Gretchen M. Lentz

Initial Events in Establishing Vaginal Entry and Infection by Human Immunodeficiency Virus Type-1

Summary Understanding the initial events in the establishment of vaginal human immunodeficiency virus type-1 (HIV-1) entry and infection has been hampered by the lack of appropriate experimental models. Here, we show in an ex vivo human organ culture system that upon contact in situ, HIV-1 rapidly penetrated both intraepithelial vaginal Langerhans and CD4+ T cells. HIV-1 entered CD4+ T cells almost exclusively by CD4 and CCR5 receptor-mediated direct fusion, without requiring passage from Langerhans cells, and overt productive infection ensued. By contrast, HIV-1 entered CD1a+ Langerhans cells primarily by endocytosis, by means of multiple receptors, and virions persisted intact within the cytoplasm for several days. Our findings shed light on the very earliest steps of mucosal HIV infection in vivo and may guide the design of effective strategies to block local transmission and prevent HIV-1 spread.

Lifetime risk of surgical management for pelvic organ prolapse or urinary incontinence.

The objective of our study was to estimate the age-specific incidence and lifetime risk of surgically managed pelvic organ prolapse (POP) and urinary incontinence (UI). Women aged 20 and older who underwent primary surgical management of POP or UI in 1993 were identified from the database of a health maintenance organization using ICD-9 codes and confirmed through chart abstraction. From a population of 147,719 women, 135 were identified who underwent prolapse surgery only, 82 incontinence only, and 34 surgery for both conditions. From the age-specific incidence, we estimated the lifetime risk of undergoing an operation by age 80 to be 11.8%. Our findings agree with a previous estimate that ∼11% of women will undergo surgery for POP or UI by age 80. POP and UI appear to be common problems, undoubtedly affecting an even larger proportion of the women than suggested by this high cumulative incidence of surgery.

Development of a bench station objective structured assessment of technical skills.

OBJECTIVE We have previously shown that objective structured assessment of technical skills performed in an animal model was an innovative, reliable, and valid method of assessing surgical skills. Our goal was to develop a less costly bench station objective structured assessment of technical skills and to evaluate the feasibility, reliability, and validity of this exam. METHODS A seven‐station examination was administered to 24 residents. The tests included laparoscopic procedures (salpingostomy, intracorporeal knot tying, closure of port sites) and open abdominal procedures (subcuticular closure, bladder neck suspension, repair of enterotomy, abdominal wall closure). All tasks were performed using life‐like surgical models. Residents were timed and assessed at each station using three methods of scoring: a task‐specific checklist, a global rating scale, and a pass/fail grade. RESULTS Assessment of construct validity, the ability of the test to discriminate among residency levels, found significant differences on the checklist, global rating scale, time for procedures, and pass/fail grade by level of training. Reliability indices calculated with Cronbachs ∞ were 0.77 for the checklists and 0.94 for the global rating scale. Overall interrater reliability indices were 0.91 for the global rating scale and 0.92 for the checklists. Total cost for replaceable parts and facilities was

Vaginal Langerhans Cells Nonproductively Transporting HIV-1 Mediate Infection of T Cells

1900. CONCLUSION The less costly and more portable bench station objective structured assessment of technical skills can reliably and validly assess the surgical skills of gynecology residents. This type of examination can be a useful tool to identify residents who need additional surgical instruction, provide remediation, and may become a mechanism to certify surgical skill competence.

Assessment of cytochrome P450 3A4 activity during the menstrual cycle using alfentanil as a noninvasive probe

ABSTRACT Although implied by other models, proof that Langerhans cells (LCs) in the human vagina participate in dissemination of infectious human immunodeficiency virus type 1 (HIV-1) has been lacking. Here, we show that LCs migrate from HIV-1-exposed vaginal epithelia and pass infectious virus to CD4+ T cells without being productively infected themselves, and we point to a pathway that might enable HIV-1 to avoid degradation in vaginal LCs. Transport by migratory LCs to local lymphatics in a nonproductive but infectious form may aid HIV-1 in evasion of topical microbicides that target its intracellular productive life cycle.

Formal teaching of surgical skills in an obstetric-gynecologic residency.

Background: Gender‐dependent differences in cytochrome P450 activity, drug metabolism, drug elimination, and their clinical consequences are increasingly apparent. P450 3A4 is the most abundant P450 isoform in the human liver and is responsible for metabolizing a vast and diverse assortment of therapeutic agents, including opioids, benzodiazepines, and local anesthetics. P450 3A4 activity is higher in women, influenced by steroid hormone levels, and is speculated to vary during the menstrual cycle. This investigation tested the hypothesis that P450 3A4 activity varies during the menstrual cycle. Alfentanil clearance was used as a metabolic probe for P450 3A4 activity. Methods: Alfentanil (20 micro grams/kg bolus) was administered to nine nonsmoking, nonpregnant female volunteers (age, 26 +/‐ 5 yr) with normal menstrual cycles on three separate occasions during the same menstrual cycle: days 2 (menstrual phase), 13 (estrogen peak), and 21 (progesterone peak). Venous plasma alfentanil concentrations were determined by gas chromatography‐mass spectrometry. Alfentanil clearance was determined by noncompartmental methods and by a three‐compartment model with both pooled population and two‐stage analysis. Results: There was no significant difference in any measure of alfentanil clearance. Noncompartmental clearances (mean +/‐ SD) were 3.62 +/‐ 0.76, 3.81 +/‐ 0.96, and 3.60 +/‐ 0.84 ml/kg/ min, respectively, on days 2, 13, and 21 of the menstrual cycle. Conclusions: Alfentanil clearances were not different on menstrual cycle days 2, 13, and 21, strongly suggesting no change in P450 3A4 activity. Menstrual cycle differences in alfentanil clearances do not contribute to interindividual variability in alfentanil disposition in women. If other P450 3A4 substrates are comparable, then menstrual cycle variability in their metabolism may not be a consideration in dosing or in the design of pharmacokinetic investigations.

Most DC-SIGNR transcripts at mucosal HIV transmission sites are alternatively spliced isoforms

OBJECTIVE To describe a formal teaching program of basic surgical skills in an obstetric-gynecologic residency program and evaluate its effectiveness. METHODS A surgical skills program was developed for all residents. Using bench and animal laboratory sessions, residents were given instruction and performed both laparoscopic and open abdominal procedures. All were given a pretest and were tested again 6 months later. Residents also evaluated their experiences. RESULTS To date, the formal teaching sessions have been given to 24 residents, all of whom believed their confidence and technical skills improved as a result of the sessions. On a scale of 1 to 5, the median rating of the bench laboratory experience was 5 (range 4-5), and the pig laboratory was 5 (range 4-5). All residents believed the surgery sessions should be continued and the number of sessions increased. Preliminary evaluation indicated that time to suture a 10-inch incision decreased by 28%, from an average of 225 (standard deviation [SD] 51) seconds to 171 (SD 43) seconds (P < .001), and evaluation of surgical technique significantly improved at the second pretest (P = .013). Laparoscopic placement of pegs on a board in 2 minutes increased from an average of 5 (SD 2.5) to 7.3 (SD 2.6; P = .001). The cost of the bench laboratory sessions was minimal. Each pig was approximately

Optimizing Viable Leukocyte Sampling from the Female Genital Tract for Clinical Trials: An International Multi-Site Study

100, and the facility charge for each 4-hour laboratory session was

Ex Vivo Comparison of Microbicide Efficacies for Preventing HIV-1 Genomic Integration in Intraepithelial Vaginal Cells

1500. CONCLUSION When formal surgical training was given to obstetric-gynecologic residents, their surgical skills improved subjectively and objectively.

Targeting Temporomandibular Disorder Pain Treatment to Hormonal Fluctuations: A Randomized Clinical Trial

The repeat region of DC-SIGNR (CD209L) is polymorphic on the genomic level, and, in a separate study, we observed a correlation between the DC-SIGNR genotype and HIV-1 susceptibility during sexual contact. However, previous investigations using immunohistochemistry failed to detect membrane-bound DC-SIGNR on cells in the genital and rectal mucosa. We therefore explored the presence of DC-SIGNR in these compartments with a more sensitive limiting dilution RT-PCR, which also allowed for quantification of alternatively spliced mRNA isoforms. DC-SIGN (CD209) and DC-SIGNR mRNA transcript isoforms were found in all 12 vaginal and two rectal biopsies obtained from 14 healthy individuals. For DC-SIGNR, we detected significantly more isoform than full-length transcripts (mean copy numbers/μg RNA: 602 vs 26; P=0.0009). Four mucosal samples lacked full-length DC-SIGNR transcripts entirely. Cloning and sequencing of DC-SIGNR mRNA in three additional individuals revealed a diverse repertoire of DC-SIGNR isoforms, many of which encoded for proteins predicted to be soluble and secreted. Indeed, in one vaginal sample, we detected only soluble isoforms. In conjunction with our prior observation that the DC-SIGNR genotype has an effect on HIV-1 transmission in vivo, these findings emphasize that DC-SIGNR, in addition to DC-SIGN, should be considered as a cofactor in sexual HIV-1 transmission. Soluble isoforms, in particular, may modulate the efficiency of viral transmission and dissemination.