In the ovine pituitary, CXCR4 is localized in gonadotropes and somatotropes and increases with elevated serum progesterone.

The pituitary is the central endocrine regulator of reproduction and in addition to various hormones regulating its actions, other molecules, such as chemokines, influence pituitary physiology as well. Despite reports over 2 decades ago that chemokines regulate the pituitary, much of the basic biology discerning chemokine action in the pituitary is unclear. A small number of chemokines and their receptors have been localized to the pituitary, yet chemokine ligand 12 (CXCL12) and its receptor, CXCR4, have received the most attention as both are increased in human pituitary adenomas. This chemokine duo was also reported in normal human and rat pituitary, suggestive of a functional role and that this chemokine axis might function in pituitaries from other mammalian species. To date, reports of CXCL12 and CXCR4 in pituitary from livestock are lacking, and research on pituitary during pregnancy in any mammalian species is limited. Moreover, progesterone regulates CXCR4 expression in a tissue-dependent manner, but whether differing concentrations of progesterone reaching the pituitary modulate CXCL12 or CXCR4 is not known. To address these gaps, our first objective was to determine if CXCL12 and CXCR4 expression and protein abundance differ in sheep pituitary during early gestation (days 20, 25, and 30 of gestation) compared to nonpregnant ewes. The second objective was to determine if CXCL12 or CXCR4 production was altered in the ovine pituitary when circulating progesterone concentrations are elevated. The expression of CXCL12 messenger RNA decreased on day 20 of gestation compared to nonpregnant ewes; CXCL12 protein was similar across all days tested. In nonpregnant and pregnant ewes, CXCR4 was localized to somatotropes and gonadotropes on all days tested. Abundance of CXCR4 increased in the pituitary tissue of pregnant ewes with elevated circulating progesterone compared with pregnant ewes with normal circulating progesterone concentrations (control). The present study details CXCL12 and CXCR4 in normal ovine pituitary and reveals that gonadotropes and somatotropes may be regulated by CXCL12/CXCR4, underscoring this signaling axis as a potential new class of modulator in endocrine functions.

Effect of acyclovir treatment of primary genital herpes on the antibody response to herpes simplex virus.

Sera from patients with first episode primary genital herpes infections who were treated with the antiviral drug acyclovir were studied to determine the effect of therapy on the immune response to herpes simplex virus (HSV) glycoproteins and polypeptides. 63 patients were evaluated, 35 patients received acyclovir: 11 intravenously, 12 orally, and 12 topically, while 28 received placebo. Topical application of acyclovir had no effect on the immune response to HSV infection. However, both oral and intravenous acyclovir were associated with later development of antibodies to two glycoproteins (of 80,000 and 60,000 mol wt [IIg80 and gD, respectively]) and one nonglycosylated polypeptide of 66,000 mol wt (vp66). Antibody to IIg80 was present in convalescent phase serum in 13/23 systemic acyclovir recipients vs. 18/19 placebo recipients (P = 0.01) and antibody to gD was detected in 8/23 oral or intravenous acyclovir recipients vs. 11/19 placebo recipients (P = 0.06). The mean time to seroconversion to IIg80 (39.0 d) and gD (55.5 d) was significantly longer for systemic acyclovir recipients than for the placebo controls, 23.4 and 18.5 d, respectively (P less than 0.05 for each comparison). 7 (30%) of 23 systemic acyclovir recipients compared with 100% of the placebo recipients had antibody to vp66 by 30 d after onset of the primary episode (P less than 0.001). Subsequent untreated recurrences of genital herpes were associated with seroconversion to gD, IIg80, and vp66. Patients who lacked antibody to both gD and vp66 in sera taken before their first clinical recurrence of disease experienced a longer duration of the recurrent episode (10.8 d) than those who possessed antibody to both vp66 and gD (6.3 d) (P less than 0.05). In addition, the mean duration of lesions, number of lesions, and mean lesion area were greater in patients who lacked antibody to vp66 but had anti gD, as compared with those who had anti-p66 but lacked anti-gD; suggesting that antibody to vp66 correlated more closely with subsequent disease severity than did antibody to gD. Acyclovir therapy appears to influence the frequency and time of development of antibody to a number of different HSV-specific polypeptides. Further studies of the effects of antiviral therapies on the immune response to these proteins may help clarify the role of these polypeptides in the pathogenesis of disease.

Herpes simplex virus-2 (HSV-2) type-specific antibody correlates of protection in infants exposed to HSV-2 at birth.

Western blot analysis was used to compare the herpes simplex virus (HSV)-2 antibody profiles of 40 infants less than 2 wk of age who had been exposed to maternal genital HSV-2 at birth. 4 mothers were HSV seronegative at delivery and seroconverted to HSV-2 ("primary infection"), 9 had HSV-1 antibodies and seroconverted to HSV-2 ("nonprimary first episode infection"), and 27 were HSV-2 seropositive ("recurrent infection"). Neonatal herpes infections developed in 1 of 4 infants of women with primary infection, in 3 of 9 infants of women with nonprimary first episode infection, and in none of the 27 infants of women with recurrent HSV-2. Antibodies to HSV-2 proteins gG-2, VP5, and ICP35 were detected in 83, 89, and 72% of the 36 uninfected infants, respectively. None of the four infected infants had detectable antibodies to gG-2 and only one (25%) had antibodies to VP5 or ICP35. The more limited profiles of the 13 infants born to mothers with first episodes of HSV-2 were then analyzed separately; these profiles were similar among infected and uninfected infants except for gG-2, which elicits antibodies that are type specific for HSV-2. None of the infected infants versus seven of nine (78%) uninfected infants were gG-2 seropositive. These comparisons suggest that maternal type-specific antibodies may play a role in preventing neonatal infection after exposure to HSV-2.

Inhibition of chemokine (C-X-C motif) receptor four (CXCR4) at the fetal-maternal interface during early gestation in sheep: alterations in expression of chemokines, angiogenic factors and their receptors.

Chemokine (C-X-C motif) ligand 12 (CXCL12) and its receptor, chemokine (C-X-C motif) receptor 4 (CXCR4), are involved in significant biological processes associated with early pregnancy including increasing trophoblast invasion and stimulating placental vascularization. To further elucidate functions of CXCL12-CXCR4 signaling during early gestation, our objective was to inhibit CXCR4 in vivo using a CXCR4 antagonist, AMD3100. We hypothesized that inhibition of CXCR4 would negatively affect chemokine and angiogenic factor regulation imperative for placental development in sheep. Osmotic pumps containing PBS (control) or AMD3100 (CXCR4 antagonist) were surgically installed ipsilateral to the corpus luteum on d 12 of gestation and administered treatments directly into the uterine lumen. Maternal (caruncle and intercaruncle) and fetal membrane tissues were collected on d 23 of gestation and mRNA and protein expression were analyzed for vascular endothelial growth factor (VEGF), kinase insert domain receptor (KDR), fms related tyrosine kinase 1 (FLT1), fibroblast growth factor 2 (FGF2), angiopoietin 1 (ANGPT1), hypoxia inducible factor 1 ɑ subunit (HIF1A), CXCL12, and its corresponding receptors (CXCR4 and CXCR7). Immunohistochemical procedures were performed for analysis of CXCL12 and cell proliferation. In caruncle tissue ipsilateral to the pump, mRNA for KDR, ANGPT1, HIF1A, and CXCL12 increased (P < 0.05) in treated ewes compared to control, whereas caruncle tissue contralateral to the pump had increased expression (P < 0.05) of KDR, and CXCL12 in treated ewes. In fetal membrane, CXCR4 mRNA and protein decreased (P < 0.05), while VEGF protein decreased (P < 0.05) in caruncle and fetal membrane tissue from treated ewes. Results from this study highlight the importance of CXCL12-CXCR4 signaling at the fetal-maternal interface. Inhibiting this axis may disrupt typical regulation of angiogenic factors needed for placental development and embryo growth.

Cloning and characterization of an ovine intracellular seven transmembrane receptor for progesterone that mediates calcium mobilization.

Classically, progesterone has been thought to act only through the well-known genomic pathway involving hormone binding to nuclear receptors (nPR) and subsequent modulation of gene expression. However, there is increasing evidence for rapid, nongenomic effects of progesterone in a variety of tissues in mammals, and it seems likely that a membrane PR (mPR) is causing these events. The objective of this study was to isolate and characterize an ovine mPR distinct from the nPR. A cDNA clone was isolated from ovine genomic DNA by PCR. The ovine mPR is a 350-amino acid protein that, based on computer hydrophobicity analysis, possesses seven transmembrane domains and is distinct from the nPR. Message for the ovine mPR was detected in hypothalamus, pituitary, uterus, ovary, and corpus luteum by RT-PCR. In CHO cells that overexpressed a mPR-green fluorescent protein fusion protein, the ovine mPR was localized to the endoplasmic reticulum and not the plasma membrane. Specific binding of 3H-progesterone to membrane fractions was demonstrated in CHO cells that expressed the ovine mPR but not in nontransfected cells. Furthermore, progesterone and 17 alpha-hydroxy-progesterone stimulated intracellular Ca2+ mobilization in CHO cells that expressed ovine mPR in Ca2+-free medium (P < 0.05) but not in CHO cells transfected with empty vector. This rise in intracellular Ca2+ is believed to be from the endoplasmic reticulum as intracellular Ca2+ mobilization is absent when mPR transfected cells are first treated with thapsigargin to deplete Ca2+ stores from the endoplasmic reticulum. Isolation, identification, tissue distribution, cellular localization, steroid binding, and a functional response for a unique intracellular mPR in the sheep are presented.

Reevaluation of the effect of mouse mammary tumor virus infection on the BALB/c mouse hyperplastic outgrowth.

The BALB/c (C-) mouse hyperplastic outgrowth line (D1) was used to study murine mammary tumor virus (MuMTV) expression in both D1 and tumors derived from D1. D1 was transplanted into virus-infected BALB/cfC3H (C+) and virus-uninfected C- animals. In duplicate studies, tumor incidence was the same in both groups. However, the tumor latency period was longer for D1 transplanted into C+ mice (D1/C+) than for D1 transplanted into C- mice (D1/C-). MuMTV was detected in 85% of D1/C+ outgrowths and in 29% of D1/C+ tumors but was never detected in D1/C- outgrowths or tumors. D1/C- outgrowth and tumors and most of the D1/C+ tissues expressed little or no MuMTV RNA. Some D1/C+ tumors expressed substantial levels of MuMTV RNA. These same tumors also had MuMTV antigen and contained the exogenously acquired C3H-MuMTV provirus in the cellular DNA as shown by DNA fragment patterns following cleavage with Pst I and Eco RI endonucleases. D1/C+ tumors containing no viral RNA were also antigen-negative and lacked the acquired C3H provirus. These studies indicate that D1 has substantially changed in its incidence and in its response to MuMTV. MuMTV infection was not tumorigenic in the traditional sense, a finding that has led to a reevaluation of our current models of virus-host relationships and the biology of precancerous conditions.

Long-term primary culture of mouse mammary tumor cells: production of virus.

Long-term primary cultures of mouse mammary tumor cells proved an excellent source of mouse mammary tumor virus (MMTV). Virus purified from these primary cultures had the same morphologic biochemical, immunologic, and biologic characteristics as MMTV. Quantitation of MMTV-protein equivalents released into the medium was measured by the radioimmunoassay for MMTV. Peak production levels were 20-40 mug MMTV protien equivalents/75-cm-2 flask/24 hours. These cultures produced MMTV for as long as 90 days. MMTV cultivation depended on the initial cell-plating density and hormones. Maximal MMTV release was obtained at a plating density of 1 times 10-6 cells/cm-2 in the presence of insulin and hydrocortisone. Insulin alone gave basal levels of MMTV, and hydrocortisone alone increased MMTV release only three-fold, but insulin and hydrocortisone together effected an eightfold increase in MMTV release. This suggested that hydrocortisone had a primary effect on MMTV release and insulin acted synergistically with hydrocortisone to maximize MMTV release.

A high-tumor-incidence subline of the D1 mouse mammary hyperplastic outgrowth line: effect of carcinogens.

Mammary tumorigenesis in some mouse strains is characterized by the appearance of a preneoplastic lesion, the hyperplastic alveolar nodule (HAN). The biology of the HAN has been characterized primarily through the study of stable outgrowth lines of serially transplanted HAN. One outgrowth line, Dl, which was developed and carried in female BALB/c mice, has been described as a low-tumor-incidence line that does not express murine mammary tumor virus (MuMTV) and is susceptible to hormonal, chemical, and viral carcinogens. In this report, a high-tumor-incidence subline of Dl, Dl/UCD, is described. Although Dl/UCD, like Dl, is susceptible to the chemical carcinogen 7,12-dimethylbenz[a]anthracene, an increase in tumor incidence was not observed when Dl/UCD outgrowth was exposed to hormones by means of pituitary isografts. Unlike Dl, Dl/UCD is refractory to the carcinogenic action of MuMTV. Both Dl and the Dl/UCD subline contained the endogenous MuMTV provirus but did not contain exogenous MuMTV provirus sequences. MuMTV antigen was not detected in Dl/UCD outgrowths or tumors. RNA hybridizable to MuMTV complementary DNA was detected in some Dl/UCD outgrowths and tumors but did not appear to correlate with tumorigenesis.

History of circumcision, medical conditions, and sexual activity and risk of penile cancer.

BACKGROUND: Epidemiological evidence suggests lack of neonatal circumcision as the strongest risk factor for penile cancer, but the role of sexually transmitted diseases in the etiology of penile cancer has remained unclear. PURPOSE: To further clarify risk factors for penile cancer, we examined the role of circumcision, personal characteristics and habits (such as smoking), sexually transmitted diseases, past sexual activity, and medical conditions of the penis. METHODS: A population-based, case-control study was conducted in western Washington state and in the province of British Columbia. We interviewed 110 men with penile cancer diagnosed from January 1979 to July 1990 and 355 control subjects from the general population, frequency matched to case subjects on age and date of diagnosis. Tumor tissue from 67 case subjects was tested for human papillomavirus (HPV) DNA by polymerase chain reaction. Results of blood tests from 69 case subjects and 208 control subjects were available for study. STATISTICALLY SIGNIFICANT RESULTS: Relative to men circumcised at birth, the risk for penile cancer was 3.2 times greater among men who were never circumcised and 3.0 times greater among men who were circumcised after the neonatal period. For current smokers, the risk was 2.8 times that of men who never smoked. The risk among men reporting a history of genital warts was 5.9 times that of men reporting no such history. Of 67 tumors tested for HPV DNA, 49% were positive; the majority of these positive tumors (70%) were type 16, which has been associated with anogenital carcinoma. Relative risks (RRs) associated with a reported history of penile rash or penile tear were 9.4 and 3.9, respectively. Among men not circumcised at birth, RRs associated with presence of smegma and difficulty in retracting the foreskin were 2.1 and 3.5, respectively. Twenty-eight percent of case subjects, compared with only 10% of control subjects, reported 30 or more sexual partners, and men with HPV-positive tumors were more likely to report a greater number of sexual partners. CONCLUSIONS: These results suggest that the absence of neonatal circumcision and potential resulting complications are associated with penile cancer. Additionally, medical conditions of the penis, sexual activity, infection with HPV, and smoking may increase the risk for penile cancer. IMPLICATIONS: A larger study would allow examination of interrelationships of circumcision, infection with HPV, and smoking as risk factors.

Development and characterization of mouse hyperplastic mammary outgrowth lines from BALB/cfC3H hyperplastic alveolar nodules.

Five hyperplastic outgrowth lines were developed by serial transplantation of hyperplastic alveolar nodules from BALB/cfC3H mice into BALB/c hosts. The lines were used to study the biology, morphology, and virology of premalignant tissue originating in mouse mammary tumor virus (MuMTV)-positive animals. The five lines differed with respect to tumor potential and growth characteristics, corroborating the previous evidence that MuMTV-positive hyperplastic alveolar nodules are biologically heterogeneous. Subgross and microscopic examination of outgrowths and tumors revealed that each line had unique morphological characteristics. The presence of atypical lobules within the hyperplastic outgrowth appeared to be correlated with tumor risk, and a morphological continuum of atypical lesions ending in overt cancer was suggested. Viral expression was detected by nucleic acid hybridization and immunoperoxidase staining for MuMTV structural antigens. While the MuMTV RNA in certain tissues appeared to vary qualitatively with tumor potential of the outgrowth line, no correlation between viral antigens detected by immunoperoxidase staining and tumor potential was observed.

Restriction endonuclease studies of hyperplastic outgrowth lines from BALB/cfC3H mouse hyperplastic mammary nodules.

The DNA's isolated from five mouse hyperplastic mammary gland outgrowth lines from BALB/cfC3H mice were digested with the restriction endonucleases PsTI, BamHI, or EcoRI; electrophoresed; and analyzed by Southern blotting and autoradiography. Proviral DNA sequences from the acquired C3H mouse mammary tumor virus were detected in the DNA of all five lines, indicating that they were infected. The DNA of the five hyperplastic lines contained more EcoRI and BamHI mouse mammary tumor virus proviral DNA fragments than did DNA from normal organs, suggesting that the hyperplastic tissues were composed of more homogeneous cell populations than was lactating mammary gland. Each hyperplastic line had unique and reproducible BamHI and EcoRI restriction (integration) patterns which were stable over as many as seven transplant generations. Three sublines, which originated from the same hyperplastic alveolar nodule, had unique integration patterns but also shared several fragments. On the basis of these observations, we propose that mouse mammary "hyperplasias" are clonal dominant premalignant neoplasms.

Placental development during early pregnancy: Effects of embryo origin on expression of chemokine ligand twelve (CXCL12).

The aim was to localize chemokine ligand twelve (CXCL12) in sheep placental tissues during early gestation and after assisted reproductive technologies (ART). Uteri were collected from naturally (NAT) mated ewes and ewes receiving embryo transfer (ET), in vitro fertilization (IVF) or in vitro activation (IVA). CXCL12 was immunolocalized to endometrial stroma, glands, and trophoblast. Greater CXCL12 immunoreactivity was present in trophoblast on day 22 and 24 and in NAT ewes compared to IVF and IVA. Increased CXCL12 expression suggests CXCL12 promotes implantation and placentation. Decreased CXCL12 in IVF and IVA embryos, may compromise pregnancy establishment when utilizing ART methods.

Maternal environment and placental vascularization in small ruminants.

Uteroplacental development is a crucial step facilitating conceptus growth. Normal placental development comprises extensive placental angiogenesis to support fetoplacental transport, meeting the metabolic demands of the fetus. Compromised pregnancies due to maternal stressors such as over or undernutrition, maternal age or parity, altered body mass index, or genetic background result in altered vascular development of the placenta. This negatively affects placental growth and placental function and ultimately results in poor pregnancy outcomes. Nonetheless, the placenta acts as a sensor to the maternal stressors and undergoes modifications, which some have termed placental programming, to ensure healthy development of the conceptus. Sex steroid hormones such as estradiol-17ß and progesterone, chemokines such as chemokine ligand 12, and angiogenic/vasoactive factors such as vascular endothelial growth factors, placental growth factor, angiopoietins, and nitric oxide regulate uteroplacental development and hence are often used as therapeutic targets to rescue compromised pregnancies. Interestingly, the presence of sex steroid receptors has been identified in the fetal membranes (developing fetal placenta). Environmental steroid mimetics known as endocrine disrupting compounds disrupt conceptus development and lead to transgenerational impairments by epigenetic modification of placental gene expression, which is another area deserving intense research efforts. This review attempts to summarize current knowledge concerning intrinsic and extrinsic factors affecting selected reproductive functions with the emphasis on placental development.

A prospective study of tender points and fibromyalgia during and after an acute viral infection.

BACKGROUND: Tender points (TPs) and fibromyalgia (FM) may be precipitated by infections, but the frequency, associated characteristics, and predictors of these outcomes are unknown. OBJECTIVES: To determine if acute infectious mononucleosis (AIM) is associated with the development of TPs or FM acutely or during the subsequent 6 months; if demographic, clinical, or psychosocial features predict TPs or FM; and if TPs or FM correlate with nonrecovery. METHODS: A total of 150 subjects diagnosed as having AIM were assessed with physical examinations (including palpation of 18 TPs), laboratory tests, and measures of psychosocial and somatic functioning at enrollment and at 2 and 6 months. Subjects also completed a structured psychiatric interview at the initial evaluation. RESULTS: At presentation and at 2 and 6 months, the mean TP counts were 7.5, 4.6, and 3.0, respectively; at these time points, 19%, 3%, and 1% of subjects also met modified criteria for FM. Tender points and degree of pain diminished over time following AIM. Acutely, TPs were associated only with higher temperature (P<.001). Baseline features that predicted more TPs at 2 and 6 months were female sex, older age, less family social support, and more TPs at presentation. Neither initial laboratory tests nor psychiatric disease or distress predicted TPs. Differences between those who had and had not recovered at 6 months were found for the mean number of TPs (P<.008), the proportion of subjects with 11 or more TPs (P<.002), and the degree of pain. CONCLUSIONS: Tender points are a common, transient finding associated with AIM, but FM is an unusual long-term outcome. Demographic, social, and physical examination features predicted TPs.

Placental development during early pregnancy in sheep: estrogen and progesterone receptor messenger RNA expression in pregnancies derived from in vivo-produced and in vitro-produced embryos.

Sex steroids are important regulators of angiogenesis and growth in reproductive tissues, including the placenta. In experiment (exp.) 1, to examine the expression of a suite of sex steroid receptors throughout early pregnancy, maternal (caruncular [CAR]) and fetal (fetal membranes [FM]) placental tissues were collected on days 14 to 30 after mating and on day 10 after estrus (nonpregnant controls). In exp. 2, to examine the hypothesis that assisted reproductive technology would affect the expression of the same suite of sex steroid receptors, pregnancies were achieved through natural mating (NAT) or transfer of embryos from natural mating (NAT-ET), in vitro fertilization (IVF), or in vitro activation (IVA), and CAR and FM were collected on day 22. In exp. 1, for CAR messenger RNA (mRNA) expression of estrogen receptors (ESR) 1 and 2, nuclear (n) progesterone receptors (PGR) and membrane (m) PGRα, ß, and γ were affected (P < 0.02) by pregnancy stage, as were ESR1, nPGR, and mPGRα, ß, and γ for FM (P < 0.03). In exp. 2, for CAR, mRNA expression of ESR1 and nPGR was decreased (P < 0.001) in NAT-ET, IVF, and IVA groups compared with NAT. For FM, mRNA expression of ESR1 tended to be greater (P = 0.10) in the IVA group compared with NAT and NAT-ET, and GPER1 was greater (P < 0.05) in NAT-ET and IVF compared with NAT. These data establish the normal pattern of sex steroid receptor mRNA expression in maternal and fetal placenta during early pregnancy in sheep, and in addition, suggest that altered expression of placental sex steroid receptors may be an early event leading to poor placental vascularization and growth after assisted reproductive technology.

Genital ulceration as a risk factor for human immunodeficiency virus infection.

Among 115 heterosexual men who presented with genital ulcers to a sexually transmitted disease clinic in Nairobi, Kenya, the prevalence of serum antibody to HIV was 16.5%. A past history of genital ulcers was reported by 12 (63%) of 19 men with antibody to HIV versus 30 (31%) of 96 without antibody (P = 0.008). HIV infection was also positively associated with lack of circumcision, but was not associated with the etiology of the current genital ulcer. Logistic regression analysis (adjusted for age, number of recent sex partners, recent prostitute contact, circumcision, tribal ethnic identity, past history of urethritis, and current diagnoses) confirmed only the association between prior history of genital ulcer disease and HIV infection; (P = 0.04, odds ratio 2.35, 95% confidence limits, 1.01-5.47). The incidence of genital ulcers, particularly chancroid, is much higher in parts of Africa than in Europe or North America. This may contribute to the increased risk of heterosexual transmission of HIV in Africa. Aggressive control of chancroid and syphilis may offer one very feasible approach to reducing transmission of HIV in this region.

Comparative analysis of methods for collection and measurement of immunoglobulins in cervical and vaginal secretions of women.

The purpose of this study was to systematically compare 3 collection methods, Sno-strips, wicks and cervical-vaginal lavage, for analysis of immunoglobulin concentrations in female genital secretions. In each of 8 women, absorbent wicks and Sno-strips were applied at 4 locations: the lateral wall of the vagina; the posterior vaginal fornix; the surface of the exocervix; and the endocervical canal. Cervical-vaginal lavage was then performed in 4 women with 5 ml PBS. Immunoglobulin and protein concentrations in lavage samples were generally over 100 times lower than in the secretions captured directly from mucosal surfaces with either Sno-strips or wicks. Capture of undiluted secretions with either wicks or Sno-strips allowed calculation of actual immunoglobulin concentrations at specific mucosal sites: for example, median IgA levels were consistently highest in the endocervix and lowest in the vagina. Such information may be crucial in evaluating the correlates of protective immunity against micro-organisms that infect or invade discrete regions of the genital mucosa.

Acute infectious mononucleosis: characteristics of patients who report failure to recover.

PURPOSE: We sought to determine how often acute mononucleosis precipitates chronic illness, and to describe the demographic, clinical, and psychosocial features that characterize patients who report failure to recover. SUBJECTS AND METHODS: We enrolled 150 patients with infectious mononucleosis during the acute illness and asked them to assess their recovery at 2 and 6 months. At baseline, we performed physical and laboratory examinations; obtained measures of psychological and somatic functioning, social support, and life events; and administered a structured psychiatric interview. RESULTS: Self-assessed failure to recover was reported by 38% of patients (55 of 144) at 2 months and by 12% (17 of 142) at 6 months. Those who had not recovered reported a persistent illness characterized by fatigue and poor functional status. No objective measures of disease, including physical examination findings or serologic or laboratory markers, distinguished patients who failed to recover from those who reported recovery. Baseline predictors for failure to recover at 2 months were older age (odds ratio [OR] = 1.4, 95% confidence interval [CI]: 1.1 to 1.8, per 5-year increase), higher temperature (OR = 1.5, 95% CI: 1.1 to 2.2, per 0.5 degrees C increase), and greater role limitation due to physical functioning (OR = 1.5, 95% CI: 1.2 to 1.9, per 20-point decrease in Short Form-36 score). At 6 months, baseline predictors for failure to recover included female sex (OR = 3.3, 95% CI: 1.0 to 12), a greater number of life events more than 6 months before the disease began (OR = 1.7, 95% CI: 1.1 to 2.5, per each additional life event), and greater family support (OR = 1.9, 95% CI: 1.1 to 4.2, per 7-point increase in social support score). CONCLUSIONS: We were not able to identify objective measures that characterized self-reported failure to recover from acute infectious mononucleosis. The baseline factors associated with self-reported failure to recover at 2 months differed from those associated with failure to recover at 6 months. Future studies should assess the generalizability of these findings and determine whether interventions can hasten recovery.

Serological responses to cytomegalovirus during renal transplant rejection.

We studied the role played by CMV in kidney transplant rejection by recording serological responses to CMV replication in cadaver graft recipients and recording clinical graft rejection by monitoring acute changes in renal function and the appearance of antidonor lymphocyte antibody (anti-Dab). If CMV plays a significant role in rejection, clinical rejection should correlate with CMV activity; if CMV does not play a role, clinical rejection would be likely to correlate with anti-Dab but not necessarily with CMV activity. We selected retrospectively 18 rejectors and 18 nonrejectors by clinical criteria and assayed for anti-Dab (by fluorescence-activated flow cytometry) and CMV antibody (by complement fixation and Western blot for IgG and IgM) over a 3-6-month period after transplantation. Primary CMV infection occurred in 8 of 12 (67%) CMV seronegative graft recipients and reactivation or reinfection occurred in 16 of 24 (67%); 12 of 14 (86%) rejectors developed anti-Dab compared with 2 of 18 (11%) nonrejectors (P less than 0.00001). Active CMV infection occurred in 11 of 18 (61.1%) rejectors and 13 of 18 (72.2%) nonrejectors (P = 0.36), and in 8 of 15 (53.3%) of those who developed anti-Dab and 12 of 17 (70.6%) of those who did not (P = 0.26). The results show no evidence to link CMV activity with kidney graft rejection.

A novel method to assay herpes simplex virus neutralizing antibodies using BHKICP6LacZ-5 (ELVIS) cells.

A novel method for determining neutralizing serum antibody titers to herpes simplex virus type 2 (HSV-2) was developed based on reduction of infectivity in BHKICP6LacZ-5 (ELVIS) cells; baby hamster kidney (BHK) cells that have been genetically engineered to contain the Escherichia coli LacZ gene under the control of an inducible herpes simplex virus type 1 (HSV-1) promoter. The test has a semiautomated, colorimetric readout resulting in rapid, objective readings of infectivity reduction. Extent of neutralization is calculated against a calibration curve of virus infectivity generated in each run. HSV-2 neutralizing activity can be detected with serum dilutions in excess of 1:5120.