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PubMed1996

Simulation of cardiac activity and the ECG using a heart model with a reaction-diffusion action potential.

A computerized model of the heart for the simulation of the electrical cardiac activity is described. The cardiac cells are arranged in a three-dimensional cubic lattice and their action potential is governed by modified FitzHugh-Nagumo reaction-diffusion state equations system which exhibits properties such as oscillations, variable excitability and refractoriness. The modifications of the FitzHugh-Nagumo equations system account for asymmetric action potential regarding the fast depolarization and slow repolarization rate and for rotational anisotropic propagation. An isolated cell is tested for reproduction of the strength-duration curves and restitution. The structure basic unit cell is assigned with an individual set of control parameters that creates inhomogeneity and anisotropy to simulate the various cardiac components such as pacers, muscle cells and conduction fibers. The spatial resolution of the structure is 1 mm. The collective activity of the cells generates a realistic ECG waveform that scales the simulated temporal step unit to 0.2 msec. The effective diffusion coefficient ranges between 0.055 mm2/msec to 1 mm2/msec. The propagation velocity of the myocardial activation is calculated at normal direction to the wavefront surface and values obtained are 1.17 mm/msec at the muscle cells and 2.5 mm/msec at the main conduction fibers. An ischemia is induced to verify the capability of the model to account for abnormalities. The developed model can give an insight into the local and global complex dynamics of the heart's electrical activity in the transition from normal to abnormal myocardial activity and may help to estimate the effects of myocardial properties on the ECG rhythm.

Action Potentials↗
PubMed1987

Identification of Mortierella wolfii, a causative agent of mycotic abortion in cattle.

Of several media tested for their ability to induce asexual reproduction in confirmed or suspected isolates of Mortierella wolfii obtained from cattle in various geographical locations, only silage extract agar worked consistently and rapidly. Its use should provide a simple reliable culturing procedure to assist with identification of clinical isolates of this fungus. Temperature growth response curves and electrophoretic patterns of soluble protein extracts of these isolates with characteristic morphological features of M. wolfii indicate substantial variation amongst them.

Abortion, Veterinary↗
PubMed1992

Estimates of gene flow among populations, geographic races, and species in the Ipomopsis aggregata complex.

Interpopulational gene flow within a species can reduce population differentiation due to genetic drift, whereas genetic exchange among taxa can impede speciation. We used allozyme data to estimate gene flow within and among geographic races and species of perennial herbs in the Ipomopsis aggregata complex (Polemoniaceae). Estimates of interpopulational gene flow within taxa from two methods (F statistics and private alleles) were correlated with one another. Gene flow among populations within each geographic race (subspecies) of I. aggregata was relatively high (Nm greater than approximately 1.0). Gene flow was also high among populations of I. arizonica and among four northern populations of I. tenuituba. However, gene flow was low (Nm less than 1.0) for I. tenuituba when a population representing subsp. macrosiphon was included. This is consistent with previous findings that subsp. macrosiphon has had an independent origin and is reproductively, as well as geographically, isolated. A recently developed model, based on hierarchical F statistics, was employed to estimate genetic exchange among taxa. Gene flow estimates were generally high among races of I. aggregata (dNmrace greater than 1.0) but were low among subspecies of I. tenuituba (dNmrace less than 1.0). Consistent with morphological evidence, estimates of interspecific gene flow were moderate between I. aggregata and I. tenuituba, which hybridize in several areas. However, contrary to morphological evidence, we estimated relatively high levels of interspecific gene flow involving I. arizonica. Our results suggest that I. arizonica has hybridized with other species without the transfer of morphological traits.(ABSTRACT TRUNCATED AT 250 WORDS)

Gene Frequency↗
PubMed2002

Localization of a proteolytic enzyme within the efferent and deferent duct epithelial cells of the turkey (Meleagris gallopavo) using immunohistochemistry.

Turkey seminal plasma contains a serine protease found to be distinct from the spermatozoal acrosin. However, the origin and biological roles of this enzyme are unknown. Our experimental objective was to identify the cellular source of this protease within the male reproductive tract. The enzyme was isolated from seminal plasma using benzamidine-Sepharose 6B chromatography. A synthetic substrate, Nalpha-benzoyl-DL-arginine p-nitroanilide, was used to detect fractions containing the enzyme. The affinity chromatography technique yielded a 150-fold increase in amidase activity. Analysis of the protease by SDS-PAGE revealed two protein bands with relative molecular masses of 37 000 and 61 000. Proteolytic activity was detected within the smaller band as evidenced by casein digestion. Further analysis of the purified protein revealed that the smaller protein band was glycosylated. To determine the cellular source of the protease, a panel of mouse monoclonal antibodies was then developed against the purified protease, and used in immunohistochemistry. Frozen tissue sections from the liver, testis, epididymal region, and deferent duct were fixed in 4% (w/v) paraformaldehyde, permeabilized with 0.2% (v/v) (octylphenoxy)polyethoxyethanol followed by routine immunohistochemistry procedures. Monoclonal antibodies did not bind to tissue sections from either the liver or testis, or to blood plasma proteins. Both the distal portion of the efferent duct and the deferent duct were immunoreactive. We concluded that the protease found in turkey seminal plasma is concentrated to the distal efferent duct and the deferent duct epithelial cells.

Animals↗
PubMed1996

Effect of ATP and platelet-activating factor on intracellular calcium concentrations of cultured oviductal cells from cows.

Oviductal endosalpingeal cells were mechanically isolated from heifers at different reproductive stages (cyclic: days 8-14, at oestrus: day 0 and pregnant: day 7) and cultured until 100% confluent. The cells were loaded with the Ca(2+)-sensitive fluorochrome fura-2/AM, and cytosolic free calcium ([Ca2+]i) was monitored spectrofluorimetrically and by use of a microscope image analysis system. ATP (400 mumol l-1) evoked a prompt increase in [Ca2+]i in all cell preparations in both the presence or absence of extracellular Ca2+ when measured with the cuvette method. Single cell measurements using oviductal cells from cyclic heifers revealed a heterogeneous [Ca2+]i pattern in response to ATP, with some cells either failing to respond or reacting very slowly. Platelet-activating factor (PAF, 30 nmol l-1) induced a rapid increase in [Ca2+]i especially in cells derived from cyclic and pregnant animals, but the effect was significantly less in cells from heifers at oestrus. The increase in [Ca2+]i in bovine cells induced by PAF was reduced when extracellular calcium was depleted, indicating that the effect of PAF on [Ca2+]i involves an influx from the extracellular space. Voltage-sensitive calcium channels do not appear to be involved in the influx of extracellular Ca2+ since verapamil had no effect on the PAF-induced increase in [Ca2+]i. The PAF receptor antagonist WEB 2086 inhibited the PAF-mediated effects on [Ca2+]i.

Adenosine Triphosphate↗
PubMed1986

Plasma testosterone-binding globulin-binding capacity in the male lesser mouse lemur (Microcebus murinus): relationship to seasonal and social factors.

Plasma testosterone-binding globulin (TeBG)-binding capacity was determined by equilibrium dialysis in the lesser mouse lemur (Microcebus murinus), a prosimian which exhibits a clear photoperiod-dependent sexual cycle. Plasma TeBG-binding capacity showed significant seasonal changes which were inversely correlated to those of plasma testosterone concentrations. The capacity of TeBG was at its maximum (1.72 +/- 0.4 mumol/l) during the sexual rest period and decreased to 0.98 +/- 0.24 mumol/l during the breeding season when testosterone concentrations were about 200-280 nmol/l. In addition, when males developed a social hierarchy in groups, the decreased sexual function in non-dominant animals was associated with a higher TeBG-binding capacity. However, although TeBG-binding capacity increased when testosterone concentrations decreased either during sexual rest or under social dominance, a positive correlation was found between TeBG-binding capacity and testosterone levels during the breeding season in males with a normal reproductive cycle, i.e. in both isolated and dominant males. This suggests that factors other than sex steroids are involved in the changes in TeBG-binding capacity occurring under seasonal and social influences.

Animals↗
PubMed1998

Biphasic effects of genistein on bone tissue in the ovariectomized, lactating rat model.

The ovariectomized (OVX), lactating rat model has been used to investigate the skeletal effects of the plant estrogen, genistein, over a 14-day period. The OVX, lactating rat on a low-calcium diet loses slightly more than 50% of its bone mineral mass during the first 2 weeks of lactation, and we have demonstrated that estrogen treatment can significantly reduce the loss of femoral mass (ash weight). Following OVX, the rats were assigned to treatment or control groups (both placebo and positive control with estrogen replacement). The treatment groups received one of three doses of a genistein-rich preparation each day via the feed for 2 weeks, after which time the pups began to have an interest in solid feed. A positive control group received conjugated estrogen in the feed. The genistein doses were: low (0.5 mg/d); intermediate (1.6 mg/d); and high (5.0 mg/d). Measurements included ash weights of the femur, scanning electron microscopy (SEM) of the proximal tibia, and uterine weights. SEM results were as follows: (1) at the low dose genistein was approximately equally effective to estrogen in the retention of cancellous bone tissue, as reflected in the number and density of trabeculae in hemisections of the tibial subepiphyseal region, but at high doses genistein was less effective; and (2) rats treated with low-dose genistein, like estradiol, had rougher endosteal surfaces and smaller pores on these surfaces than untreated control rats. Mean ash weights of the entire femur were highest in the rats treated with the low dose compared to control rats (P < 0.05), and they were higher than ash weights of rats administered the intermediate or high doses of genistein. The mean ash weights of the femurs were consistent with the genistein effects on the tibias observed by SEM. In summary, a biphasic response to the genistein preparation was found in this OVX rat model. Interpretation of the results suggests that, at the low dose, genistein appears to be an agonist at the estrogen receptor locus, whereas at higher doses the genistein is less effective and may even have adverse effects on bone cells. These findings of a biphasic effect of genistein (i.e., an inverted U effect) are consistent with those of other recent reports in the literature on isolated bone cells and on reproductive tissues. In summary, lower doses of genistein from soy foods would be expected to act similarly to estrogens with a beneficial effect on bone tissue, but at high doses that are unlikely to be consumed in human diets, this soy derivative may have potentially adverse effects on bone cell functions and thereby on bone tissue.

Animals↗
PubMed2001

The intersection of relational and cultural narratives: women's abortion experiences.

Although a great deal has been written on women's personal deliberations in deciding to have and seeking an abortion, little has been written on the experience of living with this decision or on the meanings an abortion generates in the context of a particular life. In this hermeneutic phenomenological study, 14 women aged 19-44 expressed a need for professionals and others to accept the reality of unplanned pregnancy and to acknowledge that the choice of abortion cannot be fully understood in isolation from women's other reproductive choices. Using a feminist analysis of the tensions and contradictions in the women's accounts, the authors explore the intersections between their experiences and the cultural narratives in which they are situated, in a search for new possibilities for women and new understandings of women's experience of abortion.

Abortion, Legal↗
PubMed2005

[Karyotype and relationships of the endemic char Salvelinus krogiusae from Dal'nee Lake (Kamchatka)].

Resident (lacustrine) chars, inhabiting many lakes of Kamchatka, Chukotka, and continental coast of the Sea of Okhotsk, are of particular interest for understanding the mechanisms of speciation and evolution of Salvelinus chars. Since one of these, the char from Dal'nee Lake (Paratunka River basin, southeastern Kamchatka), is substantially different from lacustrine-riverine and anadromous chars from the Paratunka River basin in several morphological traits, hemoglobin spectrum, ecology, and reproduction, it is considered an isolated species Salvelinus krogiusae. The karyotype of the resident char from Dal'nee Lake was shown to be variable in most individuals, containing 76 to 80 chromosomes in different cells, with NF = 98. This suggests that the variability of the chromosome number in this char form is explained by Robertsonian translocations. In females and males, cells with respectively 2n = 78 and 2n = 77 prevailed. Heteromorphic sex chromosomes were not found in the karyotype of this char species. Nucleolar-organizing regions (NORs) were mostly detected in the telomeric regions of short arms of large submetacentric chromosomes. These features of the Krogius char karyotype demonstrate that this species is isolated from other Kamchatka chars, being similar to Taranetz char S. taranetzi from Chukotka Peninsula, which is in good agreement with morphological data.

Animals↗
PubMed1996

Matrix-induced liver cell aggregates (MILCA) for bioartificial liver use.

Ex vivo reproduction of liver microstructure using isolated hepatocytes is critical for bioartificial liver use. We have developed a method of producing matrix-induced liver cell aggregates (MILCA) using a small number of collagen-coated beads as a nidus for formation of hepatocyte aggregates. Porcine hepatocytes were obtained by EDTA/collagenase digestion. Cell viability was assessed by trypan blue exclusion and LDH release. Cytochrome P-450 activity was determined at 4 and 24 hours by measuring the formation of 7-hydroxycoumarine (7-HC) from 7-ethoxycoumarine (7-EC). At 4 hours, the viability of MILCA was 92 +/- 2%, LDH release was 100 +/- 22 U/L and 7-HC formation was 140 +/- 34 nM/g cells. At 24 hours, MILCA viability remained greater than 90%, but 7-HC formation was lower than that of parallel control monolayer hepatocyte cultures (194 +/- 43 vs 481 +/- 78 nM/g cells; p < 0.002). On transmission electron microscopy, MILCA ultrastructure resembled that of a normal liver (maintenance of cell polarity, gap junctions, bile canaliculi, intact organellae, glycogen granules). MILCA were subsequently inoculated into hollow-fiber bioreactors which were perfused for 6 hours with plasma recovered from patients with fulminant hepatic failure (n = 6; 5 x 10(9) cells/cartridge, recirculation of 350 ml of plasma at 400 ml/min). In these studies, lidocaine (20 micrograms/ml) was cleared in less than 3 hours and 7-HC production at 6 hours was 71 +/- 8 nM/g cells. Other MILCA effects noted in this system included lowering of plasma lactate, bilirubin and ammonia and increase in the level of several non-essential amino acids.

Alginates↗
PubMed1988

The role of JH III and ecdysterone in the regulation of the left colleterial gland activities in Periplaneta americana during the reproductive cycle.

We studied the in vitro effects of JH III and ecdysterone on RNA synthesis and secretory activities of the left colleterial gland (LCG) in mature females of Periplaneta americana during the reproductive cycle. We found that RNA synthesis is a cyclic and highly dynamic process. The major increases in poly(A)- RNA synthesis occurred 8 hr before the peak leucine incorporation into colleterial polypeptides, whereas high poly(A)+ RNA synthesis coincided with high leucine incorporation. Depending on the stage of the cycle JH III exhibited stimulatory or inhibitory effects, or caused no change in RNA synthesis. JH III stimulated RNA synthesis in glands from stages in the reproductive cycle with high endogenous RNA synthesis. The pattern of protein synthesis was not affected by JH III. Ecdysterone lowered RNA synthesis in the LCG and induced secretory activity in LCGs isolated at stages far removed from ovulation. JH III reduced the inhibitory effect of ecdysterone on RNA synthesis and inhibited the ecdysterone-induced secretory activity. The presence of ovarioles isolated at 62 hr of the reproductive cycle (onset of in vivo LCG secretory activity) in the incubation medium inhibited RNA synthesis in 32 hr LCG but induced secretory activity. We suggest that changes in synthetic and secretory activities are coordinated with ovulation and ootheca formation through the interactions between JH III and ovarioles, and ecdysterone may also play a role.

Animals↗
PubMed1988

HLA sharing and fertility in Hutterite couples: evidence for prenatal selection against compatible fetuses.

Antigenic differences between mother and fetus (i.e., blood group incompatibilities) were traditionally considered deleterious for viviparous reproduction. Recently, evidence has accumulated suggesting that maternal response to paternally derived fetal antigens, paradoxically, may facilitate maintenance of pregnancy. Thus, fetuses whose paternally derived antigens do not differ from maternal antigens (i.e., histocompatible pregnancies) may be at a selective disadvantage during pregnancy. Parents sharing histocompatibility antigens (i.e., HLA) may produce compatible fetuses and show overall reduced fertility. Indeed, increased HLA sharing has been reported in some couples experiencing repetitive spontaneous abortion. However, the effects of HLA sharing in couples not selected because of previous pregnancy losses have not been assessed. To elucidate the reproductive effects of maternal-fetal histocompatibility, we initiated prospective population-based studies of parental HLA sharing and reproductive outcome in the Hutterites, a population isolate that lives communally and proscribes contraception. The relationship between HLA-A, -B, and -DR sharing and reproductive outcome was examined in 111 Hutterite couples. Intervals from marriage to each birth were no longer among couples sharing antigens; differences were significant at the second birth and remained significant through the sixth birth (P less than .05). When the effects of sharing at individual loci were examined, HLA-DR was the only individual locus that was a significant predictor of birth interval length (P = .025). Completed family sizes were 6.5 and 9.0 among couples sharing and not sharing HLA-DR, respectively (P = .082, 2-tailed). However, recognized fetal loss rates did not differ among couples sharing and not sharing antigens.(ABSTRACT TRUNCATED AT 250 WORDS)

Abortion, Habitual↗
PubMed2004

Lost in time, lonely, and single: reproductive asynchrony and the Allee effect.

Identifying linkages between life-history traits and small population processes is essential to effective multispecies conservation. Reproductive asynchrony, which occurs when individuals are reproductively active for only a portion of the population-level breeding period, may provide one such link. Traditionally, reproductive asynchrony has been considered from evolutionary perspectives as an advantageous bet-hedging strategy in temporally unpredictable environments. Here, we explore the dynamic consequences of reproductive asynchrony as a density-dependent life-history trait. To examine how asynchrony affects population growth rate and extinction risk, we used a general model of reproductive timing to quantify the temporal overlap of opposite-sex individuals and to simulate population dynamics over a range of initial densities and empirical estimates of reproductive asynchrony. We also considered how protandry, a sexually selected life-history strategy that often accompanies asynchrony, modulates the population-level effects of reproductive asynchrony. We found that asynchrony decreases the number of males a female overlaps with, decreases the average probability of mating per male/female pair that does overlap, and leaves some females completely isolated in time. This loss of reproductive potential, which is exacerbated by protandry, reduces population growth rate at low density and can lead to extinction via an Allee effect. Thus reproductive asynchrony and protandry, both of which can be evolutionarily advantageous at higher population densities, may prove detrimental when population density declines.

Animals↗
PubMed2003

Reproduction of postweaning multisystemic wasting syndrome in pigs experimentally inoculated with a Swedish porcine circovirus 2 isolate.

In recent years, porcine circovirus type 2 (PCV2)-associated postweaning multisystemic wasting syndrome (PMWS) has been reported worldwide. However, to date, PMWS has not been reported in Sweden despite the demonstration of serum antibodies to a PCV2-like virus in Swedish pigs. This communication reports the experimental reproduction of clinical PMWS after inoculation of colostrum-deprived (CD) pigs, derived from a Northern Ireland herd, with an isolate of PCV2 virus recovered from a clinically normal Swedish pig that was necropsied in 1993. The clinical disease and histological lesions observed in CD pigs inoculated with this virus were indistinguishable from those observed in previous studies on CD pigs inoculated with a PCV2 virus isolate recovered from pigs with PMWS. These results highlight the disease potential of PCV2 isolated from regions apparently free of PMWS and suggest that the status of the host and its environment is an important factor in the development of clinical PMWS.

Animals↗
PubMed2001

Comparison of porcine reproductive and respiratory syndrome virus growth in media supplemented with fetal bovine serum or a serum replacement.

Commercially available serum replacements are often used in cell culture as a cheaper and less variable substitute for fetal bovine serum (FBS). The growth of porcine reproductive and respiratory syndrome virus (PRRSV) isolates in CRL11171 cells maintained in a medium supplemented with FBS was compared with virus propagation in the same cell line maintained in the same medium with a serum replacement. The PRRSV replicated significantly better when the cell culture medium was supplemented with FBS. The results of this study have implications for the use of serum replacement-supplemented medium for PRRSV diagnosis by virus isolation.

Animals↗
PubMed2005

Economy of mate attraction in the Cassin's finch.

Mate attraction can be costly. Thus, individuals should modulate it according to its probable benefits. Specifically, individuals should modulate mate-attraction efforts based on their need for, the probability of attracting, and the reproductive competence of prospective mates. We tested these predictions by monitoring song output in laboratory-housed male Cassin's finches (Carpodacus cassinii) before, during and after brief female exposure following variable periods of isolation from females. We inferred individual reproductive competence from the product of season and reproductive schedule, the latter estimated from moult progress. Males produced little song in the presence of a female but robustly elevated song output in response to female loss. However, mere absence of a female did not elevate song output in males unaccustomed to female proximity. Furthermore, song output in response to female loss increased with her reproductive competence. We suggest that individuals modulate mate-attraction effort based on the benefits such efforts are likely to yield.

Animals↗
PubMed1978

Significance of appropriate techniques and media for isolation and identification of Ureaplasma urealyticum from clinical specimens.

Controversy over the association of Ureaplasma urealyticum with reproductive failure may be due to methods used to isolate the microorganism. U. urealyticum isolations from clinical material should be done simultaneously in broth and on Shepard's differential agar medium (A7) containing manganese sulfate. Urine sediments result in a 9% (P = 0.0002) higher rate of isolation than than cervical and urethral swabs. Primary isolations may not display standard textbook morphology. Isolated colonies may be present, but brown streaks in cervical mucus or a coalescent haze around epithelial cells in urine sediment may also be seen in areas of concentrated growth. The broth and agar media used, method of incubation, type of specimen, and method of storing specimens before culture are all factors which influence the recovery of U. urealyticum.

Bacteriological Techniques↗