Evaluation of a new biosensor-based mushroom (Agaricus bisporus) tissue homogenate: investigation of certain phenolic compounds and some inhibitor effects

A biosensor based on mushroom tissue homogenate for detecting some phenolic compounds (PCs) and usage of the biosensor for quantifying certain substances that inhibit the polyphenol oxidase activity in mushroom (Agaricus bisporus) tissue homogenate is described. The mushroom tissue homogenate was immobilized to the top of a Clark-type oxygen electrode with gelatin and glutaraldehyde. Optimization of the experimental parameters was done by buffer system, pH, buffer concentration, and temperature. Besides, the detection range of eight phenolic compounds were obtained with the help of the calibration graphs. Thermal stability, storage stability, and repeatability of the biosensor were also investigated. A linear response was observed from 20 x 10(-3) to 200 x 10(-3) mM phenol. The biosensor retained approximately 74% of its original activity after 25 days of storage at 4 degrees C. In repeatability studies, variation coefficient (C.V.) and standard deviation (S.D.) were calculated as 2.44% and +/-0.002, respectively. Inhibition studies revealed that the proposed biosensor was applicable for monitoring benzoic acid and thiourea in soft drinks and fruit juices.     

Thermal requirement for development of Sancassania rodionovi (Acari: Acaridae) on mushrooms

The free-living mite species Sancassania rodionovi (Zachvatkin) (Acari: Acaridae), is a serious pest of mushrooms in Iran. Studies were conducted to examine the development of this mite in relation to temperature on two mushroom species: Agaricus bisporus Lange (button mushroom) and Pleurotus ostreatus Kummer (oyster mushroom). The developmental time of this acarid mite was studied at eight constant temperatures, ranging from 5 to 40 degrees C, and developmental rates were modeled as a function of temperature. Sancassania rodionovi completed immature development in 17.35 +/- 0.58 and 20.17 +/- 0.88 d at 25 degrees C on button and oyster mushrooms, respectively. When the mite fed on button mushroom, the rate of development increased gradually from 10 to 35 degrees C. Using a linear model, the developmental zero was estimated to be 3.50 degrees C with a thermal constant of 357.14 degree-days. The Logan 10, Briere 1, and Thermodynamic models adequately described the data for this mite and yielded R2 values >0.95; these models provided estimates of optimum temperature for development of 33.244, 32.145, and 32.148 degrees C, respectively. Understanding the influence of temperature on development of S. rodionovi is discussed with respect to pest management in mushroom production.     

Inhibitory effects of cis- and trans-isomers of 3,5-dihydroxystilbene on the activity of mushroom tyrosinase

The effects of cis- and trans-isomers of 3,5-dihydroxystilbene on the activity of mushroom tyrosinase have been studied. The results show that both cis- and trans-isomers of 3,5-dihydroxystilbene can inhibit the diphenolase activity of the enzyme and the inhibition type was reversible. The IC(50) values were estimated as 0.405+/-0.013 and 0.705+/-0.017 mM, respectively. Kinetic analysis showed that the inhibition of cis-3,5-dihydroxystilbene and trans-3,5-dihydroxystilbene on the diphenolase activity of the enzyme belonged to competitive type, and the inhibition constants (K(I)) were determined to be 0.232+/-0.015 and 0.395+/-0.020 mM, respectively. In this investigation, the inhibitory effects of cis-3,5-dihydroxystilbene and trans-3,5-dihydroxystilbene on the diphenolase activity of mushroom tyrosinase were compared. The inhibitory capacity of cis-isomer was stronger than that of corresponding trans-isomer. Nevertheless, the trans-3,5-dihydroxystilbene was used more frequently than its corresponding cis-form compound. This research may offer some references for designing and synthesizing some novel and effective tyrosinase inhibitors. Furthermore, it may improve the use of stilbenes on the field of food preservation and depigmentation.     

Influence of stage of oestrous cycle on time of oestrus following cloprostenol treatment in the bovine

The first of 2 injections of 0.5 mg cloprostenol (PG1 and PG2) eleven days apart was given to 19 Friesian-Hereford cross heifers between days 8-14 of their cycle (Treatment A) and 16 similar animals between days 0-4 (Treatment B). Oestrus show was monitored by Kamar Heat Mount detectors and vasectomised bulls with chin-ball markers. Blood samples taken at PG1, six days later, at PG2 and four days later were assayed for progesterone to confirm that luteolysis had occurred as expected. Four hourly rectal examinations of the ovaries were carried out from 56-112 hours after PG2 and four hourly blood samples from 36-96 hours after PG2 were collected for FSH and LH assay. Mean time in hours from PG2 to oestrus onset, LH peak and ovulation respectively was 57.4 +/- 2.9, 60.2 +/- 2.0, 91.7 +/- 1.8 for Treatment A and 64.9 +/- 4.1, 68.9 +/- 2.4, 96.7 +/- 1.3 for Treatment B. Treatment A animals showed significantly higher (p<0.01) FSH levels at PG2 than Treatment B. Time from PG2 to LH peak was significantly shorter in animals treated either on days 7 and 8 (p<0.01) or days 15-16 (p<0.05) of their cycle compared with treatment on days 12-14 and it is suggested that these shorter response times correspond to an early and late cycle wave of follicular growth. Secondary FSH peaks some 28 hours after that occurring synchronously with the pre-ovulatory LH peak were observed to be significantly (p<0.01) higher at oestrus associated with the early cycle follicular growth wave as compared with that later in the cycle which may argue a difference in endocrine control of the two periods of follicular maturation.     

Hindlimb unloading induces a collagen isoform shift in the soleus muscle of the rat

To determine whether hindlimb unloading (HU) alters the extracellular matrix of skeletal muscle, male Sprague-Dawley rats were subjected to 0 (n = 11), 1 (n = 11), 14 (n = 13), or 28 (n = 11) days of unloading. Remodeling of the soleus and plantaris muscles was examined biochemically for collagen abundance via measurement of hydroxyproline, and the percentage of cross-sectional area of collagen was determined histologically with picrosirius red staining. Total hydroxyproline content in the soleus and plantaris muscles was unaltered by HU at any time point. However, the relative proportions of type I collagen in the soleus muscle decreased relative to control (Con) with 14 and 28 days HU (Con 68 +/- 5%; 14 days HU 53 +/- 4%; 28 days HU 53 +/- 7%). Correspondingly, type III collagen increased in soleus muscle with 14 and 28 days HU (Con 32 +/- 5%; 14 days HU 47 +/- 4%; 28 days HU 48 +/- 7%). The proportion of type I muscle fibers in soleus muscle was diminished with HU (Con 96 +/- 2%; 14 days HU 86 +/- 1%; 28 days HU 83 +/- 1%), and the proportion of hybrid type I/IIB fibers increased (Con 0%; 14 days HU 8 +/- 2%; 28 days HU 14 +/- 2%). HU had no effect on the proportion of type I and III collagen or muscle fiber composition in plantaris muscle. The data demonstrate that HU induces a shift in the relative proportion of collagen isoform (type I to III) in the antigravity soleus muscle, which occurs concomitantly with a slow-to-fast myofiber transformation.     

Oestrogen and seasonal effects on the production of an oestrus-associated glycoprotein in oviducal fluid of sheep

In 14 cyclic ewes, the oestrus-associated glycoprotein in the oviducal fluid was never detected between Days 7 and -2 of the oestrous cycle and it was present in 5% of fluid samples collected on Day -1, 59% on Day 0, 96% on Day 1, 100% on Day 2, 79% on Day 3, 31% on Day 4, 16% on Day 5, and 4% on Day 6. Its presence generally coincided with the period of high flow rate of oviducal fluid which occurs around oestrus. The duration of detectable levels of the oestrus-associated glycoprotein did not vary significantly during the breeding season from a mean (+/- s.d.) of 3.9 +/- 1.0 days. However, the peak flow rate of oviducal fluid dropped from 1.63 +/- 0.50 (early) to 1.38 +/- 0.40 (mid-) and to 0.85 +/- 0.21 ml/day late in the season. Anoestrous ewes (3) induced to ovulate by treatment with progesterone implants and gonadotrophin showed low peak fluid flow rates (0.92 +/- 0.30 ml/day) and the presence of the oestrus-associated glycoprotein for a shorter period (2.7 +/- 0.7 days). Pregnancy (N = 3) did not appear to prolong the production of the protein. The injection of 25 micrograms oestradiol benzoate into 3 anoestrous, 2 mid-cycle and 9 ovariectomized ewes caused an increase in fluid flow rate and appearance of the glycoprotein 1-2 days later. The glycoprotein was present for a longer period in response to the exogenous oestrogen--6.8 +/- 1.6 days in the ovariectomized ewes, 7.9 +/- 1.3 days in anoestrous ewes, and 8.4 +/- 0.8 days in the dioestrous ewes.(ABSTRACT TRUNCATED AT 250 WORDS)     

Photoperiodic alteration of postpartum reproductive function in suckled cows

Three experiments were conducted using a total of 41 cows to determine if photoperiod modulates the establishment of postpartum estrous cycles and conception. Cows calving in the autumn and winter were exposed to either 18 hr light/day (18L:6D) or natural photoperiods. In Exp. 1, cows receiving 18L:6D had shorter (P<0.025) intervals from calving to estrus (61 +/- 3.8 days) than cows not receiving supplemental light (154 +/- 23.9 days). The same was true for primiparous cows in Exp. 2 (76 +/- 5.5 days vs 153 +/- 38.0 days; P<0.06) but not for the multiparous cows in Exp. 2 (56 +/- 5.2 days vs 40 +/- 7.4 days) or for all cows in Exp. 3 (60 +/- 10.1 days vs 70 +/- 13.5 days). Because conception rate was higher for cows exposed to 18L:6D for the multiparous cows in Exp. 2 and all cows in Exp. 3, interval to conception was significantly shorter for animals exposed to 18L:6D in all experiments. Generally, interval from calving to uterine involution was also reduced by exposure to 18L:6D. No effects of photoperiod were observed on body weight changes, serum levels of luteinizing hormone (LH), follicle stimulating hormone (FSH) or estradiol or on estradiol-induced release of gonadotropins. It was concluded that in certain situations day length can hasten estrus and conception in postpartum cows. The effect of photoperiod interacts with other conditions, one of which is parity. The endocrine basis for these effects are not known.     

Reproductive and developmental biology of the oriental cockroach Blatta orientalis (Dictyoptera)

At 27 degrees C and 45% r.h. in the laboratory, the oriental cockroach Blatta orientalis (L.) developed to adulthood in seven to nine instars for males (66% had eight instars) and eight to ten instars for females (67% had nine instars) in mixed groups, with up to twelve instars for isolated females. Nymphal development lasted 185 +/- 2 days for males, 216 +/- 4 days for females, with 89% survival to adulthood. Adult longevity was significantly more for males than females in mixed groups. Virgin females lived for 135 +/- 6 days compared with 87 +/- 9 days for females kept with males. After an initial maturation time of 12.2-13.5 +/- 0.4 days for mated and unmated females, oothecae were produced, on average, every 6-7 (range 2-29) days. Ootheca viability was 68% from females kept with males, 32% from females kept apart from males. Numbers of nymphs emerging were 14.1 +/- 0.26 after 45 days from mated female oothecae 8.2 +/- 0.3 after 49 days from unmated females. With sexual reproduction the sex ratio of progeny reaching adulthood was 1.1 males per female (n = 443), whereas unmated females produced only female progency, which is consistent with parthenogenetic reproduction. Drawings of the ventral aspect of the terminalia are given to show features useful for instar determination and for distinguishing between male and female nymphs and adults of B. orientalis.     

Effect of grafted PEG on liposome size and on compressibility and packing of lipid bilayer

The aim of this study was to elucidate the effect of various mole percentages (0-25 mol%) of 2000 Da polyethylene glycol-disteroylphosphoethanolamine (PEG-DSPE) in the presence or absence of 40 mol% cholesterol and the effect of degree of saturation of phosphatidylcholine (PC) on the size and the lipid bilayer packing of large unilamellar vesicles (LUV). Egg PC (EPC, unsaturated) LUV and fully hydrogenated soy PC (HSPC, saturated) LUV partial specific volume, specific compressibility, size, and packing parameter (PP) of lipids were characterized by measurements of density, ultrasonic velocity, specific turbidity, and dynamic light scattering. Liposome size and specific turbidity decreased with increase in temperature and PEG-DSPE mol%, except at 7+/-2 mol%. At this PEG-DSPE mol%, an anomalous peak in liposome size of 15+/-5 nm was observed. We attribute this effect mainly to the change in the spatial structure of the PEG-DSPE molecule, depending on whether the grafted PEG is in the mushroom or brush configuration. In the mushroom regime, i.e., when the grafted PEG is up to 4 mol% in LUV, the PEG moiety did not affect the additive PP of the lipids in the bilayer, and the PP value of PEG-DSPE is 1.044; while in the brush regime, i.e., when the grafted PEG is higher than 4 mol%, the PP of PEG-DSPE decreases exponentially, reaching the value of 0.487 at 30 mol% of grafted lipopolymer. The specific compressibility and additive PP values for the mixture of matrix lipid (EPC or HSPC), cholesterol, and PEG-DSPE for all liposome compositions investigated reached their maximum at 7+/-2 mol% PEG-DSPE, the concentration of PEG-DSPE at which the highest biological stability of the LUV is achieved.     

Evaluation of beta-D-glucan density in blood after drinking an extraction element of Agaricus blazei murill]

beta-D-Glucan (beta-G) measurement in blood is useful for prompt diagnosis, selection of treatment and therapeutic evaluation for deep fungal infection. A patient with high beta-G blood was not improved by strong anti-fungal treatment in our hospital, and the beta-G levels in the blood fell immediately after discontinuing ingestion of the agaricus mushroom extracted element (sennseiro). To verify the elevation of beta-G concentration in the blood after intake of an agaricus mushroom extraction element, beta-G concentration in the blood was measured 4 days after 9 volunteers drank 2 g of agaricus mushroom extraction element, "SSG plus 35" twice a day for 3 days. A low value of beta-G in blood was detected in one person (11.1%), which demonstrated that beta-G concentration in the blood increased by oral ingestion of the agaricus element although the reason was unclear why it was detected in only one person. Taken together, when high beta-D-glucan is identified by uncertain cause or in spite of enough anti-fungal medication, it is necessary to confirm whether a patient has ingested health food containing an agaricus mushroom element or other fungus elements to avoid needless treatment with anti-fungal medicine.     

Effect of recombinant alpha interferons on fertility and interestrous interval in sheep

In Experiment 1, 12 unmated cyclic ewes received twice-daily intrauterine injections on Days 12 to 14 of one of the following treatments: 1) ovine conceptus secretory proteins (oCSP) containing 25 mug of ovine trophoblast protein-1 (oTP-1) as determined by RIA; 2) 25 or 50 mug recombinant human interferon alpha1 (rhlFN); or 3) 1500 ug of serum proteins (oSP) from a Day-16 pregnant ewe (estrus = Day 0) per uterine horn. Ewes receiving oCSP had longer interestrous intervals (27 +/- 2 days; P<0.05) than ewes receiving oSP (17 +/- 2 days). Ewes receiving either dose of rhlFN had an interestrous interval of 16 +/- 2 days which did not differ (P>0.10) from that of oSP-treated ewes. In Experiment 2, 59 normally cycling ewes, mated on Day 0, received twice-daily intramuscular injections of either 2 mg recombinant bovine interferon alpha1 (rblFN) or placebo on Days 12 to 15 post estrus. On Day 16, pregnancy was confirmed by flushing a morphologically normal conceptus from the uterus. Pregnancy rates for rblFN-treated (80%) and placebo-treated (62%) ewes were not different (P>0.10). Uterine flushings and conceptus-conditioned medium were assayed for oTP-1. Total oTP-1 in conceptus-conditioned culture medium was higher (P<0.02) when conceptuses were from placebo-treated (104 +/- 14 mug/conceptus) than from rblFN-treated (56 +/- 12 mug/conceptus) ewes; while total oTP-1 in uterine flushings was similar (P>0.10) for placebo-treated (132 +/- 15 mug/conceptus) and rblFN-treated (147 +/- 17 mug/conceptus) ewes. The interval from mating to subsequent estrus following conceptus removal was 31 +/- 1 and 28 +/- 1 days for pregnant ewes treated with rblFN and placebo, respectively. Interestrous intervals for nonpregnant ewes were longer (P<0.02) for rblFN-treated (27 +/- 3 days) than for placebo-treated (18 +/- 2 days) ewes.     

河西走廊绿洲灌区循环模式"农田-食用菌"生产系统氮素流动特征

河西走廊绿洲灌区在农业产业化进程加快的同时,也带来了水资源和生态环境的压力.本文利用养分流动和模型分析的方法,分析河西走廊绿洲灌区典型“农田-食用菌”生产系统(农田-食用菌集约生产模式、农田-食用菌单户生产模式、农田单作)的氮元素流动特征.结果表明:农田作物秸秆通过食用菌体系还田使氮素利用率提高了10％左右,秸秆还田氮输入量(165.6kg· hm-2·a-1)占农田氮素总输入量的37.1％,使化肥氮输入量减少,因此秸秆氮的合理循环利用可作为减少化肥投入的有效途径.但农田氮素仍有盈余,单位面积氮盈余量高达217.0 kg·hm-2· a-1,未能实现循环模式内养分平衡的理想效果,因此优化氮素管理、确定合理的大田作物和食用菌面积、调整农业产业结构是解决该问题的关键.     

T4-induced cardiac hypertrophy and the perfused and total microvasculature of the heart

The purpose of the present investigation was to determine the effects of thyroxine (T4), which induces myocardial hypertrophy, on the number per square millimetre and volume per cubic millimetre of both the total and perfused portions of the arteriolar and capillary beds of the heart. Studies were conducted in the subendocardial and subepicardial regions of the left ventricle of anesthetized open-chest rabbits. Fluorescein isothiocyanate-dextran (i.v.) or radioactive microspheres (intra-atrial) were injected to label the perfused microvessels or to determine coronary flow in three groups of rabbits: controls, and rabbits given 0.5 mg/kg T4 for 3 days and for 16 days. Fluorescent photography was used to identify the perfused microvessels. An alkaline phosphatase stain was employed to locate the total microvascular bed. There were 2369 +/- 638 (SD) capillaries/mm2 and 4 +/- 3 arterioles/mm2 in control hearts. These decreased significantly to 1380 +/- 199/mm2 and 1 +/- 1/mm2, respectively, after 16 days of T4. In controls, 60 +/- 5% of the capillaries and 59 +/- 21% of the arterioles were perfused. This increased significantly to 90 +/- 5 and 86 +/- 18%, respectively, by 16 days of T4 treatment. Similar changes, although smaller, were observed after 3 days of T4. Coronary blood flow increased to 1.7 times control after 3 days and 2.9 times after 16 days of T4. No significant subepicardial versus subendocardial differences were observed in any condition or measurement. Thus, the physiological response to the increased work and increase in anatomic minimum diffusion distance is to increase flow and the proportion of the capillary bed perfused to at least maintain physiological diffusion distances.     

Resumption of oestrous behaviour and cyclic ovarian activity in suckling and non-suckling ewes

In Préalpes de Sud ewes after an autumn lambing, the mean post-partum interval to first LH surge was 10 +/- 1 days and 17 +/- 1 days for non-suckling and suckling ewes, respectively. Post-partum interval to first luteal phase, estimated from plasma progesterone concentrations, was similar in non-suckling and suckling ewes (27 +/- 1 days and 28 +/- 5 days, respectively). Interval to first oestrus was shorter in non-suckling (22 +/- 2 days) than in suckling ewes (35 +/- 2 days) but these first oestrous periods were followed by short luteal phases in 60% (12/20) of non-suckling ewes and in only 7% (2/29) of suckling ewes. Finally, suckling slightly postponed the resumption of the first oestrus followed by a normal oestrous cycle (37 +/- 1 days versus 31 +/- 2 days) because progesterone, essential for oestrus expression, was secreted mainly during normal luteal phases in 70% (21/30) of suckling ewes and during short cycles in 95% (21/22) of non-suckling ewes. Therefore, the primary consequence of suckling is to regulate the conditions of resumption of cyclic ovarian activity after parturition.     

Changes in composition of harvested mushrooms (Agaricus bisporus)

Post-harvest changes in the biochemical composition of the mushroom were studied. Non-structural polysaccharide was found at levels greater than 10% dry wt in the fresh mushroom. After 4 days storage, the level had decreased to below 5% dry weight. The polysaccharide appeared to contain only glucose residues joined by α-1,4 and α-1,6 linkages. The chitin content of cell walls increased by ca 50% during 4 days storage, while cell wall glucan decreased. There was a large increase in urea content.     

Effect of repeated laparoscopic surgery on the bovine estrous cycle

The effects of repeated laparoscopic surgery on the length of the bovine estrous cycle, estrus, ovulation and corpus luteum function were determined after one estrous cycle of normal duration (18 to 24 days). Five, Angus x Hereford cows were subjected to laparoscopy on days 5, 13, 18 and 20 (estrus = day 0) of the subsequent cycle. Blood was collected daily during the cycle in which laparoscopy was performed (surgical cycle) and during the next cycle (postsurgical cycle). Lengths of the surgical and postsurgical cycles (22.3 +/- .5 days and 21.5 +/- .6 days, respectively) did not differ (P>.05) from that of the presurgical cycle (21.8 +/- .2 days). Average concentrations (ng/ml) of LH and progesterone in serum were similar during the surgical and postsurgical cycles (1.2 +/- .1, 2.2 +/- .2 vs 1.3 +/- .2 and 2.3 +/- .1). Progesterone concentrations remained above 1 ng/ml for 17 and 16 days during the surgical and postsurgical cycles, respectively. A pre-ovulatory rise in LH, along with estrus and ovulation was confirmed in all animals. Follicular development, characterized by follicular volume, increased progressively from days 5 to 20, with the largest increase occurring between days 13 and 18. These results indicate that laparoscopy, used at the times and frequency specified, does not alter reproductive function of cyclic cows and can provide information on ovarian activity.     

The effect of prolonged hypobaric hypoxia on growth of fetal sheep

The effect of prolonged hypobaric hypoxia on fetal sheep was studied. Pregnant ewes were subjected to an atmospheric pressure of 429 torr from 30 days to 135 days gestation (long-term study). Average fetal weight for the hypoxaemic group (3.35 +/- 0.53 kg; n = 4; mean +/- SD) was significantly lower than for the controls (4.23 +/- 0.29 kg; n = 7; P less than 0.05). A short-term study was undertaken with fetuses (n = 8) which were catheterized at 110 days gestation and whose dams were subjected to hypobaric hypoxia from 120 to 141 days gestation. The mean carotid PO2 of fetuses in the hypoxic group was 12.7 +/- 0.7 torr compared to 22.7 +/- 0.7 torr for the control group (n = 9; P less than 0.001) throughout the period of treatment. Fetal arterial oxygen content fell from 6.5 +/- 1.7 to 4.9 +/- 0.4 ml/dl (P less than 0.05), but rose to control values after 7 days due to an increase in fetal haemoglobin concentration (9.6 +/- 1.1 to 13.0 +/- 1.9 g/dl, P less than 0.001) and packed cell volume (33 +/- 3 to 45 +/- 4%, P less than 0.001). In the hypoxaemic fetuses, pH fell initially from 7.34 +/- 0.02 to 7.28 +/- 0.03 (P less than 0.05) and then recovered to 7.32 +/- 0.03 within 24 h. Mean fetal weight of the short-term hypoxic group was 3.46 +/- 0.72 kg compared to 4.15 +/- 0.51 for the control group (P less than 0.05). Both long- and short-term hypoxia produced a similar reduction in fetal body weight. The adrenal glands were significantly heavier in the hypoxic fetuses than in controls. Placental weight was not effected by hypoxia, but exposure from 30 days gestation reduced the average size of cotyledons (P less than 0.05). It is concluded that the fetal sheep increases its ability to acquire and transport oxygen in response to chronic hypoxia, but this compensation is not sufficient to prevent growth retardation or changes to the pattern of tissue growth.     

The effects of RU486 on the luteal phase of the rhesus monkey

RU486 is a steroid which possesses great affinity for the progesterone (P) receptor, but which has no P activity. It has been shown to be, as a result, a potent P antagonist. In the present study, we investigated the effect of this compound on the luteal phase of the rhesus monkey. The day of ovulation was diagnosed with a +/- 12 h accuracy, using serial laparoscopies and serum estradiol (E2) determinations, in regularly cycling rhesus monkeys. RU486 was administered by gavage (10 mg daily) in different regimens during the luteal phase: Group 1, days 1-5; Group 2, days 5-9; Group 3, days 9-13; and Groups 4, days 9-13, plus hCG (30, 60, 90, 180 and 360 IU i.m. on days 6-10). RU486 induced vaginal bleeding within 24-72 h after the initial administration in Groups 1-3. Animals of Group 4 presented luteal lengths ranging from 9-12 days. Progesterone concentrations at the onset of vaginal bleeding were 2.1 +/- 0.3, 4.9 +/- 0.6, 2.6 +/- 0.4 and 11.2 +/- 1.5 ng/ml (x +/- SEM) for animals of Groups 1-4, respectively. Serum follicle stimulating hormone (FSH), luteinizing hormone (LH), E2 and P levels were not altered during treatment. The availability of a compound such as RU486, that consistently induces vaginal bleeding due to its action at the target level (endometrium) without affecting the hormonal events of the menstrual cycle, opens a new approach to post-coital and interceptive contraception.     

The basalis of the primate endometrium: a bifunctional germinal compartment

Radioautographic analysis of epithelial and stromal cell proliferation in the primate endometrial functionalis and basalis (rhesus monkey) has identified horizontal zonal patterns of mitotic activation and inhibition during natural menstrual cycles. At 1 h after a single i.v. injection of [3H]thymidine, mitotic activity in endometrial biopsies (hysterotomy) was determined on 9 days from the late proliferative to the late luteal phase (-2 days to + 14 days relative to the estrogen [E2]peak). Labeling indices (LIs) were determined within glandular segments of the 4 horizontal endometrial zones: Transient functionalis Zone I (luminal epithelium) and Zone II (uppermost gland); Germinal basalis: Zone III (middle gland) and Zone IV (basal gland). The size of the dividing epithelial populations (LI) differed zonally. During E2 dominance (-2 days to +3 days), the epithelial LIs of functionalis I (10 +/- 0.3%) and II (9.8 +/- 1.0%) were greater than those of basalis III (5.8 +/- 0.2%) and basalis IV (3.7 +/- 0.8%). During progesterone (P) dominance (+5 days to +14 days), epithelial mitosis was strongly inhibited in functionalis I (4.3 +/- 1.9%), functionalis II (0.8 +/- 0.2%), and basalis III (1.4 +/- 0.5%). Thus germinal basalis III was linked functionally with transient functionalis I and II by periovulatory uniformity in epithelial proliferation and postovulatory mitotic inhibition. A unique mitotic pattern set basalis IV apart from other zones by a steady rise in LI from 1% (-2 days) to 11% (+10 days). The LIs for stromal fibroblasts remained quite uniform in basalis IV but varied in other zones. Thus the postovulatory primate basalis was a distinct bipartite compartment in which the mitotic rate in basalis IV glandular epithelium increased steadily whereas that of basalis III was strongly inhibited. The remarkable enhancement of epithelial mitotic activity in basalis IV may reflect expansion of the stem-progenitor cell population for gestational growth or for post-menstrual regeneration.     

Increased expansion of the lung stimulates calmodulin 2 expression in fetal sheep

Obstruction of the fetal trachea causes the lungs to expand with accumulated liquid. Although this is a potent stimulus for lung growth, the mechanisms involved are unknown. Our aim was to identify genes that are differentially expressed as a result of increased fetal lung expansion. Using differential display RT-PCR, we isolated a cDNA fragment partially encoding calmodulin 2 (CALM2) and identified the remainder of the coding region by 5'-rapid amplification of cDNA ends. Differential expression of CALM2 was confirmed by Northern blot analysis; CALM2 mRNA levels were increased to 161 +/- 5% of control at 2 days of increased lung expansion, induced by tracheal obstruction (TO), and had returned to control levels at days 4 and 10. Using in situ hybridization analysis, we found that the proportion of CALM2-labeled cells increased from 10.3 +/- 1.0% to 21.4 +/- 6.8% by 2 days of TO. This increase in CALM2 expression was reflected by a tendency for calmodulin protein levels to increase from 122.7 +/- 17.3 to 156.5 +/- 17.7 at 2 days of TO. Thus increases in fetal lung expansion result in time-dependent changes in CALM2 mRNA levels, which closely parallels the changes in lung DNA synthesis rates. As calmodulin is essential for cell proliferation, increased CALM2 mRNA levels may reflect an important role for calmodulin in expansion-induced fetal lung growth.