Two spinefoot colour morphs: mottled spinefoot Siganus fuscescens and white-spotted spinefoot Siganus canaliculatus are synonyms

Mottled spinefoot Siganus fuscescens and white-spotted spinefoot Siganus canaliculatus are two similar species that differ subtly in colouration and morphology. Three major mtDNA clades were identified for these species, but individuals were clustered by amplified fragment length polymorphism (AFLP) according to geography rather than morphology, suggesting that the colour morphs are interbreeding.     

Resolution of the Acanthopagrus black seabream complex based on mitochondrial and amplified fragment-length polymorphism analyses

In this study, DNA analyses were employed to verify the identity of six morphologically similar species that occur in the coastal waters of Taiwan: the black seabream complex (Acanthopagrus latus, Acanthopagrus schlegelii, Acanthopagrus sivicolus, Acanthopagrus taiwanensis, Acanthopagrus chinshira and Acanthopagrus pacificus). Amplified fragment-length polymorphism (AFLP) analyses clearly distinguished the same six species that are morphologically diagnosable based on subtle differences in scale counts and anal-fin colouration. In contrast, mitochondrial DNA analyses based on cytochrome b gene sequences did not distinguish individuals of A. schlegelii and A. sivicolus, reflecting either historical introgression or recent speciation and incomplete sorting of their mitochondrial lineages. Phylogenetic relationships among these six north-west Pacific Ocean species of Acanthopagrus analysed using AFLP data were consistent with scale rows above the lateral line (TRac), sperm ultrastructure and geographical distribution. The study provides molecular tools for future research relevant to improved management of these resources, and an increased understanding of the evolutionary history of this radiation.     

Ethyl pyruvate attenuates ventilation‐induced diaphragm dysfunction through high‐mobility group box‐1 in a murine endotoxaemia model

Mechanical ventilation (MV) can save the lives of patients with sepsis. However, MV in both animal and human studies has resulted in ventilator‐induced diaphragm dysfunction (VIDD). Sepsis may promote skeletal muscle atrophy in critically ill patients. Elevated high‐mobility group box‐1 (HMGB1) levels are associated with patients requiring long‐term MV. Ethyl pyruvate (EP) has been demonstrated to lengthen survival in patients with severe sepsis. We hypothesized that the administration of HMGB1 inhibitor EP or anti‐HMGB1 antibody could attenuate sepsis‐exacerbated VIDD by repressing HMGB1 signalling. Male C57BL/6 mice with or without endotoxaemia were exposed to MV (10 mL/kg) for 8 hours after administrating either 100 mg/kg of EP or 100 mg/kg of anti‐HMGB1 antibody. Mice exposed to MV with endotoxaemia experienced augmented VIDD, as indicated by elevated proteolytic, apoptotic and autophagic parameters. Additionally, disarrayed myofibrils and disrupted mitochondrial ultrastructures, as well as increased HMGB1 mRNA and protein expression, and plasminogen activator inhibitor‐1 protein, oxidative stress, autophagosomes and myonuclear apoptosis were also observed. However, MV suppressed mitochondrial cytochrome C and diaphragm contractility in mice with endotoxaemia (P < 0.05). These deleterious effects were alleviated by pharmacologic inhibition with EP or anti‐HMGB1 antibody (P < 0.05). Our data suggest that EP attenuates endotoxin‐enhanced VIDD by inhibiting HMGB1 signalling pathway.     

Ultrastructure of the sperm of blue sprat, Spratelloides gracilis; Teleostei, Clupeiformes, Clupeidae

The general sperm structure of the investigated clupeoids possess an oliviform head with a distinct deep nuclear fossa, a midpiece with one mitochondrion and a posterior flagellum. They are characterized by two apomorphies: an annular or C-shaped mitochondrion and ITDs (intratubular differentiation) in the A tubules of the axonemal doublets. The fine structure of the spermatozoa of blue sprat, Spratelloides gracilis, was investigated to see if resulting data conformed to the current hypotheses of the taxonomy of the Clupeidae. The mature sperm is characterized by the following features: (1) the nucleus is oliviform; its prominent deep nuclear fossa encloses the initial portion of flagellum, (2) a proximal centriole has not been identified, (3) a single spherical mitochondrion is located laterally in relation to the flagellum and (4) no cytoplasmic canal is present. Our blue sprat-spermatozoan morphology data suggest that the blue sprat has Clupeomorpha affinities, while indicating no close affinity with Elopomorpha. Clupeoid sperm exhibit morphological variations with the nucleus and mitochondrion being particularly variable. This study provides useful systematic characters to the existing knowledge of comparative spermatology and may provide additional clue to Euteleost phylogeny.     

Induced ovulation in Atlantic croaker (Sciaenidae) using hCG and an LHRH analog: A preliminary study

A single dose of LHRHa (D-Ala(6), des Gly(10)- LHRH-ethylamide; 100 mug/kg body weight); administered by intramuscular injection, effectively induced ovulation in the Atlantic croaker (Micropogonias undulatus ); 37 of 40 (92%) females were successfully hand-stripped and there was no mortality. A single dose of human chorionic gonadotropin (hCG; 500 IU/kg body weight) was successful in inducing oocyte hydration. However, only 16 of 30 (53%) females ovulated successfully and could be hand-stripped. Another 8 females became extremely bloated and died. Thus LHRHa was shown to be the more reliable method of inducing ovulation in the Atlantic croaker. The response interval between hormone treatment and ovulation at different water temperatures was also determined.     

Cryopreservation of Atlantic croaker spermatozoa: evaluation of morphological changes.

The spermatozoon of the Atlantic croaker (Micropogonias undulatus) is a primitive type in that it lacks an acrosome. The kidney-shaped head has a diameter of about 1.5 microns and is occupied by a granular and electron-dense nucleus. The short midpiece contains 3 spherical mitochondria and encircles the basal body of the flagellum but is separated from it. The flagellum consists of the typical 9 + 2 axoneme and surrounding plasma membrane but lacks a lateral ridge. Spermatozoa of Atlantic croaker diluted in either NaCl or sodium citrate solutions with or without DMSO were examined with the electron microscope before freezing in liquid nitrogen and after thawing. Damage following cryopreservation appeared to be greater to the mitochondria, plasma membrane, and 9 + 2 axoneme than to the nucleus. The incidence of postthaw damage in spermatozoa diluted with NaCl solutions containing DMSO was remarkably lower than that with either pure NaCl solutions, pure sodium citrate solutions, or sodium citrate solutions containing DMSO.     

A spin-label study of the effect of gamma radiation on erythrocyte membrane. Influence of lipid peroxidation on membrane structure.

Gamma-irradiation of bovine erythrocyte membranes (0.1-4 Mrad) resulted in a decrease in the degree of order of membrane lipids, as measured by spin-labelled fatty acid esters, at the depth of C12 but not at the depth of C5. Dose dependence of this phenomenon corresponded to dose dependence of malondialdehyde formation in the membranes. On this basis a mechanism for the effect of lipid peroxidation on the membrance structure is proposed. Membrane proteins underwent radiation-induced conformational transitions revealed by maleimide spin label which could be also connected with lipid peroxidation.     

Semen cryopreservation of small abalone (Haliotis diversicolor supertexa)

Methods for cryopreserving spermatozoa and maximizing fertilization rate in Taiwan small abalone, Haliotis diversicolor supertexa, were developed. The gametes (spermatozoa and eggs) of small abalone were viable 3 h post-spawning, with fertilization, and development rate decreasing with time. A minimum of 10(2) cell/ml sperm concentration and a contact time of 2 min between gametes is recommended for artificial insemination of small abalone eggs. Eight cryoprotectants, dimethyl sulfoxide (DMSO), dimethyl acetamide (DMA), ethylene glycol (EG), propylene glycol (PG), butylene glycol (BG), polyethylene glycol, glycerol and methanol, were tested at concentrations between 5 and 25% to evaluate their effect on motility of spermatozoa exposed to cryoprotectant for up to 60 min at 25 degrees C before freezing. The least toxic cryoprotectant, 10% DMSO, was added to artificial seawater (ASW) to formulate the extender for freezing. Semen was diluted 1:1 with the extender, inserted into 1.5 ml microtubes and frozen using a cooling rate between -3.5 and -20 degrees C/min to various transition temperatures (0, -30, -60, -90 and -120 degrees C), followed by transfer and storage in liquid nitrogen (-196 degrees C). The microtubes were thawed from +45 to +145 degrees C/min. Spermatozoa, cooled to -90 degrees C at a cooling rate of -12 or -15 degrees C/min and then immersed in liquid nitrogen, had the best post-thaw motility. Post-thaw sperm motility was markedly reduced compared to fresh sperm. More frozen-thawed spermatozoa are required to achieve fertilization rates comparable to those achieved using fresh spermatozoa.     

Evaluation of hydralazine and procainamide effects on fibroblast membrane fluidity

In this study the membrane fluidity of fibroblasts under different pharmacological treatment was investigated. Two drugs, hydralazine and procainamide, were used to treat the immortalized mouse NIH 3T3 and hamster B14 fibroblasts. Membrane lipid dynamics was measured by fluorescence spectroscopy and electron spin resonance techniques. Two kinds of fluorescent probes (TMA-DPH and 12-(9-anthroyloxy)-stearic acid (12-AS)) and two spin labels (5-doxylstearic acid (5-DS) and 12-doxylstearic acid (12-DS)) were used to monitor fluidity in the upper polar and in the hydrophobic core regions of the lipid bilayer. The drugs influenced the membrane hydrophobic core, of which hydralazine induced fluidization and procainamide increased the rigidity. The membrane fluidity at the surface of the lipid bilayer was not modified by the drugs which indicates that both drugs intercalated mainly into the inner core of the cell membrane.