Effect of Clomiphene Citrate on the Ultrastructure of Testis in Albino Rats

SUMMARY: The aim of the present work was to study the closer effect of clomiphene citrate on the ultrastructure of the testis of adult albino rats to provide a basis for optimizing this drug in the treatment of male infertility. The testes were removed from both groups under anesthesia and then prepared for examination by light using hematoxylin and eosin stains and a transmission electron microscope. Semithin sections were cut into 1 µm thick sections, stained with toluidine blue, and examined by light microscopy for a survey. The desired areas were placed in the center, and other areas were trimmed. Primary spermatocytes showed marked nuclear changes (pyknosis), and their nuclear membranes were ill-defined and disrupted. The cytoplasm showed widespread degeneration of mitochondria and lysosomes and focal degeneration of the rough endoplasmic reticulum compared with the control group. The spermatids were pale, and the two phases of spermatogenesis were distinctly identifiable in the control group but were confused in the treated group. Some spermatids had interrupted nuclear membranes, also containing degenerated mitochondria, focal fragmentation of rough endoplasmic reticulum, and free ribosomes. Spermatozoa in the treated group appeared deformed compared to the control, where they had deformed head caps. Leydig cells of the treated group have an irregularly shaped nucleus, with focal chromatin aggregation and peripheral chromatin condensation on the inner surface of the nuclear membrane. The observations of the present work indicate a possible causal relationship between testicular affection and ingestion of clomiphene citrate, which can be avoided by close medical observations using ultrasonography, semen analysis, or testicular biopsy to detect early malignant changes. Furthermore, the drug should not be used for more than three to six cycles and should be stopped for at least three cycles before reuse. When clomiphene citrate is ineffective in the treatment of male infertility, human menopausal gonadotropin (hMG) administration is typically selected. However, high-dose hMG therapy is associated with a variety of adverse effects. In this work, we report the success of a modified clomiphene citrate regimen in increasing sperm count without any hazards to the testicular tissue.

El objetivo del trabajo fue estudiar el efecto del citrato de clomifeno sobre la estructura de los testículos de la rata albina adulta, con la finalidad de determinar la mejor manera de utilizar este fármaco en el tratamiento de la infertilidad masculina. Los testículos se extrajeron bajo anestesia y para su análisis a través de microscopio de luz se tiñeron con HE. Además, las muestras fueron preparadas para su examen con microscopía electrónica de transmisión. Por otra parte, se cortaron secciones semifinas de 1 µm de espesor, se tiñeron con azul de toluidina y se examinaron mediante microscopía óptica. Los espermatocitos primarios mostraron cambios nucleares marcados (picnosis) y sus membranas nucleares estaban mal definidas y alteradas. En el grupo experimental las células presentaban el citoplasma con degeneración generalizada de las mitocondrias y de los lisosomas y una degeneración focal del retículo endoplásmico rugoso en comparación con el grupo control. Las espermátidas estaban pálidas y las dos fases de la espermatogénesis eran claramente identificables en el grupo control, pero se confundían en el grupo tratado. Algunas espermátidas tenían membranas nucleares interrumpidas, y también contenían mitocondrias degeneradas, fragmentación focal del retículo endoplásmico rugoso y ribosomas libres. Los espermatozoides del grupo tratado se presentaban deformados en comparación con el control. Las células de Leydig del grupo tratado presentaban un núcleo de forma irregular, con agregación focal de cromatina y condensación de cromatina periférica en la superficie interna de la membrana nuclear. Las observaciones del presente trabajo indican una posible relación causal entre la afección testicular y la ingestión de citrato de clomifeno, que puede evitarse mediante observaciones médicas minuciosas a través de ecografía, análisis de semen o biopsia testicular para detectar cambios malignos tempranos. Además, el medicamento no debiera ser usado durante más de tres a seis ciclos y debe suspenderse durante al menos tres ciclos antes de volver a usarlo. Cuando el citrato de clomifeno es ineficaz en el tratamiento de la infertilidad masculina, normalmente se selecciona la administración de gonadotropina menopáusica humana (hMG). Sin embargo, la terapia con hMG en dosis altas se asocia con una variedad de efectos adversos. En este trabajo, informamos el éxito de un régimen modificado con citrato de clomifeno para aumentar el recuento de espermatozoides sin riesgo para el tejido testicular.

Therapeutic effects of dimethyldiguanide combined with clomifene citrate in the treatment of polycystic ovary syndrome

SUMMARY OBJECTIVE In view of the high incidence of polycystic ovary syndrome (PCOS) and the unsatisfactory therapeutic effects of dimethyldiguanide or clomifene citrate alone, our study aimed to investigate the therapeutic effects of dimethyldiguanide combined with clomifene citrate in the treatment of PCOS. METHODS A total of 79 patients with POCS and 35 healthy females were included, and endometrial biopsies were obtained. The sterol regulatory element-binding protein-1 (SREBP1) expression in endometrial tissues was detected by qRT-PCR. POC patients were randomly divided into group A (n=40) and group B (n=39). Patients in group A were treated with dimethyldiguanide combined with clomifene citrate, while patients in group B were treated with clomifene citrate alone. The number of mature follicles and cervical mucus score, follicular development rate and single follicle ovulation rate, cycle pregnancy rate, early miscarriage rate, ovulation rate, endometrial thickness, positive rate of three lines sign, follicle stimulating hormone level and luteinizing hormone level were compared between the two groups. RESULTS The expression level of SREBP1 was higher in PCOS patients than that in the healthy control. SREBP1 expression was inhibited after treatment, while the inhibitory effects of combined treatment were stronger than those of clomifene citrate alone. Compared with clomifene citrate alone, the combined treatment improved cervical mucus score, follicle development rate, single follicle ovulation rate, endometrial thickness, positive rate of three lines sign, and follicle-stimulating hormone level. CONCLUSION The therapeutic effect of combined treatment is better than clomifene citrate alone in the treatment of PCOS.

RESUMO OBJETIVO Tendo em vista a alta incidência de síndrome dos ovários policísticos (SOP) e os efeitos terapêuticos insatisfatórios da dimetildiguanida ou do citrato de clomifeno isoladamente, nosso estudo teve como objetivo investigar os efeitos terapêuticos da dimetildiguanida associada ao citrato de clomifeno no tratamento da SOP. MÉTODOS Um total de 79 pacientes com POCS e 35 mulheres saudáveis foram incluídos, e biópsias endometriais foram obtidas. A expressão da proteína de ligação do elemento regulador de esterol-1 (SREBP1) nos tecidos endometriais foi detectada por qRT-PCR. Pacientes POC foram divididos aleatoriamente em grupo A (n=40) e grupo B (n=39). Os pacientes do grupo A foram tratados com dimetildiguanida combinada com citrato de clomifeno, enquanto os pacientes do grupo B foram tratados apenas com citrato de clomifeno. O número de folículos maduros e muco cervical, taxa de desenvolvimento folicular e taxa de ovulação, taxa de gravidez, abortamento precoce, taxa de ovulação, espessura endometrial, taxa positiva de três linhas, nível de hormônio folículo estimulante e nível de hormônio luteinizante foram comparados entre os dois grupos. RESULTADOS O nível de expressão do SREBP1 foi maior nos pacientes com SOP do que no controle normal. A expressão de SREBP1 foi inibida após o tratamento, enquanto os efeitos inibidores do tratamento combinado foram mais fortes do que os do citrato de clomifeno isoladamente. Comparado com o citrato de clomifeno sozinho, o tratamento combinado melhorou significativamente a pontuação do muco cervical, a taxa de desenvolvimento folicular, a taxa de ovulação do folículo único, a espessura endometrial, a taxa positiva de três linhas de sinal e o nível de hormônio folículo estimulante. CONCLUSÃO O efeito terapêutico do tratamento combinado é melhor do que o citrato de clomifeno isolado no tratamento da SOP.

[Metformin effects on clomifene-induced ovulation in the polycystic ovary syndrome]

Polycystic ovary syndrome [PCOS] is a common, complex endocrine disorder for women on reproductive age. A high incidence of ovulation failure is observed in PCO women and perhaps linked to insulin resistance related to metabolic features In the last few years some studies assessed hyperinsulinimea and insulin resistance attenuation effects, by insulin sensitizing agents such as metformin, in PCOS women suggesting potential scope for these drugs in CC ovulation induction quality improvement. Aim: Our prospective study aim is to compare the effectiveness of clomifene citrate plus metformin and clomifene citrate plus placebo in women with newly diagnosed polycystic ovary syndrome. From February 24 to September 29 [2007], PCOS was explored on women attending the Department of Obstetrics and Gynaecology sterility consultation unit [CHU Hedi Chaker-Sfax] according to the Rotterdam 2003 diagnostic criteria. PCOS patients were randomized to receive, in addition to clomifene citrate treatment, placebo or metformin 850 mg two times a day all ovulatory cycle for three trials maximum. Ovulation detection was done by the E2 serum measurements and ovarian transvaginal ultrasonography' evolution controlling on 7th, 11th and 13th day of the cycle. Within 7 months, 32 PCOS women were recruited in the study and equally allocated to the two groups. Baseline characteristics were similar in metformin group and placebo one. Ovulation was characterized by the presence of at least one mature follicle [> 16mm], a circulating estradiol concentration in the edge of 150-250pg and accessory an endometrial depth > 8mm. The ovulation rate in the metformin group was 62.5% compared with 37.5% in the placebo group, a non-statistically significant [small study population] but important difference [1.66 times]. Analyses show a higher mature follicle number and estradiol concentration in metformin group than in the placebo one. Metformin effect was, in our study, his only insulinosensitizer property consequence far away a 'making thinner' or Hyperandrogenism reducing ones. The ovulatory response to clomifene can be increased in polycystic ovary syndrome women by decreasing insulin secretion with metformin

Opioid antagonists as a sole or adjunctive therapy in ovulation induction in PCOS cases resistant to clomiphene citrate

Patients who have eugonadotrophic hyperandrogenic anovulation with the presence of multiple small ovarian follicles - defined as PCOS - are offered many ovulation induction regimens. Some prove to be clomiphene resistant others develop complications as ovarian hyperstimulation syndrome when treated by gonadotrophins. To evaluate naltrexone - an opioid antagonist - as a simple and safe method for ovarian induction in PCOS patients resistant to clomiphene citrate as a sole or adjunct therapy. Forty PCOS patients resistant to clomiphene citrate were studied. They were randomly divided into 2 groups. Group I [20 cases] who received clomiphene citrate and naltrexone and group II [20 cases] in which naltrexone alone was administered. Estimation of serum level of FSH, LH and prolactin as well as vaginal ultrasonographic follicular tracking was done for all cases. Fifteen cases [75%] of group I achieved ovulation, while none of the patients of group II ovulated even after 21 days of naltrexone treatment. Ovulation can be successfully induced using naltrexone in combination with clomiphene citrate in PCOS cases resistant to clomiphene citrate alone. It is safe simple and inexpensive treatment modality

Diagnosis and management of luteinized unruptured follicle [LUF] syndrome

A diagnosis of LUF syndrome in 490 women of unexplained infertility was made sonographically by failure to demonstrate a minimum of 5 mm reduction in follicular diameter with a change in the follicular and endometrial echogenecities within 48 hours after LH surge in urine. Fifty patients with LUF were categorized into three groups: 10 patients received only hCG for rupture of follicle, 30 patients were given clomiphene citrate therapy, then received hCG and 10 women having LUF syndrome did not receive any ovulation inducing drugs as a control group. The study concluded that the use of clomiphene citrate in LUF women increased the chance of ovulation by using 10,000 IU hCG with an increased chance of pregnancy

Efficacy of laparoscopic ovulation induction in polycystic ovary syndrome.

This prospective study was aimed to evaluate the efficacy of laparoscopic ovarian electrocoagulation in women with PCOS. Twenty-three PCOS women who had refractory to clomiphene citrate attending the Reproductive Endocrinology Unit, Ramathibodi Hospital between March 1995 and June 1998 were enrolled in the study. In all patients, electrocoagulation on the ovarian surface of both ovaries was performed through laparoscope under general anesthesia. Two patients were lost to follow-up for unknown reasons. The remaining 21 women had a mean age of 30.3 +/- 3.9 years (range 21-39) and mean duration of infertility of 4.1 +/- 2.8 years (range 1-11). There was no intra-operative and post-operative complication. After surgery, ovulation was documented in 16 out of 18 (88.9%) patients. Fifteen (71.4%) patients became pregnant. Fourteen pregnancies (93.3%) occurred within 9 months after surgery. Twelve women (80%) became pregnant in spontaneous cycles without any treatment. The outcomes of pregnancies were 10 live births, 3 ongoing pregnancies and 2 abortions. This study reveals the high efficacy of ovarian electrocoagulation in infertile women with PCOS. High pregnancy and low abortion rates are convincing. This surgical technique should be the treatment of choice for women with CC-resistant PCOS.

Induction Of ovulation.

Sixty-one patients with anovulation as a cause of infertility were selected for our study. Various ovulation-inducing drugs were used like clomiphene citrate, human menopausal gonadotropin (hMG), human chorionic gonadotropin (hCG), bromergocryptine and leptadene. The response of the different drugs was observed by serial sonography for ovulation. Indeed there was a good response to clomiphene citrate, but those patients who failed to respond to clomiphene citrate and were frustrated with the use of hMG and hCG due to the cost and the complications of the therapy were put on Aloe compound and leptadene - an ayurvedic drug which enhances fertility in different ways.

Goserelin and oral contraceptive combination versus laparoscopic ovarian cauterization and their effects on the endocrine changes in women with clomiphene citrate resistant polycystic ovary disease

The aim of this study was to compare the effects of combination of long acting GnRH agonist [Goserelin] and oral contraceptive [OC] therapy and laparoscopic ovarian cauterization on the endocrine changes in women with clomiphene citrate [CC] - resistant polycystic ovary disease [PCOD]. Twenty one women with CC- resistant PCOD were included randomly in the study to either laparoscopic ovarian cautery [n = 10] or GnRH-a and OC therapy for 3 months [n = 11]. Serum concentrations of LH, F.S.H, androstenedione [A], testosterone [T] and sex hormone binding globulin [SHBG] were determined in each group and during the follicular phase of first menstrual cycle after cessation of each treatment. Our results showed that the mean serum concentrations and clinical profiles were similar in both groups. Both groups showed significant decrease in LH, A and T and significant increase in FSH compared with pretreatment levels. The SHBG concentration increased in both groups after therapy, however, the increase was significant only in the goserelin and OC group. There were no significant difference in the final concentrations of LH, FSH and A between the two study groups after each treatment, whereas, T was significantly decreased and SHBG significantly increased in the goserelin and OC group. The ovulation rate after the cessation of either therapy was similar [8/10 versus 9/11, p > 0.05]. On the basis of our results, we conclude that despite similar endocrine effects, medical treatment with GnRH-a and OC may be more appropriate and effective in reversing the abnormal cascade in PCOD. Although both regimens have temporary and similar improvement on PCOD, the adhesion forming potential, invasiveness, cost and surgical complications of laparoscopic ovarian cauterization may enable GnRH-a and OC combination to be cost effective option in women with CC-resistant PCOD

Effect of the dopamine antagonist, domperidone alone and in combination with thyroxine, clomiphene, hcg or gnRH on serum sex steroid profile, gonadal maturation and spawning of nile catfish, clarias lazera

The present investigation was carried out on one hundred and twenty mature Clarias lazera fish during the prespawning season [March - April]. Fish were allocated into two main groups [sixty fish of each sex] Fish weight ranged between 200-220 g. b. wt. Each main group was subdivided into six equal groups [Ten fish each], Fish of each group; both males and females were subjected to the same regimen of intraperitoneal exogenous treatment, as follows. 1[st] Group: Saline 0.65% NaCL solution. [Control] 2[nd] Group: Domperidone 5 microg/g. b. wt. in saline. 3[rd] Group: Domperidone 5 microg/g. b. wt, plus thyroxine 1 nancg / g. b. wt. in saline. 4[th] Group; Domperidone 5 microg/g, b. wt. plus clomiphene citrate l gamma g/g. b wt. in saline. 5[th] Group: Domperidone 5 micro g/g. b. wt. plus hCG 2 lU/g, b. wt. in saline. 6[th] Group: Domperidone 5 microg/g. b. wt. plus GnRH 10 microg/kg. b. wt. in saline. The present study revealed that [Domperidone] administration alone, in each sex of Clarias fish; increased serum sex steroid hormones [testosterone and 17 beta estradiol], gonadal weights [testes and ovaries], final gonadal maturation and gonadosomatic indices of both sexes as compared to respective controls. Exogenous administration of domperidone plus gonadotropin releasing hormone [GnRH] was the most effective treatment for production of sex hormones, final gonadal maturation and spawning in both sexes of Clarias lazera fish, followed by domperidone plus thyroxine; domperidone plus clomiphene citrate and domperidone plus hCG, respectively. In conclusion, the exogenous administration of dopamine antagonist plus GnRH is the most efficient regimen for enhancement of gonads maturation [gonadal recrudescence], spawning and spermiation of Clarias lazera fish throughout the prespawning season

Inductores de la ovulación, su estado actual y experiencia institucional / Ovulation inductors, present state and instutional experience

Antecedentes. La anovulación o esterilidad por factor endócrino ovárico representa del 20 al 30 por ciento de todas las parejas estériles y aunque en la actualidad existen diversos fármacos capaces de inducir ovulación , siguen siendo el citrato de clomifeno y las gonadotropinas menopáusicas humanas los medicamentos de primera línea para el tratamiento de la esterilidad por factor anovulatorio, como hace 30 años que se inició su uso. Objetivo. En el presente trabajo se analiza el marco teórico para la selección de las pacientes candidatas a uno u otro medicamento, así como los esquemas terapéuticos utilizados, efectos secundarios y resultados tanto de ovulación como de embarazos. Método. Se compararon los resultados promedio de la literatura, con la experiencia institucional de los autores. Resultados. Con el citrato de clomifeno en casos bien seleccionados se obtiene ovulación en aproximadamente 80 por ciento de los casos y embarazo en el 40-50 por ciento; en el INPer se obtuvo en un análisis de 200 casos ovulación en el 72 por ciento y embarazo en el 58 por ciento. Con las gonadotropinas menopáusicas humanas los resultados son mas variables encontrando un promedio en la literatura de ovulación del 70 al 90 por ciento y de embarazo del 25 al 55 por ciento. En el INPer de 128 ciclos de tratamiento se obtuvo ovulación el 75.7 por ciento y embarazo en el 36 por ciento de un grupo de 100 mujeres anovulatorias tratadas

Existe vantagem em utilizar o citrato de clomifene em pacientes ovuladoras? / Is there any advantage in using clomiphene citrate in patients with ovulatory cicles?

Retrospectiva: O citrato de clomifene tem sido utilizado em pacientes ovuladoras com a intençao de melhorar os índices de gravidez. Objetivo: O objetivo do presente trabalho foi comparar as variaços hormonais (LH e androstenediona), as respostas ovariana (número de folículos dominantes e índice de ovulaçao) e endometrial (espessura) e os índices de gravidez obtidos por diferentes doses de citrato de clomifene e em ciclos espontâneos, em pacientes com ciclos ovulatórios. Casuistica e Metodos: Foram estudadas 38 pacientes com esterilidade sem causa aparente, tratadas de 4 maneiras: citrato de clomifene 25 mg/dia (1O pacientes), citrato de clomifene 50 mg/dia (9 pacientes), citrato de clomifene 100 mg/dia (3 pacientes) e ciclo espontâneo (1O pacientes). Resultados: Houve somente diferença estatisticamente significativa em relaçao à espessura do endométrio (p

Estudo comparativo entre a ultra-sonografia transvaginal e transabdominal na avaliaçäo do endométrio de mulheres com ciclos menstruais espontâneos e induzidos / A comparative study between transvaginal and transabdominal ultrasound in the evaluation of the endometrium of women with spontaneous and induced menstrual cycles

Estudamos a espessura e a textura do endométrio comparando-se US transabdominal com US transvaginal. Avaliamos 45 pacientes do ambulatório de Ginecologia e Infertilidade do Hospital de Clínicas de Porto Alegre, entre abril de 1990 e junho de 1992. As 45 pacientes foram divididas em três grupos: grupo A, com ciclos menstruais espontâneos; grupo B.1, com ciclos induzidos pelo citrato de clomifeno; grupo B.2, com ciclos induzidos pelo citrato de clomifeno/gonadotropina humana da menopausa/gonadotropina coriônica humana. No grupo A foram estudados o oitavo, décimo, 12( e 21( dias do ciclo menstrual. Nos grupos B.1 e B.2 estudaram-se os mesmos dias, excluindo-se o 21(. Nos grupos A e B.1, a espessura endometrial medida pela US transvaginal foi significantemente maior quando comparada com a transabdominal, excetuando-se o primeiro dia do exame, quando os resultados sugerem igualdade. No grupo B.2, nao houve diferença significante de resultados, excetuando-se o primeiro dia do exame, quando os resultados sugerem igualdade. A textura do endométrio foi absolutamente coincidente quando se comparou a via transabdominal com a transvaginal.

Comportamiento biológico en pacientes que lograron embarazo por GIFT o FIVTE: experiencia institucional / Biological behaviour in patients succeding in pregnancy by GIFT or FIVTE: institutional experience

Fueron analizados en forma retrospectiva los resultados de las determinaciones hormonales pre-captura así como el número de ovocitos capturados, ovocitos maduros y embriones transferidos en tres grupos de pacientes (2 GIFT, 1 FIV-TE) que lograron embarazo, y comparados con grupos aleatorizados, con los mismos esquemas de estimulación pero que no consiguieron embarazo. No hubo diferencias estadísticamente significativas en la mayoría de los parámetros analizados, por lo que podemos elucubar que el embarazo depende principalmente de la calidad del endometrio lo cual aún no podemos modificar en una forma satisfactoria.

Comparison of four different ovarian stimulation protocols in an in vitro fertilisation & embryo transfer programme.

Four different ovarian stimulation protocols were evaluated in an in vitro fertilisation and embryo transfer programme in 208 women (228 treatment cycles). In the rigid protocol (RP), 100 mg of clomiphene citrate (CC) was given from day 3 to day 7 of the menstrual cycle and 300 IU of human menopausal gonadotropin (hMG) was given from day 5 of the menstrual cycle. In the individualised protocol (IP) the same drugs and doses were used as in RP, but the day of initiation of CC depended on the length of the individual's menstrual cycle and hMG was administered from the last day of CC. In the programmed protocol (PP), ovarian function was suppressed with oral contraceptive pills (ethinyl estradiol 30 micrograms and norethisterone 1 mg) started on day 5 of the menstrual cycle for 45 to 70 days. Considering the last day of pill intake as day 0, CC was given for 5 days from day 5 and hMG (300 IU) from day 7. In the alternate day protocol (ADP), 100 mg of CC was administered from day 2 to day 6 and hMG (300 IU) was given on alternate days from day 2 to day 8 or day 10 of the cycle. In all the women, hCG (5000 IU) was administered when the diameter of at least 2 follicles was greater than or equal to 16 mm and estradiol levels were 300 pg/ml/dominant follicle. Patients not showing such a response were not treated further. The cardinal events of IVF-ET such as number of good responders, incidence of oocytes harvested, fertilised and embryos transferred per cycle were compared and it was concluded that the pregnancy rates were highest in women treated by the PP.

Efeito do citrato de clomifene sobre a liberaçäo hipofisária de LH e FSH induzida pelo GnRH / Effect of clomiphene citrate on pituitary liberation of LH and FSH induced by GnRH

Com o objetivo de avaliar o mecanismo de açäo do citrato de clomifene (CO), dez mulheres hipogonádicas foram submetida à infusäo endovenosa contínua de GnRH nas seguintes condiçöes: sem qualquer medicaçäo (Teste I), após uso de CC (Teste II), após uso de etinil-estradiol (EE) (Teste III) e após uso de EE e CC (Teste IV). Os resultados mostraram que em condiçöes de hipoestrogenismo o CC näo modifica a resposta hipofisária de LH e FSH ao GnRH, mostrando uma leve açäo estrogênica. Entretanto, o uso de CC após estrogenioterapia promoveu diferentes respostas para o LH e FSH quando comparadas ao uso de EE isoladamente. Para o LH houve uma significante inibiçäo na sua liberaçäo, com menor sensibilidade, reserva e capacidade hipofisária. Para o FSH observou-se o oposto, com um incremento na sua liberaçäo, com maior sensibilidade e reserva hipofisária