Importance of timely diagnosis and intervention in uterine artery pseudoaneurysm caused by hysteroscopic surgery

This manuscript explores the case on the occurrence of uterine artery pseudoaneurysm(UAP)during hysteroscopy endometrial polypectomy and the subsequent successful treatment via uterine artery embolization(UAE).Moreover,we focus on the management and treatment options for UAP in patients of advanced maternal age.A pseudoaneurysm is an extraluminal blood collection with a disrupted flow that communicates with the parent vessel via a defect in the arterial wall.The reported case involved a 48-year-old primiparous woman who developed a UAP after uterine polyp removal.The study enhances the understanding of UAP,a rare but potentially life-threatening condition,by providing a detailed and well-documented account of the comprehensive case presentation,effective use of medical imaging techniques for diagnosis,successful postoperative patient management following UAE,and practical clinical recommendations for clinicians managing similar cases.Overall,this study highlights the importance of considering UAP as a differential diagnosis in patients with abnormal vaginal bleeding following hysteroscopic surgery.Additionally,this manuscript recommends that clinicians with a high index of suspicion for UAP promptly request ultrasonography and computed tomography to facilitate early diagnosis.UAE is suggested as a primary treatment due to its effectiveness and safety,particularly in facilities capable of avoiding hysterectomy.

Small Cell Neuroendocrine Carcinoma of the Vulva: Case Report and Literature Review

Background: Neuroendocrine neoplasms are those that develop from a neuroendocrine cell. They most commonly affect the lungs, gastrointestinal tract, and pancreas, being rare conditions in the female genital tract. When present, these neoplasms often manifest with nonspecific signs and symptoms such as pain, itching, swelling, single-focus lesions, bleeding, and enlargement of inguinal lymph nodes, in addition to the presence of progressively enlarging vulvar nodules. Consequently, the diagnostic investigation involves histopathological examination and confirmation through immunohistochemistry. Objective: To present a comprehensive understanding of this rarely studied pathology. The primary objective is to provide valuable insights that could aid in the future development of universally applicable treatment guidelines. Case Presentation: A 57-year-old female, with no prior comorbidities, menopause at 36, who presented with a left vulvar nodule accompanied by intense pain and swelling, later diagnosed with small cell neuroendocrine carcinoma in the vulva. Conclusion: This case report highlights the importance of enhancing our knowledge regarding small cell neuroendocrine carcinoma in the vulva, given its scarcity in medical literature. The information presented here underscores the need for standardized diagnostic and treatment approaches, paving the way for future consensus on managing this uncommon but challenging neoplasm.

Examining the Types and Consequences of Female Genital Mutilation in Somalia: A Cross-Sectional Study on the Burden and Impact of Acute and Long-Term Complications

Background: In Africa, female genital mutilation (FGM) is a widespread and harmful practice, with Somalia having the highest incidence at 98%. This practice has severe and long-lasting physical and mental health consequences. The aim of the study was to evaluate the different types of FGM;immediate and long-term complications associated with the practice, and identify factors related to its prevalence. Method: In a one-year cross-sectional hospital-based study, data were collected from females aged 18 - 50 with a history of FGM. Face-to-face semi-structured questionnaires were used for data collection. Data analysis was performed using SPSS version 26, utilizing univariate and bivariate analyses to identify correlations between variables, which were presented in tables. Results: In a study involving 255 female participants, 65.5% were between 18 and 30 years old. The majority (82.7%) were married, while 47.1% were illiterate. FGM cases were more prevalent in rural areas (61.6%). Among the participants, 45.1% were employed. Type 3 FGM was the most common (44.3%), followed by Type 2 (32.2%) and Type 1 (23.9%). In terms of short-term complications experienced by circumcised women, the most common were bleeding, reported by 29.8% of participants, followed by infection (25.1%), and urinary retention (19.2%). Among the long-term complications observed, recurrent vaginal infections were the most prevalent, affecting 29.8% of the surveyed females. This was episiotomy during delivery (22.3%) and genital scarring (10.2%). Conclusion: The study reveals that Type 3 FGM is highly prevalent among the surveyed females, contributing to an increased risk of recurrent vaginal infections, prolonged second stage of labor, higher likelihood of episiotomy during delivery, and genital scarring. These findings emphasize the urgent need for effective preventive strategies and implementation from both international and local organizations to eliminate the harmful practice of FGM.

A Content Analysis Study Describing Information Regarding Female Genital Mutilation Present in US Medical Textbooks

This study examines the depth of information regarding Female Genital Mutilation (FGM)/Female Cutting (FC) present in US medical textbooks. According to Klein et al. (2018) [1], despite substantial attention regarding the effects of FGM, this practice is still ongoing all over the world. Healthcare providers are very important actors who need to understand FGM in order to provide care to girls and women subjected to it, but the small amount of existing work in this area shows a low level of knowledge and training about FGM/FC among U.S. physicians. The Center for Disease Control and Prevention (CDC) first published the estimated number of girls at risk of FGM/FC in the U.S. to be about 168,000 in 1997;that number had increased to 513,000 girls in the US by 2012 (according to Goldberg et al.) In this study, a small sample of medical textbooks were analyzed for the degree to which the textbooks include information regarding FGM/FC. The results of this study show that US medical textbooks do not include information regarding FGM/FC, suggesting that most medical providers don’t have knowledge and training about FGM/FC. The inadequate information on FGM/FC in medical textbooks reflects a culturally biased lack of recognition of the number of girls and women affected in the US. This maintains an ongoing pattern of silence that reinforces gender inequality. Considering the inadequate information covered in US medical textbooks on FGM/FC, and the concordant lack of provider training to provide advice and care to affected girls and women, incorrect and missed diagnoses may lead to harm to victims of FGM/FC. Providers would be better equipped to care for women and girls if their medical training included information regarding FGM/FC.

Urological Complications of Female Genital Mutilation: A Case Report of 15 Patients

Objective background: This was a retrospective study of the clinical and therapeutic aspects of urological complications of female genital mutilation in the urology department of Bamako’s Gabriel Touré University Hospital. Methods: Our study took place from 2002 to 2017 in the urology department of the Gabriel Touré University Hospital in Bamako and concerned cases of complications of female genital mutilation. Our study included all girls or women presenting with urinary disorders related to complications of genital mutilation and received in urological consultation and surgical management. Results: The average age of our patients was 2.5 years, with extremes ranging from 3 months to 35 years. The oldest patient had presented with dystocia during both deliveries. Acute urine retention (n = 7) was the most frequent reason for consultation. Other signs included urinary incontinence, pollakiuria and burning. Physical examination revealed clitoridectomy and abrasion of the labia minora in 13 patients. Type III of the WHO classification of female genital mutilation was the most frequent and corresponded to infibulation.

Clinical and Therapeutic Aspects of Genital Prolapse at Hôpital du Mali about 100 Cases

To study the epidemio-clinical aspects, and the therapeutic attitudes of genital prolapse (GP) in the gynecology department of Hôpital du Mali (HDM). This is a descriptive retro-prospective study over five (5) years from January 2015 to December 2019, conducted in the gynecology department of HDM. We had collected 100 cases of GP out of 989 surgeries, with a frequency of 9.89%. The age group of 60 years and over accounted for 33% of our patients, with a mean age of 50 years. Multiparous were the most affected (89%). The notion of obstructed labor was observed in 52% of patients. The most found reason for consultation was the feeling of lump in a vagina, with 65%. Grade III according to BADEN-WALKER classification system (BWCS) concerned 72% of our patients. Triple perineal surgery and hysterectomy involved 56% of our patients. Spinal anesthesia was performed in 96% of cases. Per and postoperative complications were dominated by urinary retention in 4% and by infection of the surgical site in 2%. The average hospital stay was 3.2 days. We recorded 88% of satisfied patients after the intervention. No deaths were recorded during the study. The management of genital prolapse remains essentially surgical;it requires a semiological analysis and a mastery of the surgical technique.

The Prevalence of Female Genital Bilharziasis (FGB) in the Northern Region of Senegal

Introduction: Female Genital Bilharzia (FGB) is a pathology secondary to infection by Schistosoma haematobium. It is one of the neglected tropical diseases, capable of causing infertility, difficulties in childbirth and even cervical cancer. It represents a real public health problem. We therefore conducted a study in the maternity ward of the Saint-Louis regional hospital, with the overall aim of determining the prevalence of FBG in patients undergoing colposcopy. The specific objectives were to define the patient profile, and to compare colposcopic images with the World Health Organization (WHO) atlas. Methodology: We conducted a descriptive, analytical cross-sectional study of all colposcopic images taken at the maternity ward of the Saint-Louis regional hospital from August 1, 2018, to September 30, 2020, i.e. 25 months. The images were compared with the BGF images described in the WHO atlas. Results: We collected 178 colposcopy images. FBG images numbered 50, or 28%. The mean age of the patients was 44.5 years ±11.4 at the extremes of 18 and 78 years. Mean gestational age was 4.69 ± 2.72. Among patients with a bilharzian lesion on the cervix, visual acid inspection (VIA) was negative in 60% of cases (p = 0.007, Odd Ratio = 3.6 (1.49 - 9.07)). Conclusion: the results of our study show that FBG is a reality in our daily practice. It occurs in adult, multigestating, non-occupational women whose examination reveals a negative visual inspection with acetic acid (VIA). Healthcare providers therefore need to be trained in the recognition of lesions associated with genital bilharziasis, and public awareness needs to be raised.

Culture, Beliefs, Attitude and Peer Group Influence on Female Genital Mutilation in Southeast Nigeria

Background: Female Genital Mutilation is still practiced in Ebonyi State in Southeast Nigeria, despite the complications that follows it and government legislation against the practice. Aim: To determine the impact of Culture, Beliefs, attitude and Peer Group Influence on the persistence Female Genital Mutilation practice in the State especially in the rural areas. Materials and Methods: Qualitative study that used Focused Group Discussion and In-depth interviews for data collection. Those willing and given consent were recruited into the group discussion according to age, marital status, educational level and their location in the state. In-depth interviews were used with the Stake Holders, Opinion Leaders, Traditional Rulers and the Clergy. Result: A total of 454 participants were recruited from the 13 local government areas of Ebonyi State but only 420 (92.5%) participated. The age ranges of participants were 25 to 35 years for single males and females while the married participants male and female were aged 35 to 70 years. One hundred single females (23.8%) and 94 single males (21.4%) participated while 97 (23.1%) married women and 95 (22.6%) married men participated. Out of the 26 health workers recruited only 22 (5.2%) participated. Four traditional rulers, 4 women leaders, 4 youth leaders and 2 clergy 12 (2.9%) in number had in-depth interviews. All the participants had knowledge of FGM and its practice. Rural health workers are getting more involved. ‘Female Genital Crushing’ is replacing actual cutting. The participants all agreed there is no benefit and the practice should stop. Conclusion: Female Genital Mutilation is secretly practiced and is getting replaced by “Female Genital Crushing” perpetrated by rural health workers as well as mothers, fathers, traditional birth attendants and the peer group playing a major role with low knowledge of the Law against Female genital mutilation.

Vaginal Cystocele Cure at Bouake University Hospital (Chu): Anatomical and Functional Results

Background: Cystocele is an anatomical form of pelvic organ prolapse. It involves herniation of the bladder through the anterior wall of the vagina. Objectives: To report the anatomical and functional results of vaginal cure of cystocele in five cases. Patients and methods: A cross-sectional descriptive study of five patients presenting with a cystocele between January 2021 and December 2022 at the Urology Department of the Bouaké University Hospital (Côte d’Ivoire). All patients underwent vaginal cure of cystocele. The results of the operation were assessed on the basis of judgment criteria. The parameters studied were: age, marital status, profession, history, reason for consultation and outcome of the operation. Results: During the study period, five patients underwent vaginal cystocele surgery. The mean age was 46.6 years with extremes of 32 and 63 years. Three patients were married. All patients had a history of obstructed labour due to fetal macrosomia. 4 patients had grade II cystocele and one patient had grade III cystocele. All patients underwent anterior colpo-perineorrhaphy. The average operating time was 56 minutes (45 - 65 minutes). There was no morbidity. The average hospital stay was 3.6 days. The average duration of the urinary catheter was 2.2 days. At three months post-op, 4 patients had a good anatomical and functional result with a very good degree of satisfaction, compared with one patient who was moderately satisfied and had an average anatomical and functional result. At six months, all five patients had a good anatomical and functional result with a very good degree of satisfaction. Conclusion: Cure of cystoceles by the vaginal route is a technique that offers several advantages, especially the significant reduction in patient morbidity. Its anatomical and functional results demonstrate its reliability.

匹多莫德抑制频发性生殖器疱疹复发的临床观察

目的观察在阿昔洛韦治疗基础上,匹多莫德间歇冲击疗法和长程抑制疗法对复发性生殖器疱疹复发频率及复发次数的抑制效果。方法 52例复发性生殖器疱疹患者按照半随机方式分配至间歇冲击治疗组(A组)和长程抑制治疗组(B组),两组均在生殖器疱疹复发就诊时开始给予常规阿昔洛韦基础治疗。A组27例,仅在疱疹复发时口服匹多莫德0.8 g,2次/d,连服10 d;B组25例,口服匹多莫德0.4 g,1次/d,连服6个月,治疗后观察随访12个月。结果在6个月治疗期间,A、B两组的复发率和平均复发次数无显著性差异,而在治疗后的12个月随访期间,B组的复发率及平均复发次数均明显低于A组。结论在阿昔洛韦治疗基础上,匹多莫德长程抑制疗法可有效降低复发性生殖器疱疹的复发率,减少复发次数,其疗效明显优于间歇冲击疗法。

Association Between Genital Ulcer Disease and HIV Seroprevalence among STD Patients in Guangzhou

Objectives: To understand genital ulcer disease(GUD) among patients attending sexually transmitteddisease (STD) clinics in Guangzhou, China, and itsassociation with HIV infection.Methods: Between September 9th, 1997 and Octo-ber 30th, 2002, 8 962 patients with STDs wereevaluated. 285 patients were diagnosed with GUD basedon clinical manifestations and microbiologic evalua-tions including dark field microscopy and serologytest for syphilis (RPR, TPPA). Swabs of each genitalulcer were processed in a multiplex PCR assay (M-PCR) for simultaneous detection of Herpes simplexvirus (HSV), Treponema pallium, and Haemophilusducreyi. Other STDs were classified by routine diag-nostic criteria, including microscopy or culture forNeisseria gonorrhoeae, Chlamydia trachomatis,Urea- plasma urealyticum, Human papillomavirus,Trichomonas, etc.Results: Of the 8 962 patients with STDs, the HIVseroprevalence in patients with and without GUD was1.75% (5/285) and 1.53% (133/8677), respectively,with no statistically significant difference (χ 2=0.09,P>0.05; OR=1.15, 95%CI=0.47-2.81) . HIVseroprevalence in patients with syphilis, genital her-pes and other STDs was 2.81% (22/784), 0.74% (6/814) and 1.49% (110/7 364), respectively. Prevalencein patients with syphilis was significantly higher thanthat in patients with genital herpes and other STDs,(χ 2=9.92, P<0.005, OR=3.89, 95%CI=1.67-9.05;χ 2=7.66, P<0.001, OR=1.90, 95%CI=1.21-3.00).Conclusions: The study shows that the HIV sero-prevalence in this population of patients with GUDis very low. The results also indicate an associationbetween syphilis and HIV infection. The relationshipbetween genital herpes and HIV infection needsfurther research.

Combination of Circumcision and Microwave in Treatment of Genital Warts in Uncircumcised Patients

Objective: To investigate the efficacy of combina-tion of circumcision and microwave on genital wartsin uncircumcised men. Methods: A randomized, prospective study of 109uncircumcised adult men with genital warts was con-ducted in a STD clinic in Zhanjiang, Guangdong. Onegroup (n=54) received microwave therapy only, whilethe other group (n=55) was taken the combination ofcircumcision and microwave therapy. The recurrenceswere observed at the end of months 3, 6 and 12, andoperative complications were also recorded. Results: There were no significant differences inthe mean age and duration of the disease between twogroups (P>0.05). No serious operative complicationswere documented. The recurrence rate in circumci-sion plus microwave group was markedly lower thanthat in microwave group (12.7% vs 29.6%, P<0.05),while the differences in early and late recurrencesbetween two groups showed no statistical significance(P>0.05). Conclusion: Circumcision can be safely performedunder local anesthesia in an outpatient setting. Com-bination of circumcision and microwave can produceexcellent effect as well as less tissue damage,therefore, it may be ideal for uncircumcised patientswith extensive condylomas.

A Study of Monocyte Excretion of TNF- α and IL-6 and Monocyte Expression of HLA Class Ⅱ Antigen In Genital Herpes

Objective: To study the role of monocytes in the pathogenesis of genital herpes. Methods: TNF- α and IL-6 levels in 27 cases of genital herpes were detected by enzyme linked immunosorbant assay (ELISA). HLA class Ⅱ antigen expression on monocytes were detected by an alkaline phosphatase anti-alkaline phosphatase method. Results: Compared with normal controls, levels of TNF- a and IL - 6 secreted by monocytes responding to LPS mitogen in vitro were significantly decreased [(3.13 ± 0.44ng/ml) vs (4.68 ± 0.54ng/ml), P<0.05 and (3.32 ± 1.06ng/ml) vs (6.46 ± 1.94ng/ ml), P<0.05, respectively]. HLA class Ⅱ antigen expression on monocytes in the genital herpes group was also significantly decreased [HLA-DR (67.48% ± 1.51%) vs (81.03% ± 1.32%), P<0.01 and HLA-DQ (29.54% ± 1.15%) vs (37.63% ± 1.79%), P <0.01 respectively]. Conclusion: These findings suggest that the decreased monocyte function may contribute to the pathogenesis of genital herpes. Augmenting or inducing monocyte function may be important in the prevention, treatment, and reduction of genital herpes cases.

Diffusion-weighted MR imaging of normal and abnorma scrotum： preliminary results

Magnetic resonance （MR） imaging of the scrotum represents an important supplemental diagnostic tool in the evaluation of scrotal diseases. Diffusion-weighted （DW） MR imaging is a developing technique, proved to improve tissue characterization. We evaluated the feasibility and diagnostic performance of DW MR imaging in the detection and characterization of scrotal lesions. We retrospectively evaluated 31 scrotal lesions （23 intratesticular and 8 extratesticular） in 26 men. All MR examinations were performed on a 1.5-T unit, using a pelvic-phased array coil. DW sequences were obtained using a single shot, multislice spin echo planar diffusion pulse sequence and a b factor of 0 and 900 s mm-2. The DW MR characteristics and the apparent diffusion coefficient （ADC） values of normal scrotal contents and scrotal diseases were evaluated. Comparison between the ADC values of normal scrotum, benign lesions and scrotal malignancies was performed. The accuracy of conventional sequences, DW images alone and DW imaging combined with conventional images in differentiating benign from malignant scmtal lesions was calculated. The ADC values of testicular malignancies were different from those of normal testis and benign intratesticular lesions, and the ADC values of benign extratesticular lesions from those of normal epididymis （P〈0.05）. The overall accuracy of conventional imaging, DW imaging alone and DW MR combined with conventional sequences in the characterization of intratesticular lesions was 91%, 87% and 100%, respectively. Our findings suggest that DW MR imaging and ADC values may provide valuable information in the diagnosis and characterization of scrotal diseases.

Age-related changes in seminal polymorphonuclear elastase in men with asymptomatic inflammation of the genital tract

Aim： To investigate age-related inflammatory events in the male genital tract. Methods： In a total of 4 265 randomly collected patients attending the andrological outpatient clinic of the Center for Dermatology and Andrology, University of Giessen, Germany, ejaculate volume, pH-value, sperm concentration, total and progressive sperm motility, concentration of polymorphonuclear （PMN） elastase, number of peroxidase-positive cells and fructose were measured and correlated with patient＇s age. Results： While ejaculate volume, motility and fructose all correlated negatively with age, sperm concentration, PMN elastase and the pH-value showed a positive correlation. The prevalence of male genital tract inflammation （as defined by PMN elastase 〉 250 ng/mL） and its severity increased significantly. PMN elastase did not correlate with sperm motility. Fructose as a marker of seminal vesicle function showed a significant negative relationship with the PMN elastase levels, the number of peroxidase-positive cells and sperm motility. Conclusion： The significant increases of PMN elastase levels as marker of male genital tract inflammation in older men appear to be indicative of age-related changes in local immunoregulatory mechanisms. Because there is no association of PMN elastase with sperm motility, a direct inhibitory effect of this enzyme can be excluded.

Evaluation of Ultrasound-guided Genitofemoral Nerve Block Combined with Ilioinguinal/iliohypogastric Nerve Block during Inguinal Hernia Repair in the Elderly

To evaluate the anesthetic effect of ultrasound-guided(USG)ilioinguinal/iliohypogastric nerve(Ⅱ/IHN)block combined with genital branch of genitofemoral nerve(GFN)block in the elderly undergoing inguinal hernia repair,54 old patients(aged 60-96years,ASAⅠ-Ⅲ)with indirect hernia were enrolled and scheduled for unilateral tensiofree herniorrhaphy.Patients were grouped randomly to receive either USGⅡ/IHN plus GFN block(Group G)or USGⅡ/IHN block alone(GroupⅠ).The intraoperative visual analogue scale(VAS)scores were recorded at skin incision,at spermatic cord/round ligament traction and at sac ligation.The resting and dynamic VAS scores were recorded postoperatively.The requirements of extra sedatives and analgesics for intra-and postoperative analgesia were assessed.Occurrence of complications of the block,postoperative nausea and vomiting and femoral nerve palsy was also reported.Both groups showed similar sensory block.When stretching spermatic cord/round ligament,the patients in group G had significantly lower VAS scores than in group.And group G used much fewer adjuvant sedatives and analgesics to achieve adequate anaesthesia.In addition,group G was presented with better intraoperative anaesthesia and lower postoperative dynamic VAS scores at all time points tested.No significant difference was found in the postoperative requirement of rescue medication.Both groups showed no complications related to the block and group G reported no femoral nerve palsy.The addition of GFN block toⅡ/IHN block improves the quality of perioperative anesthesia and analgesia in the elderly and reduces the consumption of extra sedatives and analgesics during the surgery.

Human semen hyperviscosity： prevalence, pathogenesis and therapeutic aspects

The aims of this study were （a） to determine the prevalence of subjects with semen hyperviscosity （SHV） in a large population of male partners of subfertile couples; （b） to identify any correlation between SHV and infections or inflammation of the genital tract; （c） to assess the effects of therapeutic approaches for treating SHV; and （d） to assess sperm kinetic parameters after successful treatment of SHV. A retrospective study of 1 833 male partners of subfertile couples was conducted. Next, clinical, seminal, bacteriological and ultrasound studies involving 52 subjects suffering from SHV were performed, and the SHV was classified as being mild （length of thread 〉 2 cm and ≤4 cm）, moderate （〉 4 cm and ≤ 6 cm） or severe （〉 6 cm）. The prevalence of SHV was observed in 26.2% （480） of the subjects, with 13.2% suffering from mild, 6.6% from moderate and 6.4% from severe SHV. Treatment was completely successful in only 27 subjects （52.0%）, primarily in those who had mild basal SHV with a positive semen culture. In these subjects, progressive motility percentage, straight line velocity and linearity were significantly higher than pre-treatment levels. SHV is often found in subjects with subfertility. Pathogenesis was strictly related to infective/inflammatory factors in only 48.0% of cases; therefore, it is possible that biochemical, enzymatic or genetic factors have a role in this condition.

Detection of HPV Types and Neutralizing Antibodies in Women with Genital Warts in Tianjin City,China

The serum samples and corresponding cervical swabs were collected from 50 women with genital warts from Tianjin city, China. The neutralizing antibodies against HPV-16, -18, -58, -45, -6 and -11 in serum samples were tested by using pseudovirus-based neutralization assays and HPV DNAs in cervical swabs were also tested by using a typing kit that can detect 21 types of HPV. The results revealed that 36% （18/50） of sera were positive for type-specific neutralizing antibodies with a titer range of 160-2560, of which 22%（11/50）, 12%（6/50）, 10%（5/50）, 4%（2/50）, 4%（2/50） and 2%（1/50） were against HPVs -6, -16, -18, -58, -45 and -1 l, respectively. Additionally, 60% （30/50） of samples were HPV DNA-positive, in which the most common types detected were HPV-68（18%）, HPV-16（14%）, HPV-58（12%）, HPV-33（8%） and HPV-6, HPV-11, HPV-18 and HPV-52 （6% each）. The concordance between HPV DNA and corresponding neutralizing antibodies was 56% （28/50） with a significant difference （P〈0.05）. The full-length sequences of five HPV types （HPV -42, -52, -53, -58 and -68） were determined and exhibited 98%-100% identities with their reported genomes. The present data may have utility for investigating the natural history of HPV infection and promote the development of HPV vaccines.

Sperm preparation： state-of-the-art--physiological aspects and application of advanced sperm preparation methods

For assisted reproduction technologies （ART）, numerous techniques were developed to isolate spermatozoa capable of fertilizing oocytes. While early methodologies only focused on isolating viable, motile spermatozoa, with progress of ART, particularly intracytoplasmic sperm injection （ICSI）, it became clear that these parameters are insufficient for the identification of the most suitable spermatozoon for fertilization. Conventional sperm preparation techniques, namely, swim-up, density gradient centrifugation and glass wool filtration, are not efficient enough to produce sperm populations free of DNA damage, because these techniques are not physiological and not modeled on the stringent sperm selection processes taking place in the female genital tract. These processes only allow one male germ cell out of tens of millions to fuse with the oocyte. Sites of sperm selection in the female genital tract are the cervix, uterus, uterotubal junction, oviduct, cumulus oophorus and the zona pellucida. Newer strategies of sperm preparation are founded oni （i） morphological assessment by means of＇motile sperm organelle morphological examination （MSOM E）＇; （ii） electrical charge; and （ili） molecular binding characteristics of the sperm cell. Whereas separation methods based on electrical charge take advantage of the sperm＇s adherence to a test tube surface or separate in an electrophoresis, molecular binding techniques use Annexin V or hyaluronic acid （HA） as substrates. Techniques in this category are magnet-activated cell sorting, Annexin V-activated glass wool filtration, flow cytometry and picked spermatozoa for ICSI （PICSI） from HA-coated dishes and HA-containing media. Future developments may include Raman microspectrometry, confocal light absorption and scattering spectroscopic microscopy and polarization microscopy.

Molecular and in vitro Characterization of Field Isolates of Bovine Herpesvirus-1

Bovine Herpesvirus-1 (BoHV-1) is distributed worldwide and is a major pathogen in cattle, being the causal agent of a variety of clinical syndromes. The aim of this study was to isolate and to characterize (molecular and biological characterization) BoHV-1 from 29 immunosuppressed animals. It was possible to obtain 18 isolates, each from a different animal, such as from the respiratory and reproductive tracts. In some cases the cytopathic effect was visible 12 hours post-inoculation, and became characteristic after 36-48 hours. Biological characteristics were evaluated and compared with Iowa and Colorado-1 reference strains, and differences were found in plaque size, virus titer measured by TCID50 and PFU/mL, and one step virus curves. These results showed that some isolates had a highly virulent-like behavior in vitro, compared to the reference strains, with shorter eclipse periods, faster release of virus into the supernatants, and higher burst size and viral titer. There were no differences in glycoprotein expression of BoHV-1 isolates, measured by Western blot on monolayers. Moreover, using restriction endonucleases analysis, most of the viruses were confirmed as BoHV-1.1 and just one of them was confirmed as BoHV-1.2a subtype. These findings suggest that some wild-type BoHV-1 isolates could be useful as seeds to develop new monovalent vaccines.