMP6; however, genistein raised levels of several transcripts involved in Stat3 signaling that were not up-regulated by BMP6. Chromatin immunoprecipitation and ELISA experiments revealed that genistein enhanced Stat3 binding to the Hepcidin promoter and increased phosphorylation of Stat3 in HepG2 cells. Conclusion: Genistein is the first small-molecule experimental drug that stimulates Hepcidin expression in vivo and in vitro. These experiments demonstrate the feasibility of identifying and characterizing small molecules that increase Hepcidin expression. Genistein and other candidate molecules may subsequently be developed into new therapies for iron overload syndromes.

Analyses and comparison of counter-movement jump performance and self-rated recovery in state under-18s Australian Rules Football players during a national championship

Recovery of team sport athletes during multiple competitive games is an important area for strength and conditioning coaches to monitor as it facilitates for athletes to be ready to perform (11,13). Utilising athletic performance data in conjunction with self-rated reporting measures can help determine if in fact a player or team has recovered sufficiently or shown a trend towards recovery prior to a competitive match (11). Positive improvement in recovery variables can provide confidence in the effectiveness of recovery methods used and assist in determining the training schedule in order to positively manipulate the fitness-fatigue relationship (3).

Various methods of analysing the recovery of athletes have been reported in the literature and are available to the strength and conditioning coach. These include subjective, self-rated scales and perceived level of recovery questionnaires (11,12,13). Athletic performance measures during exercises such as the counter movement jump (CMJ) have also been analysed, predominantly utilising force plates to obtain kinetic data. (5,13,14). However, such equipment can be difficult to transport, requires continual calibration and is costly to purchase. A linear transducer can provide important information on CMJ variables in the assessment of athletic movements and due to its size and portability could serve as a valuable tool to assist strength and conditioning coaches, (8,10), and potentially enable the monitoring of recovery.

Previous studies have investigated the fatigue effects of competitive games in various sports (11,13,14) including Australian Rules Football (AFL) at the senior elite league level (5, 6). To the authors’ knowledge, however, there is yet to be a study investigating the recovery response in AFL players, specifically in players 18 years and under competing in the National Under 18s Championships. Australian Rules football is an extremely physically demanding and fatiguing sport where players participate in games time exceeding 120 minutes duration, covering large distances (~12-18km, position dependent) with many high intensity efforts performed at random times throughout the game (2,6,16). Hence, it would seem pertinent to analyse the fatigue effects of competitive matches in an Australian Rules Under-18’s National Championship and the subsequent recovery from these games.

The aim of this study was to analyse and compare two self-rated subjective measures of recovery; they being muscle soreness (MS) of the lower body, overall perceived total recovery (TR), and the performance measure of peak velocity (PV) obtained from a CMJ analysed with a linear transducer. Data collection occurred between rounds four and five of the Australian Football League Under-18’s National Championship, representing a four-day recovery analysis period between matches.

A high performance position sensorless surface permanent magnet synchronous motor drive based on flux angle

A position sensorless Surface Permanent Magnet Synchronous Motor (SPMSM) drive based on flux angle is presented in this paper. The motor equations are written in rotor fixed d-q reference frame. A PID controller is used to process the speed error to generate the reference torque current keeping the magnetizing current fixed. The estimated stator flux using Recurrent Neural Network (RNN) is used to find out the rotor position. The flux angle and the reference current phasor angle are used in vector rotator to generate the reference phase currents. Hysteresis current controller block controls the switching of the 3-phase inverter to apply voltage to the motor stator. Simulation studies on different operating conditions indicate the acceptability of the drive system. The drive system only requires a speed transducer and is free from position sensor requirement. The proposed control scheme is robust under load torque disturbances and motor parameter variations. It is also simple and low cost to implement in a practical environment.

Leptin's metabolic and immune functions can be uncoupled at the ligand/receptor interaction level.

The adipocyte-derived cytokine leptin acts as a metabolic switch, connecting the body's metabolism to high-energy consuming processes such as reproduction and immune responses. We here provide genetic and biochemical evidence that the metabolic and immune functions of leptin can be uncoupled at the receptor level. First, homozygous mutant fatt/fatt mice carry a spontaneous splice mutation causing deletion of the leptin receptor (LR) immunoglobulin-like domain (IGD) in all LR isoforms. These mice are hyperphagic and morbidly obese, but display only minimal changes in size and cellularity of the thymus, and cellular immune responses are unaffected. These animals also displayed liver damage in response to concavalin A comparable to wild-type and heterozygous littermates. Second, treatment of healthy mice with a neutralizing nanobody targeting IGD induced weight gain and hyperinsulinaemia, but completely failed to block development of experimentally induced autoimmune diseases. These data indicate that leptin receptor deficiency or antagonism profoundly affects metabolism, with little concomitant effects on immune functions.