ABCC4 is a PGE2 efflux transporter in the ovarian follicle: A mediator of ovulation and a potential non-hormonal contraceptive target.

The role of prostaglandins (PGs) in the ovulatory process is known. However, the role of the ATP binding cassette subfamily C member 4 (ABCC4), transmembrane PG carrier protein, in ovulation remains unknown. We report herein that ABCC4 expression is significantly upregulated in preovulatory human granulosa cells (GCs). We found that PGE2 efflux in cultured human GCs is mediated by ABCC4 thus regulating its extracellular concentration. The ABCC4 inhibitor probenecid demonstrated effective blocking of ovulation and affects key ovulatory genes in female mice in vivo. We postulate that the reduction in PGE2 efflux caused by the inhibition of ABCC4 activity in GCs decreases the extracellular concentration of PGE2 and its ovulatory effect. Treatment of female mice with low dose of probenecid as well as with the PTGS inhibitor indomethacin or Meloxicam synergistically blocks ovulation. These results support the hypothesis that ABCC4 has an important role in ovulation and might be a potential target for non-hormonal contraception, especially in combination with PGE2 synthesis inhibitors. These findings may fill the gap in understanding the role of ABCC4 in PGE2 signaling, enhance the understanding of ovulatory disorders, and facilitate the treatment and control of fertility.

Chemical contraceptive impacts on cyclic progesterone and sexual behavior in zoo-housed Western lowland gorillas.

For regularly cycling primates in captivity, chemical contraceptives can be used to reduce the likelihood of unintentional offspring, but the effects on cyclic progesterone and sexual behavior have not been well studied in gorillas (Gorilla gorilla gorilla). As zoos are invested in maintaining naturalistic behaviors and physiology, this is an important aspect of gorilla physical and mental health. We examined how exposure to combination and progestin-only oral birth control pills altered cyclic progesterone and timing of sexual behaviors in four female gorillas at four institutions compared to cycling controls. Hormonal data were collected using fecal steroid metabolites as detected by enzyme immunoassay. Behavioral data were collected via one-zero sampling done by keeper staff. All methods of oral contraceptive eliminated the ovarian cycle, with fecal progestogens being maintained below baselines in cycling control individuals. There was also no evidence of any cyclic patterning in endogenous progesterone, suggesting that the methods tested do not allow for detection of a recovery of endogenous progesterone during a short no-pill period. We also found that the oral contraceptives altered the timing of behavioral displays when compared to cycling control individuals but was not directly related to any fluctuations in endogenous progesterone. This study reveals a need for additional research on contraceptive use in gorillas and reinforces that sexual behavior in gorillas is both hormonally and socially mediated.

Chronic treatment with hormonal contraceptives alters hippocampal BDNF and histone H3 post-translational modifications but not learning and memory in female rats.

Hormonal contraceptives prevent ovulation with subsequent reduction in endogenous levels of estradiol, progesterone and its neuroactive metabolite allopregnanolone. These neurosteroids modulate several brain functions, including neuronal plasticity, cognition and memory. We hypothesized that hormonal contraceptives might affect synaptic plasticity, learning and memory, as a consequence of suppressed endogenous hormones levels. Female rats were orally treated with a combination of ethinyl estradiol (EE, 0.020 mg) and levonorgestrel (LNG, 0.060 mg) once daily for four weeks. Decreased hippocampal brain-derived neurotrophic factor (BDNF) levels and altered histone H3 post-translational modifications (PTMs) were observed 14 days after discontinuation from chronic EE-LNG treatment. These effects were not accompanied by alterations in long-term plasticity at glutamatergic synapses, recognition memory in the novel object and novel place location tests, or spatial learning, memory, and behavioral flexibility in the Morris water maze test. Thus, decreased BDNF content does not affect synaptic plasticity and cognitive performance; rather it might be relevant for the occurrence of certain psychiatric symptoms, reported by some women using hormonal contraceptives. These results provide the first evidence of hippocampal epigenetic changes induced by hormonal contraceptives and complement previous studies on the neurobiological actions of hormonal contraceptives; the finding that effects of chronic EE-LNG treatment on BDNF content and histone PTMs are observed 14 days after drug discontinuation warrants further investigation to better understand the implications of such long-term consequences for women's health.

Contraceptive efficacy of recombinant porcine zona proteins and fusion protein encompassing canine ZP3 fragment and GnRH in female beagle dogs.

PROBLEM: To manage population of dogs (Canis familiaris), the efficacy of recombinant proteins-based contraceptive vaccines to inhibit fertility has been evaluated in female beagle dogs. METHOD OF STUDY: Female beagle dogs (n = 4) were immunized with physical mixture of Escherichia coli-expressed recombinant porcine ZP3 with promiscuous T cell epitope of tetanus toxoid (TT-KK-pZP3) and porcine ZP4 with promiscuous T cell epitope of bovine RNase (bRNase-KK-pZP4), or with a fusion protein encompassing dog ZP3 fragment and two copies of GnRH with appropriate promiscuous T cell epitopes (dZP3-GnRH2 ); control animals received only alum, the adjuvant. The immunized animals were followed-up for antibody titres by ELISA as well as for fertility status subsequent to mating with male dogs. RESULTS: Active immunization of female dogs following a three injections schedule at 4-week intervals with a physical mixture of TT-KK-pZP3 + bRNase-KK-pZP4 as well as dZP3-GnRH2 , led to generation of significant antibody titres against respective recombinant proteins. Active immunization with dZP3-GnRH2 also led to generation of antibodies reactive with both dZP3 and GnRH. A booster dose on day 383 led to an increase in antibody titres and circulating antibodies against respective recombinant proteins could be observed on day 528. Antibodies in immune serum samples from dogs immunized with TT-KK-pZP3 + bRNase-KK-pZP4 or dZP3-GnRH2 reacted with native canine ZP as assessed by an indirect immunofluorescence assay. Mating studies revealed a reduced number of pregnancies as well as a significant reduction in the number of pups born in the female dogs immunized with dZP3-GnRH2 as compared to the adjuvanted control. Curtailment of pregnancy in dZP3-GnRH2 immunized group was associated with antibody titres against dZP3-GnRH2 . However, immunization with recombinant TT-KK-pZP3 + bRNase-KK-pZP4 did not significantly decrease the number of pups born as compared to the adjuvanted control. CONCLUSION: These studies revealed the potential of recombinant dZP3-GnRH2 -based contraceptive vaccine to curtail fertility in female dogs. Large scale studies to establish the efficacy and safety of this recombinant protein for the management of community dog population are thus warranted.

Pharmacokinetics, metabolism and serum concentrations of progestins used in contraception.

Many different forms of hormonal contraception are used by millions of women worldwide. These contraceptives differ in the dose and type of synthetic progestogenic compound (progestin) used, as well as the route of administration and whether or not they contain estrogenic compounds. There is an increasing awareness that different forms of contraception and different progestins have different side-effect profiles, in particular their cardiovascular effects, effects on reproductive cancers and susceptibility to infectious diseases. There is a need to develop new methods to suit different needs and with minimal risks, especially in under-resourced areas. This requires a better understanding of the pharmacokinetics, metabolism, serum and tissue concentrations of progestins used in contraception as well as the biological activities of progestins and their metabolites via steroid receptors. Here we review the current knowledge on these topics and identify the research gaps. We show that there is a paucity of research on most of these topics for most progestins. We find that major impediments to clear conclusions on these topics include a lack of standardized methodologies, comparisons between non-parallel clinical studies and variability of data on serum concentrations between and within studies. The latter is most likely due, at least in part, to differences in intrinsic characteristics of participants. The review highlights the importance of insight on these topics in order to provide the best contraceptive options to women with minimal risks.

Biotransformation of contraceptive drug desogestrel with Cunninghamella elegans, and anti-inflammatory activity of its metabolites.

Biotransformation of an orally active contraceptive drug, desogestrel (1), with Cunninghamella elegans yielded a new metabolite, 13ß-ethyl-11-methylene-18,19-dinor-17α-pregn-4-en-20-yn-17ß-ol-3,6-dione (2), along with five known metabolites, i.e., 13ß-ethyl-11-methylene-18,19-dinor-17α-pregn-4-en-20-yn-3ß,6ß,17ß-triol (3), 13ß-ethyl-11-methylene-18,19-dinor-17α-pregn-4-en-20-yn-6ß,17ß-diol-3-one (4), 13ß-ethyl-11-methylene-18,19-dinor-17α-pregn-4-en-20-yn-17ß-ol-3-one (5), 13ß-ethyl-11-epoxy-18,19-dinor-17α-pregn-4-en-20-yn-17ß-ol-3-one (6), and 13ß-ethyl-11-methylene-18,19-dinor-17α-pregn-4-en-20-yn-10ß,17ß-diol-3-one (7). The structure of new metabolite 2 was elucidated by using 1H-, 13C-, and 2D-NMR, EI-, and HREI-MS, IR, and UV spectroscopic data. Compounds 1-7 were evaluated for anti-inflammatory activities, i.e., inhibition of T-cell proliferation, and pro-inflammatory cytokine (TNF-α). Compounds 1 (IC50 = 1.12 ± 0.03 µg/mL), 2 (IC50 = 1.15 ± 0.05 µg/mL), 3 (IC50 = 1.15 ± 0.05 µg/mL), 4 (IC50 = 1.40 ± 0.03 µg/mL), 5 (IC50 = 1.78 ± 0.08 µg/mL), and 6 (IC50 = 1.36 ± 0.07 µg/mL) were identified as potent inhibitors of T-cells proliferation, in comparison to the standard drug, prednisolone (IC50 = 3.51 ± 0.03 µg/mL). Compound 7 (IC50 = 6.18 ± 0.04 µg/mL) showed a good activity. In addition, substrate 1 (IC50 ≤ 1 µg/mL), and its metabolites 2 (IC50 = 4.1 ± 0.60 µg/mL), and 6 (IC50 = 6.8 ± 0.8 µg/mL) also showed a potent inhibition of pro-inflammatory cytokine (TNF-α) production, as compared to the standards drug, pentoxifilline (IC50 = 94.8 ± 2.1 µg/mL). Whereas compounds 3 (IC50 = 57.9 ± 7.6 µg/mL), and 5 (IC50 = 27.2 ± 6.8 µg/mL) showed a moderate inhibition of TNF-α production, while compounds 4 and 7 showed no inhibition. Compounds 1-7 were found to be non-cytotoxic to 3T3 normal cell line (mouse fibroblast).

Immunogenicity and contraceptive efficacy of recombinant fusion protein encompassing Sp17 spermatozoa-specific protein and GnRH: Relevance of adjuvants and microparticles based delivery to minimize number of injections.

PROBLEM: Requirement of multiple injections of contraceptive vaccines to achieve infertility is one of the important impediments for their application. In the present study, attempts have been made to reduce the number of injections of contraceptive vaccine. METHOD OF STUDY: Fusion protein encompassing C-terminus fragment of sperm protein Sp17 (aa residues 76-126) and two copies of gonadotropin-releasing hormone along with T-cell epitopes and dilysine linkers (abbreviated as Sp17C -GnRH2 ) was expressed in Escherichia coli. Its immunogenicity and contraceptive efficacy have been evaluated in female FVB/J mice using different adjuvants and delivery platforms. RESULTS: Immunization of female mice with recombinant Sp17C -GnRH2 (25 µg/injection/mouse) emulsified with squalene-arlacel A following two injections schedule led to failure of 88.8% immunized animals to conceive, which was not significantly different from mice immunized with same protein along with alum following three injections schedule. To make single-dose vaccine, poly d,l-lactic acid-based microparticles (PLA-MPs) entrapping Sp17C -GnRH2 were prepared. Immunization of female mice with a combination of soluble Sp17C -GnRH2 (12.5 µg/injection/mouse) along with Sp17C -GnRH2 entrapped in PLA-MPs (12.5 µg/injection/mouse) in alum showed higher antibody titres and contraceptive efficacy as compared to mice immunized with Sp17C -GnRH2 entrapped in PLA-MPs alone in alum. Immunization with recombinant Sp17C -GnRH2 led to long-term infertility as second mating (150 days after immunization) of various groups of immunized mice showed similar infertility as observed during first mating. CONCLUSION: Single-dose immunization with PLA-MPs entrapping Sp17C -GnRH2 along with soluble recombinant protein in alum generated long-lasting infertility in female mice.

O uso da pílula anticoncepcional e as alterações das principais vias metabólicas / The use of contraceptive pill and alterations of main metabolic pathways

As pílulas anticoncepcionais são esteroides que visam impedir a gravidez indesejada e regular distúrbios menstruais. Acessíveis em grande variedade no mercado e no SUS, são o método contraceptivo mais aceito pelas mulheres, entretanto trazem diversos efeitos colaterais. O objetivo deste trabalho foi analisar como a pílula anticoncepcional pode alterar as principais vias metabólicas femininas. Trata-se de uma revisão bibliográfica nas bases de dados SciELO, BVS e PubMed, com foco nas correlações entre o uso da pílula anticoncepcional e as alterações metabólicas. Os anticoncepcionais orais atuam na inibição da biossíntese de androgênios e estimulação da SHBG, o que reduz o efeito anabólico proteico. Também promovem o acréscimo dos níveis de LDL-colesterol, colesterol total, PCR-us e dímero D, e alterações na sensibilidade da insulina, no metabolismo do zinco e na hemostasia. Apesar de existirem recomendações que preconizam o uso de outros métodos contraceptivos e estudos que demonstram a satisfação feminina ao trocar os anticoncepcionais orais pelos LARCs, a pílula ainda é a mais utilizada pelas mulheres.(AU)

Contraceptive pills are steroids that prevent unwanted pregnancy and regular menstrual disorders. Accessible in a great variety in market and SUS, they are the contraceptive method most accepted by women, however, they bring several side effects. The objective of this study was analyze how the contraceptive pill can alter the main female metabolic pathways. This is a literature review in the SciELO, BVS and PubMed databases, focusing on the correlations between the use of contraceptive pill and metabolic alterations. Oral contraceptives act to inhibit androgen biosynthesis and stimulate SHBG, which reduces the protein anabolic effect. They also bring about high levels of LDL cholesterol, total cholesterol, CRP, D-dimer, changes in insulin, absence of zinc metabolism and hemostasis. Although there are recommendations that recommend the use of other contraceptive methods and studies that demonstrate the satisfaction of women in exchanging oral contraceptives with LARCs, the pill is still the most used by women.(AU)

Cunninghamella blakesleeana-mediated biotransformation of a contraceptive drug, desogestrel, and anti-MDR-Staphylococcus aureus activity of its metabolites.

Staphylococcus aureus is one of the most infectious agents among staphylococcal bacteria. Currently many strains of S. aureus have developed resistance against available antibiotics. Therefore, the treatment of infections caused by them is a major challenge. During current study, desogestrel (1), a contraceptive drug, was found to be a potent growth inhibitor of drug resistant strains of S. aureus. Therefore, in search of new and effective agents against multi-drug resistant S. aureus strains, whole-cell bio-catalytic conversion of desogestrel (1) by Cunninghamella blakesleeana ATCC 8688A at pH 7.0 and 25 °C was carried out, yielding three new metabolites, 13-ethyl-11-methylene-18,19-dinor-17α-pregn-4-en-20-yn-6ß,15ß,17ß-triol (2), 13-ethyl-11-methylene-18,19-dinor-17α-pregn-4-en-20-yn-3ß,6ß,17ß-triol (3), and 13-ethyl-11-methylene-18,19-dinor-17α-pregn-20-yn-3α,5α,6ß,17ß-tetraol (4), along with a known metabolite, 13-ethyl-11-methylene-18,19-dinor-17α-pregn-4-en-20-yn-6ß,17ß-dihydroxy-3-one (5). Among them, compounds 1-2 showed a potent activity against S. aureus EMRSA-17, S. aureus NCTC 13277 (MRSA-252), and S. aureus NCTC 13143, and clinically isolated Pakistani strain of S. aureus in an in vitro Microplate Alamar Blue Assay (MABA). Vancomycin was used as the standard drug in this assay. In addition, compound 1 also showed a significant activity against vancomycin-resistant S. aureus (VRSA) ATCC 700699. Compounds 1-5 were also evaluated against 3T3 normal cell line (mouse fibroblast) where they all were identified as non-cytotoxic. The present study thus provides new leads for the development of anti-bacterial drugs against MDR S. aureus.

Biotransformation of drospirenone, a contraceptive drug, with Cunninghamella elegans.

Biotransformation of an orally active contraceptive drug, drospirenone (1), by Cunninghamella elegans ATCC 36114 yielded four new metabolites, 6ß,7ß,15ß,16ß-dimethylene-3-oxo-14α-hydroxy-17α-pregn-4-ene-21,17-carbolactone (2), 6ß,7ß,15ß,16ß-dimethylene-3,11-dioxo-17α-pregn-4-ene-21,17-carbolactone (3), 6ß,7ß,15ß,16ß-dimethylene-3,12-dioxo-17α-pregn-4-ene-21,17-carbolactone (4), and 6ß,7ß,15ß,16ß-dimethylene-3-oxo-11ß,14α-dihydroxy-17α-pregn-4-ene-21,17-carbolactone (5), along with a known metabolite, 6ß,7ß,15ß,16ß-dimethylene-3-oxo-11α-dihydroxy-17α-pregn-4-ene-21,17-carbolactone (6). This study provides not only new analogues of orally active contraceptive drug, drospirenone, but also help in understanding the metabolism of this important drug.

Microbial transformation of contraceptive drug etonogestrel into new metabolites with Cunninghamella blakesleeana and Cunninghamella echinulata.

Biotransformation of a steroidal contraceptive drug, etonogestrel (1), (13-ethyl-17ß-hydroxy-11-methylene-18,19-dinor-17α-pregn-4-en-20-yn-3-one) was investigated with Cunninghamella blakesleeana and C. echinulata. Five metabolites 2-6 were obtained on incubation of 1 with Cunninghamella blakesleeana, and three metabolites, 2, 4, and 6 were isolated from the transformation of 1 with C. echinulata. Among them, metabolites 2-4 were identified as new compounds. Their structures were deduced as 6ß-hydroxy-11,22-epoxy-etonogestrel (2), 11,22-epoxy-etonogestrel (3), 10ß-hydroxy-etonogestrel (4), 6ß-hydroxy-etonogestrel (5), and 14α-hydroxy-etonogestrel (6). Compounds 1-6 were evaluated for various biological activities. Interestingly, compound 5 was found to be active against ß-glucuronidase enzyme with IC50 value of 13.97±0.12µM, in comparison to standard compound, d-saccharic acid 1,4-lactone (IC50=45.75±2.16µM). Intestinal bacteria produce ß-glucuronidase. Increased activity of ß-glucuronidase is responsible for the hydrolyses of glucuronic acid conjugates of estrogen and other toxic substances in the colon, which plays a key role in the etiology of colon cancer. Inhibition of ß-glucoronidase enzyme therefore has a therapeutic significance. Compounds 1-6 were also found to be non cytotoxic against 3T3 mouse fibroblast cell lines.

Medroxyprogesterone acetate increases HIV-1 infection of unstimulated peripheral blood mononuclear cells in vitro.

OBJECTIVE: Several observational studies suggest that medroxyprogesterone acetate (MPA) injectable contraceptives may increase a woman's risk of sexual HIV-1 acquisition. In-vitro studies are conflicting, mainly due to differences in the type of progestin studied or activation status of the primary cells. We sought to determine whether MPA increases infection of unstimulated peripheral blood mononuclear cells (PBMCs). METHODS: Freshly isolated PBMCs from normal blood donors were treated with physiologic MPA concentrations ranging from 0.003 to 5 ng/ml and infected with GFP-tagged R5-tropic or X4-tropic HIV-1 pseudoviruses by spinoculation. The infection was limited to a single cycle. Cells were stained with CD3, CD8 and CD14. Infection was quantified as the percentage of GFP cells by flow cytometry. RESULTS: Absolute infection was greater among unstimulated MPA-treated CD3⁺CD8⁻ T cells vs. untreated cells across MPA concentrations of 0.003-3 ng/ml using R5 (P < 0.003) and 0.03-0.3 ng/ml using X4 (P < 0.005) pseudovirus. There was increased relative infection of CD3⁺CD8⁻ T cells in MPA-treated whole PBMC cultures but not after monocytes were depleted (P < 0.02). HIV-1 infection of stimulated PBMC showed no differences in R5 or X4 infection across all MPA concentrations (P > 0.5). CONCLUSION: The CD3⁺CD8⁻ T-cell population of MPA-treated unstimulated PBMCs were more susceptible to HIV-1 infection than untreated cells. The increased infection was partly due to monocytes and was lost when PBMC were exogenously stimulated. These data provide confirmation of a biological association between MPA exposure and increased susceptibility to HIV-1 infection, particularly among women who inject drugs.

Dispositivos intrauterinos liberadores de levonorgestrel: beneficios frente a dudas, temores y creencias erróneas / Levonorgestrel releasing intrauterine devices: benefits versus doubts, fears and misperceptions

El uso inconsistente del preservativo y la falta de cumplimiento con la píldora, los dos métodos más usados en nuestro país, dan lugar cada año a un número importante de embarazos no deseados. Diversas sociedades médicas internacionales recomiendan el uso de anticonceptivos de larga duración reversibles (LARC), altamente efectivos al no depender de la participación activa de la usuaria, para evitar los embarazos no deseados en todas las mujeres, salvo que exista alguna contraindicación. Entre éstos, los dispositivos intrauterinos de cobre (DIU-Cu) y el liberador de levonorgestrel (DIU-LNG), una evolución de los anteriores, son los más populares. Pese a sus beneficios, apenas un 6 % de las mujeres los usa en nuestro país. Esto puede deberse a la existencia de dudas, temores y creencias erróneas entre los profesionales sanitarios y las mujeres que crean una barrera a su uso. En la presente revisión se analizan los beneficios que ofrece el DIU-LNG y se da respuesta a las dudas, temores y creencias erróneas más extendidas, tales como las mujeres elegibles para usarlo, la recuperación de la fertilidad, el uso tras el parto, el coste, las dificultades en la inserción o el riesgo de enfermedad inflamatoria pélvica, de infertilidad y de embarazos ectópicos. El desconocimiento de los DIU entre las mujeres, y en especial del DIU-LNG, es muy alto. Se revisan también aquellos aspectos específicos en los que es importante asesorar a la usuaria

Inconsistent use of condoms and lack of compliance with oral contraceptive pills, both the most widespread used methods in Spain, result in high rates of unintended pregnancies yearly. Several international medical societies do recommend the use of long acting reversible contraceptive methods (LARC methods), highly effective because of no need for user’s active participation, to prevent pregnancies in all women, unless specific contraindication determined. Within them, cooper intrauterine device (Cooper-IUD) and levonorgestrel releasing intrauterine device (LNG-IUD), an evolution from the first one, are the most popular ones. Despite of their proved benefits, only 6 % of women are using them in our country. A barrier has been created around the use of intrauterine devices, ought to healthcare providers and users’ fears, misperceptions and doubts. In the present review we analyse those benefits offered by LNG-IUD, giving response to the most extended myths, misperceptions and fears such as women’s adequate profile, fertility recovery, postpartum use, cost, insertion inconveniences, or risk for pelvic inflammatory disease, infertility or ectopic pregnancies. Unawareness of intrauterine devices within women, particularly of LNG-IUD, is unexpectedly high. Specific issues around contraceptive counselling are also included in this review

Regional gray matter volume differences and sex-hormone correlations as a function of menstrual cycle phase and hormonal contraceptives use.

During the menstrual cycle, hormone-driven functional and morphological changes occur in the female brain. The influence of hormonal contraceptives on these changes has received only little attention in the medical literature. The purpose of our study is to measure regional gray matter volume changes as a function of the cycle phase and use of hormonal contraceptives, in relation to blood concentrations of sex hormones. We performed a prospective study in 30 healthy young women; 15 women had a natural menstrual cycle and 15 were using monophasic combined hormonal contraceptives. MRI examinations were acquired at 2 specific time-points in the cycle (follicular and luteal phase). MRI studies included a T1-weighted, isotropic, high-resolution 3-D gradient echo acquisition, for the purpose of performing voxel based morphometry. Peripheral venous blood samples were obtained to determine concentrations of luteinizing hormone (LH), follicle stimulating hormone (FSH), estradiol, and progesterone. We found a highly significant negative correlation of regional gray matter volume in the anterior cingulate cortex with estradiol concentrations. To the best of our knowledge, this result has not been described before, and was only present in the natural cycle group, not in women using hormonal contraceptives. The anterior cingulate cortex is involved in emotion processing and there is literature describing behavioral alternations with changing hormone levels. Our findings provide a structural, morphological basis to support these data. Therefore, we advise neuroscientists to take into account the menstrual cycle phase and use of hormonal contraceptives, in order to avoid obtaining heterogeneous data sets, leading to a significant loss of accuracy and precision.

Immunogenicity and contraceptive efficacy of Escherichia coli-expressed recombinant porcine zona pellucida proteins.

PROBLEM: To overcome availability of the purified native zona pellucida (ZP) glycoproteins for immunocontraception, porcine ZP3, and ZP4 were expressed in E. coli. METHOD OF STUDY: Purified recombinant proteins were characterized by SDS-PAGE and Western blot, and immunogenicity and contraceptive efficacy determined in FvB/J female mice. RESULTS: Purified ZP3, ZP3 with promiscuous T-cell epitope of tetanus toxoid, ZP4 and ZP4 incorporating promiscuous T-cell epitope of bovine RNase revealed ~44-, ~49-, ~53-, and ~55-kDa bands by SDS-PAGE and Western blot, respectively. Immunization of female mice with recombinant proteins elicited high antibody titers as well as T-cell responses. Immune sera recognized mouse oocyte ZP and also inhibited in vitro fertilization. Immunized mice showed significant decrease in fertility. Recombinant proteins were able to recall memory antibody response in female mice primed with porcine native ZP. CONCLUSION: Availability of recombinant porcine proteins will be useful in the development of contraceptive vaccine.

Salivary testosterone: associations with depression, anxiety disorders, and antidepressant use in a large cohort study.

OBJECTIVE: Low circulating levels of testosterone have been associated with major depression, but there is more limited evidence for differences in patients with anxiety disorders. The use of selective serotonin reuptake inhibitors (SSRIs) and other antidepressants is associated with sexual side effects, warranting testing for interactions with testosterone. METHODS: Data are from 722 male and 1380 female participants of The Netherlands Study of Depression and Anxiety (NESDA), who were recruited from the community, general practice care, and specialized mental health care. Depressive and anxiety diagnoses were assessed using the DSM-IV Composite International Diagnostic Interview. To smooth the episodic secretion, the four morning saliva samples per participant and the two evening samples were pooled before testosterone analysis. RESULTS: Morning median testosterone levels were 25.2 pg/ml in men and 16.2 pg/ml in women, with lower evening levels of 18.2 and 14.1 pg/ml, respectively. Significant determinants of testosterone levels were sex, age, time of the day, use of contraceptives, and smoking status. Female patients with a current (1-month) depressive disorder (effect size 0.29; P=0.002), generalized anxiety disorder (0.25; P=0.01), social phobia (0.30; P<0.001), and agoraphobia without panic disorder (0.30; P=0.02) had lower salivary testosterone levels than female controls. Higher testosterone levels were found in male and female participants using SSRIs than in non-users (effect size 0.26; P<0.001). CONCLUSION: Salivary testosterone levels are lower in female patients with a depressive disorder, generalized anxiety disorder, social phobia, and agoraphobia as compared to female controls. SSRIs may increase salivary testosterone in men and women.

Utilización de terapias hiperestrogénicas en el lupus eritematoso sistémico / Utilization of hyperestrogenic therapies in systemic lupus erythematosus

La utilización de tratamientos que aumenten los niveles estrogénicos ha sido considerada clásicamente de riesgo en pacientes lúpicos. Estudios relativamente antiguos sobre la utilización de anticonceptivos orales y de tratamiento hormonal sustitutivo han proporcionado resultados contradictorios o inconsistentes. Más recientemente, estudios prospectivos sobre esta problemática sugieren que tanto los anticonceptivos como el tratamiento hormonal sustitutivo pueden ser utilizados en pacientes con enfermedad estable sin riesgo de un aumento en la actividad clínica de esta. Tampoco se ha observado una asociación al desarrollo de complicaciones tromboembólicas arteriales y/o venosas. Aunque, a este respecto, determinaciones y limitaciones metodológicas imposibilitan el establecer valoraciones definitivas sobre esta problemática. Con respecto a la utilización de técnicas de fertilización asistida en pacientes lúpicas, todos los datos de que se disponen son de tipo retrospectivo. Globalmente considerados, estos estudios indican que el riesgo real de reagudización de la enfermedad es relativamente bajo y que, en general, no son agudizaciones graves, siempre y cuando estas técnicas se realicen en pacientes con enfermedad estable (AU)

The use of treatments that increase the estrogenic levels has been usually deemed risky in lupus patients. Past studies about the utilization of oral contraceptive drugs and hormone replacement therapy (HRT) have shown contradictory results. More recently, prospective studies about this issue suggest that either oral anticonceptive and HRT can be used in patients with stable disease without special risk for increments in clinical activity. Neither, it has been observed an association with the development of arterial or venous thrombosis. Although, to this regard, several methodological limitations preclude to establish definitive conclusions. Regarding to the use of assisted reproduction techniques in lupus patients, only retrospective data are available. Overall they indicate that the real risk of disease worsening is quite low, being the flares mostly mild when these procedures are performed in patients with stable disease (AU)

Absorption of vaginal estrogen cream during sexual intercourse: a prospective, randomized, controlled trial.

OBJECTIVE: To determine if intercourse changed serum estradiol levels in women using vaginal E2 cream or in their male partners. STUDY DESIGN: Prospective, randomized, placebo-controlled, crossover, blinded study of 10 postmenopausal women and their male sexual partners. Subjects were randomized to estradiol or placebo cream and intercourse, then crossed over after 7-14 days. Seven to 14 days later, the woman used vaginal estradiol cream and abstained from intercourse. Serum E2 levels were obtained 10-12 hours after each exposure. RESULTS: Serum E2 levels were higher in 8 of 10 men after intercourse with vaginal estradiol cream, and this resulted in a small but significant (p = 0.03) increase in the estradiol levels as compared to placebo. Paradoxically, intercourse resulted in markedly lower estradiol levels in women as compared to abstinence (p = 0.004). CONCLUSION: Men absorb vaginal estradiol during intercourse, whereas intercourse reduces estradiol absorption in women. Although serum estradiol levels were only mildly elevated in men, it is possible that long-term exposure could cause feminizing changes. In women, estradiol levels were markedly reduced by intercourse.

Nanosilver: a nanoproduct in medical application.

Nanotechnology is a most promising field for generating new applications in medicine. However, only few nanoproducts are currently in use for medical purposes. A most prominent nanoproduct is nanosilver. Nanosilver particles are generally smaller than 100nm and contain 20-15,000 silver atoms. At nanoscale, silver exhibits remarkably unusual physical, chemical and biological properties. Due to its strong antibacterial activity, nanosilver coatings are used on various textiles but as well as coatings on certain implants. Further, nanosilver is used for treatment of wounds and burns or as a contraceptive and marketed as a water disinfectant and room spray. Thus, use of nanosilver is becoming more and more widespread in medicine and related applications and due to increasing exposure toxicological and environmental issues need to be raised. In sharp contrast to the attention paid to new applications of nanosilver, few studies provide only scant insights into the interaction of nanosilver particle with the human body after entering via different portals. Biodistribution, organ accumulation, degradation, possible adverse effects and toxicity are only slowly recognized and this review is focusing on major questions associated with the increased medical use of nanosilver and related nanomaterials.

Contraceptive counseling by gynecologists--which issues are discussed and does gender play a role?

OBJECTIVES: Explorative pilot study with the aim of gaining insight into the contraceptive counseling practices and possible gender differences of a selected group of male and female gynecologists. DESIGN: Semi-structured telephone-interviews of 48 gynecologists concerning the content and strategies of their contraceptive counseling with special focus on aspects relevant to patient adherence. RESULTS: Male and female gynecologists inform equally frequently about various methods and reproductive health aspects such as risks, the advantages and disadvantages of the methods and side effects. Male physicians speak more often about the efficiency and benefits of the methods, while their female colleagues emphasize STI and emergency contraception. Sexual health aspects were seldom mentioned as issues of discussion. For the choice of a contraceptive method efficiency was considered very important by 100%, reversibility by 83%, side effects by 85% and convenience by 79%. Naturalness and costs were more often quoted as important by female, and benefits by male gynecologists. Side effects are considered the most important factor for patient adherence by 60%, counseling and information is predominantly cited by female, and patient's character and personality by male doctors. CONCLUSIONS: While contraceptive counseling by practising gynecologists includes basic information about available methods and their efficiency, as well as some reproductive health issues, sexual health issues are often neglected. Gender differences occasionally influence the choice of the topics as well as the attitude towards the patient.