Embryotoxicity of a single dose of medroxyprogesterone acetate (MPA) and maternal serum MPA concentrations in cynomolgus monkey (Macaca fascicularis)

A single dose of MPA (Depo-Provera; Upjohn Co., Kalamazoo, Michigan) was administered intramuscularly to 12 time-mated pregnant cynomolgus monkeys on day 27 (+/- 2) of gestation at 25 mg/kg or at 100 mg/kg. Maternal blood samples were collected immediately prior to MPA injection and then at regular intervals until cesarean section at term (day 152 +/- 3). Infants in both dose groups had external genital abnormalities. Female infants in the low-dose groups had partial or complete labial fusion, prominent median raphe, and clitoral hypertrophy; at high doses (100 mg/kg), the female infants had complete labial fusion and a distinct penile urethra. MPA had an opposite effect on external genitalia of male infants. The penis was short and the scrotal swelling was absent or less conspicuous, and two males had hypospadias. The adrenal glands were significantly smaller (P less than 0.05) in infants of both sexes treated with 100 mg/kg. One of the infants treated with 25 mg/kg of MPA had a muscular ventricular septal defect. Serum concentrations of MPA were determined by radioimmunoassay in eight pregnant monkeys. In the 25 mg/kg group the patterns of MPA profiles in the serum were similar in all four animals. An initial peak occurred at 24-48 hr postinjection (2.7-9.6 ng/ml), followed by a slight decrease at 3 days postinjection (gestational day 30), and then a steady increase to maximum levels of 10-14 ng/ml occurring between gestational days 37 and 50. Serum levels gradually declined to concentrations below 5 ng/ml by midgestation in three of four monkeys. By comparison, both the patterns and magnitude of MPA concentration showed great interanimal variation in the 100 mg/kg group. MPA was present in cord blood at measurable concentrations in infants at both dose groups; the levels ranged from 0.6 to 8.3 ng/ml, corresponding to 40-72% of the maternal concentrations. These results demonstrate that a single injection of MPA during early pregnancy causes selective embryotoxicity in both male and female fetuses. Presence of high levels of MPA in maternal sera during the critical period of genital development can cause specific genital defects; however, the exact mechanism by which MPA causes these paradoxical genital abnormalities is unknown.     

南麂列岛海洋保护区浮游动物调查

主要研究南麂列岛海洋保护区浮游动物种类组成、数量分布、多样性指数、浮游动物与浮游植物动态变化及浮游动物数量变化与营养盐的关系。经鉴定共发现,浮游动物98种,主要有2个生态类群:(1)暖温带近海类群,优势种有中华哲水蚤(Calanus sinicus)、中华假磷虾(Pseudeuphausia sinicas)、五角水母(Muggiaea atlantica)、百陶箭虫(Sagitta bedoti)、拿卡箭虫(S.nagae)等;(2)暖水性外海类群,代表性种类有肥胖箭虫(S.enflata)、精致真刺水蚤(Euchaeta concinna)等。结果表明,8月份南麂列岛浮游动物生物量和丰度出现最高值,9、10月份逐渐减少,多样性指数变化范围1.78~4.38,平均3.99;保护区内浮游动物数量与浮游植物细胞密度呈良好的正相关关系,与氮含量呈负相关关系。     

中街山列岛海洋保护区鱼类物种多样性

基于2010-2012年在中街山列岛海洋保护区进行的8个航次底拖网调查数据资料,从鱼类分类学和生态类群等方面,结合多样性、资源密度和相对重要性指数,对该保护区鱼类物种多样性特征进行了分析。调查海域共采集鱼类55种,隶属2纲12目32科44属。其中,鲈形目鱼类26种,占47.3%;趋礁性鱼类41种,占74.5%;暖温性、暖水性和冷温性鱼类分别为30、24和1种,其中暖水种约占所有偶见种的70%;定居种、近海洄游种和季节性种分别为27、24和24种;底层、近底层和中上层鱼类分别占40.0%、36.4%和23.6%,其中矛尾鰕虎鱼(Chaeturichthys stigmatias)、龙头鱼(Harpodon nehereus)和鳀科鱼类分别成为各水层的绝对优势种。所有调查站点鱼类平均生物量指数夏季最高(1043.7 kg/km²),春季最低;尾数密度指数春季最高(122×10³ 尾/km²),冬季最低;生物量与尾数密度指数最高与最低季节之间均存在显著性差异(P<0.05)。对多样性的分析显示,Margalef丰富度(D)冬季最高,秋季最低,Shannon-Wiener多样性(H')与Pielou均匀度(J')均为夏季最高,秋季最低。聚类和NMDS方法分析显示不同季节鱼类群落组成格局差异极显著(P<0.01)。研究结果表明,中街山列岛海洋保护区鱼类资源结构受近海季节性洄游种类影响较大,鱼类分布季节动态多呈现洄游性更替节律,保护区内虽已出现鱼类多样性下降趋势和生态系统功能退化现象,传统经济种类的优越性在逐渐下降或边缘化,但独特的岛礁生境仍能很好地发挥着维持固有种类和提供良好栖息场所的优势,随着海洋牧场建设与一系列资源保护与修复措施,石首鱼科幼体已形成优势群体,而且保护区内出现大量小型饵料鱼类。     

In vitro functions of lymphocytes during high-dose medroxyprogesterone acetate (MPA) treatment

Summary The number and function of T and B lymphocytes were studied in endometrial cancer patients during 3 months' treatment with high-dose medroxyprogesterone acetate (MPA). No significant changes were observed in the proportions of T and B cells or in the function of B cells during MPA treatment. At 3 months, lymphocyte blastogenic responsiveness to mitogens was slightly reduced both in patients treated with standard therapy alone (intracavitary radium and surgery) and in patients receiving additional MPA therapy. These results indicate that other factors than progesterone are responsible for the suppression of lymphocyte blastogenesis induced by mitogens during high-dose progestin treatment.     

Antitumor effects of combination toremifene and medroxyprogesterone acetate (MPA) in vitro and in vivo

The estrogen (ER) and progesterone (PgR) receptor levels in various gynecological tumors were measured. The same tumors were exposed in vitro to toremifene, MPA or their combination and the growth of the tumors was followed by measuring the adenosine triphosphate (ATP) within the cells by a simple bioluminescence assay. Altogether 34 clinical samples were studied. DMBA-induced mammary tumors bearing rats were treated in vivo with toremifene, MPA and their combination. About half of the ovarian cancers and 6 out of the 7 adenocarcinomas of uteri contained ER. The ovarian tumors were PgR rich in 25% and adenocarcinomas of uteri in 6 out of the 7 cases. When compared to control toremifene (concentration 1 mumol/l) was able to decrease the number of living cells to 50% or less in 9/34 samples, MPA (concentration 10 mumol/l) in 17/34 samples, and the combination in 25/34 samples. In five cases the antitumor effect of the combination was synergistic. In two cases signs of weak antagonism were seen. In vivo the antitumor effect of toremifene and MPA was clearly synergistic against DMBA-induced cancers. The effect was dose-dependent and at sufficiently high doses it was possible to eradicate the tumors and cure the animals.     

Metabolism of medroxyprogesterone acetate (MPA) via CYP enzymes in vitro and effect of MPA on bleeding time in female rats in dependence on CYP activity in vivo

Medroxyprogesterone acetate (MPA) is a drug commonly used in endocrine therapy for advanced breast cancer, although it is known to cause thrombosis as a serious side effect. Recently, we found that cytochrome P450 3A4 (CYP3A4) mainly catalyzed the metabolism of MPA via CYP in human liver microsomes. However, the metabolic products of MPA in humans and rats have not been elucidated. In addition, it is not clear whether thrombosis could be induced by MPA itself or by its metabolites. In this study, we determined the overall metabolism of MPA as the disappearance of the parent drug from an incubation mixture, and identified the enzymes catalyzing the metabolism of MPA via CYP in rats. Moreover, the effects of CYP-modulators on MPA-induced hypercoagulation in vivo were examined. Intrinsic clearance of MPA in rat liver microsomes was increased by treatment with CYP3A-inducers. The intrinsic clearance of MPA in liver microsomes of rats treated with various CYP-inducers showed a significant correlation with CYP3A activity, but not CYP1A activity, CYP2B activity or CYP2C contents. Among the eight recombinant rat CYPs studied, CYP3A1, CYP3A2 and CYP2A2 catalyzed the metabolism of MPA. However, since CYP3A2 and CYP2A2 are male-specific isoforms, CYP3A1 appears to be mainly involved in the metabolism of MPA in liver microsomes of female rats. In an in vivo study, pretreatment of female rats with SKF525A, an inhibitor of CYPs including CYP3A1, significantly (p < 0.05) enhanced MPA-induced hypercoagulation, whereas pretreatment with phenobarbital, an inducer of CYPs including CYP3A1, reduced it. These findings suggest that CYP-catalyzed metabolism of MPA is mainly catalyzed by CYP3A1 and that MPA-induced hypercoagulation is predominantly caused by MPA itself in female rats.     

An enzyme linked immunosorbent assay (ELISA) for plasma medroxyprogesterone acetate (MPA).

A single extraction fixed antigen enzyme-linked immunosorbent assay (ELISA) that can be completed in less than 24 h is described for the measurement of medroxyprogesterone acetate (MPA) in plasma. MPA is covalently coupled to bovine thyroglobulin and passively adsorbed in guanidine hydrochloride to a standard 96-well microtitre plate where it competes with MPA in the extracted plasma sample for goat anti-MPA. Antibody binding to the solid phase is determined via binding of a horse-radish peroxidase second antibody which reacts colorimetrically with its substrate. The reaction is stopped by addition of 1.25 M H2SO4 and absorbance read at 492 nm. All steps except for sample addition and extraction can be performed on an automatic ELISA processing machine. The assay is sensitive, specific and precise, with intra- and inter-assay coefficients of variation of less than 10 and 15%, respectively. Assay sensitivity is 0.08 ng/ml. The assay follows established methodology for other assays in this laboratory which assists standardization, cost structure and sample throughput and thus is a useful alternative to radioimmunoassays for the determination of MPA in plasma.     

Immediate Effects of Experimental Human Trampling on Mid-Upper Intertidal benthic Invertebrates at the Asinara Island MPA (NW Mediterranean)

To assess potential risks of human visitation to ecological communities, the immediate effects of human trampling were investigated experimentally on small invertebrates inhabiting mid-upper intertidal hard bottoms covered by algae. Two different experimental intensities of trampling (60 and 120 footsteps) and controls (with no trampling) were applied to quadrats 20×20 cm in size (experimental area), within the two ‘no-entry, no-take’ zones of the Asinara Island MPA (Italy, Mediterranean Sea). One day after trampling ended, samples of benthic fauna were collected and the animals attributed to macrofaunal and meiofaunal components. Analyses of variance on the nine most common taxa of macrofauna identified significant higher abundance of bivalves, gammarid amphipods, polychaetes, isopods, oligochaetes in controls than in trampled plots. For nematodes, polychaetes, ostracods, oligochaetes, bivalves, acari, caprellid amphipods and tanaids a significant higher abundance of meiofaunal animals was found in controls than in trampled areas. Although no information on recovery is available, these results suggest that macrofaunal and meiofaunal taxa are vulnerable to this type of disturbance.     

Developing a scuba trail vulnerability index (STVI): a case study from a Mediterranean MPA

Scuba diving is now one of the major form of commercial use of marine protected areas (MPAs) around the world and the control of its potential impacts on the marine environment represents a fundamental key to manage this recreational activity in highly dived areas. A potential tool to tackle such issues has been thought to be the definition of a value of recreational carrying capacity of an area, but this approach has been rarely considered management-effective. Therefore, the first step for effectively managing scuba-diving should be ‘bottom-up’: characterizing the benthic communities potentially affected by diving and evaluating their vulnerability. Aim of this paper is to propose a tool to define an index of vulnerability for dive trails (STVI: scuba trail vulnerability index). This has taken into consideration both physical and biological features of each trail. All the considered features are represented by non-quantitative variables, because either they are purely qualitative or their quantitative measurement is impractical. The management of such qualitative information and its translation into a formal methodology was performed by means of fuzzy logic, which has been repeatedly proposed as a powerful technique to develop indices of environmental quality. The approach adopted in this study provided a useful tool for the preliminary assessment of the potential vulnerability of benthic assemblages to scuba-diving and may represent an alternative method to the assessment of carrying capacity. The application of this index will enable management strategies for potentially reducing the degradation of benthic organisms/assemblages, and allowing a sustainable use of MPAs.     

Interaction of the nicotinic agonist (R,S)-3-pyridyl-1-methyl-2-(3-pyridyl)-azetidine (MPA) with nicotinic acetylcholine receptor subtypes expressed in cell lines and rat cortex

The interaction of the nicotinic agonist (R,S)-3-pyridyl-1-methyl-2-(3-pyridyl)-azetidine (MPA) with different nicotinic acetylcholine receptor (nAChR) subtypes was studied in cell lines and rat cortex. MPA showed an affinity (Ki = 1.21 nM) which was higher than anatoxin-a > (−)-nicotine > (+)-[R]nornicotine > (−)-[S]nornicotine > and (+)-nicotine, but lower than cytisine (Ki = 0.46 nM) in competing for (−)-[³H]nicotine binding in M10 cells, which stably express the recombinant ₄β₂ nAChR subtype. A one-binding site model was observed in all competing experiments between (−)-[³H]nicotine binding and each of the agonists studied in M10 cells. MPA showed a 13-fold higher affinity for (−)-[³H]nicotine binding sites compared to the [³H]epibatidine binding sites in rat cortical membranes. In human neuroblastoma SH-SY5Y cells, which predominantly express the ₃ nAChR subunit mRNA, MPA displaced [³H]epibatidine binding from a single population of the binding sites with an affinity in the same nM range as that observed MPA in displacing [³H]epibatidine binding in rat cortical membranes. Chronic treatment of M10 cells with MPA significantly up-regulated the number of (−)-[³H]nicotine binding sites in a concentration dependent manner. Thus MPA appears to have higher affinity to 4-subunit containing receptor subtype than ₃-subunit containing receptor subtype of nAChRs. Furthermore MPA binds to ₄β₂ receptor subtype with higher affinity than (−)-nicotine and behaves, opposite to cytisine, as a full agonist in up-regulating the number of nAChRs. © 1998 Elsevier Science Ltd. All rights reserved.     

Ecosystem response to different management options in Marine Protected Areas (MPA): A case study of intertidal rocky shore communities

Marine Protected Areas (MPA) can be powerful coastal management tools with several specific goals, although there is debate concerning their effectiveness. There is no consensus regarding the ideal size of MPAs, and actually there is some evidence that perhaps size is not as critical as other specific factors in determining their success in terms of populations’ protection and ecological functions conservation. On the other hand, depending on the objectives, zones with different classification regimes in terms of rules and uses might enable the maintenance of the intended uses.At this light, we examined the case of the small (605 002 m²) rocky shore area of Avencas, near Lisbon, on the Atlantic western Coast of Portugal, which was classified as Biophysical Interest Zone (ZIBA) in 1998, due to its exceptional intertidal biodiversity, after what its protection status became controversial, leading to conflicts with the local population and incompliance with extant regulations. From 2010 efforts were carried out by local authorities to reclassify Avencas as Marine Protected Area, which was achieved in 2016.Monitoring intertidal communities in a MPA and adjacent areas is an effective and low-cost procedure to evaluate the evolution of the biodiversity of rocky shores. Therefore, antedating the creation of the new MPA, assessments of the ZIBA biodiversity were conducted from January 2013 to December 2015 on a monthly basis. This timeline was selected as a function of a change in visitors’ behavior induced from 2013 by several management and outreach initiatives, which increased in a certain extent the user’s compliance with regulations.A positive evolution was expected for density and/or species diversity of the different groups analysed (flora, sessile fauna and mobile fauna) in this three years period. However, a very strong storm occurred in 2014 produced a significant impact and changed large areas of the Avencas rocky shore. As a consequence, results did not display a recognizable recovery pattern of the intertidal communities, and following that extreme event are not even consistent with a hypothesized enhanced recovery capability of the ecosystem in a protected area. This suggests that longer data series are necessary to obtain more robust data regarding natural variability, since alterations caused by extreme events are always likely to occur. Additionally, results illustrate that indeed size matters because it influences the MPA openness, expressed as the ratio of periphery to area, and therefore its susceptibility to external driving forces. Such considerations must be taken into account in any management plan, which in this case should encompass an increase in the intertidal protected area, a new conditioned small-scale fishing regime, and an adequate monitoring programme to evaluate the effectiveness of the new management scheme.     

Comparison of two synthetic progesterones on ventilation in normal males: CMA vs. MPA

We administered chlormadinone acetate (CMA), medroxyprogesterone acetate (MPA), and placebo to 16 normal male subjects using a randomized double-blind crossover study. After CMA administration, minute alveolar ventilation increased by +1.04 +/- 0.22 (SE) 1/min (P less than 0.05) accompanied by decrements of arterial PCO2 (-4.0 +/- 1.0 Torr) (P less than 0.01) and [HCO3-] (-2.1 +/- 0.05 mM/l) (P less than 0.01). On the other hand, in the MPA runs the corresponding changes of the above parameters were +0.71 +/- 0.21 l/min (P less than 0.05), -2.9 +/- 0.6 Torr (P less than 0.01), and -1.3 +/- 0.3 mM/l (P less than 0.01), respectively. The slopes of hypoxic ventilatory and occlusion pressure response lines remained unchanged in hypocapnia after CMA or MPA ingestion, but they increased when entidal PCO2 was adjusted to the predrug level. The hypercapnic ventilatory and occlusion pressure response lines merely shifted to the left without changing their slopes with these agents. No significant differences in all the above parameters were found between CMA and MPA runs. We concluded that in the normal males the effect of CMA on ventilation was similar to that of MPA, despite the fact that the luteinizing activity of CMA was reported to be approximately 10 times higher than the latter.     

Singing through winter nights: Seasonal and diel occurrence of humpback whale (Megaptera novaeangliae) calls in and around the Gully MPA,offshore eastern Canada

Humpback whale use of areas off eastern Canada is poorly understood, a knowledge gap that could impact future conservation efforts. We describe the acoustic occurrence of humpback whales in and around the Gully Marine Protected Area (MPA), an eastern Scotian Shelf submarine canyon. Near‐continuous acoustic recordings sampling at 16 kHz were collected from the MPA and nearby slope areas from October 2012 to September 2014 using near‐bottom recorders. In an offshore region where humpbacks were thought to be rare, we observed calls from October to June with a peak in song and nonsong calls in December and January. This suggests that some individuals occur in Canadian waters in winter and the Gully region may be a North Atlantic humpback whale migratory corridor. Calls were predominantly songs indicating potential mating activities. Song and nonsong calls occurred more at sunset and during hours of darkness than during daylight. This study improves our understanding of the seasonal occurrence of humpback whales on the Scotian Slope and, more specifically, their use of an offshore protected area.     

Excretion of MPA in the milk of lactating ewes treated for synchronization of estrus

Ten mature lactating ewes of the Chios island breed 3.5 +/- 0.5 (Mean +/- SEM) yr of age and weighing 51.9 +/- 1.6 kg (Mean +/- SEM) were synchronized for estrus with intravaginal sponges impregnated with 60 mg 6a-methyl-17-acetoxyprogesterone (MPA). The sponges remained in place for 14 d and 500 IU im PMSG were injected at their withdrawal. Daily milk samples (3 d pretreatment, 14 d on treatment, and 5 d posttreatment) were collected and analyzed by a double antibody RIA procedure for MPA. The concentration of MPA (Mean +/- SEM) in the milk increased to 5.05 +/- 0.11 ng/ml within the first day of sponge insertion, then declined and remained at a constant level (3.08 +/- 0.26 ng/ml) while the sponge was in place, eventually dropping to the background level (0.65 +/- 0.05 ng/ml) 24 h following sponge withdrawal. The curve for the quantity of MPA excreted in the milk was identical to that of MPA concentrations, showing significant differences among experimental days and among ewes. Finally, there was a significant relationship between milk production and MPA excretion into the milk (r = +0.581( * *)). It is concluded that only a very small percentage (0.08 +/- 0.01) of MPA contained in each sponge is excreted into the milk from the moment of sponge insertion until 5 d after its removal.