Endocrine Changes after Mating in Pregnant and Non-Pregnant Llamas and Alpacas

Plasma concentrations of oestradiol-17ß, progesterone, 15-keto–dihydro–PGF2α and luteinizing hormone (LH) were monitored in llamas and alpacas after mating with an intact male. Concentrations of LH and PGF2α metabolite were high immediately after copulation. Ovulation occurred in 92% of the animals. The first significant increases in progesterone were recorded on day 4 after mating. In non-pregnant animals the lifespan of the corpus luteum was estimated to be 8–9 days. Luteolysis occurred in association with the release of PGF2α. In pregnant animals, a transient decrease in progesterone concentrations was observed between days 8 and 18 in both species. No significant changes in PGF2α secretion were registered during this period. Oes– tradiol–17ß concentrations were high on the day of mating, declined to low values on day 4, and started to increase again on day 8. Peak values after luteolysis in non-pregnant animals were significantly higher than those registered in pregnant ones. Furthermore, concentrations of oestradiol-17ß were elevated for a longer period in non–pregnant than in pregnant animals. The results suggest that progesterone from the corpus luteum exerts a negative influence on follicular activity in pregnant animals by reducing oes– tradiol-17ß secretion.     

Dynamics of Eosinophils in the Uterus after Oestrogen Administration

To investigase the role of the eosinophil leukocytes in the early oestrogenic responses in the uterus, the kinetics of oestrogen-induced uterine eosinophilia and other parameters of oestrogen stimulation were studied at very early times. Uterine eosinophils increase as early as 5 min after an intravenous injection of oestradiol to immature rats, much earlier than several other changes in the early parameters of oestrogen stimulation. Large number of uterine eosinophils are found attached to the wall of small uterine blood vessels at early times. To elucidate the mechanisms involved in the specific attraction of eosinophils to the uterus in the presence of oestrogens, the in vivo localisation of oestrogens in the rat uterus at early times was studied using a radioautographic technique. Oestrogen receptors were found in the surface of eosinophils and in the wall of small uterine blood vessels. This simultaneous presence of both oestrogen receptors is proposed to explain the specific attachment of eosinophils to uterine blood vessels in the presence of oestrogens, which is the initial step toward eosinophil penetration into the uterus.     

Plasma Steroid and Luteinizing Hormone Levels during Prostaglandin F2α Administration in Luteal Phase of Menstrual Cycle

An intravenous infusion of prostaglandin F₂α (12.5-250μg/min) was administered in four volunteers in the mid-late luteal phase and three in the early luteal phase of the menstrual cycle.Frequent measurement of plasma progesterone, oestrogens, and luteinizing hormone (LH) showed that administration of high doses depressed plasma progesterone levels in the late luteal phase and caused concomitant side effects. Levels of progesterone in the early luteal phase were unaffected. In both phases oestrogen and LH levels were little altered. In two subjects, hourly progesterone levels measured throughout the day at a similar time in a subsequent control menstrual cycle showed an appreciable variation in one but steady levels in the second. This variation may contribute to the magnitude of the fall in progesterone noted during the infusion of prostaglandins.     

IVF-ET周期取卵后血清E2的变化模式及对预测妊娠的意义

目的:通过检测IVF-ET患者取卵后血清雌激素水平的变化模式,探讨其在预测妊娠中的意义。方法:纳入因榆卵管因素或男性因素行IVF-ET的患者62例(75个周期)。对行IVF-ET的患者,在取卵后隔日监测血清雌二醇(E_2)水平,并比较其在妊娠组与未孕组的差异。结果:取卵后,血E_2水平在妊娠组与未孕组均迅速降低,在取卵后2,4,6,8 d,两组间无统计学差异。在妊娠周期,血E_2平均水平在取卵后10d降至最低,之后逐渐上升。妊娠组与未孕组之间E_2水平的差异从取卵后10d开始可以检测出(分别为816.4±537.6pg/ml和189.5±69.3pg/ml)(P<0.05)。在未孕周期,10d的E_2水平(189.5±69.3pg/ml)显著低于8d(989.2±581.5pg/ml)(P<0.05)。结论:在取卵后8d和10d连续测2次血E_2水平,有助于早期发现妊娠:妊娠患者的E_2水平在10d出现上升预示妊娠,而10d出现剧陡降时,往往预示妊娠失败。     

GnRH-a对卵巢黄体期生殖激素水平及卵泡募集的影响

目的:探讨黄体期垂体生理性抑制的情况下,应用GnRH-a后对于垂体和卵巢激素分泌水平及卵泡发育过程的影响.方法:选择体外受精(IVF)或单精子卵胞浆内注射(ICSI)治疗的不孕妇女,给予常规长方案促排卵,并随机分为短效达菲林组(14人)及长效达菲林组(6人),两组分别于黄体中期开始给予每日短效达菲林0.05mg皮下注射或长效达菲林1/3支(1.25mg)单次注射处理.测定患者注射0、2、4日(对应于排卵后第7、9、11日)以及下一月经周期第2日外周血促卵泡生成素(FSH)、促黄体生成素(LH)、雌激素(E2)及孕酮(P)水平.结果:两组FSH,LH水平在用药后第2天均达到最高值,随后呈现逐渐下降趋势,不同时间点问差异有显著性(P＜0.01).两组E2水平在用药后呈现逐渐下降的趋势,但是不同时间点间差异无显著性.短效达菲林组P水平用药前后无明显变化,长效达菲林组P水平在用药后呈现逐渐下降的趋势,但是各时间点无显著性差异.结论:自然周期黄体期或服用短效避孕药周期应用GnRH-a是否能使FSH及LH升调节,从而使得卵泡过早募集.     

GnRH—a对卵巢黄体期生殖激素水平及卵泡募集的影响

目的：探讨黄体期垂体生理性抑制的情况下,应用GnRH—a后对于垂体和卵巢激素分泌水平及卵泡发育过程的影响。方法：选择体外受精（IVF）或单精子卵胞浆内注射（ICSI）治疗的不孕妇女,给予常规长方案促排卵,并随机分为短效达菲林组（14人）及长效达菲林组（6人）,两组分别于黄体中期开始给予每日短效达菲林0．05mg皮下注射或长效达菲林1／3支（1．25mg）单次注射处理。测定患者注射0、2、4日（对应于排卵后第7、9、11日）以及下一月经周期第2日外周血促卵泡生成素（FSH）、促黄体生成素（LH）、雌激素（E2）及孕re（P）水平。结果：两组FSH,LH水平在用药后第2天均达到最高值,随后呈现逐渐下降趋势,不同时间点间差异有显著性（P〈0．01）。两组E2水平在用药后呈现逐渐下降的趋势,但是不同时间点间差异无显著性。短效达菲林组P水平用药前后无明显变化,长效达菲林组P水平在用药后呈现逐渐下降的趋势,但是各时间点无显著性差异。结论：自然周期黄体期或服用短效避孕药周期应用GnRH-a是否能使FSH及LH升调节,从而使得卵泡过早募集。     

Morphological Changes within the Rat Lateral Ventricle after the Administration of Proteasome Inhibitors

The broad variety of substances that inhibit the action of the ubiquitin-proteasome system (UPS)—known as proteasome inhibitors—have been used extensively in previous studies, and they are currently frequently proposed as a novel form of cancer treatment and as a protective factor in intracerebral hemorrhage treatment. The experimental data on the safest route of proteasome inhibitor administration, their associated side effects, and the possible ways of minimizing these effects have recently become a very important topic. The aim of our present study was to determine the effects of administering of MG-132, lactacystin and epoxomicin, compounds belonging to three different classes of proteasome inhibitors, on the ependymal walls of the lateral ventricle. Observations were made 2 and 8 weeks after the intraventricular administration of the studied substances dissolved in dimethyl sulfoxide (DMSO) into the lateral ventricle of adult Wistar rats. Qualitative and quantitative analysis of brain sections stained with histochemical and inmmunofluorescence techniques showed that the administration of proteasome inhibitors caused a partial occlusion of the injected ventricle in all of the studied animals. The occlusion was due to ependymal cells damage and subsequent ependymal discontinuity, which caused direct contact between the striatum and the lateral nuclei of the septum, mononuclear cell infiltration and the formation of a glial scar between these structures (with the activation of astroglia, microglia and oligodendroglia). Morphologically, the ubiquitin-positive aggregates corresponded to aggresomes, indicating impaired activity of the UPS and the accumulation and aggregation of ubiquitinated proteins that coincided with the occurrence of glial scars. The most significant changes were observed in the wall covering the striatum in animals that were administered epoxomicin, and milder changes were observed in animals administered lactacystin and MG-132. Interestingly, DMSO administration also caused damage to some of the ependymal cells, but the aggresome-like structures were not formed. Our results indicate that all of the studied classes of proteasome inhibitors are detrimental to ependymal cells to some extent, and may cause severe changes in the ventricular system. The safety implications of their usage in therapeutic strategies to attenuate intracerebral hemorrhagic injury and in brain cancer treatment will require further studies.     

Subcutaneous Progesterone Is Effective and Safe for Luteal Phase Support in IVF: An Individual Patient Data Meta-Analysis of the Phase III Trials

Objective

To summarize efficacy and safety data on a new progesterone compound which is available for subcutaneous administration as compared to vaginally administered progesterone for luteal phase support in patients undergoing IVF treatment.

Design

Data from two randomized phase III trials (07EU/Prg06 and 07USA/Prg05) performed according to GCP standards with a total sample size of 1435 per-protocol patients were meta-analyzed on an individual patient data level.

Setting

University affiliated reproductive medicine unit.

Patients

Subcutaneous progesterone was administered to a total of 714 subjects and vaginal progesterone was administered to a total of 721 subjects who underwent fresh embryo transfer after ovarian stimulation followed by IVF or ICSI. The subjects were between 18 and 42 years old and had a BMI <30kg/m².

Interventions

Subcutaneous progesterone 25 mg daily vs. either progesterone vaginal gel 90 mg daily (07EU/Prg06) or 100 mg intravaginal twice a day (07USA/Prg05) for luteal phase support in IVF patients.

Main outcome measures

Ongoing pregnancy rate beyond 10 gestational weeks, live birth rate and OHSS risk.

Results

The administration of subcutaneous progesterone versus intra-vaginal progesterone had no impact on ongoing pregnancy likelihood (OR = 0.865, 95% CI 0.694 to 1.077; P = n.s.), live birth likelihood (OR = 0.889, 95% CI 0.714 to 1.106; P = n.s.) or OHSS risk (OR = 0.995, 95% CI 0.565 to 1.754; P = n.s.) in regression analyses accounting for clustering of patients within trials, while adjusting for important confounders. Only female age and number of oocytes retrieved were significant predictors of live birth likelihood and OHSS risk.

Conclusion

No statistical significant or clinical significant differences exist between subcutaneous and vaginal progesterone for luteal phase support.     

Distribution of Methylene Blue after Injection into the Epidural Space of Anaesthetized Pregnant and Non-Pregnant Sheep

The aim of the study was to determine the distribution of different volumes of methylene blue solution injected into the epidural space in anaesthetized pregnant and non-pregnant sheep, to evaluate its cranial distribution and to compare between them. Fifteen pregnant and fifteen non-pregnant sheep were included in the study. Sheep were anaesthetized and received 0.05, 0.1, or 0.2 mL/kg of a lumbosacral epidural solution containing 0.12% methylene blue in 0.9% saline. Thirty minutes after the epidural injection, the ewes were euthanized. The extension of the dye within the epidural space was measured, and the correlation between the volume of the dye injected and the number of stained vertebrae was evaluated. The cranial migration of the dye between pregnant and non-pregnant sheep was also compared. The results show that the volume of methylene blue injected epidurally into pregnant and non-pregnant sheep correlated directly with its cephalic distribution into the epidural space; and a volume of 0.1 mL/kg or 0.2 mL/kg stained up to the first lumbar segment in pregnant and non-pregnant sheep, respectively. Also, the results suggest that the volume of drugs administered into the epidural space of pregnant sheep should be half the volume that would be used in non-pregnant sheep.     

Arginase,Arginine Decarboxylase,Ornithine Decarboxylase,and Polyamines in Tomato Ovaries (Changes in Unpollinated Ovaries and Parthenocarpic Fruits Induced by Auxin or Gibberellin)

Arginase (EC 3.5.3.1) activity has been found in the ovaries and Young fruits of tomato (Lycopersicon esculentum Mill. cv Rutgers).Changes in arginase, arginine decarboxylase (EC 4.1.1.19), and ornithine decarboxylase activity (EC 4.1.1.17) and levels of free and conjugated putrescine, spermidine, and spermine were determined in unpollinated ovaries and in parthenocarpic fruits during the early stages of development induced by 2,4-dichlorophenoxyacetic acid (2,4-D) or gibberellic acid (GA3). Levels of arginase, free spermine, and conjugates of the three polyamines were constant in unpollinated ovaries and characteristic of a presenescent step. A marked decrease in arginase activity, free spermine, and polyamine conjugates was associated with the initiation of fruit growth due to cell division, and when cell expansion was initiated, the absence of arginase indicated a redirection of nitrogen metabolism to the synthesis of arginine. A transient increase in arginine decarboxylase and ornithine decarboxylase was also observed in 2,4-D-induced fruits. In general, 2,4-D treatments produced faster changes than GA3, and without treatment, unpollinated ovaries developed only slightly and senescence was hardly visible. Sensitivity to 2,4-D and GA3 treatment remained for at least 2 weeks postanthesis.     

Changes in Bioenergetic Functions and Performance Capacity of Athletes after Administration of a Polyphenolic Antihypoxic Agent

A study was made of the effects of a polyphenolic antihypoxic agent on respiration in isolated mitochondria and homogenates of skeletal muscles and on the bioenergetic functions and performance capacity of runners. Polarographic analysis showed that preincubation of mitochondria isolated from skeletal muscles with the antihypoxic agent inhibited glutamate oxidation more than twofold in state 4. In homogenates of skeletal muscles, the antihypoxic agent in the presence or absence of an uncoupler strongly inhibited or slightly stimulated glutamate oxidation in the initial mitochondrial state, respectively. Thus, the antihypoxic agent inhibited mitochondrial ATPase but did not change the ATP synthase activity or exert a direct effect on the respiratory chain. The antihypoxic agent administered to runners at a concentration of about 30 mg/kg body mass 1 h prior to exercise at maximum power with a maximum duration of 1–10 min allowed higher acidification but did not affect the major bioenergetic functions such as O₂ consumption, excess CO₂ production, O₂ debt, and O₂ demand. The agent substantially increased the performance capacity of runners during exercise with a maximum duration of 3–10 min. Under competition conditions, administration of the agent to runners improved the results by 2–6% in 200- to 10000-m races.     

Changes in the Level of Peptidase Activities in Pea Ovaries during Senescence and Fruit Set Induced by Gibberellic Acid

The activities and changes in the levels of exopeptidase and endopeptidase activities were characterized in unpollinated ovaries of Pisum sativum L. cv Alaska during senescence and early fruit development induced by gibberellic acid (GA₃). Two aminopeptidases and one iminopeptidase were electrophoretically separated. These peptidases were sensitive to inhibitors of sulfhydryl proteases. Carboxypeptidase activity was inhibited by phenylmethyl sulfonyl fluoride. An azocasein-degrading endopeptidase, sensitive to thiol protease inhibitors, was also found. An increase in the specific activity of aminopeptidase during both fruit development and ovary senescence was observed. In contrast, the specific activity of carboxypeptidase and endopeptidase increased only during senescence of the ovary. Changes in exopeptidase activity in senescing ovaries could be mainly the consequence of a greater stability to proteolysis while the rise in endopeptidase activity appeared to be due to new or increased synthesis of the enzyme. These results suggest that endopeptidase, and not amino or carboxypeptidase, plays a key role in the senescence of pea ovaries and that the changes in unpollinated ovaries leading to ovary senescence or fruit development can be controlled by gibberellins.     

桃树子房发育初期山梨醇含量及其相关酶活性的变化

测定桃品种‘京玉’和‘久保’子房发育初期的子房、韧皮部、叶和花瓣中山梨醇、果糖、葡萄糖含量以及与山梨醇代谢相关酶——山梨醇脱氢酶(SDH)和山梨醇氧化酶(SOX)活性变化的结果表明:2个品种各部位的糖积累规律较为相似,子房发育初期叶中山梨醇含量较高,其他部位则较低且变化不大;各部位的果糖含量均略有升高,葡萄糖含量则有较大起伏。子房、韧皮部和叶中SDH和SOX活性变化趋势相同,SDH活性高于SOX。     

Aerobic Bacteria Occurring in the Vagina of Bitches with Reproductive Disorders

A retrospective survey was performed of aerobic bacterial species found in the vagina of 203 bitches with genital disorders, e.g. infertility, vaginitis, pyometra and puppy death. Escherichia coli, beta-hemolytic streptococci, Staphylococcus intermedius and Pasteurella multocida were the species most often isolated. From bitches with pyometra E. coli in pure culture was the most frequent isolate. In contrast, the majority of infertile bitches gave rise to mixed cultures, and no specific bacterial species was consistently associated with infertility. Thus, bacterial sampling from infertile bitches was concluded to be of low diagnostic value. Bacterial species isolated from the bitches having vaginitis were present in pure culture in 26.9% of the samples while nonspecific mixed cultures were obtained from 34.6% of the samples from these bitches. E. coli was the most frequently isolated bacterial species from bitches with dead puppies. However, in such cases it is important to relate the vaginal bacterial findings to autopsy findings and the results of bacteriological cultures of the pups.     

Proteins Deiminated by Peptidylarginine Deiminase in Mouse Uterus and Their Changes during the Estrous Cycle

Peptidylarginine deiminase (PAD) catalyzes the formation of citrullyl residues by deimination of arginyl residues of proteins in the presence of Ca²⁺. We found several deiminated proteins in mouse uterus using antibodies recognizing the citrulline residue. The levels of these proteins changed during the estrous cycle in parallel with PAD activity.     

排卵期生殖道局部细胞免疫与黄体功能不全患者不孕的关系研究

目的:探讨排卵期生殖道局部细胞免疫与黄体功能不全(LPD)患者不孕之间的关系,旨在为临床防治不孕不育提供依据及参考。方法:选择2014年10月~2015年10月本院确诊的LPD患者50例为观察组,同期筛选同期50例健康体检者为对照组,排卵后第4 d、6 d、8 d采用酶联免疫法检测两组排卵期血清孕酮(P)水平,采用放射免疫法(RIA)检测两组排卵期血清以及宫颈黏液中的肿瘤坏死因子-α(TNF-α)与白细胞介素1-β(IL-1β)的表达水平,对以上检测指标予以比较分析。结果:排卵后第4d、6d、8d观察组血清P明显低于对照组,比较差异均有统计学意义(P0.05)。排卵期观察组血清TNF-α及IL-1β水平与对照组比较,差异不具有统计学意义(P0.05),但观察组宫颈黏液中TNF-α及IL-1β水平显著高于对照组,差异有统计学意义(P0.05)。排卵期观察组宫颈黏液中TNF-α、IL-1β水平与血清P存在负相关关系(r=-0.879,-0.886,P0.05),且宫颈黏液中TNF-α、IL-1β二者存在显著正相关关系(r=0.768,P0.05)。结论:排卵期生殖道局部细胞免疫激活,宫颈黏液中TNF-α及IL-1β水平升高,对黄体发育产生影响,并导致LPD的发生。     

Changes in Activity of Choline Acetylase in Central Nervous System of Rat after Intraventricular Administration of Noradrenaline

APPLICATION of strychnine or d-tubocurarine to the exposed cerebral cortex leads after a few minutes to abnormality in somatosensory evoked potentials^1,2. This consists of a surface negative wave (peak latency, 21–23 ms) which attains an amplitude five to twenty times greater than that of the normal evoked potentials and probably reflects excessive depolarization of the apical dendrites of pyramidal neurones. We wish to report the blocking of this effect by prior application of eserine to the cortex.     

The Expression Pattern of microRNAs in Granulosa Cells of Subordinate and Dominant Follicles during the Early Luteal Phase of the Bovine Estrous Cycle

This study aimed to investigate the miRNA expression patterns in granulosa cells of subordinate (SF) and dominant follicle (DF) during the early luteal phase of the bovine estrous cycle. For this, miRNA enriched total RNA isolated from granulosa cells of SF and DF obtained from heifers slaughtered at day 3 and day 7 of the estrous cycle was used for miRNAs deep sequencing. The results revealed that including 17 candidate novel miRNAs, several known miRNAs (n = 291–318) were detected in SF and DF at days 3 and 7 of the estrous cycle of which 244 miRNAs were common to all follicle groups. The let-7 families, bta-miR-10b, bta-miR-26a, bta-miR-99b and bta-miR-27b were among abundantly expressed miRNAs in both SF and DF at both days of the estrous cycle. Further analysis revealed that the expression patterns of 16 miRNAs including bta-miR-449a, bta-miR-449c and bta-miR-222 were differentially expressed between the granulosa cells of SF and DF at day 3 of the estrous cycle. However, at day 7 of the estrous cycle, 108 miRNAs including bta-miR-409a, bta-miR-383 and bta-miR-184 were differentially expressed between the two groups of granulosa cell revealing the presence of distinct miRNA expression profile changes between the two follicular stages at day 7 than day 3 of the estrous cycle. In addition, unlike the SF, marked temporal miRNA expression dynamics was observed in DF groups between day 3 and 7 of the estrous cycle. Target gene prediction and pathway analysis revealed that major signaling associated with follicular development including Wnt signaling, TGF-beta signaling, oocyte meiosis and GnRH signaling were affected by differentially expressed miRNAs. Thus, this study highlights the miRNA expression patterns of granulosa cells in subordinate and dominant follicles that could be associated with follicular recruitment, selection and dominance during the early luteal phase of the bovine estrous cycle.