Role of matrix metalloproteinase 2 in the ovulatory folliculo-luteal transition of ewes

Tissue dissolution and remodelling are associated with the processes of rupture of the ovulatory follicle and formation of the corpus luteum. Matrix metalloproteinase 2 (MMP-2) belongs to a family of endopeptidases that cleave extracellular proteins; its primary substrate is the lattice network of basement membranes that support epithelial cells and endothelium. The aim of this study was to ascertain a putative regulatory role of MMP-2 relevant to the folliculo-luteal transformation in ewes. Luteal regression and the preovulatory surge of gonadotrophins were synchronized by administration of PGF(2alpha) and GnRH on days 14.0 and 15.5 of the oestrous cycle, respectively. Dominant antral follicles present during pro-oestrus consistently ovulate approximately 24 h after GnRH administration. Normal IgG or a bioactivity-neutralizing MMP-2 monoclonal antibody was injected into the antral cavity of preovulatory follicles at 8 h after GnRH administration. Jugular blood samples were obtained for serum progesterone analysis and ovaries were removed for light microscopic morphometry on day 8. A definitive ovulation stigma was evident in control ewes. The antra of ruptured follicles had largely been supplanted with luteal tissue. In contrast, the ovarian surface contiguous with follicles injected with anti-MMP-2 was smooth and undisturbed, which is indicative of a failure of ovulation. Luteinized unruptured follicles were filled with (entrapped) fluid. Corpora lutea of control animals contained numerous connective tissue projections that provided a framework for cellular migration and angiogenesis. Luteal tissues that surrounded the cavity of antibody-treated follicles lacked trabeculae and were deficient in blood vessels. Systemic venous progesterone concentrations were lower in ewes with a luteinized unruptured follicle compared with those with a corpus luteum. It is proposed that MMP-2 is a mediator of ovulation and luteal development.     

A portable sky luminance meter

The paper discusses the development of an instrument to collect sky luminance data over an extended period of time and store it on cassette tape. Data are collected automatically by positioning a photometer using stepping motors under microprocessor control.The instrument consists of five major components: a power supply, positioning hardware, photometer, interface hardware, and microcomputer. The elevation motor and photometer sight tube and sensor are located on the elevation support assembly. All other components are housed in a based assembly. The instrument may be carried by one person.The instrument is operational and is currently being tested on the roof of the engineering building at the University of New Hampshire.     

Roles of the ovarian surface epithelium in ovulation and carcinogenesis

Although ovarian mechanisms of ovulation have been a subject of investigation for more than a century, essential regulatory pathways remain uncertain. A role for the ovarian surface epithelium in ovulation has recently been demonstrated. Ovarian surface epithelial cells in close contact with the apical wall of preovulatory ovine follicles secrete a urokinase-type plasminogen activator in response to surge concentrations of (locally delivered) gonadotrophins. Urokinase activates latent collagenases and stimulates release of tumour necrosis factor alpha from thecal endothelium. Tumour necrosis factor alpha progressively induces matrix metalloproteinase gene expression, apoptosis and inflammatory necrosis. Collagenolysis and cellular death are a prelude to stigma formation and ovarian rupture. Epithelium exfoliated from the dome of ovulatory follicles is replenished by generative stem cell replication and migration from the wound edges. Common epithelial ovarian cancer has been related to successive bouts of ovulation and mitosis. The integrity of the DNA of surface cells circumjacent to the ovarian rupture site is compromised during the ovulatory process. Clonal expansion of an epithelial cell with damaged (unrepaired) DNA is a putative factor in carcinogenesis. Ovarian cancer is a deadly insidious disease because typically it is asymptomatic until the malignancy has reached beyond the ovaries.     

Motility activation and second messenger signalling in spermatozoa from rat cauda epididymidis

This study examined molecular mechanisms involved in the activation of motility in spermatozoa from the cauda epididymidis of rats. A 1.05-fold dilution of semen from the cauda epididymidis with 300 mmol sucrose l(-1) did not activate motility in spermatozoa. Addition of dibutyryl cAMP, pentoxifylline or Ca(2+) to the sucrose activated motility in the short term (<30-60 min). A fivefold dilution of semen from the cauda epididymidis with a modified Tyrode's medium (BWW) activated and sustained vigorous motility that could not be attenuated with kinase inhibitors. This motility was associated with a transient increase in intracellular cAMP during the first 60 s of activation. Lower motility was activated in Ca(2+)-deficient media but this was not associated with an increase in cAMP. A fivefold dilution with plasma from the cauda epididymidis did not activate motility. The addition of Ca(2+) to the sucrose induced an increase in cAMP of similar duration but lower magnitude to that associated with dilution in BWW. The results from this study indicate that the cAMP and Ca(2+) signal transduction pathways are involved in activation of sperm motility, and that the increase in intracellular cAMP in rat spermatozoa from the cauda epididymidis undergoing motility activation is Ca(2+)-dependent. This is the first study to report a Ca(2+)-dependent increase in cAMP associated with motility activation in immotile mammalian spermatozoa. In light of these data, a model is proposed whereby cAMP and Ca(2+) act as synarchic messengers, initiating a signal transduction cascade, which is independent of protein kinase A-mediated phosphorylation of flagella proteins in immotile spermatozoa from the cauda epididymidis.     

Effects of Electroosmosis on Natural Soil: Field Test

A unique configuration of horizontal sheet-like electrodes was used in the field at a site in Ohio that was underlain by silty clay glacial drift to induce electroosmotic flow and to characterize the effects of electroosmosis on soil properties (e.g., electrical conductivity and pH). The lower electrode was created at a depth of 2.2 m by filling a flat-lying hydraulic fracture with granular graphite, and the upper one was a metallic mesh placed at a depth of 0.4 m and covered with sand. The electrodes were attached to a DC power supply, creating an electrical gradient of 20–31 V∕m and a current of 42–57 A within approximately 20 m3 of soil. Total energy applied was 5,500 kW?h during approximate 4 months of operation. Electroosmotic flow rates of 0.6–0.8 L∕h were observed during tests lasting several weeks, although total flow rate (electroosmotic plus hydraulic) was strongly influenced by fluctuations of the ground-water table. The ratio of applied current to voltage decreased from 0.9 to 0.6 A∕V and was mainly due to a decrease in electrical conductivity of the soil. A low pH front developed at the anode and migrated toward the cathode. The velocity of the pH front per unit voltage gradient was 0.014 (cm∕day)/(V∕m). This was 40 times slower than what has been reported from laboratory experiments using kaolinite as a medium. These results confirm the feasibility of using horizontal electrodes at shallow depths, but they also underscore some important differences between the geochemical effects observed during field tests in natural soils and those seen in laboratory tests using ideal materials.     

Justified design evaluation for percentile standards

Regulation of pollution levels in the environment is achieved through environmental quality standards. Permits to discharge are determined by comparison of design predictions with standards. Standards are often statistical and, in consequence, the calculations are completed using Monte-Carlo techniques. In some cases, the application of Monte-Carlo is impeded by computational limitations, when only a small number of scenarios can be simulated. This can occur in complex systems, such as the marine environment, when computational fluid dynamic models are used. There is a need to relate the outcome of these simulations to the required standards in order to evaluate the design proposal. The Justified Design Evaluation (JUDE) method has been developed to meet this need. Essentially, the method identifies the set of input values required to invoke the environmental response for the percentile and standard under consideration. It has been tested successfully in a freshwater and a marine example. The method has general applicability in the sense that it is not restricted to a specific model type or environmental system. Uncertainty in applying the method is quantified.     

Charge‐Reversal Drug Conjugate for Targeted Cancer Cell Nuclear Drug Delivery

DNA‐toxin anticancer drugs target nuclear DNA or its associated enzymes to elicit their pharmaceutical effects, but cancer cells have not only membrane‐associated but also many intracellular drug‐resistance mechanisms that limit their nuclear localization. Thus, delivering such drugs directly to the nucleus would bypass the drug‐resistance barriers. The cationic polymer poly(L ‐lysine) (PLL) is capable of nuclear localization and may be used as a drug carrier for nuclear drug delivery, but its cationic charges make it toxic and cause problems in in‐vivo applications. Herein, PLL is used to demonstrate a pH‐triggered charge‐reversal carrier to solve this problem. PLL's primary amines are amidized as acid‐labile β‐carboxylic amides (PLL/amide). The negatively charged PLL/amide has a very low toxicity and low interaction with cells and, therefore, may be used in vivo. But once in cancer cells' acidic lysosomes, the acid‐labile amides hydrolyze into primary amines. The regenerated PLL escapes from the lysosomes and traverses into the nucleus. A cancer‐cell targeted nuclear‐localization polymer–drug conjugate has, thereby, been developed by introducing folic‐acid targeting groups and an anticancer drug camptothecin (CPT) to PLL/amide (FA‐PLL/amide‐CPT). The conjugate efficiently enters folate‐receptor overexpressing cancer cells and traverses to their nuclei. The CPT conjugated to the carrier by intracellular cleavable disulfide bonds shows much improved cytotoxicity.     

Mechanical design of a PERMCAT reactor module

The PERMCAT is a membrane reactor proposed for processing fusion reactor plasma exhaust gas: tritium removal is obtained by isotopic swamping operating in counter-current mode. In this work, a membrane reactor using a permeator tube of length about 500 mm produced via diffusion welding of Pd-Ag thin foils is described. An appropriate mechanical design of the membrane module has been developed in order to avoid any significant compressive and bending stresses on the very long and thin wall permeator tube: two expanded bellows have been applied to the Pd-Ag tube, so that it has been pre-tensioned before operating.The elongation of the metal permeator under hydrogenation has been theoretically estimated and experimentally verified for properly designing the membrane reactor.