Position of the Spanish Menopause Society regarding the management of perimenopause.

Perimenopause is an imprecise period in woman over 40 years of age, which comprises the time between the moment that the first changes in the menstrual cycle appear and the year following the definitive cessation of the menses. Besides irregular bleeding, many women also complain of hot flashes and other characteristic symptoms of postmenopause. Moreover, most of them are concerned about the future impact that these events may have on their health, such as needing health exams or continuing to use contraceptive methods. A panel of experts from the Spanish Menopause Society has met to establish diagnostic and therapeutic guidelines for this period based on the best available evidence.

Women who are not exposed to spermatozoa and infertile women have similar rates of stage I endometriosis.

OBJECTIVE: To compare the prevalence of endometriosis and its different stages in infertile women and women not exposed to spermatozoa. DESIGN: Prospective study. SETTING: Artificial insemination donor program at a university hospital. PATIENT(S): One hundred fifty women unable to conceive because they had not been exposed to spermatozoa (134 with azoospermic partner, 10 with an HIV-positive partner, and 6 without a male partner). Controls were 750 women in infertile couples in which the male partner had normal sperm. INTERVENTION(S): Laparoscopy was systematically performed in a blinded manner in both groups as part of the infertility work-up. MAIN OUTCOME MEASURE(S): Diagnosis of endometriosis. RESULT(S): In unexposed women and controls, the prevalence of endometriosis was similar (32% and 34.5%). Rates of stage I disease were also similar in both groups (26% and 19.3%). There was a significant trend toward higher stages of endometriosis in infertile women (stage II disease, 3.3% vs. 5.7%; stage III disease, 1.3% vs. 3.1%; stage IV disease, 1.3% vs. 6.4%). Endometriosis was not associated with the few demographic characteristics that differed between groups. CONCLUSION(S): From an epidemiologic point of view, stage I endometriosis is not more common in infertile women than in unselected women. However, stage II to IV endometriosis was more frequent in infertile women. Whereas a relation between stage I endometriosis and infertility seems unlikely, the relation between stages II to IV endometriosis and infertility seems possible.

[New observations on the structure of the lingual filiform papillae epithelium of the toad Bufo marinus L.: Light and transmission electron microscopy study]. / Nuevas observaciones en la estructura del epitelio de las papilas filiformes de la lengua del sapo Bufo marinus L.: estudio con microscopia de luz y electrónica de transmisión.

There are several inconsistencies and confusions about the microscopic structure in the anurans dorsal, non-sensory, mucosal epithelium of the tongue. The aim of this work was to differentiate the structure of the epithelium covering filiform papillae from the glandular epithelium. For this, samples of the dorsal distal mucosa of Bufo marinus tongue were processed for light microscopy (LM) and transmission electron microscopy (TEM). For LM, stained 1 micron sections were used. The results showed that the filiform papillae were covered by a biestratified epithelium formed by a basal layer of dendritic cells and an upper surface layer consisting of numerous seromucous secretory cells, dendritic cells apical processes, and some goblet cell-like and ciliated cells. LM revealed that most filiform papillae showed the seromucous secretory cells in apparent resting state. These cells were characterized by a basal, round nucleus, pale cytoplasm with small, dispersed, stained granules in the supranuclear region. With TEM, seromucous secretory cells exhibited poorly developed RER, scattered medium electron dense secretory granules and microridges at the apical surface. Dendritic cells were seen as branched cells completely surrounding the basolateral surfaces of secretory cells. In addition, some dendritic cell processes have microvilli at their abutting endings. Goblet cell-like cells theca showed a continuous mesh of medium electron dense, partially fused, mucous granules and irregular microvilli-like processes at the apical end. Ciliated cells were mostly localized at the lateral wall of filiform papillae. Results suggest that at the filiform papillae, yuxtacrine or paracrine interactions mediated by physical contacts between dendritic and secretory cells may exist.

Mechanism of vascular relaxation by thaligrisine: functional and binding assays.

In the present study we examine the mechanism by which thaligrisine, a bisbenzyltetrahydroisoquinoline alkaloid, inhibits the contractile response of vascular smooth muscle. The work includes functional studies on rat isolated aorta and tail artery precontracted with noradrenaline or KCl. In other experiments rat aorta was precontracted by caffeine in the presence or absence of extracellular Ca2+. In order to assess whether thaligrisine interacts directly with calcium channel binding sites or with alpha-adrenoceptors we examined the effect of the alkaloid on [3H]-(+)-cis diltiazem, [3H]-nitrendipine and [3H]-prazosin binding to cerebral cortical membranes. The functional studies showed that the alkaloid inhibited in a concentration-dependent manner the contractile response induced by depolarization in rat aorta (IC50 = 8.9+/-2.9 microM, n=5) and in tail artery (IC50 = 3.04+/-0.3 microM, n=6) or noradrenaline induced contraction in rat aorta (IC50 = 23.0+/-0.39 microM, n=9) and in tail artery (IC50 = 3.8+/-0.9 microM, n=7). In rat aorta, thaligrisine concentration-dependently inhibited noradrenaline-induced contraction in Ca2+-free solution (IC50 = 13.3 microM, n=18). The alkaloid also relaxed the spontaneous contractile response elicited by extracellular calcium after depletion of noradrenaline-sensitive intracellular stores (IC50 = 7.7 microM, n=4). The radioligand receptor-binding study showed that thaligrisine has higher affinity for [3H]-prazosin than for [3H]-(+)-cis-diltiazem binding sites, with Ki values of 0.048+/-0.007 microM and 1.5+/-1.1 microM respectively. [3H]-nitrendipine binding was not affected by thaligrisine. The present work provides evidence that thaligrisine shows higher affinity for [3H]-prazosin binding site than [3H]-(+)-cis-diltiazem binding sites, in contrast with tetrandrine and isotetrandrine that present similar affinity for both receptors. In functional studies thaligrisine, acted as an alpha1-adrenoceptor antagonist and as a Ca2+ channel blocker, relaxing noradrenaline or KCl-induced contractions in vascular smooth muscle. This compound specifically inhibits the refilling of internal Ca2+-stores sensitive to noradrenaline, by blocking Ca2+-entry through voltage-dependent Ca2+-channels.

Halogenated derivatives of boldine with high selectivity for alpha1A-adrenoceptors in rat cerebral cortex.

The selectivity of 3-nitrosoboldine and different halogenated derivatives of boldine (3-bromoboldine, 3,8-dibromoboldine and 3-chloroboldine) for alpha1-adrenoceptor subtypes was studied by examining [3H]-prazosin competition binding in rat cerebral cortex. In the competition experiments [3H]-prazosin binding was inhibited completely by all the compounds tested. The inhibition curves displayed shallow slopes which could be subdivided into high and low affinity components. The relative order of affinity and selectivity for alpha1A-adrenoceptors was 3-bromoboldine = 3,8-dibromoboldine = 3-chloroboldine > boldine > 3-nitrosoboldine. The competition curves for 3-bromoboldine remained shallow and biphasic following chloroethylclonidine treatment. Whereas the relative contribution of the high affinity sites increased, the 3-bromoboldine affinities at its high and low affinity sites remained similar to those obtained in untreated membranes. 3-Bromoboldine, 3,8-dibromoboldine, 3-chloroboldine and 3-nitrosoboldine did not significantly displace [3H]-(+)-cis-diltiazem binding to rat cerebral cortex membranes. This activity was lower than that shown by boldine. Compared to boldine, halogen (bromine or chlorine) substitution at position 3 increases the alpha1A-adrenoceptor subtype selectivity and decreases the affinity for the benzothiazepine binding site at the calcium channel. Further halogen substitution at position 8 did not significantly improve this activity with respect to 3-bromoboldine. In contrast, the NO substitution at position 3 of boldine (3-nitrosoboldine) gives a loss of affinity and selectivity for alpha1-adrenoceptor subtypes.

Alpha-adrenoceptor interaction of tetrandrine and isotetrandrine in the rat: functional and binding assays.

The action of 1S,1'S-tetrandrine, a bisbenzyltetrahydroisoquinoline alkaloid, on alpha1-adrenoceptors has been compared with that of its isomer 1R,1'S-isotetrandrine. The work includes binding assays to analyse the affinity of these products for the [3H]prazosin binding site of rat cerebral cortical membranes and functional studies on rat isolated aorta to examine the effects of both alkaloids on intracellular calcium processes related or not to alpha-adrenoceptor activation. A radioligand receptor-binding study showed that both compounds interacted with the alpha1-adrenoceptors displacing [3H]prazosin from the specific binding site. The Ki values (inhibition constants) were 0.69+/-0.12 and 1.6+/-0.4 microM for tetrandrine and isotetrandrine, respectively. The functional studies showed that both alkaloids concentration-dependently inhibited noradrenaline-induced contraction in Ca2+-free solution (IC50 values, i.e. the concentrations needed to induce 50% inhibition, were 252.8 and 174.9 microM for tetrandrine and isotetrandrine, respectively), the spontaneous contractile response elicited by extracellular calcium after depletion of noradrenaline-sensitive intracellular stores (increase in resting tone; IC50 values 11.6 and 19.6 microM for tetrandrine and isotetrandrine, respectively) and the refilling of intracellular Ca2+ stores sensitive to noradrenaline (IC50 values 7.4 and 14.9 microM for tetrandrine and isotetrandrine, respectively). The results show that tetrandrine and isotetrandrine interact with alpha1-adrenoceptors by displacing the [3H]prazosin binding site and that both compounds inhibit mainly the Ca2+-dependent process and have less action on alpha1-adrenoceptors. Tetrandrine is more potent than isotetrandrine.

Relationships between structure and vascular activity in a series of benzylisoquinolines.

1. In the present work, the properties of 3-methyl isoquinoline, 3,4-dihydropapaverine, tetrahydropapaverine and tetrahydropapaveroline were compared with those of papaverine and laudanosine. The work includes functional studies on rat isolated aorta contracted with noradrenaline, caffeine or KCl, and a determination of the affinity of the compounds for alpha1-adrenoceptors and calcium channel binding sites, with [3H]-prazosin, [3H]-nitrendipine and [3H]-(+)-cis-diltiazem binding to rat cerebral cortical membranes. The effects of papaverine derivatives on the different molecular forms of cyclic nucleotide phosphodiesterases (PDE) isolated from bovine aorta were also determined. 2. The three papaverine derivatives show greater affinity than papaverine for the [3H]-prazosin binding site. They are therefore more selective as inhibitors of [3H]-prazosin binding as opposed to [3H]-(+)-cis-diltiazem, while papaverine appears to have approximately equal affinity for both. [3H]-nitrendipine binding was not affected by either papaverine or papaverine derivatives in concentrations up to 100 microM. 3-Methylisoquinoline had no effect on any of the binding sites assayed. 3. Contractions evoked by noradrenaline (1 microM) in rat aorta were inhibited in a concentration-dependent manner by 3,4-dihydropapaverine, tetrahydropapaverine and with a lower potency, by tetrahydropapaveroline. In Ca2+-free solution, tetrahydropapaverine and to a lesser extent, tetrahydropapaveroline, inhibited the noradrenaline (1 microM) evoked contraction in a concentration-dependent manner and did not modify the phasic contractile response evoked by caffeine (10 mM). This suggests that these alkaloids do not act at the intracellular level, unlike papaverine which inhibits the contractile response to caffeine and noradrenaline. 4. Inositol phosphates formation induced by noradrenaline (1 microM) in rat aorta was inhibited by tetrahydropapaverine (100 microM) and tetrahydropapaveroline (300 microM), thus suggesting that alpha1D-adrenoceptors are coupled to phosphoinositide metabolism in rat aorta. 5. Unlike papaverine, which has a significant effect on all the PDE isoforms, the three alkaloids assayed did not have an inhibitory effect on the different forms of PDE isolated from bovine aorta. 6. These results provide evidence that papaverine derivatives with a partially or totally reduced isoquinoline ring have a greater affinity for alpha1-adrenoceptors and a lower affinity for benzothiazepine sites in the Ca2+-channel than papaverine. This structural feature also implies a loss of the inhibitory activity on PDE isoforms. The planarity of the isoquinoline ring (papaverine) impairs the interaction with the alpha1-adrenoceptor site and facilitates it with the Ca2+-channels and PDEs, whereas the more flexible tetrahydroisoquinoline ring increases the binding to alpha1-adrenoceptors.

A possible structural determinant of selectivity of boldine and derivatives for the alpha 1A-adrenoceptor subtype.

1. The selectivity of action of boldine and the related aporphine alkaloids, predicentrine (9-O-methylboldine) and glaucine (2,9-O-dimethylboldine) and alpha 1-adrenoceptor subtypes was studied by examining [3H]-prazosin competition binding in rat cerebral cortex. WB 4101 and benoxathian were used as selective alpha 1A-adrenoceptor antagonists. 2. In the competition experiments [3H]-prazosin (0.2 nM) binding was inhibited by WB 4101 and benoxathian. The inhibition curves displayed shallow slopes which could be subdivided into high and low affinity components (pKi = 9.92 and 8.29 for WB 4101, 9.35 and 7.94 for benoxathian). The two antagonists recognized approximately 37% of the sites with high affinity from among the total [3H]-prazosin specific binding sites. 3. Boldine, predicentrine and glaucine also competed for [3H]-prazosin (0.2 nM) binding with shallow and biphasic curves recognizing 30-40% of the sites with high affinity. Drug affinities (pKi) at the high and low affinity sites were, 8.31 and 6.50, respectively, for boldine, 8.13 and 6.39 for predicentrine, and 7.12 and 5.92 for glaucine. The relative order of selectivity for alpha 1A-adrenoceptors was boldine (70 fold alpha 1A-selective) = predicentrine (60 fold, alpha 1A-selective) > glaucine (15 fold, alpha 1A-selective). 4. Pretreatment of rat cerebral cortex membranes with chloroethylclonidine (CEC, 10 microM) for 30 min at 37 degrees C followed by thorough washing out reduced specific [3H]-prazosin binding by approximately 70%. The CEC-insensitive [3H]-prazosin binding was inhibited by boldine monophasically (Hill slope = 0.93) with a single pKi value (7.76). 5. These results suggest that whereas the aporphine structure shared by these alkaloids is responsible for their selectively of action for the alpha 1A-adrenoceptor subtype in rat cerebral cortex, defined functional groups, namely the 2-hydroxy function, induces a significant increase in alpha 1A-subtype selectivity and affinity.

Effect of divalent cations on the contractile response of rat aorta to depolarization before and after nifedipine treatment.

The influence of the divalent cations, Ca2+, Mg2+ and Ba2+, on the contractile response of the rat aorta to KCl and on the recovery of this response after nifedipine treatment was analyzed. KCl (80 mmol/l) promoted a two-phase (phasic and tonic) contractile response in Krebs solution but, as expected, no contractile response in Ca(2+)-free medium. In Mg(2+)-free medium, the phasic response to KCl was unaffected but the tonic one decreased slowly, suggesting that a long incubation time in the absence of Mg2+ (65 min) promotes a loss of or a change in the intracellular distribution of this ion that modifies Ca2+ entry through L channels or Ca2+ handling. Ba2+ (1.8 mmol/l) contracted the rat aorta in the absence or presence of Ca2+ but, when Ca2+ was not present, Ba2+ modified the contractile process so that a new addition of KCl did not reproduce the contractile response. After nifedipine treatment, no phasic response to KCl was observed and only a slow response appeared that increased as long as the KCl was present (45 min). After washing, KCl induced a phasic response similar to the standard response, but a smaller tonic one. Divalent cations accelerated the recovery of the channels involved in the tonic contraction. However, the recovery of the Ca2+ channels related to the phasic contraction was independent of divalent cations.

The sources of Ca2+ for muscarinic receptor-induced contraction in the rat ileum.

The contractile responses obtained by activation of different muscarinic receptor subtypes in the longitudinal muscle of the rat ileum and especially the responses of this muscle to acetylcholine in a Ca(2+)-free medium have been investigated. In Ca(2+)-containing solution, acetylcholine elicited similar concentration-dependent contractile responses in the duodenum, jejunum and ileum strips of the rat intestine. The response to a maximal concentration of the agonist (1 microM) consisted of a rapid phasic response followed by a slower tonic one. Nifedipine completely relaxes or inhibits the sustained response and only partially diminishes the phasic one, which suggests that the phasic contraction depends on the release of internal Ca2+ entry from the extracellular space through voltage-dependent Ca2+ channels, but that the tonic contraction only depends on the influx of the external ion. In Ca(2+)-free medium, acetylcholine (1 microM) induced phasic contractions that depend on the release of this ion from internal stores. Participation of different subtypes of receptors (M1, M2 and M3) in these responses depends on the inhibitory action shown by methoctramine, 4-DAMP and atropine but not by pirenzepine in two different experimental models.

[Histochemical characterization of glycoconjugates in the tongue of the toad Bufo marinus L. (Amphibian, anuran) with conventional techniques for light microscopy]. / Caracterización histoquímica de los glicoconjugados de la lengua del sapo Bufo marinus L. (Anfibio, anuro) con técnicas convencionales para microscopía de luz.

Histochemical stains with and without enzymatic digestions such as alpha-amylase, neuraminidase and hyaluronidase, made possible to localize and differentiate various types of glycoconjugates (GCs) in the tongue of the toad Bufo marinus. In the dorsal mucosae the covering epithelium of the filiform papillae, of the central folds and in the marginal cells of the fungiform papillae there were present large amounts of neutral GCs with little or no galactose and/or N-acetylgalactosamine and scanty carboxylic acid GCs while the superficial strata of the taste organs showed a mixture of neutral an acid GCs with a predominance of sulfated and carboxylic acid GCs. The glandular secretory cells showed neutral GCs almost exclusively with a gradient of concentrations increasing from the base to the apex being galactose or N-acetyl-galactosamine one of the component sugars. The ventral epithelium showed two types of mucous cells, one with neutral GCs and the other with neutral and acidic GCs. The connective tissue contained many mast cells showing highly acid GCs both sulfated and carboxylic with some neutral GCs. The extracellular connective matrix showed scanty neutral and acid GCs. Glycogen was present in the cytoplasm of glandular epithelial cells and of the striated muscle fibers. Additionally, the obtained results suggest the presence of a type of GC with a carboxylic acid (sialic acid) resistant to neuraminidase of Clostridium perfringens used in this study.

[Lectin histochemistry of the Bufo marinus L. toad tongue]. / Histoquímica de lectinas en la lengua del sapo Bufo marinus L.

Lectin histochemistry at light microscope level was used in the tongue of the cane toad Bufo marinus to determine the distribution of sugar residues in glycoconjugates (GCs) previously localized and characterized by conventional histochemical techniques. Five horseradish-peroxidase (HRP) labeled-lectins, namely Con A, PNA, SBA, UEA-1 and WGS were used. Additionally, neuraminidase (N) treated sections before the staining procedures were used in order to dilucidate the presence of terminal sialic acid (SA). Sugar residues in GCs of the taste organ (TO) associated mucous cells stained more intensely with WGA than with Con A and UEA-1. All the sensory cells reacted with Con A and WGA but one type of them were characteristically labeled by UEA-1. The glycocalix (gc) of the TOs resulted intensely stained with Con A and with WGA and UEA-1 before and after N treatment. The GCs in the mucous-supporting cells of dorsal mucosae filiform papillae and folds reacted intensely with WGA and weakly with Con A. The ciliated cells (cic) were intense and characteristically stained with UEA-1 and WGA and moderately with Con A. The gc reacted more intensely with WGA than with Con A. Dorsal mucosae glands secretory cells mucins were characteristically stained with PNA, SBA and WGA besides Con A, while glandular ciliated cells showed the same staining pattern as in the filiform papillae. In the ventral mucosa all epithelium cells resulted stained with WGA and Con A, while differentiated goblet cells only reacted as well with UEA-1 and PNA before and after neuraminidase treatment. Unexpectedly, ciliated ventral mucosae cells did not react with UEA-1 but only with WGA and Con A. The results have shown that lectin histochemistry is an interesting tool to characterize similarities and differences in the lingual GCs sugar residues composition and distribution, particularly those located in epithelial cells.

Skeletal growth after oral administration of demineralized bone matrix.

Oral administration of bone extracts obtained from bovine demineralized bone matrix to rats has a direct effect on bone metabolism, affecting bone proportions and some markers of bone formation such as bone malate dehydrogenase, serum alkaline phosphatase and serum osteocalcin. Furthermore collagen deposition, bone protein synthesis and nucleic acids content were significantly increased by the treatment.

Potential induction of osteogenesis by systemic administration of bovine bone proteins.

This study shows, apparently for the first time, that the administration of bone derived proteins (putative bone growth factors) obtained from bovine demineralized maxillaries has a direct effect on osteogenesis, affecting significantly some markers of bone formation such as lactate dehydrogenase activity and serum osteocalcin. Also, collagen deposition and bone protein turnover were markedly increased by the treatment, which may have important biological and clinical applications.

Spleen and thymus histology and proliferative response of splenic cells in rats fed raw and cooked Phaseolus vulgaris beans.

Histological studies of the spleen and thymus of rats fed raw black beans (Phaseolus vulgaris) show an atrophy of both lymphoid organs. Decrease in relative thymus weight was most marked. All histological organization of this organ appeared altered. An evident decrease in cell number was also observed in both organs. Proliferative response of splenic cells stimulated in vitro with Concanavalin A was increased as compared to that from animals fed the control diet. It is likely that histological changes observed in the spleen and the thymus are due mainly to a protein caloric deficiency, although the possibility that toxic factors present in the raw diet have an effect on the immune system of the rat can not be overruled.

Scanning electron microscopy study on the toad (Bufo marinus L) tongue dorsal mucosa

Empleando un método sencillo se hizo un estudio con el microscopio electrónico de barrido de la mucosa lingual dorsal del sapo, caracterizando las regiones distal, central y proximal. La región distal mostró dos tipos de proyecciones, las papilas fungiformes y filiformes; la región central presentó pliegues y papilas fungiformes acortadas, mientras que la superficie de la región proximal se presentó ciliada y con órganos gustaticos del tipo no-papilar. En las tres regiones señaladas se observaron diferencias en el aspecto superficial del disco gustativo pero siempre éste se encontró separado del epitelio adiacente mediante un surco. Las células mucosas de las papilas filiformes distales se caracterizaron por una cubierta formada por numerosas microvellosidades mientras que las células de los pliegues en la región central, mostraron células mucosas con microvellocidades o con expansiones granulares lo que sugiere que estas células se encuentran en diferentes fases de la actividad secretora. Los resultados confirman las observaciones realizadas previamente con microscopía de luz

Estructura histologica de la mucosa lingual de la rana acuatica Sudamericana: Pseudis paradoxus caribensis / Histological structure of the lingual mucosa of the aquatic South American frog: Pseudis paradoxus caribensis Gallardo

Se realiza un estudio detallado de la organización histológica de la mucosa lingual del anuro acuátioco Pseudis pardoxus caribensis mediante cortes sagitales y transversales de todo el órgano. Grupos de seis láminas consecutivas se colorearon con diferentes métodos selectivos de coloración. La lengua es redondeada y ampliamente fijada a la mandíbula inferior. La mucosa dorsal es papilar excepto cerca del borde en todo el perímetro lingual. El epitelio simple que reviste las papilas filiformes presenta tres tipos celulares distintos. Las papilas fungiformes contienen los órganos gustativos discoidales característicos de los anfibios. Ocasionalmente encuentran órganos gustativos no-papilares inmersos en la mucosa ventrolateral. El revestimento ventral es ciliado y con dos tipos de células mucosas. El tejido conectivo contiene numerosos mastocitos dispersos. Estas características indican que la organización histológica básica es similar al observado en la lengua de anuros terrestres más evolucionados, pero que presenta una combinación única de pequeñas variaciones que podrían constituir una peculiaridad de este género

Mucopolysaccharides in hypothalamic neurons of the rat.

A study of the hypothalamic nuclei of the rat, using a variety of histochemical methods, has shown the distribution of mucopolysaccharides and neurosecretory material. In all the hypothalamic nuclei there are two types of neurons and in some a third type characterized by the presence of hyaluronidase-resistant acid mucopolysaccharides in the periphery of the perikaryon. These type III neurons also contain Gomori-positive material and neutral mucopolysaccharides. It is considered that hypothalamic nuclei with type III neurons have similar neurosecretory properties.