Sublethal impact of short term exposure to the organophosphate pesticide azamethiphos in the marine mollusc Mytilus edulis.

Concern has been raised that the increased use of pesticides in intensive aquaculture practices may cause adverse sublethal effects to non-target aquatic species. Azamethiphos is an organophosphate (OP) pesticide used to combat sea lice infestations in farmed salmonids. Here, the sublethal impact on the blue mussel, Mytilus edulis, of short term exposure to azamethiphos was determined. The testing regime included biomarkers of exposure (acetylcholinesterase activity), cytotoxicity (neutral red retention), immune function (phagocytic index) and physiological condition (feeding rate). The distribution and sensitivity of M. edulis acetylcholinesterase to inhibition by azamethiphos was first determined, yielding IC(50) values of 0.736 and 1.30 mg l(-1) for gill and haemolymph, respectively. Exposure of mussels to 0.1 mg l(-1) azamethiphos for periods of up to 24h caused a significant reduction in acetylcholinesterase activity in both the haemolymph (P<0.0002) and the gill (P<0.002), alteration in cell viability (P<0.02) and decrease in phagocytic index (P<0.03). The feeding rate remained unaffected. The results support the hypothesis that, in addition to its neurotoxic effects, azamethiphos can modulate haemocyte function and immune defence in M. edulis at environmentally relevant concentrations after only a few hours.

Correlation of heel pain with body mass index and other characteristics of heel pain.

A prospective, descriptive study was performed at Oakwood Healthcare medical clinics to determine the body mass index (BMI) of patients with heel pain and of a control group of patients presenting for other reasons. A questionnaire was used to obtain information in each of the patient groups and to determine characteristics of patients with plantar fascial heel pain. Standard weightbearing lateral radiographs were taken to determine overall foot structure. The typical patient was female, had heel pain for just over 1 year, with a sedentary to moderate activity level. Although height was comparable, patients with heel pain had a higher BMI (30.4 +/- 0.7) than those without heel pain (28.2 +/- 0.7, p = .04). The BMI appears to play a greater role in heel pain than does foot structure, as the authors found no structural commonalities that would explain these patients' pain. Control patients also reported a higher level of activity. Fifty-one percent exercised three or more times per week for more than 20 minutes each time, while less than half that (25.4%) of heel pain patients did so. While half of the heel pain patients had been treated by other providers prior to visiting our clinic, fewer than 25% of these patients had been instructed to lose weight by a physician. The authors feel that a BMI of 25 (the target for decreased cardiovascular risk) represents a reasonable goal for weight loss that may reduce heel pain.

Evolutionary advances in the higher fungi.

In this report the phrase 'evolutionary advances' is used in three ways: 1. to describe monophyletic changes perceived within a lineage; 2. to describe evolutionary sequences that appear to have become parallel/convergent; 3. to describe major transitions inferred between primary taxa. In monophyletic evolution the changes occur within a specific lineage that arises from a common ancestor, e.g., modern Man, horses, rusts. In parallel/convergent evolution different lineages respond similarly over time to environmental challenges and opportunities and come to acquire a great deal of comparability (e.g., Webster, 1987). Such lineages may be designated as separate taxa, e.g., similarities in marsupial and placental carnivores, or, if the polyphyleticism is cryptic, as a collective taxon, e.g., Aves, the class of birds, the obsolete Amentiferae for catkin bearing plants, the Gasteromycetes, and lichens. In major transitions there are significant paradigm shifts in which evolutionary changes from one predominant life style pattern to another are accompanied by increases in complexity (see Smith & Szatháry, 1995), e.g., symbiosis, the water to land transition, the changes between the phyla of land fungi. Three particular terms are used in evaluating evolutionary relationships (Moore, 1996a): homology, paramology, and analogy. Homology, from Darwin's theory of common descent, is the phenomenon of having a common historical origin but not necessarily the same final structure or function (e.g., vertebrate forelimbs). Paramology (Moore, 1971) applies to inferred relationships in evolutionary schemes based on contemporary forms that lack fossil antecedents, e.g., the various phylogenetic interpretations of prokaryotes, algae, and fungi; Boekhout et al. (1993) have evaluated the taxonomic resolution of a variety of morphologic, biochemical, physiological, and molecular characters (Table 1). Analogy is generally applied to similar forms that are unrelated, e.g., insect/vertebrate wings; prokaryote/eukaryote flagella. It should also be borne in mind that, in a given taxon, biotrophism (Coffey, 1975) is an advanced character (Health, 1987) over, respectively, weaker parasitism, symbiotism, commensalism, and freeliving and that seemingly simple or less differentiated forms can be, and more than likely than not are, reduced, polyphyletic, and specialized rather than ancient and rudimentary, e.g., yeasts (Hoog et al., 1988; Kurtzman & Fell, 1996; Moore, 1988b; 1996a).

Attenuation of the magnitude of suckling-induced prolactin release with advancing lactation: mechanisms.

To study why suckling-induced plasma prolactin levels decline in magnitude with advancing lactation, we examined prolactin release in lactating rats following suckling and pharmacologic manipulations during early, mid- and late lactation. On day 2 of lactation, litters were adjusted to 8 pups. On day 3, dams were implanted with an atrial catheter and experiments were conducted on lactation days 5, 11 and 17. To examine suckling-induced prolactin release, pups were removed at 0800 h, an extension was attached to the catheter at 1100 h, and pups returned to dams at 1200 h. Blood samples were obtained before, and at 10, 30, 60, 90 and 120 min after suckling started. Prolactin responses to sulpiride and thyrotropin releasing hormone (TRH) administration were studied in lactating rats separated from their litters for 4 hours. Blood samples were obtained before, and at 10, 30, 60 and 90 min after sulpiride (10 or 40 micrograms/kg BW) and 5, 10, 20 and 30 min after TRH (1 or 4 micrograms/kg BW) in rats pretreated with sulpiride. Prolactin release in response to suckling, administration of sulpiride or sulpiride and TRH diminished as lactation advanced. From these results, we conclude that refractoriness in anterior pituitary lactrotropes to prolactin-releasing stimuli is at least partially responsible for the decline in suckling-induced prolactin release with advancing lactation.

The Rey AVLT recognition memory task measures motivational impairment after mild head trauma.

The effect of financial incentives on Rey AVLT recognition memory performance was assessed. Clinically referred mild head trauma (MHT) patients with financial incentives were compared to patients with neurologically documented brain dysfunction who did not have financial incentives. The MHT group was subdivided into a group with strong performance on a measure of motivation, the Portland Digit Recognition Test (PDRT), and a group with weak performance on the PDRT. The resulting three groups were equated on Rey AVLT acquisition, age, and education. The MHT subgroup with poor PDRT scores also was significantly impaired on recognition memory. In the total sample the prevalence of recognition scores of less than 6 correct was 27% in the MHT group and 5% in the brain dysfunction group. We conclude that severe Rey AVLT recognition memory impairment likely reflects motivation to exaggerate deficits, at least in MHT patients with no neurological evidence of brain injury.

Alcohol effects on TRH-induced prolactin response in lactating rats: in vivo and in vitro studies.

The site of action of alcohol in inhibiting suckling-induced prolactin release in lactating rats was examined by in vivo and in vitro studies. In vivo, sulpiride- and thyrotropin-releasing hormone (TRH)-induced prolactin release was studied in lactating rats separated from their litter. On day 7, dams were implanted with an atrial catheter. On day 10, pups were removed from dams at 0800 h and, after 5 h, an extension was attached to the catheter. An hour later, a baseline blood sample was removed and was followed by sulpiride (40 micrograms/kg) administration. Additional blood samples were withdrawn over 1 h. After the 60-min sample, sulpiride-administered rats were infused with 0.0, 1.0, or 2.0 g/kg b.wt. alcohol. Following alcohol, a postinfusion blood sample was removed, TRH (4.0 micrograms/kg) was administered, and subsequent blood samples were obtained 5, 10, 20, and 30 min after TRH. For in vitro studies, cells from lactating rats in midlactation were enzymatically dissociated, plated, and on culture day 5 were exposed to 0 or 10 nM TRH. Each set of cells were additionally exposed to 0, 100, or 300 mg% alcohol and media harvested after 4 h. In a subsequent study, plated cells were exposed to increasing doses of TRH in the presence of 0, 100, or 300 mg% alcohol and media harvested as above. Prolactin in plasma (in vivo studies) and medium (in vitro studies) was measured by RIA.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of thrombin inhibition on the dynamics of thrombolysis and on platelet function during thrombolytic therapy.

To evaluate the effect of thrombin on the dynamics of thrombolysis, we infused rabbits with heparin or hirudin alone or in conjunction with tissue-type plasminogen activator (t-PA) and monitored the kinetics of fibrinolysis and changes in ex vivo platelet aggregation responses over time. Both heparin and hirudin enhanced total fibrinolysis in an ex vivo arteriovenous shunt preparation: 82 +/- 2% and 79 +/- 2%, respectively, compared with 51 +/- 8% for t-PA alone (P less than 0.05) and 50 +/- 4% for t-PA plus aspirin (p less than 0.05). Heparin coadministered with t-PA significantly reduced the half-time for clot lysis compared with t-PA alone (p less than 0.05), whereas hirudin coadministered with t-PA significantly reduced the half-time for clot lysis compared with that for t-PA alone, t-PA plus aspirin, and t-PA plus heparin (5.5 +/- 0.6 versus 12.1 +/- 2.0 versus 12.6 +/- 2.2 versus 10.0 +/- 0.8 minutes, respectively; p less than 0.05). Both heparin and hirudin prevented the increase in ADP-induced platelet aggregation normally seen with t-PA alone (p less than 0.01 by t test; p less than 0.05 by two-way analysis of variance). These data demonstrate that selective, antithrombin III-independent thrombin inhibitors can enhance the efficacy of thrombolysis by modulating the dynamics of the process and preventing platelet activation associated with plasminogen activator therapy.

Cocaine inhibits mating-induced, but not human chorionic gonadotropin-stimulated, ovulation in the rabbit.

The effects of cocaine on ovulation and corpus luteum function were investigated in New Zealand White rabbits. Forty females were randomly assigned to control and cocaine-treated groups. Controls were given vehicle s.c. daily for 5 days and cocaine-treated rabbits received 40 mg/kg cocaine hydrochloride s.c. daily for 5 days. One hour after the last cocaine dose, half the control and half of the cocaine-treated groups were mated with fertile males and the other half of each group received hCG i.v. Serial blood samples were obtained over 4 h on the day of mating or hCG treatment (Day 0), and then at intervals from Days 1-18. No mated, cocaine-treated rabbits ovulated, vs. 6 of 10 controls (chi-square: p = 0.01). In contrast, all animals given hCG had comparable numbers of corpora lutea (control: 7.1 +/- 0.8; cocaine: 5.7 +/- 0.8). Peak levels of benzoylecgonine (the major cocaine metabolite) occurred between 180 and 240 min after cocaine administration. In cocaine-treated animals that were mated, Day 0 serum LH (repeated measures MANOVA, p less than 0.01) and FSH (p less than 0.03) concentrations were lower than those in pregnant controls. Serum LH and FSH levels for all hCG recipients (cocaine-treated and control) did not differ. Serum prolactin concentrations in mated, pregnant rabbits were higher than in all other groups; cocaine treatment did not affect this hormone.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of oocyte exposure to local anesthetics on in vitro fertilization and embryo development in the mouse.

The effect on fertilization and development of local anesthetics routinely used during ultrasound-guided oocyte retrieval in women undergoing in vitro fertilization was examined in a mouse in vitro fertilization system. Mouse oocytes were exposed in vitro to lidocaine, chloroprocaine, and bupivacaine at concentrations of 0 (control), 0.01, 0.1, 1.0, 10.0, 100.0 micrograms/mL for 30 min, washed, and then inseminated. In vitro oocyte fertilization at 24 and 48 h and embryo development at 72 h were determined. Bupivacaine adversely affected mouse in vitro fertilization and embryo development only at the highest exposure concentration, 100 micrograms/mL, while lidocaine and chloroprocaine produced adverse effects at concentrations as low as 1.0 and 0.1 microgram/mL, respectively. Furthermore, an adverse dose-related effect on fertilization and embryo development was shown for lidocaine and chloroprocaine, but not for bupivacaine. These data demonstrate that the local anesthetics, lidocaine (L), chloroprocaine (C), and bupivacaine (B), adversely affect mouse in vitro fertilization and embryo development in the order of C greater than L greater than B.

The gonadotropin-releasing hormone agonist leuprolide affects the thymus and other non-reproductive systems of female rats.

To evaluate the effects of a gonadotropin-releasing hormone agonist on non-reproductive systems, we administered [D-Leu6,Des-gly10]-GnRH ethylamide (leuprolide; 5 micrograms/day) for 21 days to female Sprague-Dawley rats. In Experiment 1, continuous infusion (Alzet minipumps sc) was compared to injection. Increased thymus and body weights and decreased estradiol and uterine weights were noted for both administration methods. Spleen weight increased only in rats treated by continuous infusion. Ovary, kidney and liver weights did not change. Only leuprolide-injected rats had elevated LH with decreased corticosterone and ACTH levels, possibly related to the injection process. Glucose, insulin, progesterone, FSH and corticosterone/ACTH were not different. In Experiment 2, intact and ovariectomized rats were implanted with minipumps delivering leuprolide or 0.9% NaCl. Body and thymus weights increased, whereas uterine weight and estradiol declined in both leuprolide-treated and ovariectomized rats. No synergism between leuprolide and ovariectomy was noted. Thymosin alpha 1, but not thymosin beta 4, increased in leuprolide-treated ovariectomized rats. Peripheral white blood cell count was elevated in leuprolide-treated intact rats and ovariectomized rats. In bone marrow, non-nucleated cell count declined in leuprolide-treated intact rats, contributing to the decreased total cell count in this group. Nucleated cell count was unaffected. Therefore, thymus weight gain was accompanied only in some cases by functional changes. Our results demonstrate that leuprolide affects non-reproductive systems, in a similar manner to ovariectomy. We suggest that such alterations may be due to the hypoestrogenic environment produced by leuprolide.

Alcohol inhibition of suckling-induced prolactin release in lactating rats: threshold evaluation.

Prolactin release in response to suckling was examined in primiparous lactating rats two hours after alcohol administration. Litters were adjusted to eight pups on lactation day 2 and dams were implanted with an atrial catheter on day 6. On day 10, pups were separated from the mother at 0800 h. An extension was attached to the catheter at 1100 h. Following removal of a baseline blood sample an hour later, rats were infused with alcohol doses of 0, 0.5, 1.0, 2.0 or 2.5 g/kg body weight. Two hours later, pups were returned to dams. Subsequent blood samples were obtained 10, 30, 60, 120 and 180 min after the onset of suckling. Following 10 min of suckling, plasma prolactin for groups of rats infused with alcohol at 2.0 and 2.5 g/kg body weight were lower than control, 0.5 and 1.0 g/kg groups. The blood alcohol level (BAL) for the 2.0 g/kg group was 94 +/- 8 mg% and for the 2.5 g/kg group was 162 +/- 4 mg%. After 30 min, the BAL for the 2.5 g/kg group was 134 +/- 5 mg% and plasma prolactin was suppressed in this group compared to control, 0.5 and 1.0 g/kg groups. The BAL for the 2.0 g/kg group after 30 min of suckling was 74 +/- 9 mg% but prolactin was not significantly lower than controls. We conclude that in rats, alcohol inhibition of suckling-induced prolactin release is directly correlated to the BAL. The threshold BAL which effectively inhibits this prolactin release is lower than the human legal intoxication level.

Human granulosa-luteal cell response to vinblastine exposure in vitro.

We examined the response of isolated human granulosa-luteal cells (HGLCs) to a cancer chemotherapeutic agent, vinblastine (VLB), that has been implicated in ovarian failure during treatment for Hodgkin's disease. VLB doses of 1.0 micrograms/mL for 4 h or 0.1 micrograms/mL for 24 h reduced HGLC progesterone production during exposure. The effect of high doses (10.0 and 100.0 micrograms/mL) persisted for at least 1 day after exposure. Previous 24 h, but not 4 h, high-dose VLB exposure reduced subsequent progesterone release in response to 10 IU of human chorionic gonadotropin (hCG). Without hCG stimulation, only cells previously exposed to 100.0 micrograms/mL had persistently reduced progesterone release. We conclude that HGLCs can completely recover from a short exposure to VLB, but longer exposures to 10.0 and 100.0 micrograms/mL are detrimental to their hormone secreting capacity.

Ubiquinone and urease distribution in Taphrina and Symbiotaphrina.

Identifications of ubiquinone isoprenologues are presented for isolates identified with six species of Taphrina and for isolates of the two species of Symbiotaphrina. All had Q-10 as the major ubiquinone system. The inclusion of T. populina and S. buchneri, the respective type species, establishes this as the value for these genera. Both species of Symbiotaphrina were urease positive even though, according to the literature, they are unable to utilize urea as a sole nitrogen source. The urease results for the Taphrina isolates were mixed.

The effect of cocaine on oocyte development and the follicular microenvironment in the rabbit.

We examined the effects of cocaine exposure in the rabbit on in vitro oocyte development and on steroidal content of follicular fluid (FF) and serum progesterone (P). Cocaine hydrochloride (0, 10, 20, 40, or 80 mg/kg) was administered daily subcutaneously for 5 days to New Zealand White female rabbits before superovulation. On the last day of cocaine administration, animals were given human chorionic gonadotropin intravenously, and laparotomy was performed 6 to 8 hours later. During laparotomy, ovaries were removed, the number of follicles recorded, oocytes retrieved, and FF was obtained. In vitro fertilization (IVF) was then performed on the oocytes and the rate of cleavage observed. For all cocaine dosage groups, no differences were observed in the number of follicles present, number of oocytes retrieved, or IVF and cleavage rates. Cocaine did, however, decrease periovulatory serum P, and FF P, whereas FF estradiol concentrations increased. This suggests that short-term cocaine exposure affects the follicular steroid milieu, possibly by delaying granulosa cell luteinization.

Acute alcohol infusion does not alter plasma gonadotropins or prolactin in ovariectomized rats.

We examined the effects of acute alcohol on basal plasma FSH, LH, and prolactin in ovariectomized rats. Alcohol infusion and blood sampling were done via an indwelling atrial catheter. Blood samples for alcohol and hormone determinations were collected before, and 5 to 120 min after completion of saline (control) or alcohol in saline (experimental) infusion. Plasma follicle-stimulating hormone, luteinizing hormone, and prolactin were not altered during the 2-hr period. Peak blood alcohol concentrations achieved following 1.0- and 2.0-g/kg body weight of alcohol doses were approximately equal to, and twice, the legal human intoxication levels, respectively. Alcohol clearance rates from blood for the two groups were: 130 +/- 3 mg/kg/hr for the 1.0-g/kg body weight group and 151 +/- 3 mg/kg/hr for the 2.0-g/kg body weight group. These results show that acute alcohol does not affect basal gonadotropins and prolactin secretion in ovariectomized rats.

Vinblastine reduces progesterone and prostaglandin E production by rat granulosa cells in vitro.

To elucidate the possible mechanism(s) of vinblastine-induced premature ovarian failure, we studied the effect of vinblastine (VLB) on progesterone (P) and prostaglandin E (PGE) production by rat granulosa cells in vitro. Granulosa cells obtained from immature, pregnant mares' serum gonadotrophin-primed rats were incubated for 5 h in modified Medium 199 +/- LH (10 ng/mL) with varying concentrations of VLB (0.001 to 10.0 micrograms/mL) and cells plus media were assayed for total P and PGE. VLB reduced production of both basal and LH-stimulated P in granulosa cells, with VLB at a concentration of 0.1 micrograms/mL showing the first significant difference from control. This dose also suppressed basal and LH-stimulated PGE. Granulosa cell survival was similar for all groups. Thus VLB, an agent known to disrupt microtubular function, reduced granulosa cell production of both P and PGE. Our results suggest that at a concentration (0.1 micrograms/mL) lower than that routinely achieved during chemotherapy (0.3 micrograms/mL), VLB depresses rat granulosa cell function in vitro. This system appears to be a valid model for preliminary assessment of the cytotoxic effects of chemotherapy on an ovarian component.

Ceratorhiza oryzae-sativae, a new combination for the anamorph of Cerabasidium setariae.

Rhizoctonia oryzae-sativae, the anamorph of Ceratobasidium setariae, is transferred to Ceratorhiza. A discussion of teleomorph and anamorph taxonomy and nomenclature is also given and it is concluded that, for the present, there is only one holomorph species.

Low dose alcohol contracts the human umbilical artery in vitro.

Beginning at very low concentrations (0.001 g/100 ml), alcohol elicited dose-dependent contractions of the human umbilical artery in vitro. Additionally, 16 of the 108 arteries tested had a 5-10-min spasm in response to alcohol. Alcohol (0.2 g/100 ml) also increased tension developed in response to all angiotensin II doses, but had no effect on serotonin-induced contractions. These results suggest that alcohol may increase umbilicoplacental resistance in vivo, thus decreasing fetal-placental blood flow.