Expression and Localization of Opioid Receptors in Male Germ Cells and the Implication for Mouse Spermatogenesis

The presence of endogenous opioid peptides in different testicular cell types has been extensively characterized and provides evidence for the participation of the opioid system in the regulation of testicular function. However, the exact role of the opioid system during the spermatogenesis has remained controversial since the presence of the mu-, delta-and kappa-opioid receptors in spermatogenic cells was yet to be demonstrated. Through a combination of quantitative real-time PCR, immunofluorescence, immunohistochemistry and flow cytometry approaches, we report for the first time the presence of active mu-, deltaand kappa-opioid receptors in mouse male germ cells. They show an exposition time-dependent response to opioid agonist, hence suggesting their active involvement in spermatogenesis. Our results contribute to understanding the role of the opioid receptors in the spermatogenesis and could help to develop new strategies to employ the opioid system as a biochemical tool for the diagnosis and treatment of male infertility.

Sex-change rules, stock dynamics, and the performance of spawning-per-recruit measures in protogynous stocks

Predicting and under-standing the dynamics of a population requires knowledge of vital rates such as survival, growth, and reproduction. However, these variables are influenced by individual behavior, and when managing exploited populations, it is now generally realized that knowledge of a species’ behavior and life history strategies is required. However, predicting and understanding a response to novel conditions—such as increased fishing-induced mortality, changes in environmental conditions, or specific management strategies—also require knowing the endogenous or exogenous cues that induce phenotypic changes and knowing whether these behaviors and life history patterns are plastic. Although a wide variety of patterns of sex change have been observed in the wild, it is not known how the specific sex-change rule and cues that induce sex change affect stock dynamics. Using an individual based model, we examined the effect of the sex-change rule on the predicted stock dynamics, the effect of mating group size, and the performance of traditional spawning-per-recruit (SPR) measures in a protogynous stock. We considered four different patterns of sex change in which the probability of sex change is determined by 1) the absolute size of the individual, 2) the relative length of individuals at the mating site, 3) the frequency of smaller individuals at the mating site, and 4) expected reproductive success. All four pat-terns of sex change have distinct stock dynamics. Although each sex-change rule leads to the prediction that the stock will be sensitive to the size-selective fishing pattern and may crash if too many reproductive size classes are fished, the performance of traditional spawning-per-recruit measures, the fishing pattern that leads to the greatest yield, and the effect of mating group size all differ distinctly for the four sex-change rules. These results indicate that the management of individual species requires knowledge of whether sex change occurs, as well as an understanding of the endogenous or exogenous cues that induce sex change.

Increased toxicity of Karenia brevis during phosphate limited growth: ecological and evolutionary implications

Karenia brevis is the dominant toxic red tide algal species in the Gulf of Mexico. It produces potent neurotoxins (brevetoxins [PbTxs]), which negatively impact human and animal health, local economies, and ecosystem function. Field measurements have shown that cellular brevetoxin contents vary from 1–68 pg/cell but the source of this variability is uncertain. Increases in cellular toxicity caused by nutrient-limitation and inter-strain differences have been observed in many algal species. This study examined the effect of P-limitation of growth rate on cellular toxin concentrations in five Karenia brevis strains from different geographic locations. Phosphorous was selected because of evidence for regional P-limitation of algal growth in the Gulf of Mexico. Depending on the isolate, P-limited cells had 2.3- to 7.3-fold higher PbTx per cell than P-replete cells. The percent of cellular carbon associated with brevetoxins (%C-PbTx) was ~ 0.7 to 2.1% in P-replete cells, but increased to 1.6–5% under P-limitation. Because PbTxs are potent anti-grazing compounds, this increased investment in PbTxs should enhance cellular survival during periods of nutrient-limited growth. The %C-PbTx was inversely related to the specific growth rate in both the nutrient-replete and P-limited cultures of all strains. This inverse relationship is consistent with an evolutionary tradeoff between carbon investment in PbTxs and other grazing defenses, and C investment in growth and reproduction. In aquatic environments where nutrient supply and grazing pressure often vary on different temporal and spatial scales, this tradeoff would be selectively advantageous as it would result in increased net population growth rates. The variation in PbTx/cell values observed in this study can account for the range of values observed in the field, including the highest values, which are not observed under N-limitation. These results suggest P-limitation is an important factor regulating cellular toxicity and adverse impacts during at least some K. brevis blooms.

Examining the possibilty of multiplication and breeding of (Schizothorav [sic] zarudnyi) in order to restoration of regional aquatics reservations of Hamoon Lake and taking its advantag [sic] for aquaculture

Schizothorax zarudnvi, is an endemic fish of east country waters. (Triple lagoons of Hamoon and relevant water resources) that in the world it is reported in this resource specially. This fish named Hamoon mahi is one of the most economically valuable species in this region. Because of the recent years droughts, Hamoon logoon has been drive since 2000. Also, semi-wells (a semi natural resource) were affected drastically by recent drought years and their volume reduced to nearly one third of their real volume and resulted in changing at growth and reproduction physiology process in Schizothorax zanidnyi, brood stocks. Beginning of this project was done from October 2003. It's field studies begun (brood catching) since November 2001 by two methods including entangling gairs and at semi wells of Sistan that (Beach seine) had maximum rate of preparing qualified brood stocks. Broods transferred to Cyprinidea reproduction work shop of Zahak and after taking primary measures they stored in to the edaphic pools. Increasing the success safety factor (coefficient) for artificial reproduction of Sthizothorax zarudnyi , identifying the appropriate tune for Hormonal acceptance (physiological preparation of broods) is needed , so this important work was done regularly by histological studies and GSI measurements since November. Highest GSI rates of females (%80.51) and highest IV stage abundance of sexual maturity (%l 00) were observed an march. On the base of this date, Hormone therapy was done on broods on march. The used hormones are as follows Hypophysis. extraction, GnRHa and Anti Dopamin at the dozes of 3-6 ml, 20-30kg and 10-15 ml per kg body weight respectively and 2-3 times from 11-12-80 they were injected. Injected broods kept in to two circumstances, flow-through (rounded pool) and stagnant systems. In stagnant system 14 and 19 individuals of female and male (Schizothorax zauiulnri) broods, respectively injected in 11th, 15111, 19th, and 24th of march 1380. Non of the injected broods in 11 and 15 and 19th march (in stagnant Condition) answered to Hormone therapy. After final injection broods had general less activity and a few of them died. Mean temperature of brood pond waters (daily) which were injected. Fluctuated between 10-25-13. 63°c but injected broods on 24th march had different characteristics. They had pale color and had few fecundity. In this stage of injection they hadn't any successful vulation. After injection, Mean daily water temperature was 15, 88-17, 54°c. In Flowing system, 13-16 individual of males and females respectively were injected on 15th, 19th, 22th and 23th march. None of injected producers on 15th and 19th march with mean daily water temperature of 10, 25-12°c were prepared for spawning but injected producers on 22nd an 23th march with mean daily water temperature of 13.5-1 rc responded about 75-100 percent. (Schizothorax zarudnyi) brood stocks were prepared for spawning after 353-428 hours/day from final injection. Diameter of obtained eggs (before fertilization) was between 1.9-2.3 min and of fertilized eggs was 3.8mm. Fertilized eggs of (Schizothorax zarudnyi) were hatched after 6-7 days with mean water temperature of 17.08°c. Mean length of on one day larvae was 9.47 mm. Larvae was 9.47 mm. Larvae adsorbed the whole yolk sac after , 5-6 days at 17- 1°c and were prepared for releasing in to edaphic pools. Because of the lack of necessary and complementary facilities in the region , they had to release them in to veniros and growing them for 8 days. At the end of 18th day , 35000 larvae (at first) released into an edaphic pond with a volume of 150m2. After growing them for one moth , mean length and weight of new hatched larvae was 29.41 mm and 1.12►r , respectively. With respect to results of this investigation , artificial reproduction of (Schizothorax zarudnyi) Can be possible at 14-17°C and flowing water with Hormonal treatment. It -s breeding has increased development than other cultural specious in the region. Due to high economical value of this specious in Sistan and ti-s specialization east waters of Iran and having high resistance and proper growth There is a need of it's development and reproduction and culture in fish culture fanns (edaphic ponds• two-purpose pools) at the region and country.

Field and experimental studies on the combined impacts of cyanobacterial blooms and small algae on crustacean zooplankton in a large, eutrophic, subtropical, Chinese lake

Field and experimental studies were conducted to evaluate the combined impacts of cyanobacterial blooms and small algae on seasonal and long-term changes in the abundance and community structure of crustacean zooplankton in a large, eutrophic, Chinese lake, Lake Chaohu. Seasonal changes of the crustacean zooplankton from 22 sampling stations were investigated during September 2002 and August 2003, and 23 species belonging to 20 genera were recorded. Daphnia spp. dominated in spring but disappeared in mid-summer, while Bosmina coregoni and Ceriodaphnia cornuta dominated in summer and autumn. Both maximum cladoceran density (310 ind. l(-1)) and biomass (5.2 mg l(-1)) appeared in autumn. Limnoithona sinensis, Sinocalanus dorrii and Schmackeria inopinus were the main species of copepods. Microcystis spp. were the dominant phytoplankton species and formed dense blooms in the warm seasons. In the laboratory, inhibitory effects of small colonial Microcystis on growth and reproduction of Daphnia carinata were more remarkable than those of large ones, and population size of D. carinata was negatively correlated with density of fresh large colonial Microcystis within a density range of 0-100 mg l(-1) (r = -0.82, P < 0.05). Both field and experimental results suggested that seasonal and long-term changes in the community structure of crustacean zooplankton in the lake were shaped by cyanobacterial blooms and biomass of the small algae, respectively, i.e., colonial and filamentous cyanobacteria contributed to the summer replacement of dominant crustacean zooplankton from large Daphnia spp. to small B. coregoni and C. cornuta, while increased small algae might be responsible for the increased abundance of crustacean zooplankton during the past decades.

Population dynamics and maturation cycle of Camallanus cotti (Nematoda : Camallanidae) in the Chinese hooksnout carp Opsariichthys bidens (Osteichthyes : Cyprinidae) from a reservoir in China

The seasonal population dynamics and maturation cycle of the nematode Camallanus cotti in the posterior intestine of Chinese hooksnout carp Opsariichthys bidens have been studied in the Danjiangkou Reservoir of the Hubei Province in central China from September 2004 to November, 2005. The overall prevalence, mean abundance and intensity of C cotti among fish sampled (n = 700 fish) were 47%, 2.29 +/- 12.38 ( +/- S.D.) and 1-307 (average 4.89 +/- 17.74), respectively. The overall sexual ratio of female to male nematodes (excluding L3 and L4 juveniles) was 1.17:1. Statistical results showed weakly positive correlations betweerl fish length and the number of nematodes per host. The dynamics of infection of the nematode exhibited significant seasonal pattern in changes in mean abundance. A similar pattern was found for changes in nematode prevalence, although this was not statistically significant. Higher levels of infection were observed among fish sampled in summer months and the lower in the winter. Neither the prevalence nor the abundance of the parasite was significantly different between male and female hosts. The pattern of frequency distribution of the parasite in the host was found to be over-dispersed throughout the sampling period. In addition, studies on the development and maturation of the parasite in O. bidens revealed that development (maturation), recruitment of the next generation, and reproduction may be continuous year-round, although reproduction may peak during the winter. (c) 2007 Elsevier B.V. All rights reserved.

Comparative studies on in vitro sperm decondensation and pronucleus formation in egg extracts between gynogenetic and bisexual fish

A cell-free system based upon the egg extracts from gynogenetic gibel carp (Carassius auratus gibelio) or bisexual red common carp (Cyprinus carpio red variety) was developed to investigate developmental behaviors of the demembranated sperm nuclei. Both red common carp and gibel carp sperm nuclei could decondense fully and form pronuclei in the red common carp egg extracts. Gibel carp sperm nuclei could also decondense fully and form pronuclei in the gibel carp egg extracts, but red common carp sperm nuclei could not decondense sufficiently in the same extracts. The significant differences of morphological changes were further confirmed by ultrastructural. observation of transmission electron microscopy. The data further offer cytological evidence for gonochoristic reproduction in the gynogenetically reproducing gibel carp. In addition, the sperm nuclei in vitro decondensation is dependent on the pH in the extracts, and the decondensed efficiency is optimal at pH 7. However, no DNA replication was observed in the two kinds of egg extracts during the incubation period of the sperm nuclei. It is suggested that the egg extracts prepared from the gynogenetic gibel carp should be a valid in vitro system for studying molecular mechanism on gynogenesis and reproduction mode diversity in fish.

Cyclin A2 is differentially expressed during oocyte maturation between gynogenetic silver crucian carp and gonochoristic color crucian carp

Silver crucian carp (Carassius auratus gibelio) is a unique gynogenetic fish. Because of its specific genetic background and reproduction mode, it is an intriguing model system for understanding regulatory mechanism of oocyte maturation division. It keeps its chromosomal integrity by inhibiting the first meiotic division (no extrusion of the first pole body). The spindle behavior during oocyte maturation is significantly different from that in gonochoristic fish. The chromosomes are first arranged in a tripolar spindle, and then they turn around and are reunited mutually to form a normal bipolar spindle. A new member of the fish A-type cyclin gene, cyclin A2, has been isolated by suppression of subtractive hybridization on the basis of its differential transcription in fully-grown oocytes between the gynogenetic silver crucian carp and gonochoristic color crucian carp. There are 18 differing amino acids in the total 428 residues of cyclin A2 between the two forms of crucian carps. In addition, cDNAs of cyclin A1 and cyclin B have also been cloned from them. Thus two members of A-type cyclins, cyclin A1 and cyclin A2, are demonstrated to exist in fish, just as in frog, humans, and mouse. Northern blotting reveals that cyclin A2 mRNA is more than 20-fold and cyclin A1 mRNA is about 2-fold in fully grown oocytes of gynogenetic silver crucian carp compared to gonochoristic color crucian carp. However, cyclin B does not show such a difference between them. Western blot analysis also shows that the cyclin A2 protein stockpiled in fully grown oocytes of gynogenetic crucian carp is much more abundant than in gonochoristic crucian carp. Moreover, two different cyclin A2 expression patterns during oocyte maturation have been revealed in the two closely related crucian carps. For color crucian carp, cyclin A2 protein is translated only after hormone stimulation. For silver crucian carp, cyclin A2 protein can be detected throughout the process of maturation division. The different expression of cyclin A2 may be a clue to understanding the special maturation division of gynogenetic silver crucian carp.

Experimental and field studies of Mesocyclops notius (Copepoda : Cyclopoida) from a subtropical Chinese lake, Lake Donghu

This paper reports experimental and field studies on the cyclopoid Mesocyclops notius from subtropical Lake Donghu close to the Yangtze river. Mesocyclops notius, a dominant crustacean Zooplankter throughout tropical Australia, was previously considered to be endemic to Australia, but recently, Mesocyclops leuckarti in Lake Donghu was re-identified as M. notius. Laboratory culture experiments were conducted to reveal the effect of temperature (15, 20, 25 and 30 degreesC) on the development, growth and reproduction of M. notius. Temperature was inversely related to development times of eggs, nauplii and copepodites, body length and physiological longevity of adults, and brood size. Body length and physiological longevity of females were greater than those of males at the same temperature. No reproduction took place at 15 degreesC. Production and seasonal cycles of M. notius during 1980-1982 were studied at two sampling stations of Lake Donghu. At the mid-lake station, the annual production and production/biomass (P/B) ratio of M. notius varied between 6.0 and 18.0 g dry wt m(-2) year(-1) and 74.6 and 95.5, respectively. Mesocyclops notius reached their highest density peaks in the warm months (July-October), with a maximum density of 1256 individuals l(-1) at a littoral site. No reproduction and recruitment by AL notius took place during the cold months (December March) when the temperature of the lake water was < 15 degreesC. Mesocyclops notius were more abundant at a littoral station than at a pelagic station, possibly due to different food availability. The higher male:female sex ratio of M. notius at the littoral station was most likely caused by size-selective fish predation on larger females.

The combined effects of temperature and food supply on Calanus sinicus in the southern Yellow Sea in summer

The effects of temperature and food availability on the life history strategy of the planktonic copepod Calanus sinicus in the southern Yellow Sea in summer were studied in this paper. The fifth copepodite stage (CV) dominates the population in the central part of the southern Yellow Sea, where the Yellow Sea Cold Water Mass (YSCWM) occurs below the thermocline. Incubation experiments were conducted on CV C. sinicus caught from the YSCWM to examine the effects of temperature and food availability. Temperature at the surface (27degreesC) is lethal to CVs regardless of food availability. At the temperature in the middle of the thermocline (18degreesC), survival time of the specimens depends on food availability, being similar to20 days in treatments without extra food supply. At the temperature in the YSCWM (9degreesC), most animals survive at the end of 27 day incubation even in treatments without food supply. Developmental rate of CVs at 9degreesC without extra food supply is extremely low. The increase of either temperature or food supply promotes the developmental rate of CVs. According to these results, the surface layers with high temperature and low food abundance are detrimental for the survival and reproduction of C. sinicus. Low temperature and low food availability in the YSCWM help CV to maintain a much lower developmental rate and higher survival rate. The ecological trait of C. sinicus in the southern Yellow Sea in summer cannot be sufficiently explained solely by the effects of temperature.

Seasonal and spatial variation in abundance and egg production of Paracalanus parvus (Copepoda: Calanoida) in/out Jiaozhou Bay, China

The spatial distribution of stage-specific abundance and reproduction of the copepod Paracalanus parvus were studied from October 2005 to September 2006 in the Jiaozhou Bay. This copepod occurred continuously in this bay throughout the year. The species reached the lowest abundance in April and peaked in June. From October to December, distribution center mainly occurred in offshore water and at the mouth of the bay. In winter, early copepodites and adults gradually decreased and till February, most of the population was only comprised of CIV-CV stages. Overwintering copepodites matured in March and males tended to mature before female. From May to September, each stage occurred in the population and gradually reached high abundance. Temperature and chlorophyll a (Chl-a) concentration in the three stations can't clearly explain the seasonal variation in stage-specific abundance, so we surmised the important effect of the Yellow Sea. Egg production rate (EPR) reached its lowest in winter and peaked in June at 60.8 eggs female(-1) day(-1) in nearshore water. In the warming period, EPR in nearshore water was statistically higher and EPR > 10 eggs female(-1) day(-1) lasted longer than that in offshore water, showing the importance of nearshore water for recruitment of R parvus. Our study showed that EPR was positively related to temperature and total chlorophyll a in offshore water and mouth of the bay. In nearshore water, the relationships between EPR and temperature and Chl-a in three size fractions were not the same as those in offshore water, suggesting complicated ecosystem in such a eutrophic area in warming period. (C) 2008 Elsevier Ltd. All rights reserved.

Program: a record of the first 40 years of electronic library and information systems

Tedd, L.(2006). Program: a record of the first 40 years of electronic library and information systems. Program: electronic library and information systems,40(1), 11-26.

Training for professional librarians in Slovakia by distance-learning methods: an overview of the PROLIB and EDULIB projects

Tedd, L.A., Dahl, K., Francis, S.,Tet?evov?, M.& ?ihlavn?kov?, E.(2002).Training for professional librarians in Slovakia by distance-learning methods: an overview of the PROLIB and EDULIB projects. Library Hi Tech, 20(3), 340-351. Sponsorship: European Union and the Open Society Institute

Depolarization of sperm membrane potential is a common feature of men with subfertility and is associated with low fertilization rate at IVF

STUDY QUESTION. Are significant abnormalities in outward (K+) conductance and resting membrane potential (Vm) present in the spermatozoa of patients undertaking IVF and ICSI and if so, what is their functional effect on fertilization success? SUMMARY ANSWER. Negligible outward conductance (≈5% of patients) or an enhanced inward conductance (≈4% of patients), both of which caused depolarization of Vm, were associated with a low rate of fertilization following IVF. WHAT IS KNOWN ALREADY. Sperm-specific potassium channel knockout mice are infertile with defects in sperm function, suggesting that these channels are essential for fertility. These observations suggest that malfunction of K+ channels in human spermatozoa might contribute significantly to the occurrence of subfertility in men. However, remarkably little is known of the nature of K+ channels in human spermatozoa or the incidence and functional consequences of K+ channel defects. STUDY DESIGN, SIZE AND DURATION. Spermatozoa were obtained from healthy volunteer research donors and subfertile IVF and ICSI patients attending a hospital assisted reproductive techniques clinic between May 2013 and December 2015. In total, 40 IVF patients, 41 ICSI patients and 26 normozoospermic donors took part in the study. PARTICIPANTS/MATERIALS, SETTING, METHODS. Samples were examined using electrophysiology (whole-cell patch clamping). Where abnormal electrophysiological characteristics were identified, spermatozoa were further examined for Ca2+ influx induced by progesterone and penetration into viscous media if sufficient sample was available. Full exome sequencing was performed to specifically evaluate potassium calcium-activated channel subfamily M α 1 (KCNMA1), potassium calcium-activated channel subfamily U member 1 (KCNU1) and leucine-rich repeat containing 52 (LRRC52) genes and others associated with K+ signalling. In IVF patients, comparison with fertilization rates was done to assess the functional significance of the electrophysiological abnormalities. MAIN RESULTS AND THE ROLE OF CHANCE. Patch clamp electrophysiology was used to assess outward (K+) conductance and resting membrane potential (Vm) and signalling/motility assays were used to assess functional characteristics of sperm from IVF and ICSI patient samples. The mean Vm and outward membrane conductance in sperm from IVF and ICSI patients were not significantly different from those of control (donor) sperm prepared under the same conditions, but variation between individuals was significantly greater (P< 0.02) with a large number of outliers (>25%). In particular, in ≈10% of patients (7/81), we observed either a negligible outward conductance (4 patients) or an enhanced inward current (3 patients), both of which caused depolarization of Vm. Analysis of clinical data from the IVF patients showed significant association of depolarized Vm (≥0 mV) with low fertilization rate (P= 0.012). Spermatozoa with electrophysiological abnormities (conductance and Vm) responded normally to progesterone with elevation of [Ca2+]i and penetration of viscous medium, indicating retention of cation channel of sperm (CatSper) channel function. LIMITATIONS, REASONS FOR CAUTION. For practical, technical, ethical and logistical reasons, we could not obtain sufficient additional semen samples from men with conductance abnormalities to establish the cause of the conductance defects. Full exome sequencing was only available in two men with conductance defects. WIDER IMPLICATIONS OF THE FINDINGS. These data add significantly to the understanding of the role of ion channels in human sperm function and its impact on male fertility. Impaired potassium channel conductance (Gm) and/or Vm regulation is both common and complex in human spermatozoa and importantly is associated with impaired fertilization capacity when the Vm of cells is completely depolarized.

Stability of time-delay systems via Lyapunov functions

In this paper, a Lyapunov function candidate is introduced for multivariable systems with inner delays, without assuming a priori stability for the nondelayed subsystem. By using this Lyapunov function, a controller is deduced. Such a controller utilizes an input-output description of the original system, a circumstance that facilitates practical applications of the proposed approach.