Decreasing herpes simplex viral infectivity in solution by surface-immobilized and suspended N,N-dodecyl,methyl-polyethylenimine.

PURPOSE: To explore surface-immobilized and suspended modalities of the hydrophobic polycation N,N-dodecyl,methyl-polyethylenimine (DMPEI) for the ability to reduce viral infectivity in aqueous solutions containing herpes simplex viruses (HSVs) 1 and 2. METHODS: Surface-immobilized (coated onto surfaces) and suspended DMPEI were incubated with aqueous solutions containing HSV-1 or -2 to measure the antiviral effect of the hydrophobic polycation's formulations on HSVs. RESULTS: DMPEI coated on either polyethylene slides or male latex condoms dramatically decreases infectivity in solutions containing HSV-1 or -2. Moreover, DMPEI suspended in aqueous solution markedly reduces the infectious titer of these HSVs. CONCLUSION: Our results suggest potential uses of DMPEI for both prophylaxis (in the form of coated condoms) and treatment (as a topical suspension) for HSV infections.

Microbicide excipients can greatly increase susceptibility to genital herpes transmission in the mouse.

BACKGROUND: Several active ingredients proposed as vaginal microbicides have been shown paradoxically to increase susceptibility to infection in mouse genital herpes (HSV-2) vaginal susceptibility models and in clinical trials. In addition, "inactive ingredients" (or excipients) used in topical products to formulate and deliver the active ingredient might also cause epithelial toxicities that increase viral susceptibility. However, excipients have not previously been tested in susceptibility models. METHODS: Excipients commonly used in topical products were formulated in a non-toxic vehicle (the "HEC universal placebo"), or other formulations as specified. Twelve hours after exposure to the excipient or a control treatment, mice were challenged with a vaginal dose of HSV-2, and three days later were assessed for infection by vaginal lavage culture to assess susceptibility. RESULTS: The following excipients markedly increased susceptibility to HSV-2 after a single exposure: 5% glycerol monolaurate (GML) formulated in K-Y® Warming Jelly, 5% GML as a colloidal suspension in phosphate buffered saline, K-Y Warming Jelly alone, and both of its humectant/solvent ingredients (neat propylene glycol and neat PEG-8). For excipients formulated in the HEC vehicle, 30% glycerin significantly increased susceptibility, and a trend toward increased HSV-2 susceptibility was observed after 10% glycerin, and 0.1% disodium EDTA, but not after 0.0186% disodium EDTA. The following excipients did not increase susceptibility: 10% propylene glycol, 0.18%, methylparaben plus 0.02% propylparaben, and 1% benzyl alcohol. CONCLUSIONS: As reported with other surfactants, the surfactant/emulsifier GML markedly increased susceptibility to HSV-2. Glycerin at 30% significantly increased susceptibility, and, undiluted propylene glycol and PEG-8 greatly increased susceptibility.

Knowledge of genital herpes infection among antenatal clinic attendees in South-Eastern Nigeria.

BACKGROUND: Herpes simplex virus (HSV) is a major cause of genital ulcer disease worldwide and a significant factor for increased risk of acquisition and transmission of the Human Immune Deficiency Virus (HIV). The determination of the level of knowledge of genital herpes is necessary for the design and implementation of its specific preventive strategies as well as the reduction of the contribution of genital herpes to HIV transmission. OBJECTIVE: To determine antenatal women's knowledge on genital herpes infection. DESIGN: A cross sectional descriptive study. SETTING: Antenatal clinic of Abia State University Teaching, Hospital, Aba, Nigeria. SUBJECTS: Three hundred and fifty consecutive and consenting antenatal clinic attendees of Abia State University Teaching Hospital (ABSUTH), Aba, South Eastern, Nigeria. RESULTS: Seventy nine respondents (22.6%) had ever heard of genital herpes whilst sixty two (17.7%) had ever had recurrent blisters around their genitals. Two hundred and sixteen respondents (61.7%) reported having had cold sores or blisters around the lips or mouth following an episode of fever. Seventy four (21.1%) of the respondents knew that the virus that causes cold sores or blisters can be sexually transmitted. Higher educational levels attained and occupations other than being a housewife or farmer were associated with a greater awareness of genital herpes (p < 0.05) among the subjects. CONCLUSION: The antenatal attendees showed a poor knowledge of genital herpes infection. Discussion of genital herpes should be considered in the antenatal clinic setting along with the counselling of pregnant women regarding genital herpes, HIV infection and maternal-to-child transmission of both HSV and HIV.

Vaginal microbicides: detecting toxicities in vivo that paradoxically increase pathogen transmission.

BACKGROUND: Microbicides must protect against STD pathogens without causing unacceptable toxic effects. Microbicides based on nonoxynol-9 (N9) and other detergents disrupt sperm, HSV and HIV membranes, and these agents are effective contraceptives. But paradoxically N9 fails to protect women against HIV and other STD pathogens, most likely because it causes toxic effects that increase susceptibility. The mouse HSV-2 vaginal transmission model reported here: (a) Directly tests for toxic effects that increase susceptibility to HSV-2, (b) Determines in vivo whether a microbicide can protect against HSV-2 transmission without causing toxicities that increase susceptibility, and (c) Identifies those toxic effects that best correlate with the increased HSV susceptibility. METHODS: Susceptibility was evaluated in progestin-treated mice by delivering a low-dose viral inoculum (0.1 ID50) at various times after delivering the candidate microbicide to detect whether the candidate increased the fraction of mice infected. Ten agents were tested - five detergents: nonionic (N9), cationic (benzalkonium chloride, BZK), anionic (sodium dodecylsulfate, SDS), the pair of detergents in C31G (C14AO and C16B); one surface active agent (chlorhexidine); two non-detergents (BufferGel, and sulfonated polystyrene, SPS); and HEC placebo gel (hydroxyethylcellulose). Toxic effects were evaluated by histology, uptake of a 'dead cell' dye, colposcopy, enumeration of vaginal macrophages, and measurement of inflammatory cytokines. RESULTS: A single dose of N9 protected against HSV-2 for a few minutes but then rapidly increased susceptibility, which reached maximum at 12 hours. When applied at the minimal concentration needed for brief partial protection, all five detergents caused a subsequent increase in susceptibility at 12 hours of approximately 20-30-fold. Surprisingly, colposcopy failed to detect visible signs of the N9 toxic effect that increased susceptibility at 12 hours. Toxic effects that occurred contemporaneously with increased susceptibility were rapid exfoliation and re-growth of epithelial cell layers, entry of macrophages into the vaginal lumen, and release of one or more inflammatory cytokines (Il-1beta, KC, MIP 1alpha, RANTES). The non-detergent microbicides and HEC placebo caused no significant increase in susceptibility or toxic effects. CONCLUSION: This mouse HSV-2 model provides a sensitive method to detect microbicide-induced toxicities that increase susceptibility to infection. In this model, there was no concentration at which detergents provided protection without significantly increasing susceptibility.

The Low-Cost Compound Lignosulfonic Acid (LA) Exhibits Broad-Spectrum Anti-HIV and Anti-HSV Activity and Has Potential for Microbicidal Applications.

OBJECTIVES: Lignosulfonic acid (LA), a low-cost lignin-derived polyanionic macromolecule, was extensively studied for its anti-HIV and anti-HSV activity in various cellular assays, its mechanism of viral inhibition and safety profile as potential microbicide. RESULTS: LA demonstrated potent inhibitory activity of HIV replication against a wide range of R5 and X4 HIV strains and prevented the uptake of HIV by bystander CD4+ T cells from persistently infected T cells in vitro (IC50: 0.07 - 0.34 µM). LA also inhibited HSV-2 replication in vitro in different cell types (IC50: 0.42 - 1.1 µM) and in rodents in vivo. Furthermore, LA neutralized the HIV-1 and HSV-2 DC-SIGN-mediated viral transfer to CD4+ T cells (IC50: ~1 µM). In addition, dual HIV-1/HSV-2 infection in T cells was potently blocked by LA (IC50: 0.71 µM). No antiviral activity was observed against the non-enveloped viruses Coxsackie type B4 and Reovirus type 1. LA is defined as a HIV entry inhibitor since it interfered with gp120 binding to the cell surface of T cells. Pretreatment of PBMCs with LA neither increased expression levels of cellular activation markers (CD69, CD25 and HLA-DR), nor enhanced HIV-1 replication. Furthermore, we found that LA had non-antagonistic effects with acyclovir, PRO2000 or LabyA1 (combination index (CI): 0.46 - 1.03) in its anti-HSV-2 activity and synergized with tenofovir (CI: 0.59) in its anti-HIV-1 activity. To identify mechanisms of LA resistance, we generated in vitro a mutant HIV-1 NL4.3LAresistant virus, which acquired seven mutations in the HIV-1 envelope glycoproteins: S160N, V170N, Q280H and R389T in gp120 and K77Q, N113D and H132Y in gp41. Additionally, HIV-1 NL4.3LAresistant virus showed cross-resistance with feglymycin, enfuvirtide, PRO2000 and mAb b12, four well-described HIV binding/fusion inhibitors. Importantly, LA did not affect the growth of vaginal Lactobacilli strains. CONCLUSION: Overall, these data highlight LA as a potential and unique low-cost microbicide displaying broad anti-HIV and anti-HSV activity.

Management of neonatal herpes simplex virus infection.

Herpes simplex viruses (HSV) are ubiquitous pathogens which can be transmitted vertically causing significant morbidity and mortality in neonates. Neonatal HSV infection is infrequent with an incidence ranging from 1 in 3,500 to 1 in 20,000, depending on the population. Neonatal HSV infection is much more frequent in infants born to mothers experiencing a primary HSV infection with an incidence approaching 50%, while infants born to mothers experiencing recurrent HSV infection have an incidence of less than 3%. Neonatal infections are clinically categorised according to the extent of the disease. They are: (i) skin, eye and mouth (SEM) infections; (ii) central nervous system infection (encephalitis)--neonatal encephalitis can include SEM infections; and (iii) disseminated infection involving several organs, including the liver, lung, skin and/or adrenals. The central nervous system may also be involved in disseminated infections. Caesarean section, where the amniotic membranes are intact or have been ruptured for less than 4 hours, is recommended for those women who have clinical evidence of active herpes lesions on the cervix or vulva at the time of labour. This procedure significantly decreases the risk of transmission to the infant. Diagnosis of neonatal infection requires a very high level of clinical awareness as only a minority of mothers will have a history of genital HSV infection even though they are infected. Careful physical examination and appropriate investigations of the infant should accurately identify the infection in the majority of cases. Treatment is recommended where diagnosis is confirmed or there is a high level of suspicion. The current recommendation for treatment is aciclovir 20 mg/kg 3 times daily by intravenous infusion. Careful monitoring of hydration and renal function as well as meticulous supportive care of a very sick infant is also required. The newer anti-herpes agents, valaciclovir and famciclovir, offer no advantage over aciclovir and are not recommended for neonatal HSV infection. Prognosis is dependent upon the extent of disease and the efficacy of treatment, with highest rates of morbidity and mortality in disseminated infections, followed by central nervous system infection and the least in SEM infection. However, SEM infection is associated with poor developmental outcome even in infants who do not have encephalitis. Studies to improve the outcome of SEM infection are in progress. Neonatal HSV infections, although being relatively uncommon, are associated with significant morbidity and mortality if unrecognised and specific treatment is delayed. Diagnosis relies on a high level of clinical suspicion and appropriate investigation. With early therapy, the prognosis for this infection is considerably improved.

Neonatal herpes simplex virus infections. Presentation and management.

Neonatal herpes simplex virus (HSV) infections are recognized to be severe because of their association with significant morbidity and mortality. Through ongoing studies performed by the National Institute of Allergy and Infectious Diseases Collaborative Antiviral Study Group, the presentation, natural history, outcome and value of antiviral chemotherapy have been considered. Infants developing neonatal HSV infections can be classified according to the extent of disease, disseminated or localized. Localized infection can be subdivided into either central nervous system (CNS) disease, occurring in 35% of infected infants, or skin, eye and mouth (SEM) disease, in 41% of infants. Disseminated disease accounts for 24% of neonatal HSV infection. Therapeutic outcome depends upon disease classification. Administration of either 15 or 30 mg/kg/day of vidarabine resulted in significantly decreased mortality for infants with life-threatening disseminated and CNS disease as compared to placebo recipients. Approximately one-third of children developed normally following disseminated disease or CNS infection. When disease was localized to the SEM, no death occurred, and 88% of treated infants developed normally. While these data indicate that therapy is effective for management of infants with neonatal HSV infection, improvements are necessary. Hopefully, a study in progress will demonstrate improved outcome with acyclovir treatment.

Adolescents' and young women's use of a microbicide surrogate product when receiving oral sex.

STUDY OBJECTIVE: Genital herpes, which can be spread through oral sex, is an important target for microbicides. We examined episode-specific predictors of young women's receptive oral sex and of microbicide surrogate use. DESIGN: Longitudinal study. SETTING: Participants were recruited to participate in a microbicide acceptability study from adolescent clinics and local colleges and through snowballing. PARTICIPANTS: Young women (ages 14 to 21 y) who reported sexual contact on at least 1 weekly phone interview (n = 181) were included from the larger sample of 208 young women. MAIN OUTCOME MEASURES: On weekly diary phone interviews, participants reported whether or not their last sexual contact included receptive oral sex and whether or not their last sexual contact included use of a microbicide surrogate. RESULTS AND CONCLUSIONS: Participants reported a total of 1042 episodes of sexual contact of which 311 included receptive oral sex and 354 included microbicide surrogate use. Being older, having sex for the first time with a partner, and having given oral sex were associated with having received oral sex during a sexual episode. Being older, being African American, and having discussed the microbicide surrogate with their partner were associated with having used the microbicide surrogate use during a sexual episode. These results indicate that oral sex should be considered in the design of clinical trials. Future studies need to evaluate ways to promote consistent microbicide use in the context of receiving oral sex as well as those factors (eg, taste, pleasure) which may serve as a barrier.

Inactivation of microbial infectiousness by silver nanoparticles-coated condom: a new approach to inhibit HIV- and HSV-transmitted infection.

Recent research suggests that today's condoms are only 85% effective in preventing human immunodeficiency virus (HIV) and other sexually transmitted diseases. In response, there has been a push to develop more effective ways of decreasing the spread of the disease. The new nanotechnology-based condom holds the promise of being more potent than the first-generation products. The preliminary goal of this study was to develop a silver nanoparticles (Ag-NPs)-coated polyurethane condom (PUC) and to investigate its antimicrobial potential including the inactivation of HIV and herpes simplex virus (HSV) infectiousness. The Ag-NPs-coated PUC was characterized by using ultraviolet-visible spectrophotometry, Fourier transform-infrared spectroscopy, high-resolution scanning electron microscopy, and energy-dispersive analysis of X-ray spectroscopy. Nanoparticles were stable on the PUC and not washed away by water. Morphology of the PUC was retained after coating. The NP binding is due to its interaction with the nitrogen atom of the PUC. No significant toxic effects was observed when human HeLa cells, 293T and C8166 T cells were contacted to Ag-NPs-coated PUC for three hours. Interestingly, our results demonstrated that the contact of the Ag-NPs-coated PUC with HIV-1 and HSV-1/2 was able to efficiently inactivate their infectiousness. In an attempt to elucidate the antiviral action of the Ag-NPs, we have demonstrated that the anti-HIV activity was primarily mediated by the Ag-NPs, which are associated with the PUC. In addition, the data showed that both macrophage (M)-tropic and T lymphocyte (T)-tropic strains of HIV-1 were highly sensitive to the Ag-NPs-coated PUC. Furthermore, we also showed that the Ag-NPs-coated PUC was able to inhibit the growth of bacteria and fungi. These results demonstrated that the Ag-NPs-coated PUC is able to directly inactivate the microbe's infectious ability and provides another defense line against these sexually transmitted microbial infections.

A comprehensive murine model to evaluate topical vaginal microbicides: mucosal inflammation and susceptibility to genital herpes as surrogate markers of safety.

A critical gap in microbicide development is the absence of surrogate safety markers. The objective of the present study was to develop a murine model to examine the mucosal response to microbicides and to assess the functional implication of observed changes. Mice received 14 daily intravaginal doses of nonoxynol-9, PRO 2000, or placebo gel. Nonoxynol-9 induced an inflammatory response characterized by increases in levels of cytokines and chemokines, recruitment of neutrophils and monocytes into the genital tract, and activation of the transcription factors NF- kappa B and activator protein-1. Minimal inflammation was observed in response to 2% PRO 2000. Nonoxynol-9-treated mice were significantly more susceptible to challenge with a low dose of herpes simplex virus type 2; the response of PRO 2000-treated mice was similar to the response to placebo. These findings suggest that PRO 2000 has little deleterious effect on mucosal immunity and, if validated by clinical experiences, support the inclusion of this model in the preclinical evaluation of future candidate microbicides.

Interrupting herpes simplex virus type 2 transmission: the role of condoms and microbicides.

Condoms act as mechanical barriers to genital infection. In vitro models demonstrate that condoms are almost impermeable to viruses. A small amount of virus may cross the condom, but the condom can still reduce the level of virus exposure by several orders of magnitude. However, in vivo factors, such as condom failure during intercourse, can potentially limit their effectiveness. The weight of available evidence suggests that consistent and correct use of the male condom protects against herpes simplex virus (HSV) infection, as well as other sexually transmitted infections. Female condoms should theoretically protect against HSV infection as they cover a large surface area of potentially infected or susceptible tissues, but research is needed to confirm this. The lack of acceptance, or lack of consistent use, of condoms is the biggest barrier to their effectiveness. There is a need for effective counselling of individuals whose sexual behaviour increases their risk of HSV acquisition, such as young adults. Pregnant women are likely to be receptive to counselling due to the risk of neonatal herpes. No commercially available microbicides specifically inhibit HSV, although many inactivate the virus. More potent and specific microbicides are in development, but may be more expensive than those currently available. Although the effectiveness of condom and microbicide use is not completely proven, there is sufficient evidence to support the promotion of their use by healthcare professionals as an important part of safe sex counselling.

Oral sex and the transmission of viral STIs.

OBJECTIVE: To review the literature on the role of oral sex in the transmission of viral sexually transmitted infections (STIs). METHOD: A Medline search was performed using the keywords oro-genital sex, and those specific to each infection. Further references from each article identified by Medline were also included, as were relevant references from "Current contents". CONCLUSIONS: Oral sex is a common sexual practice among both heterosexual and homosexual couples. The evidence suggests that HIV transmission can take place through oro-genital sex from penis to mouth and vagina to mouth. Case reports describe apparent transmission from mouth to penis although this appears less likely. The risk of oro-genital transmission of HIV is substantially less than from vaginal and anal intercourse. Receptive oro-genital sex carries a small risk of human papillomavirus infection and possibly hepatitis C, while insertive oro-genital contact is an important risk factor for acquisition of HSV 1. Oro-anal transmission can occur with hepatitis A and B. The transmission of other viruses may occur but is unproved. The relative importance of oral sex as a route for the transmission of viruses is likely to increase as other, higher risk sexual practices are avoided for fear of acquiring HIV infection.

[Genital herpes with special emphasis on perinatal herpes simplex virus infection]. / Genitalni herpes s posebnim osvrtom na perinatalnu infekciju virusom herpes simpleksa.

INTRODUCTION: The incidence of genital herpes is increasing worldwide and at present herpes simplex virus type 2 is the most common cause of genital ulceration all over the world. CLASSIFICATION: The International Herpes Management Forum (IHMF) was established in 1993, suggesting a new classification of genital herpes: primary genital herpes, non-primary genital herpes, recurrent genital herpes, first episode genital herpes, atypical genital herpes and asymptomatic HSV (herpes simplex virus) infection. DIAGNOSIS: Clinical diagnosis of genital herpes should be confirmed by laboratory techniques, whereas a positive HSV culture is the best test for confirming the clinical diagnosis. Serological testing, including Western blot assay, is not the method of choice for diagnosis genital herpes. THERAPY: Management of patients with genital herpes must include various antiviral drugs (acyclovir, valacyclovir, famiclovir), but also must take into consideration the patients' clinical and emotional issues. Patients with few recurrences are best managed with episodic antiviral therapy, but those with more frequent recurrences may find a long term suppressive therapy more beneficial. Herpes simplex virus is acquired during labor in about 90% of neonatal herpes virus cases with direct contact with infected maternal genital secretions, in 5% of cases in utero (ascending infection or transplacentary) and in another 5% of cases HSV is acquired post partum. Herpes simplex virus infection includes skin infection, eye and mouth manifestations, CNS diseases and disseminated disease with multiorgan involvement. CONCLUSION: In order to reduce the risk of HSV transmission to the infant IHMF has suggested management of pregnant women with primary genital herpes: delivery by Caesarean section between 34th week and term. Acyclovir treatment may reduce the viral load at delivery, but before this can generally be recommended, more data are still required.

Nonoxynol-9 protects mice against vaginal transmission of genital herpes infections.

A vaginal application of a commercially available contraceptive jelly containing nonoxynol-9 (N9) prevented vaginal transmission of herpes simplex virus type 2 (HSV-2) infections in the mouse. When N9 jelly was delivered to the vagina with the virus inoculum, 20 s before inoculum, or 5 min before inoculum, mice were completely protected from visible infection (P < .001). Protection lasted for at least 30 min (P < .03), and significant protection occurred even when the N9 jelly was delivered 15 min after the HSV-2 inoculum (P < .05). These results are consistent with results of studies using N9 products in other animal models and suggest that N9-based contraceptive products can provide significant protection against vaginal transmission of enveloped virus infections in animals.

Tests of Buffergel for contraception and prevention of sexually transmitted diseases in animal models.

BACKGROUND: BufferGel is a novel spermicidal and microbicidal gel formulated to maintain the natural protective acidity of the vagina by acidifying semen, which otherwise alkalinizes the vagina. GOAL: To test the efficacy of BufferGel for preventing sexually transmitted infections and pregnancy in animal models. STUDY DESIGN: Animals were challenged with pathogens or sperm after pretreatment with both test and control agents, or after no pretreatment, then evaluated for infection or pregnancy using standard methods. RESULTS: BufferGel provided significant contraceptive efficacy in the rabbit, and significant protection against vaginal and rectal transmission of herpes simplex virus type 2 (HSV-2) in the mouse, vaginal transmission of Chlamydia trachomatis in the mouse, and skin transmission of cottontail rabbit papillomavirus in the rabbit. It did not protect against vaginal transmission of Neisseria gonorrhoeae in the mouse. CONCLUSIONS: The protective efficacy of BufferGel in five of the six animal models suggests that this microbicide warrants clinical evaluation for both contraception and disease prevention.

Microbicides for preventing transmission of genital herpes.

Available technologies for preventing sexual transmission of genital herpes infection are limited. This article focuses on the ongoing development of a new technology, topical microbicides, for preventing sexually transmitted infection. Recent data evaluating detergent-based spermicides as potential microbicides are reviewed. The first generation of broad-spectrum, non-detergent microbicides that are currently in clinical development, including the sulphated polymer-based inhibitors and acid buffers, are discussed. Finally, the potential of monoclonal antibodies as an example of a specific microbicide in late pre-clinical development is considered.

Herpes study and resources.

AIDS: Herpes is caused by a virus that causes recurring bouts of cold sores or genital lesions. The differences between herpes simplex virus 1 (HSV-1) and herpes simplex virus 2 (HSV-2) are explained. Herpes outbreaks can become harder to treat when the immune system is damaged by HIV. Acyclovir and famciclovir are safe and effective treatments, but preventing infection is especially important in HIV-infected individuals, as the amount of HIV in the blood increases during a herpes outbreak. World Wide Web addresses are provided for alternative herpes treatment information. A current trial is studying the effectiveness of acyclovir used with an antiviral gel, SP-303. Participants will be treated with acyclovir alone or with acyclovir and SP-303 gel. Call the Network for referral information.^ieng