GnRH immunocontraception of male cats

The development of nonsurgical contraceptives for cats may facilitate population control of the species. The purpose of this study was to investigate the utility of GnRH for immunocontraception of male cats. Male cats (n=12) were divided into groups of three and were immunized once with 0 (sham), 50, 200, or 400 microg synthetic GnRH coupled to keyhole limpet hemocyanin and combined with a mycobacterial adjuvant to enhance immunogenicity. GnRH antibody titer, serum testosterone concentration, and scrotal size were determined monthly. At 6 months, semen was collected by electroejaculation and testes were examined histologically. GnRH antibodies were detected in all cats receiving GnRH vaccine by 1 month post-treatment and persisted throughout the study. No dose effect of GnRH was observed; titers were not different among cats treated with 50, 200, or 400 microg GnRH (P=0.5). Six of nine treated cats were classified as responders based on high GnRH antibody titers (>32,000). By 3 months post-treatment, responder cats had undetectable testosterone concentrations and testicular atrophy. Nonresponder cats had GnRH titers of 4000-32,000 and testosterone concentrations intermediate between responder and sham-treated cats. At 6 months, total sperm counts were similar for sham-treated cats (3.1+/-1.8 x 10(6) sperm) and nonresponder cats (3.4+/-1.6 x 10(6) sperm; P=0.7). Only one of the six responder cats produced sperm, none of which were motile. Combined testicular weights of responder cats (1.3+/-0.1 g) were lower than sham-treated controls (5.3+/-1.3 g; P=0.02) and nonresponder cats (2.9+/-0.3 g; P=0.02). Histologic evaluation of the testes revealed that in responder cats, the interstitial cells that were present were pale and shrunken compared to the plump, polyhedral eosinophilic cells in sham-treated cats. GnRH responder cats had marked tubular atrophy with vacuolated Sertoli cells and a paucity of germ cells. Single-dose GnRH treatment resulted in testosterone concentrations and semen quality consistent with immunocastration in a majority of cats treated.     

Predicting the efficacy of virally-vectored immunocontraception for managing rabbits

Models were developed to examine the efficacy of immunocontraception as an alternative control method for pest rabbits. The models simulated the dynamics of rabbit populations structured by age and sex, and helped identify the benefits of an integrated pest management strategy that includes immunocontraception and lethal control. Virally vectored immunocontraception (VVIC) using a sterilising myxoma virus reduced the long-term density of rabbits. However, our models indicated that the efficacy of VVIC is much less than using lethal methods of management currently available (e.g. poison baiting). Nevertheless, in areas where lethal control cannot be used, a sterilising strain of myxoma may be a useful tool for rabbit management, but competition between sterilising and non-sterilising strains will reduce the overall efficacy of VVIC. Regardless of how effective poisoning is at reducing rabbit numbers, the benefits in terms of increased pasture availability and wool growth may not outweigh the costs incurred by a poisoning campaign. In our analysis the most cost-effective control remains natural epizootics of myxomatosis since they do not incur a cost to the landholder. There is likely to be some small additional benefit to releasing a sterilising strain of myxoma, and landholders may need to supplement this with baiting under some circumstances (e.g. above average breeding season or areas where myxomatosis occur infrequently).     

Survey of zona pellucida antigens for immunocontraception of cats

The purpose of this study was to screen a panel of native zona pellucida (ZP) antigens isolated from five mammalian species for immunocontraceptive activity in the cat (Felis catus). Native soluble-isolated ZP (SIZP) was prepared from the ovaries of cows (bZP), cats (fZP), ferrets (feZP), dogs (cZP), and mink (mZP). Vaccines were constructed using SIZP from each of the above species encapsulated in liposomes suspended in saline and emulsified with Freund's complete adjuvant (SpayVac). Female cats were immunized once (n = 3 cats per group). Serum was collected for determination of antibody titers against SIZP and for binding of antibodies to feline ovaries. All cats responded to immunization by producing anti-SIZP antibodies. The most immunogenic SIZP in cats was from mink, followed by feZP, cZP, and fZP in descending order. Antibodies had low reactivity for fZP, and no reactivity against feline ovaries was detected by immunohistochemistry. A breeding trial was commenced 20 weeks after immunization. All cats became pregnant, averaging 4.1 +/- 0.7 viable kittens per litter. We have previously shown that porcine SIZP is not an effective antigen for immunocontraception of cats. In this study, SIZP from five other mammalian species were immunogenic in the cat, but ZP antibodies failed to bind to fZP in situ, and fertility was not impeded.     

Antigenic cross-reactivity between human and marmoset zonase pellucidae, a potential target for immunocontraception.

Cross-reactivity between marmoset, chimpanzee, human and pig zona pellucida antigens was demonstrated by immunofluorescence and zona precipitation. In marmosets, anti-zona antibody prevented sperm attachment to eggs in vitro, and the antibody could be detected on zonae of ovarian oocytes following passive immunization. Use of the marmoset as an animal model in testing feasibility of the zona approach to immunocontraception is discussed.     

精囊镜联合非那雄胺治疗慢性精囊炎反复血精的疗效分析

目的:探讨精囊镜联合非那雄胺治疗慢性精囊炎反复血精的临床有效性及安全性。方法:2012年1月至2014年8月于我院住院治疗患者中选取慢性精囊炎反复血精患者62例,应用随机数字表法,其中实验组31例,采取精囊镜联合非那雄胺治疗,对照组31例,采取单纯精囊镜手术治疗。对比两组患者有效率、复发率、症状改善及精浆果糖改善情况。结果:62例慢性精囊炎反复血精患者均顺利完成手术。实验组有效率96.77%、复发率10%,对照组有效率64.52%、复发率50%,两组比较差异有统计学意义;两组患者治疗后精浆果糖均显著升高,实验组升高程度更为明显(P0.05);实验组患者症状如会阴及下腹部不适、尿道烧灼感、性欲减退、早泄及头晕四肢乏力改善(90.32%)明显好于对照组(64.52%),差异有统计学意义。结论:精囊镜联合非那雄胺治疗慢性精囊炎反复血精安全、有效,值得临床上推广应用。     

Identification of human sperm peptide sequence involved in egg binding for immunocontraception

Development of a vaccine based on sperm antigens represents a promising approach to contraception. The sperm-zona pellucida (ZP) interaction constitutes the most important event in the fertilization process, and the molecular sequences involved at this site may provide the most attractive candidates for immunocontraception. In the present study, using the phase peptide display technique, a novel dodecamer sequence, designated as YLP(12), was identified that is involved in sperm-ZP recognition/binding. The synthetic 12-mer peptide based on this sequence and its monovalent Fab' antibodies specifically and significantly (P < 0.05) inhibited human sperm-ZP binding. In Western blot and immunoprecipitation procedures, the YLP(12) peptide recognized the ZP3 component of solubilized human ZP proteins. In the Western blot procedure involving 10 different human tissue extracts, the anti-YLP(12) Fab' antibodies recognized a protein band of approximately 72 +/- 2 kDa only in the testis lane. The peptide sequence was localized on the acrosomal region of the human sperm cell. These findings indicate that the novel testis-specific 12-mer YLP(12) that is present in the acrosomal region and is involved in human sperm-ZP interaction may find applications in contraceptive vaccine development, as well as in diagnosis and treatment of male infertility mediated through sperm dysfunction.     

Seminal plasma gamma-glutamyltransferase activity and fertility.

The γ-glutamyltransferase activity of human seminal plasma does not correlate with the usual laboratory indicators of fertility. Overlapping enzyme activities in euspermic samples and aspermic post-vasectomy specimens probably excludes both sperm and epididymis as the major sources of the seminal plasma enzyme.     

不育男性精浆锌与其它精液参数的回归分析

对２２３例男性不育患者的精浆锌进行了测定,并用回归分析的方法研究了精浆锌与其它精液参数的关系．结果显示：精浆锌与精液ｐＨ、精子存活率之间存在显著的线性回归（Ｐ＜０．０１）,而精浆锌与精子密度、前向运动精子百分率、精子畸型率具有显著的曲线回归（Ｐ＜０．０１）．     

Evaluation of two sperm antigens, rSMP-B and YWK-II, as targets for immunocontraception

To determine whether sperm membrane components, rSMP-B and YWK-II, are suitable candidates as immunocontraceptives in humans, antifertility activities of the antibodies to the peptide fragments, rSMP-229 and rSMP-230 of rSMP-B and YAL-198 of YWK-II, were examined. In a previous report, anti-rSMP-230 antibody was shown to immobilise human sperm and to block human fertilisation, and the antigen (rSMP-230) to interact with antisperm antibodies found in sera of infertile women. Antibody to the second synthetic peptide, rSMP-229, corresponding to a different segment of rSMP-B, mimicked the biological activities of the anti-rSMP-230 antibody. Anti-YAL-198 antibody significantly, although weakly, inhibited human fertilisation. In the murine model, the anti-rSMP-B antibodies blocked in vitro fertilisation of mouse eggs but had no influence on embryo growth. Anti-YAL-198 antibody, however, arrested the growth of zygotes. In conclusion, rSMP-B, a human sperm protein, is a promising candidate in the development of an immunocontraceptive for human application. A second sperm protein, YWK-II, is effective as an antifertility immunogen in experimental animals.     

Application of a gene vaccine targeting HER-2/neu in immunocontraception

HER-2/neu was widely used as a target for tumor prevention and therapy because of its overexpression in many tumors. However, it also plays an important role in proliferation of endometrium, embryo implantation, and development. Here, HER-2/neu was used in immunocontraception. A gene vaccine encoding the extracellular domain of human HER-2/neu was constructed. After immunization, it especially elicited both humoral and cellular responses in mice. Embryo implantation was interfered by intravenous and intraluminal injection of anti-HER-2/neu serum or lymphocytes. Lower fertility was induced after vaccination when compared with the control groups, while injuries to the uterus and ovary were not observed. Our results suggested a new and impactful target for contraceptive vaccines development.     

Effects of immunocontraception on behavior in fallow deer (Dama dama)

Immunocontraceptives can control growth in wild and captive populations; however, in doing so, they should not disrupt species‐typical behavior patterns. The presence of treated females could disrupt social interactions in a population; yet, few studies have examined effects of immunocontraception on behavior. The goal of this project was to determine whether behavior in a fallow deer (Dama dama) population vaccinated with porcine anti‐zona pellucida (pZP)‐immunocontraceptives differed from behavior in a population of unvaccinated deer. We predicted that pZP‐treated females would spend a higher percentage of time exhibiting mating and dominance behavior than nontreated females, and that males interacting with pZP‐treated females would spend a higher percentage of time demonstrating mating and aggressive behavior than males interacting with nontreated females. We recorded activity budgets of males and females in two fallow deer populations, one pZP‐treated and the other untreated, before, during, and after rut. Females did not differ in time spent displaying mating or dominance behavior compared to nontreated females. Males coexisting with nontreated females spent more time exhibiting aggressive behavior during rut than males living with pZP‐treated females, but males did not differ in time spent in mating behavior. Thus, immunocontraception did not seem to affect behavior adversely. However, sample sizes, living conditions, and sex ratios may have affected the results. Given these limitations, future research is needed to confirm our findings. Zoo Biol 27:49–61, 2008. © 2007 Wiley‐Liss, Inc.     

Use of a murine cytomegalovirus K181-derived bacterial artificial chromosome as a vaccine vector for immunocontraception

Cytomegaloviruses (CMVs) are members of the Betaherpesvirinae subfamily of the Herpesviridae, and their properties of latency, large DNA size, gene redundancy, and ability to be cloned as bacterial artificial chromosomes (BACs) suggest their utility as vaccine vectors. While the K181 strain of murine CMV (MCMV) is widely used to study MCMV biology, a BAC clone of this virus had not previously been produced. We report here the construction of a BAC clone of the K181(Perth) strain of MCMV. The in vivo and in vitro growth characteristics of virus derived from the K181 BAC were similar to those of wild-type K181. The utility of the K181 BAC as a method for the rapid production of vaccine vectors was assessed. A vaccine strain of BAC virus, expressing the self-fertility antigen, murine zona pellucida 3, was produced rapidly using standard bacterial genetics techniques and rendered female BALB/c mice infertile with a single intraperitoneal inoculation. In addition, attenuated vaccine strains lacking the open reading frames m07 to m12 exhibited no reduction in efficacy compared to the full-length vaccine strain. In conclusion, we describe the production of a K181-based BAC virus which behaved essentially as wild-type K181 and allowed the rapid production of effective viral vaccine vectors.     

Evaluation of a porcine zona pellucida vaccine for the immunocontraception of domestic kittens (Felis catus)

With a seasonally polyestrus breeding structure, the unwanted domestic cat population has proven difficult to control. Various lethal methods have been used in an attempt to lower this population of cats. Recently, humane attempts to control "pest species," such as the feral cat, have focused on immunocontraception. SpayVacTM is a vaccine that uses antibodies raised against porcine (ZP) antigens to prevent fertilization of the ovum. SpayVac, delivered in a single dose, has been evaluated in fallow deer and several species of seals with >90% reduction in fertility and no adverse reactions. This study evaluated the effectiveness of SpayVac in reducing fertility in domestic kittens. Thirty female kittens were treated with SpayVac containing either Freund's complete adjuvant (FCA) or alum, or with a control vehicle. Kittens were monitored for side effects, estrus cycling at maturity, and fecundity. Anti-porcine ZP antibodies were quantified by ELISA. Immunohistochemical assays measured the species specificity of the antibodies produced and IgG binding in vivo. Despite high anti-porcine ZP antibody titers, neither formulation of SpayVac prevented estrus cycling at maturity or reduced fecundity. Immunohistochemical assays indicated that antibodies produced by cats treated with SpayVac recognized porcine ZP, but not feline ZP.     

The Transurethral Seminal Vesiculoscopy in the Diagnosis and Treatment of the Seminal Vesicle Disease

To develop a transurethral endoscopy technique of the transurethral seminal vesiculoscopy to examine and treat seminal vesicle disease. A total of 61 patients with seminal vesicle disease were diagnosed and treated with the transurethral seminal vesiculoscopy through the distal seminal tracts and vesicles. 58 cases were successfully treated using transurethral seminal vesiculoscopy via the seminal vesicles. The operation took 25 ~ 85 min, with an average of (35.6) mins. In this group, seven cases were diagnosed as ejaculatory orifice cyst, 14 cases had blood clots in the seminal vesicles, and nine patients had stones in the seminal vesicles. All patients were treated properly. Follow-up occurred at 3 months, with two cases showing post-operative discomfort in perineal region. One patient had recurrence with seminal vesiculitis, which improved with treatment. Four infertile patients had a significant increase in sperm count and ejaculation volume and two of these patients were able to naturally inseminate within seven to 18 months post-surgery. This approach enables a new endoscopic technique with the transurethral seminal vesiculoscopy to diagnose and treat seminal vesicle disease through the normal anatomic pathway which can be easily performed with few post-operative complications.