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One aspect of the present invention relates to a method of hormone replacement in mammals, which method comprises the oral administration of an estrogenic component and a progestogenic component to a mammal in an effective amount to prevent or treat symptoms of hypoestrogenism, wherein the estrogenic component is selected from the group consisting of substances represented by the above formula in which formula R1, R2, R3, R4 independently are a hydrogen atom, a hydroxyl group or an alkoxy group with 1-5 carbon atoms; each of R5, R6, R7 is a hydroxyl group; and no more than 3 of R1, R2, R3, R4 are hydrogen atoms; precursors capable of liberating a substance according to the aforementioned formula when used in the present method; and mixtures of one or more of the aforementioned substances and/or precursors. Another aspect of the invention concerns a pharmaceutical kit comprising oral dosage units that contain the aforementioned estrogenic component and a progestogenic component as well as an androgenic component.
1. A method of hormone replacement in mammals comprising orally administering an estrogenic component and a progestogenic component to a mammal in an effective amount to treat symptoms of hypoestrogenism, the estrogenic component being selected from the group consisting of estetrol;
precursors capable of liberating estetrol when used in the present method, which precursors are derivatives of estetrol, wherein the hydrogen atom of at least one of the hydroxyl groups has been substituted by an acyl radical of a hydrocarbon carboxylic, sulfonic or sulfamic acid of 1-25 carbon atoms; tetrahydrofuranyl; tetrahydropyranal; or a straight or branched chain glycosidic residue containing 1-20 glycosidic units per residue; and
mixtures of estetrol and the precursors.
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12. A pharmaceutical kit comprising at least 20 oral dosage units that contain the estrogenic component as defined in
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The present invention relates to a method of hormone replacement in mammals. More particularly the invention is concerned with a method of hormone replacement that comprises the oral administration to a mammal of a combination of an estrogenic component and a progestogenic component in an effective amount to prevent or treat symptoms of hypoestrogenism.
In hormone replacement therapy (HRT), sometimes also referred to as estrogen replacement therapy, estrogens are administered to prevent or treat symptoms resulting from estrogen deficiency or hypoestrogenism. Hypoestrogenism can occur in both females and males, and can lead to disorders and ailments such as osteoporosis (loss of bone mass), arteriosclerosis, climacteric symptoms such as hot flushes (flashes), sweats, urogenital atrophy, mood disturbances, insomnia, palpitations. Estrogen deficiency has also been associated with cognitive disturbances and Alzheimer's disease.
Hypoestrogenism, and in particular chronic hypoestrogenism, is frequently observed in (peri-)menopausal and post-menopausal women. However, it can also result from hypogonadism or castration, as well as from primary ovarian failure, treatment of e.g. breast cancer with aromatase inhibitor and gonadotropin-releasing hormone analogue treatment of benign gynaecological diseases such as endometriosis, adenomyosis, uterine fibroids (leiomyomas), dysmenorrhoea, menorrhagia and metrorrhagia.
HRT employs continuous administration of effective amounts of an estrogen for prolonged periods of time. The administration of estrogens has been associated, however, with endometrial proliferation in women and it is now widely accepted that "unopposed" estrogen therapy substantially increases the risk of endometrial cancer (Cushing et al., 1998. Obstet. Gynecol. 91, 35-39; Tavani et al., 1999. Drugs Aging, 14, 347-357). There is also evidence of a significant increase in breast cancer with long-term (10-15 years) use of estrogen therapy (Tavani et al., 1999. Drugs Aging, 14, 347-357; Pike et al., 2000. Steroids, 65, 659-664).
In order to counteract the negative effects of unopposed estrogen therapy, adjunctive progestogen treatment is nowadays commonly applied. Regular progestogen administration is believed to inhibit the continual estrogen stimulation of the endometrium through an anti-proliferative effect and appears to reduce the incidence of endometrial carcinoma in post-menopausal women receiving estrogen replacement therapy (Beral et al., 1999. J. Epidemiol. Biostat., 4, 191-210). Such an adjunctive treatment, generally using synthetic progestogens, is given either in continuous combined regimens with estrogen, or added sequentially, typically for about 14 days each month, to continuous estrogen treatment.
Endogenous and exogenous estrogens fulfil important central nervous and metabolic functions in the female organism: normal estrogen levels make a decisive contribution to a woman's well-being. Notwithstanding the widespread use of estrogens in HRT methods, there are still some unsolved problems. Known estrogens, in particular the biogenic estrogens (i.e. estrogens naturally occurring in the human body), show serious pharmacokinetic deficits. Biogenic estrogens such as estradiol, estrone, estrone sulphate, esters of estradiol and estriol become bioavailable only to a very low degree when taken orally. This degree may vary so much from person to person that general dosage recommendations cannot be given. Fast elimination of these estrogens from the blood is another related problem. For instance, for the main human biogenic estrogen 17β-estradiol the half-life is around 1 hour. As a result, between separate (daily) administration events, blood serum levels of such biogenic estrogens tend to fluctuate considerably. Thus, shortly after administration the serum concentration is usually several times higher than the optimum concentration. In addition, if the next administration event is delayed, serum concentrations will quickly decrease to a level where the estrogen is no longer physiologically active.
The most important synthetically altered estrogenic steroid is 17α-ethinyl estradiol (EE). This estrogen is hardly used in HRT methods because prolonged administration of EE has been associated with an increased risk of thromboembolism, which is deemed to be particularly detrimental in menopausal and post-menopausal females. Apart from EE, mestranol has been used in a few cases; mestranol is a "prodrug" that is metabolised to EE in the organism. When applied orally to humans, EE has a much better bioavailability than the biogenic estrogens mentioned above, but its oral bioavailability varies to a large extent from individual to individual. Several authors have pointed to this as well as to the fact that concentrations in the blood proved to be highly fluctuating after oral application of this substance.
In addition to pharmacokinetic problems, the known estrogens also show pharmacodynamic deficits. After resorption from the intestinal lumen, orally applied active ingredients enter the organism via the liver. This fact is of specific importance for estrogenic agents as the liver is a target organ for estrogens; oral intake of estrogens results in strong estrogenic effects in the liver. The secretion activity that is controlled by estrogens in the human liver includes increased synthesis of transport proteins CBG, SHBG, TBG, several factors that are important for the physiology of blood clotting, and lipoproteins. If biogenic estrogens are introduced to the female organism while avoiding passage through the liver (e.g. by transdermal application), the liver functions mentioned remain largely unchanged. Therapeutically equivalent doses of biogenic estrogens, when applied orally, result in clear responses of hepatic parameters, such as increase of SHBG, CBG, angiotensinogen and HDL (high density lipoprotein). These hepatic effects of estrogens are also observed when equine estrogen formulations (so-called conjugated estrogens) are used. Ethinyl estradiol and diethylstilbestrol (DES) have an even greater hepatic estrogenicity. Elger et al., J. Steroid Biochem. Molec. Biol. (1995), 55(3/4), 395-403, have reported that EE or DES have much higher hepato-cellular than systemic estrogenicity: in relation to FSH-secretion inhibitory activity these estrogens are 4-18 times more active in the liver than estrone sulfate.
The aforementioned deficits are of considerable clinical significance when commonly known biogenic and synthetic estrogens are applied. Consequently, there is an as yet unmet need for estrogens that do not display these deficits and which can suitably be administered orally in HRT methods to effectively replace endogenous ovarian secretion of estradiol, i.e. to treat or prevent symptoms of hypoestrogenism.
The inventors have surprisingly found that these objectives are met by estrogenic substances that are represented by the following formula
A known representative of this group of estrogenic substances is 1,3,5(10)-estratrien-3,15α,16α,17β-tetrol, also known by the names of estetrol, oestetrol and 15α-hydroxyestriol. Estetrol is an estrogen that is produced by the fetal liver during human pregnancy. Unconjugated estetrol levels in maternal plasma peak at about 1.2 ng/ml at term pregnancy and are about 12 times higher in fetal than in maternal plasma (Tulchinsky et al., 1975. J. Clin. Endocrinol. Metab., 40, 560-567).
In 1970, Fishman et al., "Fate of 15α-hydroxyestriol-3H in Adult Man", J Clin Endocrinol Metab (1970) 31, 436-438, reported the results of a study wherein tritium labeled 15α-hydroxyestriol (estetrol) was administered intravenously to two adult women. It was found that the estetrol was rapidly and completely excreted in urine as the glucosiduronate and that virtually no metabolism except for conjugation took place.
Between 1975 and 1985 several researchers have investigated the properties of estetrol and reported on its estrogenic potency and uterotrophic activity. The most relevant publications that were issued during this period are mentioned below:
All of the above publications have in common that the authors have investigated the estrogenic potency of estetrol. Without exception they all conclude that estetrol is a weak estrogen. In some of the cited articles the estrogenic potency of estetrol has been found to be lower than that of another biogenic estrogen, namely, 17β-estradiol, which is considered to be a relatively weak estrogen (e.g. compared to ethinyl estradiol). With these findings in mind, it is not surprising that the interest in estetrol has dwindled since the early eighties and that no publications on the properties of estetrol have been issued since.
U.S. Pat. No. 5,468,736 (Hodgen) describes a method of hormone replacement therapy involving the administration of estrogen together with an amount of antiprogestin (antiprogestogen), which inhibits estrogen-induced endometrial proliferation in women. In Example 3 the combined use of estetrol and lilopristone is mentioned. No clues are given in the examples as to the mode and frequency of administration or regarding the dosage level employed. A disadvantage associated with the use of antiprogestogens, such as lilopristone, is the risk of inducing abnormal endometrial morphology, i.e. cystic hyperplasia, as has been observed in women who received an antiprogestogen treatment against endometriosis (Murphy et al., 1995. Fertil. Steril., 95, 761-766).
U.S. Pat. No. 5,340,586 (Pike et al.) is concerned with compositions and methods which are effective to treat oophorectomised women, wherein an effective amount of an estrogenic composition and an androgenic composition are provided over a period of time. In the U.S. patent it is stated that natural and synthetic estrogenic compositions that can be used include natural estrogenic hormones and congeners, including but not limited to estradiol, estradiol benzoate, estradiol cypionate, estradiol valerate, estrone, diethylstilbestrol, piperazine estrone sulfate, ethinyl estradiol, mestranol, polyestradiol phosphate, estriol, estriol hemisuccinate, quinestrol, estropipate, pinestrol and estrone potassium sulfate, and furthermore that equine estrogens, such as equilelinin, equilelinin sulfate and estetrol, may also be employed. Except for the exhaustive inventory of known estrogens, no other reference to estetrol (which is erroneously referred to as an equine estrogen) is made in this U.S. patent.
The same exhaustive list of estrogens is found in the following patent documents:
The benefits of the present invention may be realised without the co-administration of anti-progestogens, LHRH compositions, GnRH compositions and/or antisense oligonucleotides that are complementary to the nucleotide sequence of the follicle stimulating hormone (FSH) receptor as proposed in the aforementioned patents. Also, the present invention may suitably be applied in individuals who have not been oophorectomised, or in whom the risk of endometrial stimulation by estrogenic compositions is not minimised or absent, other than through the co-administration of a progestogen. Furthermore the present method does not require the use of a slow release formulation as is dictated by most of the aforementioned publications.
It is noted that none of the aforementioned publications describe the oral administration of estetrol. The only modes of administration described therein are intravenous and subcutaneous (depot) administration. For each of these modes of administration it can be concluded that the performance of estetrol is very much inferior to that of e.g. 17β-estradiol. Given that there was no reason to assume that a different outcome might be obtained in case of oral administration, it is not surprising that oral administration of estetrol has not been pursued and that no reports to this effect can be found in the prior art.
In view of the low estrogenic potency of the estetrol-like substances that are employed in accordance with the invention, it is surprising that these substances can effectively be used in HRT methods, particularly in HRT methods that employ oral administration of such substances. Although the inventors do not wish to be bound by theory, it is believed that the unexpected efficacy of orally administered estetrol-like substances results from the combination of unforeseen favourable pharmacokinetic (ADME) and pharmacodynamic properties of these substances.
As regards the pharmacokinetic properties of the present estrogenic substances the inventors have discovered that their oral bioavailability is surprisingly high and that their in vivo half-life is considerably longer than that of other biogenic estrogens. Thus, even though estetrol and estetrol-like substances have relatively low estrogenic potency, they may effectively be employed in an oral HRT method because their low potency is compensated for by a relatively high oral bioavailability in combination with a high metabolic stability, as demonstrated by a long half-life.
An important advantage of oral administration of estetrol and estetrol-like substances resides in the fact that the hepatic effects of estetrol-like substances are deemed to be minimal since they are hardly metabolised during the so called "first pass". The first-pass effect of drugs given orally, refers to the process of drug degradation by the liver during a drug's transition from initial ingestion to circulation in the blood stream.
Another advantageous property of the present estrogenic substances resides in the fact that sex hormone-binding globulin (SHBG) hardly binds these estrogenic substances, meaning that, in contrast to most known estrogens, serum levels are representative for bio-activity and independent of SHBG levels.
Yet another important benefit of the present estrogenic substances is derived from their relative insensitivity to interactions with other drugs (drug-drug interactions). It is well known that certain drugs may decrease the effectiveness of estrogens, such as ethinyl estradiol, and other drugs may enhance their activity, resulting in possible increased side-effects. Similarly estrogens may interfere with the metabolism of other drugs. In general, the effect of other drugs on estrogens is due to interference with the absorption, metabolism or excretion of these estrogens, whereas the effect of estrogens on other drugs is due to competition for metabolic pathways.
The clinically most significant group of estrogen-drug interactions occurs with drugs that may induce hepatic microsomal enzymes which may decrease estrogen plasma levels below therapeutic level (for example, anticonvulsant agents; phenyloin, primidone, barbiturates, carbamazepine, ethosuximide, and methosuximide; antituberculous drugs such as rifampin; antifungal drugs such as griseofulvin). The present estrogenic substances are less dependent on up- and downregulation of microsomal liver enzymes (e.g. P450's) and also are less sensitive to competition with other P450 substrates. Similarly, they do not interfere significantly in the metabolism of other drugs.
The conjugates of most estrogens, as formed in the liver, are excreted in the bile and may be broken down by gut bacteria in the colon to liberate the active hormone which can then be reabsorbed (enterohepatic recirculation). There are clinical reports that support the view that enterohepatic recirculation of estrogens decreases in women taking antibiotics such as ampicillin, tetracycline, etc. Conjugated forms of the present estrogenic substances are hardly excreted in the bile, meaning that they are substantially insensitive to drugs that do influence the enterohepatic recirculation of other estrogens.
The above observations serve to explain why the estrogenic substances of the invention hardly suffer from drug-drug interactions and thus produce a very consistent, i.e. predictable, impact. Thus, the efficacy of the estrogenic substances of the invention is highly reliable.
Accordingly one aspect of the present invention relates to a method of hormone replacement in mammals, which method comprises the oral administration of an estrogenic component and a progestogenic component to a mammal in an effective amount to prevent or treat symptoms of hypoestrogenism, wherein the estrogenic component is selected from the group consisting of:
substances represented by the following formula
The HRT method according to the invention may advantageously be used to treat all known forms of hypoestrogenism, e.g. hypoestrogenism associated with (peri-)menopausal and post-menopausal women, hypoestrogenism resulting from hypogonadism or castration, as well as hypoestrogenism resulting from primary ovarian failure, treatment of e.g. breast cancer with aromatase inhibitor and gonadotropin-releasing hormone analogue treatment of e.g. benign gynaecological diseases. Examples of manifestations of hypoestrogenism that can effectively be treated or prevented with the present method in both females and males include osteoporosis, arteriosclerosis, cognitive disturbances and Alzheimer's disease. The method may also advantageously be used in the (prophylactic) treatment of climacteric symptoms such as hot flushes (flashes), sweats, urogenital atrophy, mood disturbances, insomnia and palpitations. The present method is particularly suited for treating or preventing osteoporosis and climacteric symptoms.
The term "estrogenic component" as used throughout this document encompasses substances that are capable of triggering an estrogenic response in vivo, as well as precursors that are capable of liberating such an estrogenic component in vivo when used in accordance with the present invention. In order for estrogenic components to trigger such a response they normally have to bind to an estrogen receptor, which receptors are found in various tissues within the mammalian body. The term "progestogenic component" is defined as a substance that is capable of triggering an progestogenic response in vivo or a precursor which is capable of liberating such a substance in vivo. Usually progestogenic components are capable of binding to a progestogen receptor.
It is noted that the present invention not only encompasses the use of estrogenic and progestogenic components specifically mentioned in this application, but also metabolites of these hormones that display comparable in vivo functionality. In this context it is observed that, for instance, levonorgestrel is a metabolite of norgestimate and that estriol is a metabolite of 17beta-estradiol. Both these progestogens and estrogens have found application in contraceptive formulations and/or hormone replacement therapy. The term "estrogenic substances" as used in this document does not encompass tritium (3H) labeled estrogenic substances such as tritium labeled estetrol.
The present estrogenic substances are distinct from both the biogenic and synthetic estrogens that are commonly applied in pharmaceutical formulations in that they contain at least 4 hydroxyl groups. The present substances are special in that the 5 membered ring in the steroid skeleton comprises 3 hydroxyl substituents rather than 0-2.
Known estrogens that contain at least 4-hydroxyl groups and derivatives thereof are:
1,3,5(10)-estratrien-2,3,15α,16α,17β-pentol 2-methyl ether
1,3,5(10)-estratrien-2,3,15β,16α,17β-pentol 2-methyl ether
1,3,5(10)-estratrien-3,4,16α,17β-tetrol 4-methyl ether
1,3,5(10)-estratrien-3,15α,16α,17β-tetrol tetra acetate
1,3,5(10)-estratrien-3,15β,16β,17β-tetrol tetra acetate
Preferably, the estrogenic substance applied as the active component in the present composition is a natural estrogen, i.e. an estrogen that is found in nature and especially in mammals. Even more preferably, the estrogenic substance is a so called biogenic estrogen, i.e. an estrogen that occurs naturally in the human body, a precursor of a biogenic estrogen or mixtures thereof. Because biogenic estrogens are naturally present in the fetal and female body, side-effects are not expected to occur, particularly not if the serum levels resulting from the exogenous administration of such estrogens do not substantially exceed naturally occurring concentrations. Since estetrol serum levels in the fetus are several times higher than those found in pregnant females and knowing that the fetus is particularly vulnerable, estetrol is deemed to be a particularly safe biogenic estrogen. Side-effects are not expected to occur, particularly not if the serum levels resulting from the exogenous administration of such estrogens do not substantially exceed naturally occurring (fetal) concentrations. With synthetic estrogens such as ethinyl estradiol there is a (dose dependent) risk of undesirable side-effects, such as thromboembolism, fluid retention, nausea, bloating, cholelithiasis, headache and breast pain.
In a preferred embodiment of the present invention the estrogenic substance contains 4 hydroxyl groups. Also, in the aforementioned formula, R1 preferably represents a hydrogen atom. In said formula preferably at least 2, more preferably at least 3 of the groups R1, R2, R3 and R4 represent a hydrogen atom.
The estrogenic substances according to the formula encompass various enantiomers since the carbon atoms that carry hydroxyl-substituents R5, R6 and R7 are chirally active. In one preferred embodiment, the present estrogenic substance is 15α-hydroxy substituted. In another preferred embodiment the substance is 16α-hydroxy substituted. In yet another preferred embodiment, the substances is 17β-hydroxy substituted. Most preferably the estrogenic substances are 15α,16α,17β-trihydroxy substituted.
In another preferred embodiment of the present invention R3 represents a hydroxyl group or an alkoxy group. In another preferred embodiment the groups R1, R2 and R4 represent hydrogen atoms, in which case, if R3, R5, R6 and R7 are hydroxyl groups, the substance is 1,3,5(10)-estratrien-3,15,16,17-tetrol. A preferred isomer of the latter substance is 1,3,5(10)-estratrien-3,15α,16α,17β-tetrol (estetrol).
The invention also encompasses the use of precursors of the estrogenic substances that constitute the active component in the present method. These precursors are capable of liberating the aforementioned estrogenic substances when used in the present method, e.g. as a result of metabolic conversion. These precursors are preferably selected from the group of androgenic precursors as well as derivatives of the present estrogenic substances. Suitable examples of androgenic precursors include androgens that can be converted into the present estrogenic substances through in vivo aromatisation. Examples of derivatives of the present estrogenic substances that can suitably be used as precursors include such substances wherein the hydrogen atom of at least one of the hydroxyl groups has been substituted by an acyl radical of a hydrocarbon carboxylic, sulfonic acid or sulfamic acid of 1-25 carbon atoms; tetrahydrofuranyl; tetrahydropyranal; or a straight or branched chain glycosidic residue containing 1-20-glycosidic units per residue.
Typical examples of precursors which can suitably be used in accordance with the invention are esters that can be obtained by reacting the hydroxyl groups of the estrogenic substances with substances that contain one or more carboxy (M+−OOC—) groups, wherein M+ represents a hydrogen or (akali)metal cation. Hence, in a particularly preferred embodiment, the precursors are derivatives of the estrogenic substances, wherein the hydrogen atom of at least one of the hydroxyl groups in said formula has been substituted by —CO—R, wherein R is a hydrocarbon radical comprising from 1-25 carbon atoms. Preferably R is hydrogen, or an alkyl, alkenyl or aryl radical comprising from 1-20 carbon atoms.
The present method usually employs uninterrupted oral administration of the estrogenic component during a period of at least 10 days, preferably of at least 20 days. The term "uninterrupted" as used in here, means that the estrogenic component is administered at relatively regular intervals, with no (therapeutically) significant interruptions. Naturally, minor interruptions may occur that do not affect the overall effectiveness of the present method, and indeed such aberrations are encompassed by the present invention. In a preferred embodiment, and more arithmetically, the administration regimen is deemed to be continuous if the longest interval between 2 subsequent administrations is not more than 3.5 times as long as the average interval. Even more preferably said longest interval is not more than 2.5 times, most preferably not more than 1.5 times as long as the average interval.
The benefits of the present invention are most pronounced when the estrogen component is used in longer term hormone replacement therapy so as to minimise the negative effects of chronic hypoestrogenism. Therefore, the method of hormone replacement therapy, preferably, comprises administering the estrogenic component for a period of at least 1 month, more preferably of at least 3 months.
In the present method, the estrogenic and progestogenic component may be administered in separate oral dosage units. However, it is also possible and indeed very convenient to combine these two components into a single oral dosage unit.
The invention may suitably be reduced to practice in the form of a variety of HRT methods that are known to the person skilled in the art. Amongst these methods are the so called "combined" methods. The combined methods make use of preparations that contain a combination of an estrogen and a progestogen. The combined methods have in common that they are based on a regimen which involves administration of the aforementioned combined preparation, followed by an administration-free interval of about 7 days whereby withdrawal bleeding, simulating the natural menses, occurs. Thus 21 day intervals of hormone administration alternate with 7 days during which no hormones are administered.
As an alternative to the aforementioned combined methods, the so called "sequential" method has been proposed. Typical of the sequential method is that it comprises two consecutive phases, i.e. one phase during which estrogen and no progestogen is administered and another phase during which a combination of estrogen and progestogen is administered. The first sequential methods, like the aforementioned combined methods, made use of an administration free interval of about 7 days. More recently, sequential methods have been proposed which do not include an administration-free (or placebo) period, meaning that estrogen is administered throughout the full cycle and that progestogen is co-administered during only part of that cycle. WO 95/17895 (Ehrlich et al.) describes such an uninterrupted sequential method.
Yet another example of an HRT method which is encompassed by the present invention is the so called "continuous combined" method, which is a particular version of the combined method that uses uninterrupted combined administration of a progestogenic and an estrogenic component during a prolonged period of time, e.g. more than 50 days. In contrast to ordinary combined and sequential methods, no regular menses occur in the continuous combined method as the continuous administration of progestogen in the indicated amounts induces amenorrhoea.
In one embodiment of the invention, which relates to the continuous combined method, the present method comprises the uninterrupted oral administration of the combination of the estrogenic component and the progestogenic component during a period of at least 28, preferably at least 60 days.
In another embodiment of the invention, which relates to sequential and combined methods that employ a significant administration-free interval, the method of the invention comprises an interval of at least 2 days, preferably from 3-9 days, most preferably from 5-8 days, during which no progestogenic component and no estrogenic component is administered and wherein the resulting decrease in serum concentration of the progestogenic component and the estrogenic component induces menses.
Yet another embodiment of the invention, which concerns a sequential method without a significant pause, is characterised in that it comprises the uninterrupted oral administration of the estrogenic component during a period of at least 28 days, preferably at least 60 days, and in that, following the combined administration of the estrogenic component and the progestogenic component, the estrogenic component and no progestogenic component are administered during 3-18 consecutive days, preferably during 5-16 consecutive days and the resulting decrease in serum concentration of the progestogenic component should normally be sufficient to induce menses.
In the present methods uninterrupted administration of the estrogenic component may usually occur at intervals of between 6 hours and 7 days, preferably of between 12 hours and 3 days. The relatively high in vivo half-life of the present estrogenic components in comparison to most known estrogens makes it feasible to employ oral administration intervals that are significantly longer than 1 day. For practical reasons, and particularly with a view to user compliance, it is preferred to orally administer the estrogenic component as well as the progestogenic component at least once daily, most preferably once daily.
In all of the aforementioned methods it is preferred to orally administer the estrogenic component and the progestogenic component at least once daily during a period of at least 10, preferably of at least 20 days. In case of a sequential method without pause or a continuous combined method it is preferred to orally administer the estrogenic component and/or the progestogenic component at least once daily during a period of at least 30 days, more preferably of at least 60 days, most preferably of at least 150 days. Uninterrupted sequential methods, which employ continuous estrogen administration, are characterised by excellent cycle control.
The general concerns about the so called unopposed administration of estrogen, i.e. administration of estrogen without co-administered progestogen might cause hyperplasia of the endometrium, are less applicable to the estrogenic components of the present invention. Therefore, in a particularly preferred embodiment, the present HRT method is executed in accordance with a sequential method without pause.
Good results can be obtained with the present method if the estrogenic component is orally administered in an amount of less than 1 mg per kg of bodyweight per day, preferably of less than 400 μg per kg of bodyweight per day, more preferably of less than 200 μg per kg of bodyweight per day. In order to achieve a significant impact from the administration of the present estrogenic component, it is advisable to orally administer in an amount of at least 1 μg per kg of bodyweight per day. Preferably, the orally administered amount is at least 2 μg per kg of bodyweight per day. More preferably, the orally administered amount is at least 5 μg per kg of bodyweight per day.
In the present method, particularly when used in humans, the estrogenic component is usually administered in an average dosage of at least 0.05 mg per day, preferably of at least 0.1 mg per day. The maximum dosage is normally kept below 40 mg per day, preferably below 20 mg per day. The normally employed dose of the progestogenic component is equivalent to an average oral dosage of 30-750 μg levonorgestrel per day, preferably to an average oral dosage of 50-400 μg levonorgestrel per day.
In the present method, the estrogenic component is preferably administered in an amount effective to achieve a blood serum concentration of at least 1 nanogram per liter, more preferably of at least 10 nanogram per liter, most preferably at least 100 nanogram per liter. Generally the resulting blood serum concentration of the estrogenic component will not exceed 100 μg per liter, preferably it will not exceed 50 μg per liter, more preferably it will not exceed 25 μg per liter.
In accordance with the present invention the progestogenic component is advantageously administered in an amount which is equivalent to a daily oral dosage of 0.3 to 20 μg levonorgestrel per kg of bodyweight, preferably of 0.5-5 μg levonorgestrel per kg of bodyweight.
Examples of progestogens which may suitably be used in accordance with the present invention include: progesterone, levonorgestrel, norgestimate, norethisterone, dydrogesterone, drospirenone, 3-beta-hydroxydesogestrel, 3-keto desogestrel (=etonogestrel), 17-deacetyl norgestimate, 19-norprogesterone, acetoxypregnenolone, allylestrenol, anagestone, chlormadinone, cyproterone, demegestone, desogestrel, dienogest, dihydrogesterone, dimethisterone, ethisterone, ethynodiol diacetate, fluorogestone acetate, gastrinon, gestodene, gestrinone, hydroxymethylprogesterone, hydroxyprogesterone, lynestrenol (=lynoestrenol), medrogestone, medroxyprogesterone, megestrol, melengestrol, nomegestrol, norethindrone (norethisterone), norethynodrel, norgestrel (includes d-norgestrel and dl-norgestrel), norgestrienone, normethisterone, progesterone, quingestanol, (17alpha)-17-hydroxy-11-methylene-19-norpregna-4,15-diene-20-yn-3-one, tibolone, trimegestone, algestone acetophenide, nestorone, promegestone, 17-hydroxyprogesterone esters, 19-nor-17hydroxyprogesterone, 17alpha-ethinyl-testosterone, 17alpha-ethinyl-19-nor-testosterone, d-17beta-acetoxy-13beta-ethyl-17alpha-ethinyl-gon-4-en-3-one oxime and precursors of these compounds that are capable of liberating these progestogens in vivo when used in the present method. Preferably the progestogen used in the present method is selected from the group consisting of progesterone, desogestrel, etonogestrel, gestodene, dienogest, levonorgestrel, norgestimate, norethisterone, drospirenone, trimegestone, dydrogesterone, precursors of these progestogens and mixtures thereof.
The present method also encompasses the co-administration of active principles in addition to the progestogenic and estrogenic component. For instance, androgens may advantageously be co-administered in order to prevent symptoms of hypoandrogenicity. Thus, a preferred embodiment of the invention comprises the co-administration of an androgenic component. The androgenic component is suitably co-administered in an effective amount to suppress symptoms of hypoandrogenicity. Hypoandrogenicity has been associated with mood disturbances, unfavourable changes in haemostatic parameters and lack of bone mass.
The term "androgenic component" is defined as a substance that is capable of triggering an androgenic response in vivo or a precursor which is capable of liberating such a substance in vivo. Usually androgenic components are capable of binding to an androgen receptor.
Androgenic components that may suitably be employed in the present method may be selected from the group consisting of dehydroepiandrosterone (DHEA), danazol, gestrinone, testosterone esters, precursors capable of liberating these androgens when used in the present method and mixtures thereof. Preferably the testosterone esters employed comprise an acyl group which comprises at least 6, more preferably from 8-20 and preferably 9-13 carbon atoms. The androgens that can be used most advantageously in the present method are DHEA and/or testosterone undecanoate.
It is noted that, for instance, DHEA and testosterone undecanoate are precursors of testosterone and that said precursors per se exhibit virtually no affinity for androgen receptors in the female body. The effectiveness of the androgens within the method of the invention is determined by their functionally active form, which may well be different from the form in which they are administered.
In a preferred embodiment the androgen is provided in an amount equivalent to a daily oral dosage of 5 to 250 mg DHEA, which is equivalent to a daily oral dosage of 1 to 50 mg testosterone undecanoate. More preferably the androgen is provided in an amount which is equivalent to a daily oral dosage of 10-120 mg DHEA. Most preferably the androgen is administered in an amount which is equivalent to a daily oral dosage of 20-60 mg DHEA.
In order to obtain the desired impact from the present method it is advisable to administer the dosage units in an amount which leads to an increase in blood serum androgen level of no more than 5 nmole testosterone equivalent per liter, preferably less than 3 nmole testosterone equivalent per liter and most preferably less than 1.5 nmole testosterone equivalent per liter.
The present method preferably does not employ a gonadotropin hormone releasing hormone composition as described in the aforementioned patents U.S. Pat. No. 5,211,952, U.S. Pat. No. 5,340,584 and U.S. Pat. No. 5,340,585. Similarly, the present method preferably does not employ a luteinizing hormone releasing hormone composition as described in U.S. Pat. No. 4,762,717 and U.S. Pat. No. 5,130,137. Furthermore, the present method preferably does not comprise the co-administration of an anti-progestogen as described in U.S. Pat. No. 5,468,736. The method may also suitably be applied without the co-administration of an antisense oligonucleotide that is complementary to the nucleotide sequence of the follicle stimulating hormone (FSH) receptor (WO 00/73416).
The present method is preferably not used in oophorectomised females or in females in whom endometrial stimulation by estrogenic compositions is minimised or absent, other than by combined administration of a progestogen and an estrogen, e.g. as a result of hysterectomy.
Another aspect of the invention relates to a pharmaceutical kit comprising at least 20 oral dosage units that contains the estrogenic component as defined herein before and/or the progestogenic component and/or the androgenic component as described herein before, wherein at least 10 units contain between 0.01 and 20 mg, preferably between 0.05 and 10 mg of the estrogenic component, at least 10 units contain the progestogenic component in an amount equivalent to 30-750 μg levonorgestrel and at least 10 dosage units contain the androgenic component in an amount equivalent to 5-250 mg dehydroepiandrosterone.
In the present kit, the estrogenic component may conveniently be combined with the progestogenic component and the androgenic in a single dosage unit. Accordingly, the kit preferably comprises at least 10 dosage units which contain between 0.01 and 20 mg of the estrogenic component, the progestogenic component in an amount equivalent to 30-750 μg levonorgestrel and the androgenic component in an amount equivalent to 5-250 mg dehydroepiandrosterone. A pharmaceutical kit that is particularly suitable for use in combined and sequential methods will usually comprise 20-35 oral dosage units, wherein 10-35 units contain a combination of the estrogenic component and the progestogenic component in the indicated amounts, 0-25 units contain no progestogenic component and the estrogenic component in the indicated amounts, and 0-8 units contain no estrogenic component and no progestogenic component.
A pharmaceutical kit that is particularly suitable for use in a continuous combined regimen or a combined regimen comprises at least 20 oral dosage units which either contain the combination of the progestogenic and the estrogenic component or neither of these two components (placebo's) and of which dosage units at least 15, preferably at least 20 contain the combination of the estrogenic component and the progestogenic component and 0-8 contain no estrogenic component and no progestogenic component. If such a kit is to be used in a continuous combined method, the kit may advantageously comprise at least 28, preferably at least 60 dosage units, all of which dosage units contain the combination of the estrogenic component and the progestogenic component in the amounts indicated above.
In case the present kit is meant to be used in an HRT method that employs an administration free interval so as to induce menses (e.g. a combined method or a sequential method with pause) the kit will usually comprise at least 3 units, preferably at least 5 units that contain no estrogenic component and no progestogenic component.
In a particularly preferred embodiment of the invention the present kit is designed for use in a sequential method. Such a kit will usually comprise 20-35 oral dosage units wherein 10-32 units contain the combination of the estrogenic component and the progestogenic component, and 3-18 units contain the estrogenic component and no progestogenic component. Particularly preferred is a kit that is designed for use in a sequential method without a significant pause. In such a kit, which will usually comprise 20-35 oral dosage units, 10-20 units contain a combination of the estrogenic component and the progestogenic component, 10-18 units contain the estrogenic component and no progestogenic component and at most 1 unit contains no estrogenic component and no progestogenic component.
The pharmaceutical kits according to the present invention will normally contain only one or more of the following types of oral dosage units: units that contain the combination of the estrogenic and the progestogenic component; units that contain the estrogenic component and no progestogenic component; and units that effectively function as placebo's. Preferably the kit comprises at least 20 units that contain the combination of the estrogenic and the progestogenic component or the estrogenic component and no progestogenic component.
If the present kit is to be used in a combined or sequential protocol, the oral dosage units are preferably arranged within the kit in a fixed sequence corresponding to the intended order of administration. Data indications may be provided on the packaging. The packaging may be a tube or box or a strip. The box may be circular, square, or otherwise shaped with the tablets being accommodated separately therein for ease of administration. Date indications may appear adjacent to each tablet corresponding with the days on which each tablet is to be taken. Some indication of the sequence in which the tablets are to be taken preferably appears on the packaging regardless of its form.
Generally speaking, the oral dosage units in the present kit are prepared according to well known pharmaceutical procedures. The active ingredient(s) are combined with a pharmaceutically acceptable excipient and converted into a pharmaceutically acceptable form for oral administration, e.g. a tablet, capsule, cachet, pellet, pill, powder or granules. The excipient may include appropriate pharmaceutical carriers such as diluents, binders and lubricants. For example gums, starches and sugars are commonly used as pharmaceutical carriers. Tablets and other oral dosage units can suitably contain materials such binders (e.g. hydroxypropylmethyl cellulose, polyvinyl pyrrolidine, other cellulosic materials and starch), diluents (e.g. lactose and other sugars, starch, dicalcium phosphate and cellulosic materials), disintegrating agents (e.g. starch polymers and cellulosic materials) and lubricating agents (e.g., stearates and talc).
The active ingredient(s) may comprise from about 0.01% by weight to about 50% by weight of the formulation in the dosage unit, the remainder consisting of excipient. The active ingredient(s) are compounded with the chosen carrier and in for example the case of a tablet form, placed in a tablet moulding apparatus to form the tablets. Alternatively the compounded material may be incorporated as a powder or granules in a capsule. Various other options that may suitably be used in accordance with the present invention are well known to the person skilled in the pharmaceutical art.
The present invention is further illustrated by the following examples, which, however, are not to be construed as limiting. The features disclosed in the foregoing description, in the following examples and in the claims may, both separately and in any combination thereof, be material for realising the invention in diverse forms thereof.
Vaginal cornification was chosen as a tissue-specific and estrogen-sensitive endpoint to determine the estrogenicity of estetrol (E4), after oral administration, in hypoestrogenic rats. 17α-ethinylestradiol (EE) and vehicle (10% ethanol/sesame oil) served as controls in these bioassays.
Uterine weight increase in the rat is more commonly used as a measure of estrogenicity. However, uterine weight also responds to progesterone, testosterone, and other agents not characteristically regarded as estrogens. In the early 1920s it was discovered that follicular fluid from the pig ovary contained a factor(s) that caused cornification/keratinization of the vaginal epithelium in the rat (Allen and Doisy, 1923, JAMA, 81, 819-821; Allen and Doisy, 1924, Am. J. Physiol., 69, 577-588). The so-called vaginal cornification response in rats subsequently provided a bioassay for testing estrogenicity. Vaginal epithelial cornification/keratinization in ovariectomized rats can be produced only by compounds considered to be true estrogens (Jones et al, 1973, Fert. Steril. 24, 284-291). Vaginal epithelial cornification/keratinization represents, therefore, a highly selective endpoint to determine the potency of estrogens (Reel et al., 1996, Fund. Appli. Toxicol. 34, 288-305).
Adult intact female CD rats were ovariectomized to induce estrogen deficiency. Vaginal lavages were performed daily for seven days to ensure that the rats demonstrated castrate vaginal smears (predominance of leukocytes in the vaginal smear, and similar in appearance to a diestrous vaginal smear). Castrate vaginal smears are indicative that complete ovariectomy was achieved. Treatment commenced following completion of the 7 days of smearing (day 0=first day of dosing). Animals were dosed, once daily for 7 consecutive days. Daily vaginal lavages continued to be obtained for 7 days after dosing was initiated in order to detect vaginal cornification, as an indication of an estrogenic response. A drop of vaginal washings was placed on a glass slide and examined by light microscopy to detect the presence or absence of cornified epithelial cells. Vaginal lavages were obtained prior to dosing on days 0-6 and prior to necropsy on day 7.
The vaginal cornification bioassay was performed in order to determine the estrogenic profile of E4 when given orally (po) to ovariectomized adult rats. EE was used as a positive control. The vehicle (10% ethanol/sesame oil) served as the negative control. Steroids were dissolved in absolute ethanol and then brought to the final concentration with sesame oil (10% ethanol in sesame oil). A vaginal estrogenic response occurred in all rats (8/8) given 50 μg/kg/day EE po by day 7 (Table 1). Similarly, vaginal epithelial cornification was observed in all rats (8/8) treated po with either 0.1, 0.3, 1.0, or 3.0 mg/kg/day E4 by day 7 (Table 1), whereas animals treated with the vehicle did not exhibit vaginal epithelial cornification (0/8). Even in rats given relatively low doses of E4 (e.g. 0.1 and 0.3 mg/kg/day), the onset of vaginal cornification (defined as the amount of animals responding at days 1-3 of the study) was as fast as in EE-treated animals (Table 1), demonstrating estetrol's superb bioavailability characteristics after oral administration.
Table 1: Vaginal estrogenic response in ovariectomized rats treated orally (po) with 17α-ethinyl estradiol (EE) or estetrol (E4). Data are expressed as the number of rats showing vaginal cornification over the number of rats (ratio) treated.
The ovariectomized aged rat was used as a model for the human disease osteoporosis. This is an established animal model, recommended by the United States Food and Drug Administration (FDA), to evaluate and assess potential agents for osteoporosis prevention and therapy. The anti-resorptive efficacy of estetrol (E4) was tested by ex vivo measuring total and trabecular bone mineral density and bone strength after 4 weeks of treatment at necropsy. 17α-ethinyl-estradiol (EE) and vehicle (1% ethanol/arachidis oil) served as controls in this bioassay.
Three months old female Sprague-Dawley rats were either sham-operated (Sham) or ovariectomized (OVX) one day prior to commencement of the dosing study. Animals were anesthetized using a ketamine/xylazine anesthetic mixture and underwent a bilateral ovariectomy or were sham-treated. A section of hair on the dorsal surface was shaved and an incision made overlying the lumbar region of the spine. The skin was separated from the underlying fascia so that a second incision could be made through the abdominal musculature approximately caudal to the kidneys. The ovaries were then exteriorized and removed and the musculature was closed with a single suture. The skin incision was closed using surgical staples.
Ten animals per treatment group were orally dosed once per day for four consecutive weeks. The dosing commenced 1 day after surgical removal of the ovaries and was administered by oral gavage using a syringe and stainless steel gavage needle at doses of 0.1 mg/kg/day EE, or either 2.5, 0.5 or 0.1 mg/kg/day E4. Vehicle control was daily administered to one group of OVX-animals and sham-operated rats. After treatment, anesthetized rats were subjected to cardiac puncture and asphyxiated by CO2 inhalation. Tibiae and femura were removed, cleaned of soft-tissue and fixed and stored in 70% ethanol/saline at 4� C. (tibia) or saline at 4� C. (femura) until further analysis.
Ex vivo peripheral Quantitative Computed Tomography (pQCT) was performed on the excised left tibiae using a Stratec XCT-RM and associated software (Stratec Medizintechnik GmbH, Pforzheim, Germany, software version 5.40). The scans were performed at 12% of the total length from the proximal end of the tibiae. The positions were verified using scout views and one 0.5-mm slice perpendicular to the long axis of the tibial shaft was acquired from each site. The scans were analyzed using a threshold for delineation of the external boundary. The total and trabecular bone mineral content, area and density at each site were determined. Mean values are shown in Table 2. Furthermore, pQCT data for mean total bone mineral density are depicted in
Table 2: pQCT densitometry data from the proximal tibiae of Sham- and OVX-rats orally (po) treated with 17α-ethinyl estradiol (EE), estetrol (E4) or vehicle. Data are expressed as the mean values obtained for each group (n=10).
Comparison of the pQCT densitometry data from the proximal tibiae of Sham-operated and OVX-rats demonstrated a consistent loss of total and trabecular bone in the OVX-group, as expected (Table 2,
Ex vivo evaluation of bone biomechanical strength was performed with an indentation test at the distal femura. Prior to mechanical testing femura were rinsed in cold saline and carefully cleaned of any remaining adherent soft tissue. A 3-mm segment of the distal femoral metaphysis was cut directly proximal to the femoral condyle with a low-speed diamond saw under constant saline irrigation. The load was applied with a cylindrical indenter (with a flat testing face of 1.6 mm diameter) to the center of marrow cavity on the distal face of the segment. The indenter was allowed to penetrate the cavity at a constant displacement of 6 mm/min to a depth of 2 mm before load reversal.
Table 3: Indentation testing of the distal femur of Sham- and OVX-rats orally (po) treated with 17α-ethinyl estradiol (EE), estetrol (E4) or vehicle. Data are expressed as the mean values obtained for each group (n=10).
The maximum load, stiffness and energy absorbed were determined from load displacement curves. Ultimate strength was calculated by dividing the maximum load by the indenter area. Mean values of maximum load, stiffness, energy and ultimate strength are shown in Table 3. Furthermore, mean ultimate strength values are depicted in
The morphine-dependent ovariectomized (OVX) rat was used as a model for postmenopausal hot flush. The potency of estetrol (E4) to prevent tail skin temperature rises, normally accompanied by a drop in core body temperature, after naloxone-induced opiate withdrawal was tested. 17α-ethinyl-estradiol (EE) and vehicle (hydroxy propyl-beta-cyclodextrin 20% wt/vol) served as controls in this bioassay.
The most common and characteristic symptom of human menopause is the hot flush, which is experienced by over 70% of menopausal females. While the exact mechanism underlying this vasomotor instability is unknown, the characteristic features of the hot flush appear to reflect a centrally mediated adaptation to a progressive decline in the levels of estrogens. In women experiencing the hot flush the symptoms are manifested by 1) rapid, regional elevations in skin temperature; 2) a decrease in core body temperature; 3) an increased heart rate with no change in blood pressure; and 4) closely timed surges in release of luteinizing hormone (LH) and β-endorphin.
The morphine-dependent ovariectomized (OVX) rat model has been proposed by several investigators (Katovich et al, 1986, Maturitas, 67-76; Merchenthaler et al. 1998, Maturitas, 307-316) as an animal model for the hot flush. During opiate withdrawal with the morphine antagonist naloxone, tail skin temperature (TST) rises and this rise is accompanied by a drop in core body temperature. In addition, the temperature changes are accompanied by surges in LH and a transient tachycardia. These events are similar in magnitude and temporal nature to those observed in the menopausal hot flush.
8-week-old OVX rats were treated orally (po) with estetrol (E4), 17α-ethinyl estradiol (EE) or vehicle control (hydroxy propyl-beta-cyclodextrin 20% wt/vol) for seven consecutive days prior to, and on the morning of naloxone-induced opiate withdrawal in morphine-dependent animals. Three days prior to the commencement of dosing, animals were anesthetized using a ketamine/xylazine anesthetic mixture and underwent a bilateral ovariectomy. A section of hair on the dorsal surface was shaved and an incision made overlying the lumbar region of the spine. The skin was separated from the underlying fascia so that a second incision could be made through the abdominal musculature approximately caudal to the kidneys. The ovaries were then exteriorized and removed and the musculature was closed with a single suture. The skin incision was closed using surgical staples. Six rats per treatment group were dosed once per day for eight consecutive days prior to and including the day of naloxone-induced opiate withdrawal (the hot flush session). The dosing commenced three days after surgical removal of the ovaries and was administered by oral gavage using a syringe and stainless steel gavage needle. Morphine dependency was induced by implantation of subcutaneous pellets containing 75-mg morphine. The first pellet was implanted five days before the hot flush session under a light inhalation anesthesia. Three days before the hot flush session, two additional morphine pellets were implanted under the same conditions.
For the hot flush manipulations the animals were placed in a test cage. Following a 5-10 minute adaptation period, the rats were anesthetized with ketamine HCl approximately 10 minutes prior to the hot flush session. A temperature sensitive electrode was fixed to the ventral surface of the tail with tape and the electrode was connected to a multi-channel temperature recorder. The tail-skin temperature was recorded until it was stable and the animals were then injected with naloxone HCl (1 mg/kg). The temperature recordings then continued for a period of 60 minutes and the temperature was reported at 5-minute intervals. At the completion of the hot flush session, all animals were killed using CO2 asphyxiation followed by cervical dislocation.
As expected, vehicle control was ineffective in preventing the naloxone-induced TST increases in the morphine addicted OVX rats (
To evaluate the oral bioavailability of estetrol (E4) and to determine the elimination half-life, single oral (po) and subcutaneous (sc) dose studies were performed in female Sprague Dawley rats followed by frequent blood sampling over a 24 hours interval.
Female Sprague Dawley rats were equipped with a permanent silatic heart catheter, as described by Kuipers et al. (1985, Gastroenterology, 88, 403-411). Rats were allowed to recover from surgery for 5 days and were than administered 0.05, 0.5, or 5 mg/kg E4 in 0.5 ml arachidis oil. For sc administration, E4 was injected in the neck area using a 1 ml syringe and 20 g needle. For po administration of E4, rats were lightly anaesthesized with halothene/N2O/O2 and E4 was directly applied intragastrically using a plastic stomach intubator. Blood samples were subsequently collected via the heart catheter in heparinized tubes at 0.5, 1, 2, 4, 8 and 24 hours. Erythrocytes were removed by centrifugation at 5000�g for 10 minutes at 4� C. and blood plasma was stored at −20� C. After thawing the plasma samples, liquid-liquid extraction (hexane and diethyl ether) was employed to prepare the E4-containing plasma samples for HPLC analysis (Perkin Elmer 200) and tandem mass spectrometry using a PE Sciex 3000 tandem mass spectrometer and APCI interface. With each sample batch, a calibration curve with 6 calibrators was recorded. The calibration curve was calculated using linear regression (correlation coefficient >0.98), which permitted quantitation of plasma concentrations. For each rat plasma, sampled at different time intervals, data were collected.
Plasma E4 concentration data were analysed with "WinNonLin, edition 3.1" and involved pharmacokinetic parameters for Cmax, half-life and AUC0-24. Especially, using the lower and intermediate dose levels of 0.05, 0.5 mg/kg, E4 demonstrated an oral bioavailability equal to the bioavailability obtained with sc administration (80-100%). At the highest dose level tested, 5.0 mg/kg E4, absorption kinetics gave rise to an oral bioavailability approximating 30-60% of sc administered E4. Interestingly, E4 demonstrated a relatively long half-life of 2-3 hours, enabling the detection of bioactive levels of unconjugated E4 at all time points over a 24 hour interval.
An established competitive steroid-binding assay (Hammond and Lahteenmaki. 1983. Clin Chem Acta 132:101-110) was used to determine the relative binding affinity of estetrol (E4), 17α-ethinylestradiol (EE2), 17β-estradiol (E2), testosterone (T) and 5α-dihydrotestosterone (DHT) for human sex Hormone Binding Globulin (SHBG).
Human SHBG was purified from transgenic mouse serum, as described previously (Avvakumov G V et al., 2000. J Biol Chem 275: 25920-25925). The human SHBG prepared in this way was assessed to be >99% pure by polyacrylamide gel electrophoresis under denaturing conditions. Its steroid-binding characteristics are indistinguishable from SHBG in human serum (Avvakumov G V et al., 2000. J Biol Chem 275: 25920-25925). The in vitro assay involved the use of the purified human SHBG and [3H]DHT or [3H]estradiol as labeled ligands. Human SHBG was treated for 30 min at room temperature with a dextran-coated charcoal (DCC) suspension in phosphate buffered saline (PBS) to remove any steroid ligand. After centrifugation (2,000�g for 10 min) to sediment the DCC, the supernatant containing the human SHBG was diluted in PBS to a concentration of 1 nM based on its steroid binding capacity.
Duplicate aliquots (100 μl) of this human SHBG solution were then incubated with an equal volume of either [3H]DHT or [3H]estradiol at 10 nM, together with 100 μl of PBS alone or the same amount of PBS containing increasing concentrations of unlabeled steroid ligands as competitors in polystyrene test tubes. After incubation for 1 h at room temperature the reaction mixtures were placed in an ice bath for a further 15 min. Aliquots (600 μl) of an ice cold suspension of DCC were then added to each tube, and after a brief 2 seconds mixing, each tube was incubated in an ice bath for either 10 min or 5 min depending on whether [3H]DHT or [3H]estradiol were being used as labeled ligands, respectively. The unbound ligands adsorbed to DCC were then removed by centrifugation (2,000�g for 15 min at 4 C), and the amounts of [3H]labeled ligands bound to SHBG were counted in 2 ml ACS scintillation cocktail using in liquid scintillation spectrophotometer. The average amounts of [3H]labeled ligands bound to SHBG at each concentration of competitor (B) were expressed as a percentage of the average amounts of [3H]labeled ligands bound to SHBG in the absence of competitor (B0), and were plotted against the concentration of competitor in each assay tube.
The results of the competitive binding assays are depicted in
The present estrogenic components may suitably be processed, together with additives, excipients and/or flavouring agents customary in galenic pharmacy, in accordance with the conventional methods into the usual forms of administration. For oral administration, suitable are, in particular, tablets, dragees, capsules, pills, suspensions, or solutions.
Estetrol tablets: 1,000 tablets of 185 mg, containing 1.5 mg estetrol and 0.15 mg levonorgestrel, are produced from the following formulation:
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He recorded the testimony of men, women, and sometimes children who came face-to-face with the living, or at least the undead, hell of that dreadful time. World War Z is the result. Never before have we had access to a document that so powerfully conveys the depth of fear and horror, and also the ineradicable spirit of resistance, that gripped human society through the plague years.
Ranging from the now infamous village of New Dachang in the United Federation of China, where the epidemiological trail began with the 12-year-old Patient Zero, to the unnamed northern forests where untold numbers sought a terrible and temporary refuge in the cold, to the United States of Southern Africa, where the Redeker Plan provided hope for humanity at an unspeakable price, to the west-of-the-Rockies redoubt where the North American tide finally started to turn, this invaluable chronicle reflects the full scope and duration of the Zombie War.
Note: Some of the numerical and factual material contained in this edition was previously published under the auspices of the United Nations Postwar Commission.

subset: commoncrawl


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Rooney has been linked with a move away from the club after he was left out of the starting line-up for the Champions League second-leg tie with Real Madrid on Tuesday.
Ferguson said of Rooney: "He will be here next year, you have my word on that.
"The issue you're all going on about in the papers is absolute rubbish.
"There is absolutely no issue between Wayne Rooney and I. To suggest we don't talk to each other on the training ground is absolute nonsense.
"There are no issues with the player and he will be involved on Sunday.
"Wayne understood the reasons for not playing him. It was completely tactical and tactically we got it right. We don't always but we definitely did on Tuesday.
"Danny Welbeck is the best player we have in terms of operating in a double role.
"We had to choke Xabi Alonso's ability to control the game, which Danny did, and that took away Alonso's control of the game and his ability to go further forward and be an attacking player."


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High ceilings call for dramatic lighting.
Large modern pendant lights float ethereally in this vast atrium above the dining room, where nothing else competes with the beauty of the slatted-wood wall and the natural stone floors.
For more lighting options, see our Statement Lighting photo gallery.
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The 28-year-old Brazil international is currently the highest paid player on the Wolfsburg pay roll but is out of contract come the end of the season.
Playmaker Diego has featured in 10 of Wolfsburg's 12 Bundesliga clashes to date and produced performances which have attracted European attention.
German media reports suggest English Premier League and Spanish La Liga clubs have registered interest in the midfielder.
Although Diego admits he knows there are offers on the table, the player insists there is no final decision made as yet.
Speaking to Kicker-Sport magazine he said: "Many people are obviously out to find me a new club to play for.
"But there is no decision to be taken just yet. It's a fact I am receiving many, many enquiries but there is still nothing official to reveal."
While transfer speculation continues to circulate in the media, Diego remains focussed on adapting to manager Hecking's developing formation.
"Actually, I am listening carefully to the Wolfsburg manager, I am doing a good job in trying to adapt to his ideas.
"I am attempting to become a better player."
Diego will travel with Wolfsburg to face Freiburg on Sunday afternoon in the Bundesliga as they look to continue climbing the Bundesliga rankings from sixth.


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(Click for full size images)
Apple has made it clear that it isn't going to get into the megapixel race with the likes of Nokia and Sony when it comes to the camera system used in its iPhones. The new iPhone 5s has the same 8-megapixel shooter as previous generations, but Apple is touting a brand new camera system for the device that includes a new five-element lens designed by Apple, a F2.2 aperture, a sensor with a 15 percent larger active area, auto stabilization, and bigger 1.5 micron pixels. However, the big new standout feature for the camera is a dual LED "True Tone" flash that should vastly improve shooting in poor lighting conditions. The real question is, how does Apple's new camera system in the 5s stack up against some of the best smartphone cameras in the game: That's the 41 megapixel PureView camera in the Nokia Lumia 1020, and the 20.7 megapixel camera with Sony Exmor RS mobile image sensor packed into the Sony Xperia Z1.
Recombu put together the comparison above which shows how the three cameras perform under the same nighttime lighting conditions for a portrait. We'll let you decide, but, as the site points out, the iPhone 5s seems to do a much better job of producing accurate colors:
Admitedly, the dual-LEDs can't rival the illuminating clout of the Nokia Lumia 1020's Xenon flash, but with its combi warm and cool LEDs, the 5S clearly delivers a significantly more accurate shot – just look at the white canopies!
With the camera on the Nokia Lumia 1020 one of the device's big selling points, it's not surprising Nokia has gone after Apple in the past with attack ads claiming the iPhone 5 takes subpar photos.
Leave a Reply
19 Responses to "iPhone 5s camera w/ True Tone flash vs Nokia Lumia 1020 & Sony's Xperia Z1"
iPhone 5s: Natural but a bit faded
Nokia Lumia 1020: Very detailed but quite saturated
Sony Z1: A mix of 5s and 1020 but a bit blurry
Personal opinion
The photo from Nokia looks totally idiotic. 5s and Xperia, great photos but I kinda like the 5s photo better.
I do not believe this …
The z1 and iPhone 5s does fine, but the Lumia 1020 is set to a wrong whitebalance, and the Lumia 1020 is the only one with a manuel whitebalance setting.
When you take a image with a camera it knows the colortone of the flash so the whitebalance will match that.
The exif data says that Lumia 1020 was on auto whitebalance – so…
If you need to futz with a manual white balance to get a decent shot, you're a crappy cell camera.
These aren't for professional photographers. They're meant for everyday folks to be able to just snap a great pic.
Maybe that's why the Lumia bombed.
He meant that the nokia was set to manual white balance when it should have been set to auto.
Home chick checked the exif data!
The Nokia looks horrible. 5S and Experia, I like them both for different reasons. The 5S has a better skintone and the Experia does a better job in lighting out the general scene/background.
Please, check this: http://www.nbcnews.com/technology/smartphone-camera-battle-iphone-5s-faces-stiff-competition-4B11218947
Lmao! The iPhone's outdoor shot is much crisper than the Lumia's, and the Lumia's indoor shot with flash turned the guy's skin red.
Devin is a known MS slappy. His opinion is as far from gospel as one can get.
by the look ,5s -original colors
Nokia-little different(gud pic)
my bf is being a dumbass and saying your experiment in inconclusive because iPhone photo is smaller in size then the other two, like it makes a fucking difference. Seriously dude just shut the fuck up! He's been an apple hater for ever yet i give him an iPhone and it's all he plays with, but he still wants to talk shit when other people praise it. he really is fucking stupid!
So, looks like based on 1 picture comparison, everyone is jumping out of joy. I love iphone, but credit should be given where it's due. It's a known fact that Lumia 1020 is better than any phone camera as of now, including iphones.
Below is a good review of Lumia's camera. I believe, it will get even more better with future SW updates.


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Building Demolished
Updated: 02/23/2005 - A Lauderdale County crew removed an old store building that had officials said had become a safety hazard.
Updated: 02/23/2005 - A Lauderdale County crew removed an old store building that had officials said had become a safety hazard.
Updated: 02/23/2005 - As the Mississippi Medicaid debate continues, a prominent figure involved says using money from the state's tobacco settlement is not the answer.
Updated: 08/02/2005 - Details of Lauderdale County's 2005 Relay for Life were announced Wednesday.
Updated: 02/23/2005 - An academic study released Wednesday points to Newton County High School for "doing things right."
Updated: 02/22/2005 - When it comes to budget cuts, officials with Mississippi's community colleges say enough is enough. School representatives lobbied Tuesday in Jackson.
Updated: 02/22/2005 - Project ChildSafe's second safety tour made a stop in east Mississippi Tuesday as the National Shooting Sports Foundation distributes 12 million gun locks in 2005.
Updated: 02/22/2005 - Winn-Dixie Stores announced it has filed for bankruptcy reorganization after reporting increased losses from a year ago.
Updated: 02/22/2005 - The Mississippi House of Representatives has approved a bill that includes $10 million in bonds for the Mississippi Arts and Entertainment Center in Meridian.
Updated: 02/22/2005 - Former New York City Mayor Rudolph Giuliani joined U.S. Sen. Trent Lott in Jackson Tuesday to help the Republican lawmaker raise money for a 2006 reelection campaign.
Updated: 02/22/2005 - The appeals of four Mississippi death row inmates have been declined by the U.S. Supreme Court.
Updated: 02/21/2005 - Evangel Temple in Meridian has plans to build a new theatre to accommodate large audiences for its Christian productions.
Updated: 02/21/2005 - Sports at some levels may involve pushing and shoving, maybe even a little name calling, but at North Park Church in Meridian, there's a little something different going on called "Upward Basketball."
Updated: 02/21/2005 - Meridian's Northwest Junior High School saluted important contributions made by African-Americans in a special program Monday.
Updated: 02/21/2005 - Singer Faith Hill visited her home state on behalf of the Mississippi Arts and Entertainment Center, slated to be built in Meridian.
Updated: 02/21/2005 - A veteran employee of Northrop Grumman is accused of opening fire at work Monday morning, seriously injuring two other workers.
Updated: 02/21/2005 - The Kemper County Sheriff's Department has charged a suspect in a deadly shooting Sunday that claimed the life of a local high school student.
Updated: 02/21/2005 - Alabama Gov. Bob Riley says the state's long-running dispute with Medicaid is over.
Updated: 02/21/2005 - The clock is ticking for Mississippi lawmakers to come up with a solution on how to address the state's Medicaid deficit.
Updated: 02/20/2005 - Members of the Mississippi National Guard's Company G, 185th Aviation returned home to a huge welcome at the armory in Meridian.
Updated: 02/20/2005 - The fatal crash of a small aircraft in Lauderdale County recently may have raised safety concerns, but at least one Mississippi pilot defends the safety of single engine planes.
Updated: 02/20/2005 - A Mississippi man was sentenced to death for the fatal shooting and stabbing of a retired minister.
Updated: 02/20/2005 - About a dozen coyotes have been roaming around a Lamar County, Miss. neighborhood, causing fear for some residents.
Updated: 02/20/2005 - Gov. Haley Barbour's "UpGrade" education package is touted as a way to make significant changes without a hefty price tag. But some education officials wonder about its effectiveness.
Updated: 02/18/2005 - Members of the Meridian City Council discussed holiday pay for police officers and firefighters, but the controversial issue is still undecided.
Updated: 02/18/2005 - Former hostage Thomas Hamill has a new lease on life and wrote a book, but he says he's not exactly on "Easy Street."


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IT Asset Managers Need Strong Communication, Admin Skills: Gartner
By Datamation.com Staff | Article Published July 16, 2012
A new reports says communication skills are more important than technical skills for IT asset managers.
Tags: IT managers, Gartner, asset management, communication skills
Asian Recruiters on the Prowl for IT Managers
By Datamation.com Staff | Article Published April 05, 2012
A growing number of IT managers from the U.S. and the U.K. are taking work in Southeast Asia.
Tags: China, IT jobs, IT managers
9 IT Technologies that Drive Product Development
By Larry Marion | Article Published March 16, 2011
A survey of managers finds that companies working to improve their product development need these IT tools.
Tags: software, IT, IT managers, Enterprise IT, IT infrastructure
50 Android Apps for Business Travelers
By Thor Olavsrud | Article Published March 16, 2011
Android apps to help travelers get where theyre going, find a place to eat, stay in touch, and more. Many free Android apps.
Tags: Android, Android Marketplace, android apps, IT managers, business software
HP Updates Project Portfolio & Management Software
By David Needle | Article Published March 01, 2011
The latest 9.1 release of HP Project Portfolio & Management plays into the computer giant's "Instant-On Enterprise" set of solutions.
Tags: IT managers, enterprise applications, Enterprise IT, HP, project management software
Android Apps for Linux IT Managers
By Eric Geier | Article Published February 24, 2011
Android apps that help you run your data center.
Tags: Android, android apps, IT managers
Cloud Computing and Microsoft Exchange Coexistence
By Datamation.com Staff | Article Published February 16, 2011
A look at the advantages of both on-premise and off-premise Exchange hosting.
Tags: cloud computing, IT managers, Exchange, Microsoft Exchange, hosting
Horror Stories of IT Management
By Carla Schroder | Article Published February 01, 2011
There's plenty of finger pointing about problems, but who's really at fault?
Tags: IT Jobs/Salary, IT managers, Enterprise IT, IT management job, network manager
Mobile App Growth Challenges IT Managers in 2011: Survey
By David Needle | Article Published January 14, 2011
Security concerns and the proliferation of mobile operating systems pose a looming challenge.
Tags: IT, IT managers, Sybase, mobile apps, mobile security
Five Key Data Center Staffers
By Kenneth Hess | Article Published January 11, 2011
The central players in the daily drama of keeping your data center functional.
Tags: security, IT managers, data center jobs, help desk, data center facilities management
Two Top Acquisitions Execs Leaving Microsoft
By Stuart J. Johnston | Article Published January 06, 2011
Two key merger and acquisition executives are leaving Microsoft, but is it problems with the company's strategy or greener grasses that are at fault?
Tags: Microsoft, enterprise software, IT managers, Yahoo, mergers and acquisitions
What Hiring Managers Look for in IT Professionals
By Jim Sloan | Article Published December 15, 2010
IT job applicants need technical skills, but hiring managers are looking for other qualities as well. Here are five qualities that can help you rise above the competition.
Tags: IT Jobs/Salary, IT managers, IT salary, computer management, information technology
IT Jobs: What Was Hot in Online Hiring in 2010 and What's Ahead?
By David Needle | Article Published December 08, 2010
Elance report recaps the year in IT hiring and predicts what skills will be in demand in 2011. Competitor oDesk weighs in as well.
Tags: software, IT Jobs/Salary, IT managers, developer salary, IT salary
Data Center Case Study: Efficiency and Consolidation
By Drew Robb | Article Published December 07, 2010
In an effort to lower energy costs, a midwestern data center is working towards consolidating its infrastructure.
Tags: IT managers, data center management, servers, energy conservation, data center consolidation
Why Paying Attention Is Getting More Expensive
By Mike Elgan | Article Published September 23, 2010
Technology is changing how and what we pay attention to, in ways that are dramatically altering business and personal life.
Tags: IT managers, marketing, online advertising, Online Marketing, advertising tools
Major Data Center Irritations (the Short List)
By Kenneth Hess | Article Published September 10, 2010
Spending time in a modern day data center may make you wonder why, exactly, things have gone so wrong.
Tags: IT management, IT managers, Data Center Manager, data center management, data center virtualization
Why Mark Hurd is a Bad Match for Oracle
By Rob Enderle | Article Published September 09, 2010
As a turnaround CEO, Hurd could be unmatched, but he would not be the best choice if you wanted to build a company rather than sell one.
Tags: Oracle, IT management, IT managers, HP, Hurd
Obama Calls for R&D Tax Credit Extension
By Kenneth Corbin | Article Published September 09, 2010
President touts technology agenda as a centerpiece to economic recovery, drawing plaudits from often-critical IT groups for the proposal to extend the tax credit for research and development.
Tags: IT management, IT Jobs/Salary, IT spending, IT managers, IT budget
IT Managers: Managing Engineers vs. Managing Support Staff
By Scott Alan Miller | Article Published September 01, 2010
IT managers need to manage these two groups knowing that the engineers' goal of productivity contrasts with support staff's focus on availability.
Tags: support, developers, IT Jobs/Salary, IT manager, IT managers
How IT Managers Can Stay Employed
By Pam Baker | Article Published July 07, 2010
By making a carefully considered business case -- and not tooting your own horn for its own sake -- IT leaders can more safely hang on to their berth.
Tags: IT Jobs/Salary, IT job, IT manager, IT managers, CIO's job
'The IT Manager's Job Is Not Getting Easier'
By Chris Nerney | Article Published August 03, 2009
'The IT Manager's Job Is Not Getting Easier'
Tags: social networks, mobile, IT managers, networks, IT security, attacks
Coping Tips For Managers Who Lay Off Workers
By Chris Nerney | Article Published March 25, 2009
Coping Tips For Managers Who Lay Off Workers
Tags: IT jobs, IT managers, layoffs, career, advice, CIOs
For IT Workers, Wireless Is Where It's At
By Heidi Cyre | Article Published April 14, 2008
For IT Workers, Wireless Is Where It's At
Tags: wireless, IT managers, IT security, IT survey, RF


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Funding Resources
Rural Funding Sources
USDA. NAL. Rural Information Center.
Check out this page, which serves as a gateway to major funding programs, tools and assistance for rural areas from both public and private sources.
Federal Funding Sources for Rural Areas Database
USDA. NAL. Rural Information Center.
Search the database for information on rural federal programs. Full program description and contact information for each program. This database is in cooperation with the Catalog of Federal Domestic Assistance.
A Guide to Funding Resources
USDA. NAL. Rural Information Center.
This full-text publication provides a variety of information that link to helpful web sites and non-electronic publication citations that assist in identifying funding programs.
Allows organizations to electronically find and apply for Federal grants.


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Lecture series
Women Surrealists: Imaging/Imagining the Self
Women Surrealists: Imaging/Imagining the Self from Manchester Art Gallery on Vimeo.
In this lecture, Professor Elza Adamowicz explores issues relating to subjectivity, identity and gender in the works of surrealist artists, including Claude Cahun, Frida Kahlo and Leonora Carrington. It shows how they rejected some of the existing views of femininity and offered alternatives through performance art, object-making and combining materials in new and unusual ways.
Women Surrealists and the Politics of Eros
Women Surrealists and the Politics of Eros from Manchester Art Gallery on Vimeo.
With Dr Alyce Mahon, Senior Lecturer in History 20th Century Art, University of Cambridge
In this lecture, Dr Alyce Mahon explores the role of erotic desire in surrealism, and women surrealists' provocative use of the sexual body to radically awaken the viewer to gender politics. Addressing a range of women surrealists, including Meret Oppenheim, Leonor Fini, Frida Kahlo and Mimi Parent, Mahon considers how these women both offered an alternative to the dominant modes of surrealist masculinity and paved the way for later, post 1960s, generations of feminist artists.
Woman in Plural: Czech Surrealism's Women Artists
Woman in Plural: Czech Surrealism's Women Artists from Manchester Art Gallery on Vimeo.
With Krysztof Fijalkowski, Norwich School of Art & Design
This lecture will focus on Czech women surrealist artists and offer a rare insight into these less well-known key surrealist artists.
Sphinx: Leonor Fini and Surrealism
Sphinx: Leonor Fini and Surrealism from Manchester Art Gallery on Vimeo.
With Dr Peter Webb, author of Sphinx: The Life and Art of Leonor Fini
In his lecture Sphinx: Leonor Fini and Surrealism , Dr Peter Webb will investigate artist Leonor Fini's role as a female artist in the predominantly male surrealist group, and the contribution she made with her self-images as empowered sphinxes and ancient deities. Dr Webb knew the artist Leonor Fini well and interviewed her extensively in the years before her death.
Curator's Talk – Women Artists, Surrealism and Tradition
Curator's Talk – Women Artists, Surrealism and Tradition from Manchester Art Gallery on Vimeo.
With Dr Patricia Allmer, MIRIAD, Manchester Metropolitan University
In this lecture, exhibition curator Dr Patricia Allmer discusses how some women surrealist artists explore, challenge and rework conventional artistic traditions. By doing so, these artists demonstrate and reveal how art history contributes to constructing and maintaining power relations between men and women.
The talk gives further insight into a number of artists and artworks represented in the exhibition, Angels of Anarchy, including Meret Oppenheim's objects, Lee Miller's photographs of desert-scapes and Mimi Parent's strange and unsettling assemblages in boxes.
'Womb-Magic' and 'the Coral of the Mind': Eileen Agar's Surrealism
'Womb-Magic' and 'the Coral of the Mind': Eileen Agar's Surrealism from Manchester Art Gallery on Vimeo.
With Dr Neil Matheson, Senior Lecturer in Theory and Criticism of Photography, University of Westminster
Focusing on the period 1936-45, this lecture explores the work of Eileen Agar and her particular contribution to the emergence of a surrealist movement in England. The discussion embraces Agar's 'seaside surrealism', her use of natural and marine forms and her conception of female creativity, as well as considering her wartime work.
The content of these lectures is © Krysztof Fijalkowski, © Dr Peter Webb, © Dr Alyce Mahon, ©Dr Patricia Allmer and © Professor Elza Adamowicz. All images used within the presentation remain the © of the owner or licensee and are protected by copyright laws and treaties around the world. All such rights are reserved.


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Universities help drive the transformation now underway regionally, focusing on conducting research, incubating new businesses and upskilling or reskilling labor to meet technology driven changes in labor demand.
"On a positive note, embracing societal change means that people have to learn to compete with the help of smarter machines—ones that augment individual, institutional and organizational capabilities. As a result, people are gravitating toward becoming life-long learners—able to adapt to change—and able to compete with machines and not against them," states Spohrer.
To attract the "life long learner", universities are aligning their capabilities to support a new type of student—one that maintains a full-time job, has a family or has limited time, thereby requiring flexibility to meet program requirements.
"This transformation calls for doubling student enrollment via on-line courses. Some Universities such as Southern New Hampshire University are way ahead of the curve and others such as MIT and Stanford are also moving in this direction," Spohrer elaborates. Read full story.


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If you are a regular user of wordpress and keep searching for CMS themes than you must be aware of VibrantCMS theme . It is designed by WOOTHEMES and cost is around $70 .
You can download this theme legally from given source below . I just discovered this source and sharing with NB members .
VibrantCMS theme
Demo | Download
It can be a promotional offer , so just go and grab it as soon as possible . If you want to check some premium free wordpress theme then you can visit - Themetoday.com :D


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Clocking In
You have clocked in to MySA.com's business news blog about San Antonio and South Texas businesses and business people.
Yearly Archive for 2009
Local oil change masters awarded
San Antonio-based Express Lube has been recognized by lubricant manufacturer Royal Purple with its President's Award for marketing and customer service excellence. Criteria considered before receiving this awarded included sales, community involvement, marketing and advertising expertise and facilities maintenance. Chris Quinn [Read More]
Sprint launches 4G in S.A.
Beating rivals AT&T and Verizon Wireless to the punch, Sprint Nextel Corp. launched its next generation 4G wireless service today in San Antonio and Austin. The two cities join a list of about a dozen other Texas markets to gain access to Sprint's 4G wireless connection this year. Sprint says its 4G service "delivers download […] [Read More]
Kiolbassa Provision Co. is a Texas Treasure
Kiolbassa Provision Co. is the newest Texas Treasure. The company that makes the famous smoked sausage won the designation from the State Historical Commission. The announcement comes as the San Antonio-based company celebrates its 60th anniversary of doing business from its plant on San Antonio's West Side at 1325 South Brazos St., according to State […] [Read More]
Crab stuffed steaks and nearly 250 jobs coming to S.A.
Longhorn Steakhouse, a national chain with more than 300 locations, will hire roughly 240 workers from the San Antonio area to fill positions at three local restaurants the company plans to open in the next six months. The franchise opened its first location in Atlanta 28 years ago and has since expanded to about 30 […] [Read More]
Victoria Garcia named to Port San Antonio board
Victoria M. Garcia, a San Antonio lawyer, is the City Council's latest appointment to the Port San Antonio's 11-member board of directors. Garcia is a partner with the law firm of Bracewell & Giuliani LLP, specializing in the areas of domestic and international labor and employment law as well as immigration. She earned her law […] [Read More]
Spurs TV Companion launched
No, it's not a service for lonely San Antonio Spurs fans. But the new Spurs TV Companion could be the answer for Spurs fans who don't get enough information, analysis and opportunities to network with their digital contacts while watching Spurs games. Go online at spurs.com during live game broadcasts to get the new companion. […] [Read More]
NuStar nets $289 million from sale of units
Asphalt refiner and pipeline owner NuStar Energy LP has netted $289 million from the sale of 5.7 million common units. Each unit was priced at $52.45 a share. The San Antonio-based master limited partnership said it will use the proceeds for general partnership purposes, including possible acquisitions and capital expenditures. NuStar may use some or […] [Read More]
Toyota plant gears up for more hiring
In anticipation of the Tacoma's arrival, Toyota's San Antonio truck plant will hire "dozens" of team members to repair and maintain manufacturing equipment, officials said today. That comes on top of the 850 team members Toyota has already started hiring to help build the Tacoma pickup when production starts next year. So far, Toyota says […] [Read More]
Former Spurs go for pancakes
Grab a tall stack at The Vistana. A pair of former hot cakes for the San Antonio Spurs, Bruce Bowen and Robert Horry, will attend the grand opening of the new IHOP café at The Vistana condominium building from noon to 2 p.m. on Monday, Nov. 16. This café will be different from other IHOP […] [Read More]
GVTC promotes executive
Pettiette Josh Pettiette of GVTC Communications has been named that company's vice president-product development. In his new role, Pettiette will oversee managers for the company's broadband, cable and phone businesses while also managing GVTC's profitability and new product revenue Pettiette joined GVTC in 2004 and since 2005 served as its manager of installation and repair. […] [Read More]
Show More


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Oman: Pre-paid vouchers and starter kits
Provision of a distributor for pre-paid vouchers and pre-paid starter kits for a telecoms company
Subscriber access only
Sign In
- If you are a new, paid MEED subscriber accessing the site for the first time, please activate your account.
- If you are a registered user who would like to to enjoy unlimited access to MEED.com, click subscribe to purchase a subscription or contact Customer Service by email or by calling +971 (0) 4 818 0200.
- Alternatively, you may browse MEED.com as a registered user for free. Our quick registration will allow you to sample premium business intelligence which is otherwise only available to paying subscribers through our Editor's Choice section, sign up for newsletters and to set up a library of saved articles.


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Instant Markdown
Instant Markdown
Book Details
About This Book
- Learn something new in an Instant! A short, fast, focused guide delivering immediate results
- Install and configure Markdown
- Learn Markdown syntax, right from basic to complex examples
- Full of focused examples on different services that you can use with Markdown
Who This Book Is For
Instant Markdown is for people who are interested in an introduction to Markdown. You will not need any specific knowledge about programming or markup languages. This book will give you many ideas to use and take full potential of Markdown over many different services.
Table of Contents
What You Will Learn
- Install and configuring Markdown
- Learn Markdown syntax
- Participate in forums and discussions
- Write emails
- Share documents
- Create to-do lists
- Produce presentations
- Convert Markdown files to other formats, such as PDF
- Use Markdown extensions
In Detail
Markdown is becoming more and more popular. The simplicity of Markdown and the advantages that it has to offer has led to a tremendous rise in the number of sites and applications that use it. The conversion of simple human readable text format to HTML has never been so simple, thanks to Markdown.
Instant Markdown will walk you through the syntax of Markdown and will get you up to speed with the basics, which will allow you to write your own content in Markdown. It will then show you how to use different services to make the most of Markdown.
Apart from learning the basic syntax through practical and focused examples, you will also learn how to use Markdown to do many different things, such as participating in forums and discussions, writing emails, sharing documents, blogging, creating to-do lists, creating presentations, converting Markdown files to other formats such as PDF, or using Markdown extensions.
You will learn everything you need to know about Markdown, right from scratch to practical examples.


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New App from Deptartment of Education
By The Frankfort State Journal
That ringing iPhone can now notify Kentucky parents of a changing grade in science class or an absence excuse note that never made it into the teacher's hands. Detailed information about public school kids' grades, attendance and homework assignments is now available instantly from any iPhone, iPad or iPod Touch with a free app provided by the state Department of Education. The information that's available isn't new – it's just taking a new form.


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Computing is of central importance to the work of the College of Architecture and Environmental Design (CAED). Two computing labs, the digital commons and the digital fabrication and print center are all integral amenities to education at the CAED. Computing facilities, including laser cutting, plastics printing, plaster printing, large format color and black and white plotting are supported by CAED's I.T. coordinator, computing staff and faculty. A full complement of software systems are available in the labs. Network and notebook support is assisted by CAED and the university helpdesk support system.
Rendering your Revit file can be done in minutes instead of hours using the interactions listed here.
Several new policies have been implemented that will affect everyone using the Digital Fabrication and Print Center. Please read them here.
Digital Fabrication and Print Center schedule
To sign up for laser cutting or to color plot, please click here. To view the waiting list, please click here.
Networking | Print Drivers | Support
CAED computer requirements


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Check out Deadliest Catch store for DVDs, Deadliest Catch t-shirts, hoodies, and merchandise at the official Discovery Channel store including complete season DVD sets and exclusive Deadliest Catch DVDs. Get Deadliest Catch merchandise at the Discovery Channel store.
View As: grid list
- Exclusive
- Deadliest Catch: Season 6 DVD
- Catch the exclusive Deadliest Catch Season 6 DVD set and join crews of commercial fisherman as they try to haul in a year's salary in just a week. Forty-foot waves, an almost 100-percent injury rate and gale force winds make crabbing on the Bering Sea, even more dangerous. In Deadliest… [ read more ]
- Deadliest Catch: Season 1 DVD
- Catch up with a Deadliest Catch Season 1 DVD set and see the thrilling first season of the series that made a big splash! Join crabbing Captains and their crews, as they are faced with walls of water, bone-chilling winds and an incredible injury rate. In these Deadliest Catch DVDs, witness… [ read more ]
- Deadliest Catch: Season 4 DVD
- Experience one of the most dangerous professions in the world in Discovery Channel's top-rated, Emmy Award-winning hit series Deadliest Catch. Crab fishing this season on the turbulent, frigid Bering Sea proves to be exceptionally difficult as the injuries pile up. Two men get hurt by a… [ read more ]
- Exclusive
- Deadliest Catch: Season 3 DVD
- See the Deadliest Catch Season 3 DVD set and continue on with the captains and the crews that you have come to love and who brave treacherous elements on the Bering Sea to bring in the most lucrative catch. Or if you are a new fan, you will want to see another thrilling episode of the show… [ read more ]
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- Deadliest Catch Mash Up T-Shirt - Navy
- As a true Deadliest Catch fan, you could quote some of the guys best lines off and on the Bering Sea. Be an honorary crew member in this Deadliest Catch T-Shirt featuring memorable quips to monikers of fishermen and their captains that capture the experience that brings them together. [ read more ]
- Deadliest Catch: Season 7 DVD
- In the far northern reaches of the planet a rare breed of extreme fishermen hunt the Bering Sea. Adventure is their call, crab is their prey, and a one hundred million dollar plunder is their reward. Skippers and their crews endure boat killing storms and body crushing working conditions -… [ read more ]
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