Identify normal and abnormal semen via mRNA fluorescence multiplex amplification system: A preliminary study / 中国法医学杂志

Objective Construct a mRNA multiplex amplification system to identify different types of semen stains. Methods First, collect normal, oligozoospermia and azoospermia semen samples to make semen stains. Second, extract total RNA with Qiagen RNeasy Micro Kit. Then use reverse transcript PCR to amplify goal mRNA markers: 2 markers for sperm(PRM1, PRM2), 2 markers for seminal plasma(TGM4, SEMG1) and 2 housekeeping genes(TEF, UCE). Results All semen mRNA markers can be detected in normal semen samples. The RFU of sperm mRNA markers are lower in oligozoospermia semen samples than that in normal controls. No sperm mRNA markers can be detected in the azoospermia semen samples, only seminal plasma specific can be detected. Conclusion The differentiation of normal and azoospermia semen can be achieved by using multiplex mRNA fluorescence amplification system. While normal semen and oligozoospermia semen compared to no statistical difference.

Metodologia da análise seminal para pacientes azoospérmicos no laboratório fleury / Methodology of seminal analysis for azoospermicos patients in the laboratório fleury

A azoospermia é definida como a ausência de espermatozoide no líquido seminal ejaculado pelo homem depois de aplicada a técnica de centrifugação em pelo menos duas amostras. Dada a importância de um diagnóstico correto da análise seminal para os casais, toda amostra que não apresentar espermatozoides no exame a fresco deve seguir em avaliação laboratorial. Com isso, o presente estudo tem como objetivo analisar os resultados de centrifugação de uma alíquota do sêmen ejaculado ou de todo o volume ejaculado de pacientes com diagnóstico de azoospermia para determinar qual o melhor método a ser empregado na análise seminal para esse grupo de pacientes.

The azoospermia is defined as the absence of sperm in the ejaculate by the seminal fluid man after centrifugation technique conducted in at least two samples. Given the importance of a correct diagnosis of the seminal analysis for couples, all sample no sperm present in fresh examination should follow in laboratory tests. Thus the present study aims to analyze the results of a spin rate of ejaculate or all of the ejaculate volume of patients with azoospermia to determine the best method to be used in semen analysis for this group of patients.

The role of bacterial infections on male infertility in Al-Anbar province of Iraq.

Background: Bacterial infections on male infertility has always been in the field of debate due to scarce analysis tools to examine seminal fluid specimens as a result of which these infectious processes leads to deterioration of spermatogenesis, impairment of sperm function and/or obstruction of the seminal tract. Aims & Objective: In the current study we investigated the role of bacterial infections in male factor infertility in Al-Anbar Province, West of Iraq through detection of abnormal sperms and other factor pertains to male infertility. Material and Methods: Seminal fluid from six hundred volunteer males was investigated for infertility by the detection of abnormal sperms using the WLJY-9000 TYPE WEILI Color Sperm Analysis System and the Neubauer counting chamber. Results: From the six hundreds patients investigated for infertility, it was found that 408 (68%) patients had a positive culture for pathogenic bacteria, of different species. The results indicate that 32.0% had sperm density less than twenty million per millilitre. The oligospermic were 23.0%, severe oligospermic 0.17% and Azoospermia 8.83%. Asthenospermia was reported to be 76.33% and Teratospermia 86.16% respectively. Conclusion: Seminal fluid infection increases with decreasing sperm density, motility and morphology. The prevalence of abnormal sperm indices and bacterial infection is high with Klebsiella spp. infection. Hence, treatment measures should be taken properly in the management of male factor infertility.

Diferencias del estado redox entre espermatozoides y plasma seminal en adultos jóvenes / Difference of redux status between spermatozoa and seminal plasma in young adults

Introducción: Las especies reactivas del oxígeno y el estrés oxidativo desempeñan un papel fundamental en la infertilidad masculina. Sin embargo, estas especies oxidantes también han sido asociadas con los procesos de capacitación de los gametos masculinos cuando son generadas a bajos niveles y de manera controlada. Actualmente los biomarcadores redox son introducidos en el diagnóstico clínico de la infertilidad masculina en el mundo, como una herramienta complementaria a los parámetros del espermiograma. Sin embargo, en Cuba, esta metodología aún no se encuentra extendida a los servicios de salud. Objetivo: El objetivo del presente trabajo fue caracterizar el estado redox de espermatozoides y el líquido seminal de sujetos aparentemente sanos, a través de la determinación de una serie de biomarcadores de estrés oxidativo. Los niveles de malonildialdehído, la actividad de las enzimas antioxidantes superóxido dismutasa y catalasa, así como los niveles de glutatión reducido, el potencial de peroxidación y la capacidad reductora fueron determinados mediante técnicas espectrofotométricas. Métodos: Se analizaron 40 muestras de semen de sujetos aparentemente sanos, las cuales fueron obtenidas mediante masturbación sin el empleo de lubricantes y con al menos tres días de abstinencia eyaculatoria. En el estudio se incluyeron sujetos de 20 a 35 años, aparentemente sanos según exámenes de laboratorio clínico y con paternidad probada. Resultados: Los resultados demostraron que existen diferencias significativas (p< 0,05) entre los marcadores evaluados en el líquido seminal con respecto al espermatozoide, lo cual sugiere la existencia de un estado redox diferenciado entre ambos compartimentos. Conclusiones: Estos hallazgos demuestran la necesidad de evaluar el nivel de estrés oxidativo tanto en las células sexuales como en el fluido que las contiene. Ello contribuirá, sin lugar a dudas, a un diagnóstico más eficaz e integral de la capacidad fértil del hombre

Background: Reactive oxygen species and oxidative stress play a fundamental role in male infertility. However, these oxidizing species have also been associated with the process of capability of male gametes when they are generated at low levels and in a controlled manner. Currently, redox biomarkers are introduced in the clinical diagnosis of male infertility in the world as a complementary tool to the spermiogram parameters. However, in Cuba, this methodology is not yet extended to health services. Objective: The objective of this study was to characterize the redox status of spermatozoa and seminal fluid of apparently healthy subjects through the identification of a number of biomarkers of oxidative stress. Methods: 40 samples of semen of apparently healthy subjects were analyzed, which were obtained by masturbation without the use of lubricants and with at least 3 days of ejaculatory abstinence. The study included subjects from 20 to 35 years of age, who were apparently healthy according to both laboratory tests and paternity test results. Results: The results showed that there are significant differences (p<0, 05) between the markers evaluated in the seminal fluid and the spermatozoon which suggests the existence of a differentiated redux status between the two compartments. Conclusions: These findings demonstrate the need to evaluate the level of oxidative stress in both sexual cells and the fluid that contains them. It will contribute, with no doubt, to a more effective and comprehensive diagnosis of man's fertility

RP-HPLC method development for estimation of sildenafil citrate in tablets and in seminal fluid.

A RP HPLC method for estimation of sildenafil citrate (SC) in tablet dosage form and seminal fluid was developed and validated. Best resolution was obtained with column Waters Spherisorb® C18 bonded silica, (5 μm, 4.6 x 250 mm) at 230 nm with retention time of 5.01 min. The mobile phase used was TEA (0.2%) pH adjusted at 3 with OPA and ACN (60:40) with flow rate of 1.0 mL/min. The method for estimation of sildenafil citrate in tablet dosage form was found to be linear, accurate, precise, sensitive and selective. Whereas the bioanalytical estimation of SC in seminal fluid was found to be in the range of 100 ng/mL to 10μg/mL. Method was found to be highly sensitive as LOD and LOQ were found to 0.3 μg/ml and 0.9μg/ml. The repeatability and reproducibility were within the range i.e. less than 2%.The accuracy of the method was 99.3%. The percentage purity was calculated for market formulation was 102.8%. The internal standard used for bioanalytical methods was Diclofenac sodium. The drug was extracted from seminal fluid by protein precipitation using ACN as precipitating agent. The linearity range was from 100.0ng/mL to 2.0μg/mL. The LOD and LOQ were found to 0.03μg/mL and 0.1μg/mL. The repeatability and reproducibility were within the range i.e. % RSD less than 15%. The accuracy of the method was 90.36%.and the extraction efficiency was found to be 98.25%. The stability of drug was found to be within the range i.e. less the 15%.

Human seminal plasma allergy: successful pregnancy after prophylactic anti-histamine treatment

Human seminal plasma allergy is a rare phenomenon. Its clinical manifestations are diverse, and range from mild local pruritus to fatal anaphylaxis. Treatment varies with severity of the reactions: abstinence, condom usage or immunotherapy (subcutaneous or intravaginal) with seminal fluid. Local allergic reactions can be managed by prophylactic use of antihistamines or local cromolyn cream. A 33-year-old female visited the Asthma and Allergy Clinic in Seoul National University Bundang Hospital for the recurrent generalized urticarial reactions after sexual intercourse. She had been suffering from asthma, allergic rhinoconjunctivitis and atopic dermatitis for 10 years. She gave birth to a baby 6 months ago and no problem before. However, recently she began to recognize unexpected generalized urticaria that occurred after the sexual intercourse with husband. She wanted to have the second baby but hesitated because of the recurrent symptoms after the intercourse. She showed positive response to skin prick test with her husband's seminal fluid. The IgE-binding components were 15, 22, 28, and 35 kDa. Considering her moderate cutaneous reactions, we decided to try prophylactic treatments with oral anti-histamine one hour before sexual intercourse. She did not experience urticarial reactions with intercourse while oral anti-histamine was administered in advance. Finally, treatment outcome was successful, and the couple successfully gave birth to their second baby. We suppose that prophylactic antihistamine may be also applied in seminal plasma allergy patients if systemic reactions are limited to mild to moderate generalized urticaria.

Second to fourth digit ratio (2D:4D) in men attending Infertility Clinics in Akure Metropolis Nigeria: a predictive index?

The ratio of index finger length to ring finger length is called the “2D:4D digit ratio,” or more simply, the “digit ratio”. This study was to investigate if there are ignificant differences in the digit ratio (2D:4D) of infertile men attending an infertility clinic and men drawn from the general population in Akure Nigeria; to generate data locally to serve as a source for future referencing in anthropometry as it relates to male fertility assessment. A total of 84 participants were involved in this study. They include men attending an infertility clinic (n=42), and those drawn randomly from the general population (n=42) with regards to their fertility. Information on 2D:4D and the seminal fluid data from two samples were obtained. Direct digit estimates was done using digital calipers and indirectly by taking measurements from a digital image of the hand. The digit ratios were obtained by dividing the lengths, of the index finger by the ring finger. Semen was collected from each participant by masturbation and examined for count and motility to ascertain their fertility status. There was a statistically significant (p < 0.05) increase in the length of the fourth digit compared to the second digit in fertile men. The 2D:4D ratio in fertile men was significantly lower (p < 0.05) compared to infertile men. This study demonstrates an association between 2D:4D ratio and the fertility status in adult men in Akure metropolis Nigeria.

Routine Seminal Fluid Data Analysis of 128 Cases of Unsterility Seamen / 浙江中医药大学学报

[Objective] To analyse seminal fluid quality of seamen with unsterility,discuss the concerned environmental influencing factors.[Method] Take 128 seamen as research group,100 drivers as control;apply the Qinghua Tongfang Auto Analytic Instrument to routinly test the seminal fluid.[Result] For the seamen with unsterility,the important indexes of spermatozoon concentration,activity rate and level-a ratio were much reduced(P

Change in the Concentration of Cholesterol and Phospholipid of Seminal Plasma in Patients with Chronic Prostatitis / 대한비뇨기과학회지

PURPOSE: The complaint of infertility are found in patients with chronic prostatits. In vitro studies tend to show that the addition of massive micro organisms to semen results in decreased viability. Because such massive concentrations of pathogens seldom occur in the secretory fluids of infected prostate gland and chronic non bacterial prostatitis is more common, it seems that the chronic prostatitis produces infertility on the basis of the change in composition of seminal plasma besides a direct effect of the pathogen on spermatozoa. There is evidence that fluids from the accessory genital glands play an important role in sperm viability and fertility. The plasma membrane of spermatozoa is composed of cholesterol and phospholipid. The correlation exists between cholesterol to pohospholipids membrane ratio and cholesterol to phospholipid ratio in seminal plasma. We examined the change in the concentration of cholesterol and phospholipid of seminal plasma in patients with chronic prostatitis. MATERIALS AND METHODS: Semen from 14 healthy males and 23 chronic prostatitis patients were evaluated for the concentration of cholesterol and phospholipid of seminal plasma. RESULTS: The ages(mean +/- standard error) of control and patients with chronic prostatitis were 28.0 +/- 1.4 year and 27.0 +/- 0.4 year(p>0.05). The concentrations(mean +/- standard error) of cholesterol in control and patients with chronic prostatitis were 26.4+/-3.gmg/dl and 23.3+/- 1.8mg/dl(p>0.05). The concentrations(mean +/- sandard error) of phospholipid in control and patients with chronic prostatitis were 858.1 +/-23.7mg/dl and 789.9+/-6.0mg/dl(p<0.05). CONCLUSIONS: The decreased concentration of phospholipid of seminal plasma in patients with chronic prostatitis was found and this fact is helpful to understand the causes of functional change of sperm in patients with chronic prostatitis.

Intravaginal desensitization in a woman with seminal fluid allergy / 천식및알레르기

Human seminal fiuid (HSP) hypersensitivity is rare, but possibly a life-threatening disease. The pathogenesis of seminal plasma hypersensitivity and the exact nature of the HSP allergens remains to be clarified. We report a case of 25-year-old female patient who complained of severe itching sensation, flushing and edema of external genitalia, facial edema and dyspnea after sexual intercourse. The diagnosis was established by skin pr ick test with her husbands diluted semen. Intravaginal desensitization was performed by modified Matloffs method. Dilutions was made with sterile human serum albumin(0.2%) and 0.4% pheno1-0.9% saline solution. Two ml each of progressively greater concentrations of semen dilutions(1: 100,000 v/v, 1: 10,000 v/ v, 1:1,000 v/v, 1:100 v/v, 1:10 v/v) were inserted intravaginally at 45-min intervals, followed by an undiluted specimen. The patient was successfully desensitized and could have unprotected intercourse without anaphylaxis.

EXAMINATING THE SPERM ANTIGEN IN SEMINAL FLUID AND SEMINAL STAIN USING ELISA METHOD / 中国法医学杂志

The sperm antigens were examined using the method of ELISA and specific monoclonal antiboby against human sperm.The results of ELISA method of 10 fresh seminal fluid samples and 15 seminal stains showed 100% positive rate. Even diluted the fresh seminal fluid to 10~6 times of seminal stains extract to 51?0~5 times,both were positive also.However,it was negtive for testing other specimens such as saliva stains,urine stains,colostrum stains,sweat stains, vaginal secretion stains and human seminal stains after vesoligation. The result certificate that using specific antibody to the sperm,the ELISA method is high sensitive and specific for the sperm antigen assay.