Influence of gonadectomy on isoelectrofocusing profiles of pituitary gonadotropins in rhesus monkeys

Isoelectrofocusing (IEF) profiles of bioactive and immunoreactive follicle-stimulating hormone (FSH) and luteinizing hormone (LH) were studied in saline extracts of pituitary glands from rhesus monkeys. No sex differences were found in the IEF profiles of FSH and LH in intact or gonadectomized animals. Gonadectomy in both sexes resulted in a marked increase in the formation of relatively more acidic molecular species of FSH and LH in parallel with the production of an unusual strongly alkaline FSH species.     

Seasonal variation in pituitary gonadotropin in the adult male newt, Cynops pyrrhogaster pyrrhogaster, revealed by isoelectric focusing technique and radioreceptor assay

The seasonal variation in pituitary gonadotropin in the adult male newt, Cynops pyrrhogaster pyrrhogaster was investigated by means of isoelectric focusing (IEF) coupled with radioreceptor assay (RRA), which employed Anolis or Xenopus testicular homogenates as receptors and 125I-rat FSH as radioligand. In the Anolis RRA system, the standard curve was obtained with 0.125-16 ng/tube of NIAMDD rat FSH I-3. Purified preparations, chicken LH IEF-1, chicken FSH AGCHD11113A and bullfrog basic gonadotropin-IV competitively inhibited the binding of the radioligand, but NIAMDD rat LH I-4 and human chorionic gonadotropin did not crossreact. The autoradiographic study revealed that 125I-rat FSH bound to the constituent cells of the seminiferous tubules in the Anolis testis, but scarcely to Leydig cells. In the IEF pattern of gonadotropin in February obtained by Anolis RRA, distinct peaks were observed at pH 9.05 (component B) and 8.55 (component C), and less distinct peaks were observed at pH 9.80 (component A), 7.55 (component D) and 7.05 (component E). When the same fractions were assayed by Xenopus RRA, five components were found in the alkaline region, which corresponded to those observed with Anolis RRA. Similar results were obtained with pituitary extracts in May. In July, the IEF pattern obtained by Anolis RRA indicated two additional components at pH 6.30 (component F) and 5.27 (component G) in the acidic region, which were not found by Xenopus RRA. The relationship between the testicular function and the nature of pituitary gonadotropin in the reproductive cycle was discussed.     

Separation of bacterial cells by isoelectric focusing, a new method for analysis of complex microbial communities.

A simple isoelectric focusing (IEF) method for whole bacterial cells was developed. In a pH gradient of 2 to 10 and an electric field of 11.5 V cm-1, mixtures of cells from the three different bacterial strains Chlorobium limicola 6230, Pseudomonas stutzeri DSM 50227, and Micrococcus luteus DSM 20030 could be separated. A density gradient of Ficoll prevented convective currents in the system. The method was tested with a concentrated mixture of bacteria from a shallow eutrophic lake and yielded up to 10 different bands. Species composition in each IEF band was analyzed by PCR plus denaturing gradient gel electrophoresis (DGGE). Each IEF band exhibited a different species composition. After the separation of cells by IEF three times more 16S ribosomal DNA signals could be detected by DGGE than in the unfractionated natural bacterial community. It is concluded that the resolution of these molecular biological methods is significantly enhanced if cells are first separated by IEF. At the same time, the IEF fractions are enriched for certain species, which can be used in subsequent cultivation experiments.     

Characterization of various molecular species of follicle-stimulating hormone in baboon pituitary preparations

The properties of baboon (Papio hamadryas) follicle-stimulating hormone (bFSH) have been studied after isoelectrofocusing (IEF) of individual pituitary extracts from five female and three male baboons and of a partially purified bFSH preparation (CM-1). The in vitro bioactivities of the female and male pituitary extracts and of CM-1 were 6.0 (range 4.1-9.6), 10.8 (6.9-18.2), and 34.1 (30.3-38.3) mg equivalent of LER 1909-2 reference standard per mg protein, respectively. The corresponding ratios of bioactivity to immunoreactivity (B/I) were 1.72 (range 1.31-2.06), 1.82 (1.53-2.25), and 1.54 (1.51-1.58), respectively. There was a significant increase (p less than 0.05) in all B/I ratios after IEF, due to a diminished immunoreactivity without any loss in bioactivity. Several molecular forms of bFSH were observed with pI values ranging from 4.5 to 7.5, with maximum activity between pH 4.5 to 6.0. The IEF profiles of female and male pituitaries and of the purified bFSH preparation were similar. However, the B/I ratios of the different molecular species of bFSH increased with increasing pI values in every case. It is concluded that there is a significant heterogeneity of bFSH in crude as well as in purified pituitary extracts, but neither the sex of the animals nor the process of purification influenced the quantity and distribution of various molecular species.     

Subpopulations of luteinizing hormone (LH) possessing various ratios of bioactivity to immunoreactivity in the female rat pituitary glands and their changes during the estrous cycle

Twenty-four adult female Sprague-Dawley rats (3 from each of 8 litters), showing 4-day cycles, were used in the present study. Aqueous extracts of pools of 6 pituitary glands in each cycle date were fractionated with a column isoelectrofocusing (IEF) technique, pH range of 3.5-10. Biological and immunological LH activities were determined by an in vitro bioassay and a radioimmunoassay, respectively, in the original aqueous extracts of the pituitary glands and in the fractions separated by IEF. Pituitary content of LH was the highest in the proestrus before the preovulatory LH surge (1243.7 +/- 67.8 micrograms NIAMDD rat LH-RP-1/pituitary gland for the biological activity). In the estrus, after the LH surge, it was reduced to 688.9 +/- 51.2 micrograms/pituitary gland. The decreased pituitary content was recovered to the level in the proestrus during the metestrus and the diestrus (1047.0 +/- 53.8 and 1173.0 +/- 58.5 micrograms/pituitary gland, respectively). Rat LH in the pituitary aqueous extracts was separated into multiple subpopulations in terms of pI values by IEF; i.e. Subpopulations A (pI = 10.3), B (9.3), C (9.0), D (8.7), E (8.3), F (neutral LH), and G (acidic LH). Among them the most predominant one was Subpopulation A throughout the estrous cycle. Subpopulations A, B and C exhibited statistically significant cyclic changes as was observed in the pituitary LH content, whereas the remaining ones stayed at constant levels during the cycle. The highest ratio of biological to immunological LH activities (B/I ratio) was obtained in Subpopulation A (6.41), followed by G, C and B (5.15, 4.24 and 3.99, respectively). Depressed B/I ratios were revealed in D, E and F (2.59, 1.86 and 3.07, respectively). High alkaline LH subpopulations, i.e. A, B and C, preserving high biological potency and showing cyclic changes during the estrous cycle, seem to be the releasable types of the hormone and to be mainly discharged for the preovulatory LH surge. Although characteristic features of other types of the hormone are not known, it is possible that one of them, presumably the acidic LH, might be the newly-synthesized type of the hormone, which might attain releasability by certain molecular modifications involving a shift in the pI value.     

Gonadal steroids modulated hypocretin/orexin type-1 receptor expression in a brain region, sex and daytime specific manner

Orexins A and B (hypocretins A and B) are regulatory peptides that control a variety of neuroendocrine and autonomic functions including feeding and sleep-wakefulness. Previously, we described a clear relationship between the hormonal milieu of the estrous cycle and the mRNA expression of the components of the orexinergic system, in the hypothalamus, pituitary and ovary. Here, we investigate whether steroid hormones are involved in the modulation of the hypocretin/orexin type-1 receptor expression at the protein level, and its time of the day dependence, in hypothalamus and pituitary of castrated male and female rats and castrated receiving hormone replacement.Orchidectomy decreased the hypocretin/orexin type-1 receptor expression in anterior hypothalamus, but not in mediobasal hypothalamus or cortex; in pituitary this treatment resulted in an increase. Testosterone and dihydrotestosterone were able to restore receptor expression and gonadotropins.In females, pituitary and ovarian hormones increased during proestrous afternoon. Hypocretin/orexin type-1 receptor expression was higher at 19:00 of proestrus in hypothalamus and pituitary. Ovariectomized treated with estradiol or oil and sacrificed at 11:00 h showed the receptor expression similar to 11:00 h of proestrus in hypothalamus and pituitary. At 19:00 h, low expression persisted in these areas in oil-treated ovariectomized rats; in contrast, estradiol replacement increased the expression to high levels of normal cycling rats at 19:00 h.Sexual steroids modulate the orexinergic system and the anatomical regions, hormones and times of the day all have to be considered when the roles of orexins, and probably other peptides, are under consideration.     

Identification of human red cell glutamate-pyruvate transaminase (GPT) phenotypes by isoelectric focusing.

Isoenzymes of human red cell glutamate-pyruvate transaminase (GPT) were resolved by isoelectric focusing (IEF) of hemolysates in polyacrylamide gels at pH 5.0-7.0. The bands of enzyme activity required both alpha-ketoglutarate and L-alanine in the staining mixture for visualization, indicating that the bands were not lactate dehydrogenase or glutamate dehydrogenase. Phenotyping of 41 individuals by IEF, including types GPT 1, 2A, 1-2A, 1-2B, and 2A-2B, agreed with the typing results obtained by electrophoresis in starch gels and in polyacrylamide gels at acid and alkaline pH. Analysis of one kindred demonstrated autosomal codominant transmission of the rare GPT*2B gene through 3 generations. IEF facilitates phenotyping by permitting identification of the GPT types on a single gel with a considerable reduction in time and cost. Although no new variants were found in this investigation, IEF may be more powerful for the recognition of presently undetected variants of GPT.     

Partial purification and characterization of rhinoceros gonadotropins, growth hormone, and prolactin: comparison with the horse and sheep

The rhinoceros is an endangered species related to the horse family. Little is known of its reproductive endocrinology. The objectives of this study were to partially purify rhinoceros pituitary hormones, determine which assays could be used for their assessment, and to ascertain whether rhinoceros LH possesses the intrinsic FSH activity of equine LH. A single pituitary each from a White (1.3 g) and a Black (1.2 g) Rhinoceros was homogenized and extracted (pH 9.5), then subjected to pH and salt fractionation, and ion-exchange chromatography (DEAE and Sephadex SP-C50) to yield partially purified fractions of LH, FSH, growth hormone (GH), and prolactin (PRL). LH was readily measured by a rat Leydig cell assay (0.1-1% x equine LH) and an RIA using a monoclonal antibody to bovine LH (6-11% x equine LH). FSH activity detected in the LH by either an FSH RIA or a calf testis radioreceptor assay (RRA) was extremely low. No FSH activity could be detected in the White Rhinoceros pituitary "FSH" fraction, but was readily detected in the Black Rhinoceros fraction (RIA: 0.2% x equine FSH: RRA: 0.8% x equine FSH). The presence of GH and PRL was determined by SDS-PAGE and Western blots. Results showed a single immunoreactive GH band and multiple immunoreactive PRL bands. Adsorption with Concanavalin A-Sepharose indicated that some of the PRL bands are glycosylated.     

Chromogranin A, B and C: immunohistochemical localization in ovine pituitary and the relationship with hormone-containing cells

Chromogranins A, B and C, three distinct groups of proteins found in bovine chromaffin granules, were also found to be present in the pituitary using immunoblotting techniques. Their distribution was therefore studied in the normal ram pituitary using an immunoperoxidase technique applied to semithin serial sections and compared with that of some of the hormones of the anterior pituitary. Chromogranin-immunoreactivity was found in gonadotrophs (all three), thyrotrophs (A with some positive for C) and corticotrophs (a fraction with A and fewer with B and C). The mammotrophs and somatotrophs were negative. Chromogranin C was the only one of the three to be located in the pars nervosa, whilst chromogranin B was rarely found in the pars intermedia. The results suggest that chromogranins A, B and C are not always stored together, some hormone-containing cells do not contain immunohistologically detectable levels of the chromogranins.     

贵州麻阳河国家级自然保护区红外相机鸟兽监测

为进一步查清贵州麻阳河国家级自然保护区鸟兽多样性, 丰富生物编目, 我们于2018年7月至2019年8月利用红外相机进行公里网格全境布设监测。共记录兽类5目10科18属20种、鸟类7目18科34属42种, 其中新增保护区记录11种, 记录国家I级重点保护野生动物2种, 分别是黑叶猴(Trachypithecus francoisi)和白颈长尾雉(Syrmaticus ellioti), 国家II级重点保护野生动物大灵猫(Viverra zibetha)、白冠长尾雉(Syrmaticus reevesii)等7种。相对多度指数计算结果表明, 小麂(Muntiacus reevesi)、猪獾(Arctonyx collaris)和野猪(Sus scrofa)是该区域内常见兽类, 而地栖雉类则以红腹锦鸡(Chrysolophus pictus)和灰胸竹鸡(Bambusicola thoracicus)为主。分时段的相对多度指标表明, 小麂在6:00-15:00和17:00-21:00两个时段均保持活跃, 猪獾则偏好5:00-6:00和19:00-23:00活动, 野猪在6:00-7:00最为活跃, 红腹锦鸡活动高峰在12:00-13:00, 而灰胸竹鸡的活动高峰分别为14:00-15:00和16:00-17:00。在物种丰度上, 单个相机位点的平均拍摄物种数显示, 在缓冲区、灌丛及针阔混交林以及800-1,200 m海拔段上物种较多。监测结果有利于进一步掌握此区域的动物多样性资源并促进资源的保护与管理。     

Taxonomic comparison of three different groups of aflatoxin producers and a new efficient producer of aflatoxin B1, sterigmatocystin and 3-O-methylsterigmatocystin, Aspergillus rambellii sp. nov

Accumulation of the carcinogenic mycotoxin aflatoxin B, has been reported from members of three different groups of Aspergilli (4) Aspergillus flavus, A. flavus var. parvisclerotigenus, A. parasiticus, A. toxicarius, A. nomius, A. pseudotamarii, A. zhaoqingensis, A. bombycis and from the ascomycete genus Petromyces (Aspergillus section Flavi), (2) Emericella astellata and E. venezuelensis from the ascomycete genus Emericella (Aspergillus section Nidulantes) and (3) Aspergillus ochraceoroseus from a new section proposed here: Aspergillus section Ochraceorosei. We here describe a new species, A. rambellii referable to Ochraceorosei, that accumulates very large amounts of sterigmatocystin, 3-O-methylsterigmatocystin and aflatoxin B1, but not any of the other known extrolites produced by members of Aspergillus section Flavi or Nidulantes. G type aflatoxins were only found in some of the species in Aspergillus section Flavi, while the B type aflatoxins are common in all three groups. Based on the cladistic analysis of nucleotide sequences of ITS1 and 2 and 5.8S, it appears that type G aflatoxin producers are paraphyletic and that section Ochraceorosei is a sister group to the sections Flavi, Circumdati and Cervini, with Emericella species being an outgroup to these sister groups. All aflatoxin producing members of section Flavi produce kojic acid and most species, except A. bombycis and A. pseudotamarii, produce aspergillic acid. Species in Flavi, that produce B type aflatoxins, but not G type aflatoxins, often produced cyclopiazonic acid. No strain was found which produce both G type aflatoxins and cyclopiazonic acid. It was confirmed that some strains of A. flavus var. columnaris produce aflatoxin B2, but this extrolite was not detected in the ex type strain of that variety. A. flavus var. parvisclerotigenus is raised to species level based on the specific combination of small sclerotia, profile of extrolites and rDNA sequence differences. A. zhaoqingensis is regarded as a synonym of A. nomius, while A. toxicarius resembles A. parasiticus but differs with at least three base pair differences. At least 10 Aspergillus species can be recognized which are able to biosynthesize aflatoxins, and they are placed in three very different clades.     

Impact of nutrient enrichment and alkalinization on periphyton communities in the New Jersey Pine Barrens

The impact of residential and agricultural development on stream periphyton communities in the New Jersey Pine Barrens was examined by comparison with communities in undeveloped areas. Watershed disturbance resulted in stream water primarily characterized by greatly elevated pH levels and nitrate concentrations. A total of 53 periphyton species were encountered in bimonthly samples over a one year period in the three disturbed and three undisturbed study streams. Species richness was significantly greater in the disturbed streams based on three criteria: the average number of species per stream on each sampling occasion (disturbed = 6.3; undisturbed = 4.9), the average number of species per stream for the entire year (disturbed = 19.3; undisturbed = 16.0), and the total number of species found in streams within a type (disturbed = 40; undisturbed = 31). Species composition also changed significantly as the result of disturbance. There appeared to be replacement of species characteristic of undisturbed Pine Barrens streams with species peripheral to the region. The expected effects of both elevated pH and nitrate were consistent with these results.     

Effects of gonadectomy on polymorphism in stored and circulating gonadotropins in the bullfrog, Rana catesbeiana. II. Gel filtration chromatography

Marked polymorphism was revealed in both stored and circulating forms of immunoreactive follicle-stimulating hormone (FSH) and luteinizing hormone (LH) in the bullfrog, Rana catesbeiana, by exclusion chromatography on columns of Sephracyrl-S200. FSH behaved as a more homogeneous and larger molecule than LH from the same pituitary or plasma, but the properties of both hormones in the plasma were markedly affected by gonadectomy. Chromatographic profiles of FSH stored in the pituitaries were similar in intact and gonadectomized frogs, but pituitary LH in the latter was comprised of a larger proportion of early eluting activity. Previously purified preparations of bullfrog FSH and LH were more homogeneous than these extracts. Differences between pituitary hormones in intact and gonadectomized frogs were small compared with those between circulating hormones. Plasma FSH and lH from gonadectomized frogs behaved as more homogeneous and larger molecules than those from intact frogs in which plasma gonadotropins were elevated normally or by injections with gonadotropin releasing hormone (GnRH). Some differences in circulating hormones were also observed between a normal male and female and both differed from gonadectomized an GnRH-treated intact frogs. Chromatographs of plasma gonadotropins in GnRH-treated animals generally resembled those of the hormones stored in the pituitary, whereas plasma FSH and LH in gonadectomized frogs appeared more homogeneous and larger than the pituitary-stored forms. Those pronounced differences in chromatographic properties of gonadotropins in intact and gonadectomized frogs correlate with previously observed effects of gonadectomy on clearance profiles of circulating FSH and LH.     

Immunocytochemical localization of protein antigens in large sections of tissues embedded in water-soluble embedding media

JB4 and Immunobed are water-soluble embedding media used for embedding large blocks of tissue. Immunobed was specifically designed for immunocytochemistry because ethanol extraction of an additive in the monomer of the resin is reported to render tissue sections permeable to immunoglobulins. We have modified the manufacturer's protocol to accomplish localization of two protein antigens in tissues embedded in either JB4 or Immunobed. Luteinizing hormone-beta (LH beta) was localized in sections of rat and bovine pituitary tissues and bovine placental lactogen (bPL) was localized in sections of placentomes from bovine placentas. Sections received one of the following pre-treatments: absolute EtOH; NaHCO3 buffer, pH 6-10; EtOH followed by NaHCO3 buffer; one of several enzymes; EtOH followed by enzyme; NaHCO3 buffer followed by enzyme. Anti-LH beta stained only pituitary gonadotrophs and anti-bPL stained only placental binucleate cells, as assessed by absorption controls. Pre-treatment with enzyme was required for staining of sections, but an alkaline pH change (NaHCO3) had little or no effect. Ethanol pretreatment had little or no effect alone or in conjunction with NaHCO3 or enzyme. Sections were sufficiently thin (1.5 micron) to afford resolution of structure, but suitably large (approximately 2 cm2) to minimize problems of sampling.     

Separation and isolation of human apolipoproteins C-II, C-III0, C-III1, and C-III2 by chromatofocusing on the Fast Protein Liquid Chromatography system

Chromatofocusing, which separates proteins based on differences in isoelectric point, has been used on the Fast Protein Liquid Chromatography (FPLC) system (Pharmacia) to separate the C apolipoproteins from human very low density lipoproteins (VLDL). Using a Mono P column (Pharmacia), a pH gradient between pH 6.2 and pH 4.0 was generated using buffers containing 6 M urea, at a flow rate of 0.5 ml/min. Typically, runs took approximately 45 min. Chromatofocusing of delipidated whole VLDL produced sharp, well-resolved peaks for the C apolipoproteins. However, as determined by analytical isoelectric focusing (IEF), the apolipoprotein E isoforms were not separated from apoC-II, and they contaminated the other apoC species to a variable extent. In addition, apoC-II was not resolved from apoC-III0. Preliminary precipitation of VLDL with acetone prior to delipidation removed both apolipoproteins E and B. Using a start buffer of 25 mM histidine, pH 6.2, and a 1:30 dilution of the polybuffer exchanger (eluting buffer), apoC-II, C-III0, C-III1, and C-III2 were well resolved in run-times of approximately 60 min. The C apoproteins proved to be pure by analytical IEF and immunoassay with monospecific antisera against apoC-II and C-III. Recovery was over 90% of the protein chromatographed. In addition, a variant of apoC-II present in VLDL of a hypertriglyceridemic subject was clearly resolved from the other C apolipoproteins. This technique is superior to conventional methodology in terms of its time saving and high resolution. The application of this technique to the study of C apolipoprotein variants and C apolipoprotein specific radioactivity determinations is possible.     

CD46 is a cellular receptor for all species B adenoviruses except types 3 and 7

The 51 human adenovirus serotypes are divided into six species (A to F). Adenovirus serotypes from all species except species B utilize the coxsackie-adenovirus receptor for attachment to host cells in vitro. Species B adenoviruses primarily cause ocular and respiratory tract infections, but certain serotypes are also associated with renal disease. We have previously demonstrated that adenovirus type 11 (species B) uses CD46 (membrane cofactor protein) as a cellular receptor instead of the coxsackie-adenovirus receptor (A. Segerman et al., J. Virol. 77:9183-9191, 2003). In the present study, we found that transfection with human CD46 cDNA rendered poorly permissive Chinese hamster ovary cells more permissive to infection by all species B adenovirus serotypes except adenovirus types 3 and 7. Moreover, rabbit antiserum against human CD46 blocked or efficiently inhibited all species B serotypes except adenovirus types 3 and 7 from infecting human A549 cells. We also sequenced the gene encoding the fiber protein of adenovirus type 50 (species B) and compared it with the corresponding amino acid sequences from selected serotypes, including all other serotypes of species B. From the results obtained, we conclude that CD46 is a major cellular receptor on A549 cells for all species B adenoviruses except types 3 and 7.     

Alkaline phosphatases of the mosquito, Culex tarsalis Coquillett

Spectrophotometric and isoelectric focusing (IEF) electrophoretic characterization of the alkaline phosphatase (ALKP) of the mosquito, Culex tarsalis, are presented. With p-nitrophenylphosphate (Pnp) as substrate, ALKP was optimally active at 37 degrees C, pH 8.0, 30 mM MgCl2, Vmax was 35.8 mumoles/10 min and the Km was 5.7 mM, with no demonstrable requirement for Zn2+. The spectrophotometric enzyme(s) was stimulated by dithiothreitol, 2-mercaptoethanol, and poly-vinylpyrollidone (PVP); inhibited by NaF, several alternative cations (Ca2+, Ba2+, Fe2+, Cu2+), and EDTA. ALKP activity was cyclic during the 15 day post-adult emergence period of the study. No significant differences were noted between the specific activities of males and females. IEF electrophoresis revealed 6 ALKP isozymes detected with alpha-naphthylphosphate within the pH range 4.0-5.5, with a second group of 3 rather indistinct species in the pH 6.0-7.0 range. IEF ALKP isozymes were stimulated by Mg2+ and PVP and inhibited by EDTA (except ALKP5.0) and cysteine; partial inhibition with phenylalanine. IEF detection of ALKP activity with Pnp indicated that the majority of the activity was localized in the pH 4.0-5.5 range, in close agreement with the alpha-naphthylphosphate results.     

Qualitative and quantitative differences in hLH species in the first and second LH release induced by continuous stimulation with synthetic LHRH in normal menstrual cycle as assessed by isoelectrofocusing

It is known that continuous stimulation with LHRH induces a biphasic release of LH. The two pools theory is generally accepted in interpreting this phenomenon. The present study was conducted in order to elicit qualitative differences in LH included in the two pools. Five hundred micrograms synthetic LHRH was infused for 2h in 10 women in various phases of the menstrual cycle and 5 h blood sampling was performed during and after the infusion. Biphasic release of LH was demonstrated in all the cases investigated. The responsiveness to LHRH was most predominant in the midcycle, followed by the luteal phase. Pools of plasma samples at 30-105 min and 150-225 min of the LHRH infusion (plasma pools I and II; corresponding to the initial and the second LH release, respectively) were subjected to isoelectrofocusing (IEF) fractionation, pH range of 3.5-10. LH in plasma was classified in terms of isoelectric point (pI); i.e. A(pI = 9.13), B(8.60), C(8.16), D(7.67), E(7.24), and F(LH species migrating in the acidic pH area). Percentage of high alkaline LH species was significantly elevated in plasma pool I of the midcycle. No phasic difference was observed in the IEF profiles in plasma pool II. Consistent and significant increase and decrease in the percentage of acidic and high alkaline LH species, respectively, were observed throughout the menstrual cycle when the IEF profiles of plasma pool II were compared with those of pool I. The ratio of biological to immunological LH activities (B/I ratio) was markedly depressed in acidic LH species. The highest B/I ratios were obtained in the LH species migrating in the mid-alkaline region. The acidic LH species might represent the young generation of LH molecules prior to acquisition of biological potency. LH species migrating in the mid-alkaline region might be mature and bio-potent forms of the hormone. High alkaline LH species might represent LH molecules over-processed and having lost a degree of biological potency during a prolonged period of storage.     

Overgrowth in a marine epifaumal community: Competitive hierarchies and competitive networks

Summary The frequencies with which organisms of a species overgrew or were overgrown by organisms of other species in a marine epifaunal community were estimated. The ranking of the ability of the major taxonomic groups to overgrow others was basically hierarchical:ascidianssponges>bryozoans>barnacles, polychaetes, tubicolous amphipods, hydroids. In contrast, the ranking of the competitive ability of species in the community did not form a simple linear hierarchy and there was no single competitively dominant species (measured in terms of overgrowth). There were often no significant differences in the ability of species to overgrow each other within the three major taxonomic groups of sponges, ascidians and bryozoans. Such results were common also between the species of large sponges and ascidiams which dominated substrata immersed for periods longer than two years.A lack of a significant difference in the competitive ability of species was usually the result of (a) frequent formation of delay/ties or standoffs and (b) changes in the outcome of interactions due to change in the relative size of interacting colonies. In many two-species interactions the species which had the larger colony in a given encounter had a greater probability of winning.When the range of colony sizes of two species was similar there was often no significant difference between the competitive ability of each species. Such cases without a clearcut winner often represented a backloop in an otherwise hierarchical sequence of competitive ability, i.e. Species A beats Species B, Species B beats Species C, no significant differences in competitive ability between Species C and A. No examples of competitive networks of the form Species A beats Species B, Species B beats Species C, Species C beats Species A were found. Backloops in otherwise hierarchical sequences (no significant differences in competitive ability) occurred most frequently between species within the same major taxonomic groups and were the result of a very even balance in the generalised competitive mechanism of overgrowth.It seems probable that backloops in hierarchical sequences are more commonly due to the absence of clear competitive dominance in interactions between species (reversals in the outcome of overgrowth interactions and standoffs), rather than to direct backloops formed by a specialised or to a generalised competitive mechanism. Network-like arrangements of competitive ability formed by the type of processes described here are likely to contribute significantly to the high levels of species diversity observed in many marine epifaunal communities subject to low levels of physical disturbance.     

Covalent binding of benzo[a]pyrene metabolites to DNA, RNA and chromatin proteins in the AKR mouse embryo cell-line

A specific fraction from the nuclei of the AKR mouse embryo cell-line (fraction I) displayed a much greater localization of radioactivity compared to fraction II and III when the chemical carcinogen, [3H]benzo[a]pyrene (B[a]P) was incubated with the cells for 24 h. The radioactivity in fraction I consisted of both covalently and non-covalently bound metabolites. Isolation of the DNA, RNA and protein of fraction I revealed that 94% of the covalently bound radioactivity was to protein, 5% to RNA and 1% to DNA. Analysis of the fraction I proteins by SDS gel electrophoresis revealed that there was more radioactivity covalently bound to the larger proteins than to smaller proteins. Isoelectric focusing (IEF) of the purified proteins displayed two peaks of radioactivity, one at a pH of 5 and the other at 11. The former proteins bound more radioactivity per mass of protein than the latter proteins. Analysis of fraction I histones on acid urea polyacrylamide gels showed that the radioactivity coincided with histones H3 and H2B and low levels of radioactivity associated with histones H1, H2A and H4. Two significant peaks of radioactivity closely migrated near but did not co-migrate with histone H1. The distribution of the bound radioactivity is probably a reflection of the availability of the proteins to the reactive carcinogen metabolites. The possible binding of B[a]P metabolites to phosphorylated histones and to the high mobility of group (HMG) proteins 1 and 2 is discussed.