Differential expression of upstream stimulatory factor (USF) 2 variants in eutopic endometria from women with endometriosis: estradiol regulation

BACKGROUND: Endometriosis, pro-inflammatory and invasive benign disease estrogen dependent, abnormally express in endometria the enzyme P450Arom, positively regulated by steroid factor-1 (SF-1). Our objective was to study the nuclear protein contents of upstream stimulating factor 2 (USF2a and USF2b), a positive regulator of SF-1, throughout the menstrual cycle in eutopic endometria from women with and without (control) endometriosis and the involvement of nuclear estrogen receptors (ER) and G-coupled protein estrogen receptor (GPER)-1. RESULTS: Upstream stimulating factor 2 protein contents were higher in mid (USF2b) and late (USF2a and USF2b) secretory phase in eutopic endometria from endometriosis than control (p < 0.05). In isolated control epithelial cells incubated with E2 and PGE2, to resemble the endometriosis condition, the data showed: (a) significant increase of USF2a and USF2b nuclear protein contents when treated with E2, PPT (specific agonist for ERa) or G1 (specific agonist for GPER1); (b) no increase in USF2 binding to SF-1 E-Box/DNA consensus sequence in E2-treated cells; (c) USF2 variants protein contents were not modified by PGE2; (d) SF-1 nuclear protein content was significantly higher than basal when treated with PGE2, E2 or G1, stimulation unaffected by ICI (nuclear ER antagonist); and (e) increased (p < 0.05) cytosolic protein contents of P450Arom when treated with PGE2, E2, PPT or G1 compared to basal, effect that was additive with E2 + PGE2 together. Nevertheless, in endometriosis cells, the high USF2, SF-1 and P450Arom protein contents in basal condition were unmodified. CONCLUSION: These data strongly suggest that USF2 variants and P450Arom are regulated by E2 through ERa and GPER1, whereas SF-1 through GPER1, visualized by the response of the cells obtained from control endometria, being unaffected the endogenously stimulated cells from endometriosis origin. The lack of E2 stimulation on USF2/SF-1 E-Box/DNA-sequence binding and the absence of PGE2 effect on USF2 variants opposite to the strong induction that they exert on SF1 and P450 proteins suggest different mechanisms and indirect regulations. The sustained USF2 variants protein expression during the secretory phase in eutopic endometria from women with endometriosis may participate in the pathophysiology of this disease strongly associated with infertility and its characteristic endometrial invasion to ectopic sites in the pelvic cavity.

Estudo da correlação instrumental e sensorial de uma composição aromática aplicada na pele em função do ciclo menstrual / Study of instrumental and sensory correlation of a aromatic composition on the skin in function of mentrual cycle

A área de perfumaria no mundo vem se desenvolvendo a cada dia buscando maior conhecimento das matérias-primas aromáticas, desde suas reações, estabilidade até suas interações com o substrato onde é aplicado, sempre em busca do conhecimento de todas as variáveis que possam influenciar a relação perfume-substrato e a aceitação dos consumidores, medida por meio da avaliação sensorial. Apesar de muitos estudos sobre a relação perfume-pele, poucos envolveram a relação com ciclo menstrual. Neste estudo o objetivo foi correlacionar às análises sensorial e instrumental (medidas biomecânicas e cromatográficas), estudar as matérias-primas aromáticas em função do ciclo menstrual. O estudo envolveu indivíduos com idade entre 18-40 anos: 29 mulheres e 3 homens, estes usados como grupo controle. Cada voluntária teve 40 µl da composição aromática Ciclo 1910 aplicado no antebraço, onde foram feitas as medidas biomecânicas (corneometria, sebumetria e TEWL) nos tempos inicial e 6h. Nos tempos inicial, 1.5h, 3h, 4,5h e 6h se auto-avaliaram sensorialmente a intensidade de perfume por meio de escala sensorial de magnitude rotulada (LMS) e foram coletados os compostos aromáticos liberados pela técnica de headspace e analisados por espectrometria com cromatografia a gás e detetor de massa (CGMS). Realizou-se também medidas biomecânicas de corneometria, sebumetria e TEWL interescapulares em 5 voluntárias nos tempos inicial e 6h para comparação. Os resultados obtidos foram que as fases do ciclo não interferiram nas variáveis biomecânicas (p>0,05) quando analisadas isoladamente. Para as medidas realizadas no antebraço, os índices de Corneometria (p<0,001) e TEWL (p: 0,011) na região tratada apresentou média maior que a do controle. A média dos índices de Corneometria e Sebumetria da região do antebraço tratada foi menor (p<0,001) que da região interescapular. Na análise cromatográfica não houve um padrão de resposta em diferentes fases do ciclo. As correlações entre a avaliação sensorial e instrumental (cromatografia e propriedades biomecânicas) não observaram nenhum nível de correlação (p>0,05). A Cromatografia (dados cromatográficos) foi maior na fase Folicular que nas fases Menstrual e Ovulatória (p=0,003), ao considerar como variável resposta em função do ciclo menstrual e da avaliação sensorial. Houve uma forte correlação positiva entre a análise sensorial e a avaliação na pele do homem (p<0,001). No entanto, o fator intrínseco do indivíduo Mulher influenciou na resposta, ocasionando grande variabilidade, porém percebeu-se claramente que os hormônios sexuais interferiram na resposta sensorial, cromatográfica e biomecânica da pele

The perfumery in the world has been developed everyday bringing more knowledge about aromatic raw-materials, as from chemistry reactions, stability until their interactions with substrate where is applied, always looking for variables could influence in the relation perfume-substrate and consumer acceptability, measured by sensory evaluation. Despite a lot of studies were done on this subject, few involved effects as function of menstrual cycle. The aim of this study was to correlate sensory and instrumental analysis (biomechanical and chromatographic measurements), to study the olfactory profile of raw materials in function of menstrual cycle. The study involved people with 18-40 years old: 29 volunteers, three men, who were used as control group. Each volunteer had 40 µl of perfume applied on forearm, where were done Biomechanical measurements (Corneometer, Sebumeter and TEWL) at initial and 6h, At time initial, 1.5h, 3h, 4.5h and 6h; they did self-sensory assessment in perfume intensity in own forearm using labeled magnitude scale (LMS) and also where aromatic compounds released were collected by headspace technique spectrometry and gas chromatography with mass detector (CGMS). In addition, it was done biomechanical measurements (Corneometer, Sebumeter and TEWL) on interscapular region at initial and 6h for comparing. Resulting that the phases of the cycle did not affect the biomechanical variables (p > 0.05) when analyzed individually. For measurements in the forearm, Corneometry index (p < 0.001) and TEWL (p=0.011) in the treated area were higher than the control. The average of the Sebumetry, Corneometry indexes of the forearm treated was lower (p <0.001) than the interscapular region. In the analysis of chromatographic, there was a standard response at different stages of the menstrual cycle, however the analysis by individual had no a pattern response to the release of aromatic compounds. The correlations between sensory and instrumental (chromatography and biomechanical properties) did not observe any correlation (p> 0.05). But when considering chromatography as the response variable as a function of the menstrual cycle and the sensory evaluation, the follicular phase was higher than the Menstrual and Ovulatory phase (p=0.003). There was a strong positive correlation between sensory analysis and evaluation on men skin (p<0.001). However, the intrinsic factor of the individual woman influenced the response, leading to large response variability; however, see clearly that sex hormones interfere in the sensory response, chromatographic and biomechanics of the skin

Genes responsible for vaginal extracellular matrix metabolism are modulated by women's reproductive cycle and menopause

Objectives To analyze the expression of genes involved in extracellular matrix (ECM) biogenesis and remodeling in vaginal tissue of women with clinically normal pelvic floor support (defined as controls) according to the phase of menstrual cycle and postmenopausal women with and without pelvic organ prolapse (POP). Materials and Methods This study examined the expression of matrix metalloproteinases (MMPs), their tissue inhibitors (TIMPs), and the Lysyl oxidase (LOX) family genes in the anterior vaginal wall of Caucasian women by real-time RT-PCR. Initially, mRNA expression was assessed in premenopausal controls in the secretory (group 1, n = 10) vs. proliferative (group 2, n = 8) phase of menstrual cycle. In addition, we compared premenopausal controls in the proliferative phase (group 2) vs. postmenopausal controls (group 3, n = 5). Finally, we analyzed postmenopausal controls (group 3) vs. postmenopausal women with advanced POP (group 4, n = 13). Results According to the phase of menstrual cycle, MMP1 was significantly reduced (p = 0.003), whereas the expression of TIMP1 and LOXL4 was significantly up-regulated during proliferative phase (both p < 0.01) when compared to the secretory phase in premenopausal control women. Regarding menopausal status/ageing, all MMPs were down-regulated, while TIMP3, TIMP4 and LOXL2 were significantly up-regulated in postmenopausal control women when compared to premenopausal controls (p = 0.005, p = 0.01 and p < 0.001, correspondingly). TIMP4 and LOXL2 mRNA levels were significantly decreased in postmenopausal POP patients compared to asymptomatic postmenopausal controls (p < 0.01 for both). Conclusions Our results indicate that ovarian cycle and age-related changes influence the expression of genes encoding proteins responsible for ECM metabolism in human vagina. Moreover, POP is associated with alteration in vaginal ECM components after menopause. .

Anticoncepcionais orais combinados em regime estendido / Combined oral contraceptives in extended regimen

Denominam-se regimes estendidos em contracepção oral combinada a utilização de pílulas por mais de 28 dias sem pausa, visando a supressão da menstruação. Incluem o uso contínuo dos contraceptivos, bem como de regimes com intervalos trimestrais. Os questionamentos acerca da necessidade da menstruação, bem como dos intervalos mensais entre as usuárias de anticoncepcionais hormonais, motivou, nos últimos anos, o interesse crescente por regimes contraceptivos não convencionais. Nesse sentido, a conveniência e a melhora dos sintomas como cólicas, cefaleia e inchaço figuram entre as principais indicações dos regimes estendidos, além do possível efeito sobre doenças menstruais relacionadas. O propósito deste estudo foi identificar os principais aspectos referentes ao uso dos anticoncepcionais em regime estendido, com ênfase sobre as indicações, formulações disponíveis, padrão de sangramento, efeitos adversos e perfil metabólico

Extended regimens in combined oral contraception mean continuous administration, greater than 28 days of active hormone, in order to avoid menstruation. Extended regimens include some kind of contraception with no interval (as continuous) and with intervals every three months. Questions about the necessity of menstruation, as well as monthly intervals between hormonal contraceptive has motivated the growing interest in unconventional contraceptives regimens. Convenience and improved symptoms such as cramping, bloating and headache are among the main indications for extended regimens, in addition to the possible effect on menstrual-related diseases. The purpose of this study was to identify the main aspects regarding the use of contraceptives in extended regimens, with emphasis on indications, formulations available, bleeding patterns, adverse effects and metabolic profile

Endometriose: papel da superfamília do fator transformador de crescimento beta / Endometriosis: the role of transforming growth factor β superfamily

A endometriose é uma condição ginecológica, que atinge mulheres em idade reprodutiva e pode ser causa de dor e infertilidade. A patogênese da doença é multifatorial e envolve a perda da capacidade de diferenciação das células endometrióticas, moléculas de adesão celular para adesão do endométrio ao peritônio, neoangiogênese, características do fluido peritoneal e alterações do sistema imune. A superfamília do fator transformador de crescimento β (TGF-β) parece exercer papéis importantes na implantação e manutenção do tecido ectópico na endometriose. Ativinas, inibinas, folistatina, hormônio anti-mülleriano e as proteínas morfogenéticas ósseas são membros da superfamília do TGF-β. Estas moléculas são expressas no endométrio humano e apresentam ações importantes na proliferação celular, diferenciação celular, função imune, regulação da apoptose e remodelamento dos tecidos, apresentando, por conseguinte, um importante papel no ciclo menstrual, decidualização do endométrio e no início da gestação. Este artigo objetiva rever os achados sobre tais proteínas no endométrio e seus possíveis papéis na gênese e fisiopatologia da endometriose

Endometriosis is a gynecological pathological entity typical of women in reproductive age, associated with pelvic pain and infertility. The pathogenesis of the disease is multifactorial and it involves loss of the endometriotic cell differentiation, cell adhesion, neo-angiogenesis, peritoneal fluid characteristics, and changes in the immune system. The transforming growth factor β (TGF-β) superfamily seems to play important roles in the implementation and maintenance of ectopic tissue in endometriosis. Activin, inhibin, follistatin, anti-Mullerian hormone, and bone morphogenetic proteins are members of the superfamily of TGF-β. The TGF-β and family members are expressed by human endometrium and act on cell proliferation, differentiation, immune function, apoptosis and tissue remodeling, playing a role in menstrual cycle, decidualization, and early pregnancy. The aim of this study is to review the findings about these molecules in the endometrium and their possible roles in the genesis and pathophysiology of endometriosis

Biochemical evaluation in human saliva with special reference to ovulation detection.

Aim : The aim of the present study was to investigate the level of salivary sialic acids and glycosaminoglycans with reference to salivary hormones during the normal menstrual cycle. Settings and Design: Fifty women volunteers were selected for the present study. Materials and Methods : Saliva was collected from 50 women and ovulation was detected in women with normal menstrual cycles through basal body temperature (BBT), ultrasound and salivary ferning. Samples were divided into five categories, as prepubertal (6-9 years), pre-ovulatory phase (6-12 days), ovulatory phase (13-14 days), postovulatory phase (15-26 days) and menopause (above 45 years). Each sample was subjected to evaluation of the sialic acids and glycosaminoglycans along with salivary hormones. Results : The result revealed that the ovulatory phase has increased sialic acid and glycosaminoglycans during the menstrual cycle when compared with that of the other phases. Consequently, an increased level of hormones such as luteinizing hormone and estrogen during the ovulatory period when compared to that of the pre-ovulatory and postovulatory periods appeared to be noteworthy. Statistically, analysis was performed using one way-ANOVA (LSD; post hoc method) to determine the significance as P < 0.001, 0.01, 0.05 in between the reproductive phases of the menstrual cycle. Conclusion : This study concluded that saliva-specific carbohydrates in the ovulatory saliva make the possibility to develop a biomarker for detection of ovulation by non-invasive methods.

Ciclo menstrual e consumo de alimentos / Menstrual cycle and food consumption / Ciclo menstrual y el consumo de alimentos

Objetivou-se neste trabalho, avaliar o consumo alimentar de mulheres durante as fases folicular e lútea do ciclo menstrual e estabelecer as possíveis relações entre a ingestão alimentar e os sintomas da Síndrome Pré-Menstrual. Foram investigadas 30 alunas dos cursos de Pós-Graduação da Universidade Federal de Santa Catarina, por meio de registro alimentar de três dias, sendo um de final de semana, durante as fases folicular e lútea. Na lútea, aplicou-se um questionário de sintomas pré-menstruais. Os dados foram avaliados no programa SPSS 14 for Windows. O consumo energético, de carboidratos e lipídios, foi significativamente maior na fase lútea (p<0,0001). Os sintomas da Síndrome Pré-Menstrual ocorreram em 96,7% da amostra investigada, sendo que 63,4% apresentaram manifestação severa e 3,3% leve. Os sintomas emocionais foram os mais citados pela amostra (93,3%). O apetite foi maior na fase pré-menstrual em 53,3% das participantes e 43,3% relataram ter vontade de comer alimentos doces neste período. Não foram encontradas correlações entre o consumo alimentar (energético e de macronutrientes) e a manifestação de sintomas. A partir destes resultados, acredita-se que exista interferência do ciclo ovariano sobre o comportamento alimentar. Assim, sugere-se a realização de pesquisas com maior amostragem, a fim de verificar a estreita relação entre o consumo alimentar e a manifestação de sintomas da Síndrome Pré-Menstrual.

The objective of this study was to evaluate the dietary intake of women during the follicular and luteal phases of the menstrual cycle and establish the possible relationship between food intake and symptoms of Premenstrual Syndrome. We investigated 30 female students of graduate courses at the Federal University of Santa Catarina, through three days of food records, one weekend during the follicular and luteal phases. In the luteal, applied a questionnaire to premenstrual symptoms. The data were analyzed using SPSS 14 for Windows. The energy consumption of carbohydrates and lipids, was significantly higher in the luteal phase (p <0.0001). The symptoms of premenstrual syndrome occurred in 96.7% of the investigated sample, and 63.4% had severe outbreak and 3.3% mild. The emotional symptoms were the most cited by the sample (93.3%). The appetite was higher in the premenstrual phase in 53.3% and 43.3% of participants reported having cravings for sweet foods in this period. No correlation was found between dietary intake (energy and nutrients) and the manifestation of symptoms. From these results, we believe that there is interference of the ovarian cycle on feeding behavior. Thus, it is suggested to conduct research using larger samples to verify the close relationship between food consumption and manifestation of symptoms of premenstrual syndrome.

El objetivo de este estudio fue evaluar la ingesta alimentaria de las mujeres durante las fases folicular y lútea del ciclo menstrual y establecer la posible relación entre la ingesta de alimentos y los síntomas del Síndrome Premenstrual. Se investigaron 30 alumnas de los cursos de postgrado en la Universidad Federal de Santa Catarina, a través de tres días a partir de los registros de alimentos, un fin de semana durante las fases folicular y luteínica. En la fase lútea, aplicó un cuestionario a los síntomas premenstruales. Los datos fueron analizados utilizando el programa SPSS 14 para Windows. El consumo de energía de los carbohidratos y lípidos, fue significativamente mayor en la fase lútea (p <0,0001). Los síntomas del síndrome premenstrual se produjo en el 96,7% de la muestra investigada, y 63.4% tenían brote severo y el 3,3% leve. Los síntomas emocionales son los más citados por los encuestados (93,3%). El apetito es mayor en la fase premenstrual en el 53,3% y 43,3% de los participantes reportó haber antojos por alimentos dulces en este período. No se encontró correlación entre el consumo de la dieta (energía y nutrientes) y la manifestación de los síntomas. A partir de estos resultados, creemos que hay interferencia del ciclo ovárico en la conducta alimentaria. Por lo tanto, se sugiere llevar a cabo la investigación con muestras más grandes para comprobar la estrecha relación entre el consumo de alimentos y la manifestación de los síntomas del síndrome premenstrual.

Positive correlation between serum and peritoneal fluid CA-125 levels in women with pelvic endometriosis / Correlação positiva entre os níveis séricos e no fluido peritonial de CA-125 em mulheres com endometriose pélvica

CONTEXT AND OBJECTIVE: One of the diagnostic markers of endometriosis is CA-125, and elevated levels of this are caused by high concentrations in the ectopic endometrium. The objective of this study was to correlate CA-125 levels in serum and peritoneal fluid from women with and without pelvic endometriosis. DESIGN AND SETTING: This was a prospective, cross-sectional, controlled study of consecutive pa-tients undergoing laparoscopy for infertility, pelvic pain or tubal ligation, during early follicular phase, at the university hospital of Faculdade de Medicina de Ribeirão Preto. METHODS: Fifty-two patients were divided into two groups: endometriosis group, consisting of 35 patients with biopsy-confirmed pelvic endometriosis, and control group, consisting of 17 patients without endometriosis. CA-125 levels in serum samples and peritoneal fluid were determined by chemiluminescence. RESULTS: CA-125 levels in serum and peritoneal fluid were higher in patients with advanced pelvic endometriosis (means of 39.1 ± 45.8 U/ml versus 10.5 ± 5.9 U/ml in serum, p < 0.005; 1,469.4 ± 1,350.4 U/ml versus 888.7 ± 784.3 U/ml in peritoneal fluid, p < 0.05), and showed a positive correlation between each other (correlation coefficient (r) = 0.4880). Women with more advanced degrees of endometriosis showed higher CA-125 levels in both serum and peritoneal fluid (p = 0.0001). CONCLUSION: There is a positive correlation between serum and peritoneal fluid values of CA-125 in women with and without endometriosis, and their levels are higher in peritoneal fluid. Advanced endometriosis is related to higher levels in both serum and peritoneal fluid.

CONTEXTO E OBJETIVO: Um dos marcadores diagnósticos de endometriose é o CA-125, e seus níveis elevados são devidos à alta concentração no endométrio ectópico. O objetivo deste estudo foi correlacionar os níveis de CA-125 no soro e fluido peritonial de mulheres com e sem endometriose pélvica. TIPO DE ESTUDO E LOCAL: Estudo prospectivo, longitudinal, controlado, de pacientes consecutivas submetidas a laparoscopia por infertilidade, dor pélvica ou laqueadura tubária, durante a fase folicular precoce no Hospital Universitário da Faculdade de Medicina de Ribeirão Preto. MÉTODOS: Cinqüenta e duas pacientes foram divididas em dois grupos: grupo endometriose, com 35 pacientes com biópsia confirmada de endometriose pélvica, e grupo controle, com 17 pacientes sem endometriose. Níveis de CA-125 em amostras no soro e fluido peritonial foram determinadas por quimiluminescência. RESULTADOS: Os níveis de CA-125 no soro e fluido peritonial foram mais altos nas pacientes com endometriose pélvica avançada (média 39,1 ± 45,8 U/ml versus 10,5 ± 5,9 U/ml no soro, p < 0,005, 1469,4 ± 1350,4 U/ml versus 888,7 ± 784,3 U/ml no fluido peritonial, p < 0,05), e o estudo mostrou uma correlação positiva entre eles (coeficiente de correlação = 0,4880). Mulheres com estágios mais avançados de endometriose mostraram níveis de CA-125 maiores em ambos soro e fluido peritonial (p = 0,0001). CONCLUSÃO: Há uma correlação positiva entre os valores de CA-125 no soro e no fluido peritonial em pacientes com e sem endometriose e seus níveis são maiores no fluido peritonial. Endometriose avançada é relacionada com níveis mais altos de CA-125 em ambos soro e fluido peritonial.

Is the menstrual cycle affecting the skin prick test reactivity?

Allergen skin prick tests (SPT) are very sensitive and specific tests to detect allergic sensitization in atopic patients. Certain factors like antihistamines, antidepressant therapies or circadian rhythms can alter the results of SPT. In women, the changes in endogenous hormone levels throughout the menstrual cycle may affect the allergic responses and natural course of allergic diseases. The aim of this study was to investigate the probable influence of the phases of the menstrual cycle on SPT reactivity to allergen extracts and histamine. Forty-two female patients with seasonal allergic rhinoconjunctivitis were enrolled in the study. Skin prick test reactivities to allergens and histamine were measured at the beginning of the menstrual cycle (3rd or 4th day), mid-cycle (14th or 15th day) and end-cycle (27th or 28th day) consecutively. Serum estradiol, progesterone, luteinizing hormone (LH), and follicle stimulating hormone (FSH) levels were determined simultaneously. We observed the most significant reactions to allergens when SPT is performed at mid-cycle. However, SPT reactivity to histamine did not vary throughout the menstrual cycle. Serum estradiol and LH levels showed positive correlation with SPT reactivity to allergens at mid-cycle. Our results suggest that SPT give the best results when they are performed at mid-cycle. Additionally, allergens seem to cause mast cell degranulation to a greater extent in subjects in which endogenous hormones like estradiol and LH are elevated.

Immunohistochemical localization of macrophage CD68+, HLA-DR+, L1+ and CD44+ subsets in uterine endometrium during different phases of menstrual cycle.

Different tissue macrophage subsets were immunohistochemically examined in normal endometrial samples collected from proliferative (n=4), peri-ovulatory (n=6) and secretory (n=8) phases of menstrual cycles in women. The different macrophage subsets, namely CD68 (pan macrophage marker), CD44 (transmembrane adhesion molecule), HLA-DR (transmembrane heterodimeric protein involved in antigen presentation) and L1 (calprotectin)-positive cells, as well as, CD45 (common leucocytic antigen)-positive cells were examined on the basis of immunohistochemical staining, and areas of immunoprecipitation were analyzed morphometrically using computer-assisted video imaging system. The stage-specific distribution of receptors for estrogen (ER) and progesterone (PR) in endometrial cells were examined and morphometrically analyzed. There was an increase in the number of CD45+ cells (P < 0.01) and CD68+ cells (P < 0.05) in secretory phase endometrium compared with proliferative and peri-ovulatory phases. There was no remarkable cycle dependent pattern in HLA-DR+ and L1+ cells. However, there was an increase in CD44 immunopositive area in peri-ovulatory (P < 0.05) and in secretory (P < 0.01) phases of endometrium compared with proliferative phase endometrium. A higher (P < 0.01) degree of immunopositivity for ER was observed during peri-ovulatory phase, and for PR, during peri-ovulatory (P < 0.05) and secretory (P < 0.01) phases compared with proliferative phase of cycle. Positive correlations between areas occupied by (i) CD68+ cells and PR (P < 0.01), (ii) HLA-DR+ and L1+ cells (P < 0.05), (iii) CD45+ and CD68+ cells (P < 0.01), (iv) CD45+ and L1+ cells (P < 0.05), and (v) PR and L1+ cells (P < 0.05) were obtained. It appears that the recruitment of different macrophage subsets in human endometrium involves a complex set of endocrine and paracrine factors.

Cycle modulation of insulin-like growth factor-binding protein 1 in human endometrium

Endometrium is one of the fastest growing human tissues. Sex hormones, estrogen and progesterone, in interaction with several growth factors, control its growth and differentiation. Insulin-like growth factor 1 (IGF-1) interacts with cell surface receptors and also with specific soluble binding proteins. IGF-binding proteins (IGF-BP) have been shown to modulate IGF-1 action. Of six known isoforms, IGF-BP-1 has been characterized as a marker produced by endometrial stromal cells in the late secretory phase and in the decidua. In the current study, IGF-1-BP concentration and affinity in the proliferative and secretory phase of the menstrual cycle were measured. Endometrial samples were from patients of reproductive age with regular menstrual cycles and taking no steroid hormones. Cytosolic fractions were prepared and binding of 125I-labeled IGF-1 performed. Cross-linking reaction products were analyzed by SDS-polyacrylamide gel electrophoresis (7.5 percent) followed by autoradiography. 125I-IGF-1 affinity to cytosolic proteins was not statistically different between the proliferative and secretory endometrium. An approximately 35-kDa binding protein was identified when 125I-IGF-1 was cross-linked to cytosol proteins. Secretory endometrium had significantly more IGF-1-BP when compared to proliferative endometrium. The specificity of the cross-linking process was evaluated by the addition of 100 nM unlabeled IGF-1 or insulin. Unlabeled IGF-1 totally abolished the radioactivity from the band, indicating specific binding. Insulin had no apparent effect on the intensity of the labeled band. These results suggest that IGF-BP could modulate the action of IGF-1 throughout the menstrual cycle. It would be interesting to study this binding protein in other pathologic conditions of the endometrium such as adenocarcinomas and hyperplasia

Localization of estrogen and progesterone receptors in the endometrium of common marmosets Callithrix jacchus.

In the present study, changes in the immunohistochemical localization of endometrial estrogen receptor (ER) and progesterone receptor (PR) during various stages of the ovarian cyclicity in common marmoset, have been reported. Ovarian cyclicity was monitored by estimating plasma estradiol and progesterone. During the early follicular phase, weak ER immunolocalization was observed in the endometrial stroma. During the late follicular phase under the influence of rising estradiol levels, stromal ER localization was intense. During the luteal phase, ER localization was absent in the stroma indicating that high concentrations of progesterone suppressed ER. PR localization was not observed in the stroma during the early follicular phase, while weak staining was seen in the stroma during the late follicular phase. PR localization was maximum during the mid luteal phase. However in marmoset, endometrial ER and PR localization was restricted only to the stroma. This unique feature may be due to the characteristic reproductive profile of this nonmenstruating species and needs to be studied further. Thus it can be hypothesized that in the marmoset endometrium, steroid hormone mediated effects possibly occur directly in the stroma and are then transmitted to the epithelium by autocrine/paracrine action of growth factors and cytokines.

Immunohistological localisation of vascular endothelial growth factor in human endometrium.

Several polypeptide growth factors regulate epithelial and stromal development in endometrium under the influence of estrogen and progesterone, and thereby regulate growth and differentiation of endometrium during menstrual cycle. However, little is known about the angiogenic growth factors that may affect endometrial vasculature throughout each menstrual cycle. Vascular endothelial growth factor (VEGF) is suggestively an important angiogenic growth factor in the female reproductive tract. The aim of the present study was to immunolocalize and assess semi-quantitatively VEGF immunostaining in cells of proliferative phase (n = 3), secretory phase (n = 6) and hyperplastic (n = 6) human endometrial samples. VEGF concentrations were significantly higher in glandular (P < 0.001) and stromal (P < 0.01) compartments of proliferative stage endometrium compared with those in secretory stage and hyperplastic endometrial samples, with no difference in the scores for glandular and stromal compartments between secretory stage and hyperplastic endometrial samples. Generally, glandular expression of VEGF was higher as compared to stromal compartment. Thus, it appears that endometrial VEGF expression and concentration are enhanced by estrogen, and may be correlated with neovascularization and increased vascular permeability during late proliferative period. Additionally, there was no enhancement in VEGF expression in hyperplastic glands, suggesting that regulation of glandular growth and that of angiogenesis in human endometrium operate through different mechanisms.

Integrin cell adhesion molecules in endometrium of fertile and infertile women throughout menstrual cycle.

Integrins (ITGs) are ubiquitous cell adhesion molecules that undergo dynamic alterations during the normal menstrual cycle in human endometrium. The distribution of four different subunits, viz. alpha 4, alpha 6, beta 3 and beta 4 in human endometrial tissue at different stages of the menstrual cycle was studied using immunohistochemical, enzyme immunoassay and SDS-PAGE/Western blot techniques. The specificity of each mAb to their respective ligands viz., laminin (Ln), fibronectin (Fn) and vitronectin (Vn) was done by cell adhesion assays. Both alpha 6 and beta 4 subunits (Ln receptors) expressed primarily on the glandular epithelium, while glandular, stromal and luminal cells expressed predominantly alpha 4 (Fn receptor) and beta 3 (Vn receptor). The appearance of alpha 4 and beta 3 ITG subunits was found to be cell and cycle specific. The levels of both alpha 6 and beta 4 increased throughout the menstrual cycle, while beta 3 subunit appeared abruptly on cycle day 19/20. The immunostaining for alpha 4 and beta 3 was absent in 90% of infertile women. The timing of expression of alpha 4 and beta 3, the two cycle--dependent ITGs framed the putative window of implantation and suggests a role in the diagnosis of infertility. In conclusion, the absence of alpha 4 and beta 3 ITG expression in the endometrium of infertility subjects during mid luteal phase may be associated with defects in uterine function. The defective uterine receptivity may be an unrecognised cause of infertility in these group of women.

Hormonal-induced changes on lipidi composition and DPH fluorescence anisotropy of erithrocyte ghost from pre-and postmenopausal women

Fluorescence anisotropy determination were performed on hypotonically lysed red blood cells from 12 healthy, free living, premenopausal and 10 postmenopausal women fed a self-selected diet. Fatty acid composition of phospholipids and cholesterol esters, lipid/lipid and lipid/protein ratios were also determined in erythrocyte ghosts along the menstrual cycle or during hormonal replacemente therapy. The postmenopausal women were treated with transdermal 17-beta estradiol (approx. delivery rate of 50 mug/day) for a 5-week period and with a combination of percutaneous administration of estradiol and orally administered medroxyprogesterone acetate (5 mg/day) for 5 weeks more. Premenopausal women were studied in the mid-follicular and mid-luteal phases of their menstrual cycles. Membrane fluidity in cells from the luteal phase was greater than that observed in the follicular phase. After estradiol treatment, postmenopausal women exhibited a significant raise in the fluorescence anisotropy. The association with progesterone prevented the estrogen-induced increase of the anisotropy values. Estrogen administration also produced an increment in the relative content of cholesterol and membrane proteins that was reverted by progesterone addition. Changes induced by estradiol treatment in the fatty acid pattern of phospholipids and cholesterol esters were characterized by an increment in the monoenoic/saturated fatty acid ratio and linoleic relative content, with a concomitant decrease in the proportion of arachidonic and docosatetraenoic acids. These results suggest that the modification of the desaturase activities may be involved in a mechanism by which these hormonal-induced changes take place. The present study demonstrated that human erythrocyte fluidity is under hormonal control subjected through changes in the proportion and quality of the membrane lipids.

Metallic ion concentration during menstrual cycle in normally menstruating women.

Plasma concentration of metallic ions levels during menstrual cycle of twenty normally menstruating women were observed in four phases i.e. menses, follicular, ovulatory and luteal. The concentration of magnesium, zinc, selenium and manganese was highest during menses and lowest at ovulatory phase. There was rise in ionic levels of magnesium and selenium, while fall in zinc and manganese during luteal phase. Findings demonstrate changes in metallic ions (Magnesium, zinc, selenium and manganese) level in relation to hormonal status during menstrual cycle in women.

Lysine clonixinate in the treatment of primary dysmenorrhea

The efficacy tolerance of Lysine Clonixinate (LC), a NSAID with prostaglandin synthesis inhibiting mechanism was studied in 24 patients with primary dysmenorrhea according to a double-blind randomized crossover Placebo (P) controlled design with patients serving as their own controls. Treatment consisted in administering 1 tablet of LC or P q6h as from onset of menstrual pain during 5 days and 6 menstrual cycles. Patients were controlled monthly as from the 5th day of the cycle rating changes in pain intensity according to a 4-point scale, presence of pain during pre-, post-and menstrual periods: possible intracycle changes, amount of bleeding, tolerance and related total and general signs and symptons. Intensity of baseline menstrual pain amounted to 2.9. Menstrual, intramenstrual and postmenstrual pains were observed in 19 out of 24,24/24 and only 2 out of the patients, respectively. Concomitant symptons consisted in headache (12), mastalgia (14) and disconfort (12). Results were obtained by averaging the data from the 3 tratment periods with each drug. Menstrual pain was reduced from 2.9 + 0.7 to 1.9 + 0.7 with P administration and to 0.66 + 0.4 with the administration of LC, a highly significant difference between tratments (p<0.0001). Premenstrual pain was reduced nonsignificantly from 0.79 per cent to 0.58 per cent with P administration and significantly to 0.29 per cent with administration of LC (p<0.001). Intramenstrual pain affection all patients at baseline was reduced significantly by 8 per cent with P and also significantly by 50 per cent with LC p<0.001). No differences were encontered in encomitant symptoms during P treatment periods while the incidence was significantly reduced with LC (p<0.0001). No changes in cycle duration or amount of bleeding were observed between treatments. No adverse events were reported.