Andrology: Results of retroactive testing of human semen donors for cystic fibrosis and human immunodeficiency virus by polymerase chain reaction

Retroactive testing for cystic fibrosis carrier status was performedon 149 semen donors and 100 donor applicants; 162 human immunodeficiencyvirus (HIV-1) sero-negative non-active semen donors and 45 HIV-1sero-negative actively donating semen donors were also retroactivelytested for HIV-1 by polymerase chain reaction (PCR). The 249individuals tested for cystic fibrosis were examined for sevenmutations including F508, G542X, S5491, S549N, G551D, R553X,and W1282X. Of 149 retroactively tested donors, five (3.4%)were determined to be heterozygous carriers of the F508 mutationfor cystic fibrosis. One of the 100 donor applicants was alsoheterozygous for F508. The 207 HIV-1 sero-negative donors hadan average of 15.4 (range 2–45) HIV antibody tests duringan average of 15.6 (range 1–68) months of donations. Nodonors had positive seroconversion. These donors also testednegative for HIV by PCR. Cystic fibrosis testing of donor applicantsprevents known carriers from being used for artificial inseminationby donor therapy and reduces substantially the risk of an offspringbeing born with the disease. With stringent donor selectionand exclusion of high-HIV-risk applicants, the occurrence ofHIV infection and subsequent seroconversion in donor populationsshould be very infrequent.     

The personality and motivation of semen donors: a comparison with oocyte donors.

Seventeen consecutively recruited candidates for semen donation were evaluated by a psychologist with testing and a structured interview. Most men (71%) were motivated by financial compensation. Only 29% would donate semen if records were open to potential offspring. Fifty-nine per cent of the men were rated as excellent candidates from a psychological perspective and 35% were rated as acceptable with slight reservations. One was excluded as a donor. Psychological testing revealed mildly abnormal subscale scores for 35% of donors. Forty-seven per cent had histories of minor depressive or anxiety episodes and 35% had had periods of heavy alcohol use. Compared to oocyte donors at the same institution, the men were less altruistic, more affluent, and more likely to have abused alcohol. Women had more traumatic family and reproductive histories. Psychological evaluation can be a valuable tool in gamete donor selection.     

The incidence of human herpes virus‐8 expression in lymph node biopsies from human immunodeficiency virus‐positive patients

Beukes C A & Thiart J
(2012) Histopathology  61, 942–944 The incidence of human herpes virus‐8 expression in lymph node biopsies from human immunodeficiency virus‐positive patients Aims: Human immunodeficiency virus (HIV)‐related lymphadenopathy is characterized by a wide spectrum of histological changes. Three patterns have been described which correspond to clinical stages of HIV/acquired immune deficiency syndrome (AIDS). Castleman disease is a heterogeneous group of disorders. A recently described variant, multicentric Castleman disease (MCD), of which some cases are associated with human herpes virus‐8 (HHV‐8), has been reported in both HIV‐seropositive and ‐negative patients. Considerable morphological overlap occurs between one of the patterns of HIV lymphadenopathy and this variant. Methods and results: This retrospective histopathological study on 95 cases of HIV‐reactive lymphadenopathy assessed the incidence of the different patterns and HHV‐8 on immunohistochemistry (IHC). Of the 95 cases, 78 (82.1%) were HHV‐8‐negative, of which 46 (59.0%) were classified as pattern A, 20 (25.6%) as pattern B and 12 (15.4%) as pattern C. Nine (31.0%) of 29 cases with pattern B and 8 (40.0%) of 20 cases with pattern C were HHV‐8 positive. In total 15 cases of MCD were diagnosed in this series. Conclusion: This study draws attention to the overlap between HIV lymphadenopathy and MCD. We recommend that cases of HHV‐8‐associated MCD should be investigated for HIV infection.     

Asymptomatic seminal infection of herpes simplex virus:impact on male infertility

In more than half of infertile men,the cause of their infertility is unknown.Several studies revealed the role of viral infections in male infertility.The aim of the present study was to determine the prevalence of herpes simplex virus-1 (HSV-1) and HSV-2 in semen from asymptomatic infertile male patients,and its association with altered semen parameters.A total of 70 semen samples were collected from infertile men who attended the Research and Clinical Center for Infertility in Yazd,Iran.Semen analysis and diagnostic real-time PCR using specific primers and probes for HSV-1 and HSV-2 DNA were performed.Comparison of semen parameters between virally infected and non-infected samples were performed with independent t-test and Mann-Whitney test.Semen analysis showed that infertile men fell into two groups,the male factor group and the unexplained group.HSV-1 and HSV-2 DNA was detected in 16 (22.9%) and 10 (14.3%) of 70 semen samples,respectively.All HSV-positive samples had abnormal semen parameters (the male factor group).Although HSV infection was not associated with sperm motility and morphological defects,it was correlated with lower sperm count in the seminal fluid.The findings suggest that asymptomatic seminal infection of HSV plays an important role in male infertility by adversely affecting sperm count.     

Short communication: previous semen donors and their views regarding the sharing of information with offspring

BACKGROUND: The UK government has decided to introduce, from 2005, rules that will allow donor-conceived persons to have access to identifying information concerning their donor. This has led to many concerns regarding future gamete donor recruitment. METHODS: Semen donors who had been recruited between 1988 and 2002 were invited to take part in a telephone interview. The interview sought these previous donors' views on issues associated with recruitment, attitudes regarding information sharing and views concerning the offspring. Responses regarding information sharing were compared with their views recorded at the time of recruitment. RESULTS: All 32 donors were recruited altruistically. Eighteen (56%) held the same views concerning the provision of identifying information as they did at the time of recruitment. Of those who had changed their views, eight (25%) expressed a willingness to be more open and four (12%) now wished to be anonymous having previously been unsure. Half of the donors would still have donated if they had been required to be identified to offspring, one-quarter would not have and one-quarter were undecided, although the majority of these said they may have donated under an open system. CONCLUSION: The study shows that it is possible to recruit identifiable donors at this clinic and this suggests that it may be possible for other clinics to do likewise.     

Variations in semen parameters from fathers.

Semen volume, pH, sperm characteristics and the ejaculate content of six compounds secreted by the epididymides, prostate and seminal vesicles are presented for several semen samples from 25 men (mean age 31 years) who, without clinical assistance, had fathered children within the previous 29 months. There was a large variation both within and between individual's samples for concentrations and amounts per ejaculate of all parameters except pH. The range including 96% of all values from these fertile men are presented as standards against which samples from infertile men could be compared. Lower limits (combined arithmetic means minus twice the combined within- and between-father standard deviation) were: for semen volume, 1.9 ml; for semen pH, 7.4; for total sperm count, 39 X 10(6); for normal morphological forms, 42%; for motility (WHO grades) a, 4%; b, 19%; c, 1%; d, 19%; for curvilinear velocity, 27 microns/s; for total glucosidase, 27 mU/ejaculate; for neutral alpha-glucosidase, 17 mU/ejaculate; for L-carnitine, 0.4 mumol/ejaculate; for glycerophosphocholine, 2.4 mumol/ejaculate; for fructose, 15 mumol/ejaculate; for citrate, 30 mumol/ejaculate; for zinc, 2.8 mumol/ejaculate.     

Andrology: The screening for cytomegalovirus antibody in semen donors and recipients within a donor insemination programme

A total of 67 semen donors (62 Caucasian and five non-Caucasian)were tested for the presence of immunoglobulin G antibodiesto cytomegalovirus (CMV). Ten (16%) of the Caucasian donorstested positive for CMV, while four (80%) of the non-Caucasiandonors were positive. A total of 131 women receiving donor insemination(114 Caucasian and 17 non-Caucasian) were tested. Of these,43 (38%) of the Caucasian recipients tested positive for CMV,while 16 (94%) of the non–Caucasian recipients were positive.There was a significant association between the incidence ofpositive tests and the age of the Caucasian recipients.     

Semen parameters of a semen donor before and after infection with human immunodeficiency virus type 1: case report

Semen samples from a donor who seroconverted for human immunodeficiency virus type 1 (HIV-1) during the period that he was donating at our clinic were stored before and after infection. Semen analysis was done on all of these samples before cryopreservation. Retrospectively, both qualitative and quantitative HIV-1 testing was performed on the cryopreserved semen samples to determine the time of primary HIV-1 infection. After HIV-1 infection, semen volume, sperm motility and the percentage of spermatozoa with normal morphology were reduced compared with the same parameters before HIV-1 infection. HIV-1 RNA was intermittently detectable in semen. HIV-1 infection led to a reduction in semen volume, sperm motility and normal sperm morphology in this donor. However, the clinical significance of these findings is unclear. A longitudinal cohort study on the effects of HIV-1 infection on semen quality is necessary to confirm these findings.     

British Andrology Society guidelines for the screening of semen donors for donor insemination (1999)

The British Andrology Society (BAS) guidelines for the screeningof semen donors have undergone a recent review, and followingconsultation with members of the Society and with experts inthe allied professions, the following revised guidelines havebeen issued. Major changes include the introduction of an upperage limit for semen donors (<40 years old) and the generalexclusion of men who are seropositive for cytomegalovirus asdonors. The BAS recommends the screening of prospective semendonors for chromosomal abnormalities and for cystic fibrosiscarrier status. Following the report of cross-contaminationof human cells with hepatitis B virus within a liquid nitrogenstorage vessel, the BAS recommends that steps be taken to ensurethe safe cryopreservation of donor gametes.     

Human herpesvirus 8 and human herpesvirus 2 infections in prison population

Incarcerated persons have high rates of infectious diseases. Few data on the prevalence of sexually transmitted diseases in prisoners are available. This multi-center cross-sectional study enrolled 973 inmates from eight Italian prisons. Demographic and behavioral data were collected using an anonymous standardized questionnaire and antibodies to HIV, HCV, HBV, HSV-2, and HHV-8 were detected in a blood sample obtained from each person at the time of the enrollment in the study. Two hundred and two out of the 973 subjects (20.7%) had antibodies against HHV-8. HHV-8-seropositive subjects were more likely to be older than 30 years with a higher educational level. HHV-8 infection was associated significantly with HBV (P < 0.001) and HSV-2 (P = 0.004) seropositivity and with previous imprisonments. Multivariate analysis showed that HHV-8 infection in Italian inmates was associated with HBV (P < 0.001) and HSV-2 (P = 0.002) seropositivity otherwise among foreigners inmates HHV-8 was significantly associated with HBV infection (P = 0.05). One hundred and eighty-six (21.2%) prisoners had anti-HSV-2 antibodies. At multivariate analysis HSV-2-positivity was significantly associated with HIV (P < 0.001) and HHV-8 infections (P = 0.003), whereas it was inversely associated with HCV infection (0.004). A relatively high seroprevalence of HHV-8 and HSV-2 among Italian prison inmates was found. The association of HHV-8 and HSV-2 infections suggest sexual transmission of these viruses among Italian prison inmates.     

Effect of herpes simplex virus on the DNA of human papillomavirus 18

Some types of human papillomaviruses (HPV) appear to be associated with carcinoma of the cervix or other tissues, but patients infected with HPV do not necessarily develop carcinoma. Some epidemiological studies of risk factors for cervical carcinoma have indicated the involvement of herpes simplex virus (HSV). To study the effect of HSV on the genome of HPV, total DNAs were extracted and analyzed after HeLa cells, or A431 cells, transiently transfected with HPV18 DNA, were infected with HSV-1 or -2 for 24 hours. In HeLa cells, integrated HPV18 DNA was amplified almost threefold. In A431 cells, HPV 18 DNA fragments, sensitive to the restriction enzyme Mbo I, indicated newly replicated DNA. Replication intermediates were detected when the DNA was resolved by two-dimensional gel electrophoresis. This study showed that HSV caused some amplification of HPV and indicated the possibility of HSV involved in the integration and amplification of HPV in host cells. J. Med. Virol. 53:4–12, 1997. © 1997 Wiley-Liss, Inc.     

Ultrastructural characterization of human herpesvirus 8 (Kaposi's sarcoma- associated herpesvirus) in Kaposi's sarcoma lesions: electron microscopy permits distinction from cytomegalovirus (CMV)

Kaposi's sarcoma (KS) has been shown by molecular techniques to be associated with infection with human herpesvirus 8 (HHV8/KSHV), but specific ultrastructural characterization of the virus has been impaired by the frequent presence in these lesions of other herpesviruses, particularly cytomegalovirus (CMV). Since the ultrastructural appearance of HHV8/KSHV has been studied in the cell line KS-1 uninfected with other viruses including CMV, it was possible to undertake a comparative study of CMV and HHV8/KSHV in KS lesions. HHV8/KSHV was sparsely present and lytic infection was restricted to endothelial cells. The following specific ultrastructural features allowed distinction between HHV8/KSHV and CMV: the viral particles were more delicate and less numerous in cases of HHV8/KSHV infection; the viral tegument was more electron-dense in CMV than in HHV8/KSHV; dense bodies characteristic of CMV were absent in HHV/KSHV; complete CMV viral particles were more variable in size and generally larger (150–200 nm) than HHV8/KSHV (120–150 nm); and finally, the viral envelope was more pleomorphic in CMV than in KSHV/HHV8. Similarities between CMV and HHV8/KSHV included the basic structure of the nucleocapsids and the presence of capsids lacking central DNA cores (so-called non-infectious enveloped particles). These observations show that electron microscopy can be used to identify HHV8/KSHV and confirm the relationship between HHV8/KSHV and KS. © 1997 John Wiley & Sons, Ltd.     

Human herpes virus 8‐unrelated primary effusion lymphoma‐like lymphoma: report of a rare case and review of the literature

Primary effusion lymphoma (PEL) is a very rare type of lymphoma usually confined to the body cavities predominantly in immunosupressed patients infected with human herpes virus 8 (HHV‐8). The new term for HHV‐8 independent PEL is HHV8‐unrelated PEL‐like lymphoma. We describe an 89‐year‐old human immunodeficiency virus (HIV)‐negative male patient with HHV8‐unrelated PEL‐like lymphoma in the pleura. No hepatosplenomegaly or lymphadenopathy was detected. Chest radiography and computed tomography revealed right pleural effusion, but no evidence of tumor mass or lymph node enlargement. Cytological analysis of the pleural effusion revealed a high‐grade lymphoma with round nuclei, prominent nucleoli and abundant cytoplasm with immunophenotypes positive for CD45, CD30, CD38, CD7 and CD71. Because of the advanced age, no chemotherapy was given. Effusion resolved spontaneously. One year after the diagnosis, a new pleural effusion developed at the left side. Following thoracentesis and pleurodesis, the patient remained in complete remission for 40 months. To date, 30 cases of HHV8‐unrelated PEL‐like lymphoma/HIV negative have been reported in the literature. The outcome of the HHV8‐unrelated PEL‐like lymphoma patients who were HIV negative seems to be better than HIV‐ and HHV‐8‐positive PEL.     

The relationship between fertility potential measurements on cryobanked semen and fecundity of sperm donors

Sperm penetration assay (SPA) scores obtained from cryobanked semen were correlated with therapeutic insemination (TI) fecundity in a group of established sperm donors, thereby evaluating the efficacy of the SPA in screening donors for sperm banking. While the SPA has been used to separate fertile from infertile males, we altered assay conditions to use frozen semen and to distinguish performance among fertile donors. Three frozen ejaculates from 11 pregnancy-proven donors were analysed. Of 905 TI cycles, 275 recipients achieved 95 pregnancies. There were no significant relationships between fecundity and donor semen, washed sperm parameters, sperm recoveries or recipient age. A significant relationship was revealed between mean SPA scores (range 8.7-66.6 penetrations/ovum) and donor fecundity (range 0.04-0.16, P < 0.03). Sperm concentration was varied in an effort to establish the most sensitive test condition. Using 0.25x10(6) motile spermatozoa/ml, a highly significant relationship was observed (P < 0.002). The four donors with the lowest SPA scores achieved the four lowest fecundities. It is concluded that a modified SPA can be used on frozen donor semen to estimate donor fertility potential. If applied routinely in donor semen banking, poor quality applicants could be excluded, thereby increasing pregnancy rates while decreasing donor screening costs.     

Relationship between cervical disease and infection with human papillomavirus types 16 and 18, and herpes simplex virus 1 and 2

Persistent infection with high‐risk HPV, particularly Type HPV 16 and 18, is necessary in the development of cervical cancer, but apart from HPV infection, other causative factors of most cervical cancers remain unknown. The aim of this study was to determine the prevalence of HPV 16 and HPV 18 and HSV 1 and HSV 2 in cervical samples, and to assess the role of HSVs in cervical carcinogenesis. Two hundred thirty‐three healthy controls and 567 cases (333 of cervicitis, 210 of cervical intraepithelial neoplasia, and 24 of squamous cell carcinoma) in cervical exfoliative cells were tested for HPV 16, HPV 18, HSV 1, and HSV 2 DNA using the triplex real‐time polymerase chain reaction method. In contrast to healthy women, positive rate of HPV is related significantly to cervical lesions (odds ratios (ORs) = 4.1, P < 0.01 for cervical intraepithelial neoplasia; ORs = 24.9, P < 0.01 for squamous cell carcinoma), but not cervicitis (ORs = 2.3, P > 0.05). HSV 2 prevalence in cervical intraepithelial neoplasia and squamous cell carcinoma was higher than in healthy women (ORs = 4.9, P < 0.05 for cervical intraepithelial neoplasia; ORs = 4.7, P < 0.05 for squamous cell carcinoma). HSV 2 coinfection with HPV in cervical intraepithelial neoplasia and squamous cell carcinoma was strongly higher than in healthy women (ORs = 34.2, P < 0.01 for cervical intraepithelial neoplasia; ORs = 61.1, P < 0.01 for squamous cell carcinoma). The obtained results indicated that the presence of HPV is associated closely with cervical cancer, and that HSV 2 infection or co‐infection with HPV might be involved in cervical cancer development, while HSV 1 might not be involved. J. Med. Virol. 84:1920–1927, 2012. © 2012 Wiley Periodicals, Inc.     

Leukocyte detection in human semen using flow cytometry

This study set out to establish a new method, using flow cytometry, to evaluate leukocytes in semen. Ejaculates of 59 males, asymptomatic for genitourinary infections, were examined. Routine semen analyses were carried out as well as peroxidase and polymorphonuclear granulocyte-elastase detection. Leukocytes were detected combining flow cytometry and monoclonal antibodies (anti-CD45, anti-CD53). This technique reliably assessed the total number of leukocytes and differentiated subpopulations even at low concentrations. The peroxidase test and elastase determination showed good specificity, but only moderate sensitivity versus flow cytometry combined with monoclonal antibodies. No significant association was observed between semen parameters and leukocytospermia whether evaluated by conventional methods or flow cytometry except for a moderate correlation between spermatozoa and CD53-positive cell concentrations. A first comparison of data from patients grouped on the basis of leukocytospermia (>10(6) white blood cells, WBC/ml) or non-leukocytospermia revealed no significant differences in semen parameters; lowering the threshold value for leukocytospermia to 2x10(5) WBC/ml, sperm concentration was reduced in the group with a low number of WBC identified by monoclonal antibodies. Flow cytometry using monoclonal antibodies was seen to be a simple, reproducible method that enables leukocytes in semen to be accurately detected and to identify WBC subpopulations without preliminary purification procedures.     

Immunohistological detection of human herpes virus 6 in formalin-fixed,paraffin-embedded lung tissues

Human herpes virus type 6 (HHV-6) infection is widespread in healthy individuals. The only definite disease association is with exanthem subitum in infants though the virus has been linked with a variety of other diseases including interstitial pneumonitis in bone marrow allograft recipients. In order to investigate the role of HHV-6 in the latter disease we have developed an optimised staining method for the demonstration of specific antigen in routinely processed post-mortem tissues. Formalin-fixed, paraffin-embedded lung tissue from 8 immunocompromised patients who died from interstitial pneumonitis was subjected to immunoperoxidase staining with monoclonal antibodies against HHV-6, cytomegalovirus (CMV), and adenovirus, using a modified avidin-biotin complex (ABC) method. Staining for HHV-6 was obtained in 6 of the 8 patients studied and was present in pneumocytes and macrophages. CMV and adenovirus antigens were identified in 4 and 6 patients, respectively. Whilst the lung tissue of 6 patients contained more than one virus, there was no evidence of cross-reactivity between the monoclonal antibodies. We demonstrated that accurate localisation of HHV-6 using monoclonal antibodies is possible in post-mortem lung tissue and conclude that either HHV-6 alone or in combination with other viruses may play a role in the development of interstitial pneumonitis following bone marrow transplantation or chemotherapy.     

Isolation of the predominant coagulum protein of human semen before liquefaction

The predominant protein of freshly ejaculated human semen andseminal coagulum in reducing sodium dodecyl sulphate polyacrylamidegel electrophoresis was purified for the first time prior tocleavage of coagulum proteins during liquefaction. The methodinvolves three steps using semen immediately after ejaculation.Washed seminal coagulum was first collected from fresh semensamples in acetate buffer at pH 4.5 at 0°C, solubilizedimmediately in deionized urea followed by reduction and carboxymethylationof disulphide bonds among the aggregating coagulum proteins.Then using cation exchange (CM-Sephadex) chromatography, elutedwith stepwise ionic strength gradient and finally SephacrylS-300 high resolution chromatography in the presence of urea,the predominant coagulum protein was highly purified (99%) witha yield of 6.3%. The mol. wt of the purified protein was 57000 Da. At physiological pH, the protein is rich in positivecharge.