GC Cultures Research Articles

1534. Gonorrhea Testing and Ceftriaxone Resistance in the Department of Veterans Affairs (VA), 2010-2019

BackgroundGonorrhea (GC) infections caused by Neisseria gonorrhoeae are an ongoing public health issue and have shown resistance to nearly all antibiotic classes, including cephalosporins which are a CDC recommended treatment. We investigated national GC testing and Ceftriaxone resistance in VA.MethodsGC cases identified from VA data sources from 1/1/2010-12/15/2019 by either molecular laboratory testing or GC culture with sensitivity testing. Patients were reviewed for positive results, whether culture testing was performed after 2 positive molecular GC results in < 90 days, and sensitivity patterns.Results10,642 of 644,968 (2%) GC molecular results were positive with annual number of positive cases (1,365 to 3,225), number of tests performed (97,636 to 164,085) and percent positive for GC (1.4% to 2%) increasing over the time period studied. 2,358/10,642 (22%) of positive molecular test results had repeat testing < 3mo with 351 (15%) positive on repeat testing which is concerning for possible resistance. 2,624 GC cultures were performed with 2,179 (83%) positive. 1,480/2,179 (68%) positive GC culture tests had some sensitivity testing performed. Culture testing remained stable with 287 in 2010 to 289 in 2019 with percent positive ranging from 78-89%. Of the 351 patients with repeat positive GC molecular testing done < 3mo from their positive molecular test, only 18 (5%) had GC culture testing performed proximal to the second positive test. Among all cultured isolates, resistance to Ceftriaxone was noted in 1 sample in 2017 in Missouri (also intermediate resistance to Tetracycline) and 1 in 2019 in New Jersey (also resistant to Penicillin, Tetracycline, and Ciprofloxacin).ConclusionIn VA, GC infections have increased from 2010 to 2019 and GC culture testing has remained stable despite increasing molecular testing. Only 2 samples were identified with Ceftriaxone resistance. However, the low percentage of GC culture testing after persistent positive molecular testing in VA could mask treatment failures and possible resistance. Culture testing with sensitivity testing should be increased in VA in cases where patients may have resistance to initial treatments (i.e., – repeat positive testing less than 3 months after a positive test).Disclosures All Authors: No reported disclosures

Myostatin is expressed in bovine ovarian follicles and modulates granulosal and thecal steroidogenesis

Myostatin plays a negative role in skeletal muscle growth regulation but its potential role in the ovary has received little attention. Here, we first examined relative expression of myostatin (MSTN), myostatin receptors (ACVR1B, ACVR2B and TGFBR1) and binding protein, follistatin (FST), in granulosa (GC) and theca (TC) cells of developing bovine follicles. Secondly, using primary GC and TC cultures, we investigated whether myostatin affects steroidogenesis and cell number. Thirdly, effects of gonadotropins and other intrafollicular factors on MSTN expression in GC and TC were examined. MSTN, ACVR1B, TGFBR1, ACVR2B and FST mRNA was detected in both GC and TC at all follicle stages. Immunohistochemistry confirmed follicular expression of myostatin protein. Interestingly, MSTN mRNA expression was lowest in GC of large oestrogen-active follicles whilst GC FST expression was maximal at this stage. In GC, myostatin increased basal CYP19A1 expression and oestradiol secretion whilst decreasing basal and FSH-induced HSD3B1 expression and progesterone secretion and increasing cell number. Myostatin also reduced IGF-induced progesterone secretion. FSH and dihydrotestosterone had no effect on granulosal MSTN expression whilst insulin-like growth factor and tumour necrosis factor-alpha suppressed MSTN level. In TC, myostatin suppressed basal and LH-stimulated androgen secretion in a follistatin-reversible manner and increased cell number, without affecting progesterone secretion. LH reduced thecal MSTN expression whilst BMP6 had no effect. Collectively, results indicate that, in addition to being potentially responsive to muscle-derived myostatin from the circulation, myostatin may have an intraovarian autocrine/paracrine role to modulate thecal and granulosal steroidogenesis and cell proliferation/survival.

Direct Inoculation and Human Transferrin Supplementation Increases the Survivability of Neisseria gonorrhoeae in the Upper Reproductive Tract of Estrogen‐Treated Female BALB/c Mice

ObjectiveThis study intended to investigate methods of increasing Neisseria gonorrhoeae (Gonococcus; Gc) survivability in the upper reproductive tract (URT) of female mice with the aim of establishing a previously undescribed murine model of Gc URT infections ‐ also known as pelvic inflammatory disease (PID) ‐ in women. Such a model would allow for further investigations into the natural history and pathogenesis of ascending Gc infections.MethodsTwo main experimental arms were used in this study: 1. Gc‐infected mice treated with estrogen alone vs estrogen and human transferrin (hTF) and 2. Gc‐infected mice treated with long‐acting progesterone (Medroxyprogesterone) alone vs progesterone and hTF. Appropriate uninfected, negative controls were used in all experiments. All mice were pre‐treated with antibiotics to suppress commensal flora and then given either estrogen or progesterone prior to inoculation, according to their study groups. Diestrous or anestrous stage BALB/c mice were then inoculated with Neisseria gonorrhoeae (strain FA1090) using a novel, trans‐cervical catheterization technique and the appropriate groups were treated with hTF up to 5 days post‐inoculation. Daily vaginal swabs were taken for polymorphonuclear white blood cell (PMN WBC) enumeration and quantitative Gc cultures up to day 5. In addition, part of each study group was sacrificed at days 3 and 5 post‐inoculation for Gc quantitative cultures of endometrial scrapings. Lastly, whole URTs from a portion of each group were fixed and stained for WBCs and Gc on days 3 and 5 post‐inoculation.ResultsDirect inoculation via trans‐cervical catheter proved successful in introducing and establishing Gc infection in the URT of all groups of estrogen‐treated mice for at least five days as determined by culture of endometrial scrapings and Gc‐specific IHC staining at day 5 post‐inoculation. Mice treated with progesterone showed no infection at any time point by either culture of daily vaginal swabs or of endometrial scrapings. Higher numbers of Gc were recovered from vaginal swabs (p&lt;.05) and endometrial tissue on day 3 and 5 post‐inoculation from mice treated with estrogen and hTF compared to those treated with estrogen alone (comparisons made by geometric mean of Gc colonies recovered on each day).Conclusions and Future DirectionsThis project established a novel method of direct, trans‐cervical inoculation of Gc into the female mouse URT thereby creating a usable model of Gc URT infection (PID) in women for further study. Future projects will utilize this model to better understand the natural history of Gc infections and to investigate potential methods for treating these infections. Currently, this laboratory is investigating host immune responses by measuring serum antibodies and vaginal chemokines/cytokines using this model. In addition, GFP‐labeled FA1090 strain Gc have been created and will be used for better visualization of Gc in histologic samples, which in turn could be used to track Gc movement through the mouse URT, as well as cellular responses to infection using sequentially fixed tissue samples.Support or Funding InformationThe Uniformed Services University of the Health Sciences and NIH/NIAID grant # R01 AI42053.

P167 Managing rectal GC : room for improvement

Background/introduction Most (~60%) rectal gonorrhoea (GC) occur in MSM with 25% symptomatic at presentation. Those with rectal GC are at increased risk of other STIs. BASHH provide guidelines on GC management and targets to be achieved in testing, treatment and partner notification. Aim(s)/objectives To compare our clinic’s performance in managing rectal GC compared to the national recommendations. Methods Retrospective case-note review of confirmed cases of rectal GC on NAAT between 1 st November 2011 and 31 st March 2015. Data were obtained from clinic notes, the clinic database and laboratory results. Audit standards were based on BASHH guidelines in managing GC. Results 184 cases from 156 men: 61% White, 12% Black, median age 31 (IQR 26,37) years, 71% MSM 29% bisexual, 58% symptomatic. Triple site testing was done in 91%. Rectal GC cultures were taken in 55%. Adequate treatment was given to 94%. Quinolone resistance occurred in 31%. Partner notification was done in 43%. 14% had other STIs (syphilis, LGV, chlamydia and HSV). There were 2 new diagnoses of HIV at the time of GC diagnosis, and 2 further cases at 3 months follow-up. Discussion/conclusion Management of rectal GC did not reach the BASHH targets on any recommendation, suggesting that improvements in managing rectal GC are needed within our clinic. Re-testing and re-attendance were poor. Staff has received further training and a re-audit in 2017 will assess improvement. We have established a robust call/recall system to enable early diagnosis of HIV which was significant in our cohort of men with rectal GC.

P9 Low false positive rate of gonorrhoea cases referred to gum from a low prevalence chlamydia screening population

<h3>Background/introduction</h3> Recent national guidance for the detection of gonorrhoea (GC) has raised concerns that the majority of initial positive GC test results are likely to be false positives when a low prevalence population is screened using a nucleic acid amplification test (NAAT). Our local chlamydia screening programme uses a dual NAAT for chlamydia and GC and has a low reactive GC rate &lt;1%. All GC reactive cases are referred to GUM for further investigation. <h3>Aim(s)/objectives</h3> To determine whether GC reactive cases referred from our local chlamydia screening programme were true GC infections or likely to be false positive cases. <h3>Methods</h3> A retrospective case notes review of 13 consecutive GC reactive cases seen at our clinic referred from the local chlamydia screening programme. <h3>Results</h3> 10/13 were women. 10/13 cases were confirmed as true GC infections by positive genital GC cultures. In 1 case genital culture was negative but screening and supplementary NAAT with a different target confirmed a true GC infection. In 1 case genital cultures were negative however pharyngeal culture was positive indicating a true GC infection. In 1 case GC cultures were all negative but NAAT tests were reactive from genital and pharyngeal sites and equivocal from the rectum. <h3>Discussion/conclusion</h3> Only 1/13 GC reactive cases seen in our GU referred from a low prevalence screening population might have been a false positive. Contrary to recent publications, in our area, using a dual NAAT is unlikely to lead to high numbers of false positive GC results.

P116 An audit of gonorrhoea management in a UK sexual health clinic

<h3>Background/introduction</h3> Gonorrhoea (GC) is a prevalent sexually-transmitted infection in the UK. The British Society for Sexual Health and HIV (BASHH) published guidelines in 2011 for the management of GC. <h3>Aim(s)/objectives</h3> To audit the management of all patients who tested positive for GC at our centre between 1 May 2013 and 1 May 2014 against BASHH guidelines. <h3>Methods</h3> A proforma was developed and a retrospective notes review performed for all patients who tested positive for GC from 1 May 2013 to 1 May 2014. <h3>Results</h3> 115 patients tested positive for GC during the audit period. The prevalence of GC in this cohort was 0.9%. 46% of patients had symptom (s), 54% were truly asymptomatic, 18% of patients presented as contacts. Microscopy was performed on 80% of symptomatic patients and intracellular gram-negative diplococci were seen in 66% of these. 97% of patients were diagnosed by PCR testing and one by culture. 2 patients were diagnosed elsewhere with negative testing at our clinic. 83% had GC cultures prior to treatment. The sensitivity of culture compared to PCR testing was 59%. 93% of patients were treated as per BASHH guidelines. 65% were documented to have received written information about their diagnosis. All patients were offered test of cure, of which 61% attended within 2 weeks. 92% of patients saw health advisers for partner notification. <h3>Discussion/conclusion</h3> Management of GC was largely in line with BASHH guidance. However, this study highlighted a need to increase written information offer and to encourage attendance for test of cure.

P3.121 Are We Seeing a True Rise in Neisseria Gonorrhoeae and Chlamydia Trachomatis in England in Men Who Have Sex with Men?

BackgroundThe Health Protection Agency (HPA) in England have reported the number of sexually transmitted infections (STIs) diagnosed in men who have sex with men (MSM) continues to rise. Of note,...

The effect on Neisseria gonorrhoeae screening rates in an integrated clinic following the introduction of dual nucleic acid amplification tests

Sirs: We read with interest the findings of Killick et al. 1 and agree that Neisseria gonorrhoeae (GC) nucleic acid amplification testing (NAAT) can provide a valuable addition to culture for identifying GC in low prevalence settings. Following the introduction of GC NAAT into our integrated family planning (FP) and genitourinary (GU) medicine service, we evaluated whether the use of NAAT increased the rates of GC screening and of GC detection. 2 Prior to the introduction of GC NAAT, prevalence of diagnosed GC within the clinic as assessed by culture alone was 0.87%. We sought to investigate concordance between NAAT and culture when both were carried out, within this low prevalence population. Dual-testing for GC and chlamydia (CT) using Aptima Combo 2 (AC2; Gen-Probe Inc., San Diego, CA, USA) was per- formed using self-taken vaginal swabs for asymptomatic women and urine for asymptomatic men. Those patients with clinical symptoms or signs, or contacts of GC underwent GC culture in addition to NAAT. When using AC2, all positive GC results were confirmed according to the manufacturer's instructions using the single analyte NAAT targeting a different RNA sequence: Aptima GC (Gen-Probe). All patients attending the clinic and undergoing GC screen- ing during a four-month period prior to, and following GC NAAT introduction were identified using laboratory databases; data were collected retrospectively. Follow-up attendances within an episode were excluded from analysis. Individuals diagnosed with GC were identified and case-notes were reviewed. We found that in the four months prior to the introduction of NAAT, 2307 symptomatic and asymptomatic patients were screened using GC culture alone, of which 20 (0.87%) were positive. Following the introduction of GC NAAT during a similar four-month period, a significant increase in screening was observed, with 3444 symptomatic and asymptomatic patients being screened with AC2. Of these, 43 (1.25%) were confirmed positive for GC. GC culture was also performed in 217 of the symptomatic individuals; of these 18 (8.3%) were culture posi- tive for GC. GC culture and NAAT showed concordance of positivity in 20/43 (46.5%) of cases. Eleven patients were NAAT positive, culture negative; 10/11 of these individuals had symptoms, signs or were high risk for GC. One of 11 cul- tured Neisseria meningitidis although subsequent GC NAAT testing using a different platform was also positive at the Sexually Transmitted Bacteria Reference Laboratory in Colindale, making it unlikely this was a false-positive. Twelve patients who were AC2 positive had no culture performed. Similarly to the findings by Killick et al., all patients with posi- tive GC cultures were also GC NAAT positive. In conclusion, following the introduction of GC NAAT we observed increased rates of screening and a doubling in the number of GC diagnoses during a similar time period. While it is possible there was an actual rise in the local GC incidence, the increase in diagnoses might also be attributable to a more sensitive test, acceptability of self-screening and a less time- consuming approach for the clinicians. The use of GC NAAT identified more infections than if culture alone had been used. Despite concern over the positive predictive value of GC NAAT in low prevalence settings, of those patients with posi- tive NAAT/negative cultures all had documented risk factors or symptoms of GC. We believe both these studies provide reassuring evidence for the use of GC NAAT in this setting, provided a confirmatory strategy is in place.

O2 Who should have gonorrhoea cultures in addition to gonorrhoea and chlamydia nucleic acid amplification tests? Cost effectiveness study

BackgroundNucleic acid amplification tests (NAATs) can now detect chlamydia (CT) and gonorrhoea (GC) but cultures are needed in GC NAAT positive people prior to treatment. This necessitates re-evaluation, examination and...

P161 Routine three-site NAAT testing in MSM increases pharyngeal and rectal diagnoses of chlamydia and gonorrhoea

BackgroundBASHH and Health Protection Agency guidelines acknowledge the increased sensitivity of Gonorrhoea (GC) NAAT testing compared to culture at extragenital sites. Chlamydia (CT) NAAT testing is also the test of...

Neisseria gonorrhoeae and Chlamydia trachomatis among human immunodeficiency virus-infected women.

Neisseria gonorrhoeae and Chlamydia trachomatis among human immunodeficiency virus-infected women.

Incidence of HIV and sexually transmitted diseases (STD) in a cohort of HIV-negative men who have sex with men (MSM).

To determine the prevalence of sexually transmitted diseases (STD) and incidence of and risk factors for STD, including HIV-1, among a cohort of HIV-negative men who have sex with men (MSM). Seattle, Washington, United States. Prospective cohort of 578 HIV-negative MSM in which risk factors for acquiring a STD over 12 months follow-up were evaluated using a cumulative incidence analysis. Baseline tests obtained were: herpes simplex virus types 1 and 2 (HSV-1 and HSV-2) Western blots, hepatitis B, and syphilis serologies; anorectal and pharyngeal Neisseria gonorrhoeae (GC) cultures; first-catch urine for leukocyte esterase (LE) and Chlamydia trachomatis (CT) ligase chain reaction (LCR). Men with a positive urine LE had urethral GC cultures obtained. The following outcomes were measured over 12 months follow-up: incident symptomatic bacterial STD (urethritis, proctitis, epididymitis), HSV-1 and HSV-2 seroconversion, and HIV-1 seroconversion. The 31 incident cases of STD (men with bacterial STD) were compared with those 489 men without symptomatic bacterial STD or seroconversion to HSV-1, HSV-2 or HIV-1 infection. Bacterial STD were found in nine participants at enrollment; there were two cases of nonchlamydial urethritis, two cases of nonchlamydial epididymitis, and five cases of asymptomatic GC infection. At enrollment, HSV-2 antibodies were detected in 149 (26.0%) of 572 men and prior hepatitis B infection in 145 (34.8%) of 417 unvaccinated men. During the 1-year of follow-up, 31 men (5.7/100 person-years) had 34 episodes of a symptomatic bacterial STD syndrome (urethritis, epididymitis or proctitis). Urethritis was the most common STD syndrome, detected in 29 men, of whom 10 had GC and 19 had nongonococcal urethritis. In the 1-year of follow-up, five participants seroconverted to HIV-1 (1.3/100 person-years), four to HSV-2 (1.0/100 person-years), and seven to HSV-1 (4.3/100 person-years). Unprotected insertive anal sex [odds ratio (OR) 2.6; 95% confidence interval (CI) 1.2-5.6]; and nitrite inhalant ('poppers') use (OR, 2.3; 95% CI, 1.0-5.0) were independently associated with incident STD. STD and HIV infection continue to be acquired even in a city with an overall low bacterial STD prevalence and among educated MSM receiving regular HIV screening and risk-reduction. Urethritis was the most common STD detected, and public health messages aimed at MSM need to emphasize safe insertive as well as receptive sexual practices.

Hybrid structural analogues of 1,25-(OH)2D3 regulate chondrocyte proliferation and proteoglycan production as well as protein kinase C through a nongenomic pathway

1,25-(OH)2D3 and 24,25-(OH)2D3 mediate their effects on chondrocytes through the classic vitamin D receptor (VDR) as well as through rapid membrane-mediated mechanisms which result in both nongenomic and genomic effects. In intact cells, it is difficult to distinguish between genomic responses via the VDR and genomic and nongenomic responses via membrane-mediated pathways. In this study, we used two hybrid analogues of 1,25-(OH)2D3 which have been modified on the A-ring and C,D-ring side chain (1 alpha-(hydroxymethyl)-3 beta-hydroxy-20-epi-22-oxa-26,27-dihomo vitamin D3 (analogue MCW-YA = 3a) and 1 beta-(hydroxymethyl)-3 alpha-hydroxy-20-epi-22-oxa-26,27-dihomo vitamin D3 (analogue MCW-YB = 3b) to examine the role of the VDR in response of rat costochondral resting zone (RC) and growth zone (GC) chondrocytes to 1,25-(OH)2D3 and 24,25-(OH)2D3. These hybrid analogues are only 0.1% as effective in binding to the VDR from calf thymus as 1,25-(OH)2D3. Chondrocyte proliferation ([3H]-thymidine incorporation), proteoglycan production ([35S]-sulfate incorporation), and activity of protein kinase C (PKC) were measured after treatment with 1,25-(OH)2D3, 24,25-(OH)2D3, or the analogues. Both analogues inhibited proliferation of both cell types, as did 1,25-(OH)2D3 and 24,25-(OH)2D3. Analogue 3a had no effect on proteoglycan production by GCs but increased that by RCs. Analogue 3b increased proteoglycan production in both GC and RC cultures. Both analogues stimulated PKC in GC cells; however, neither 3a nor 3b had an effect on PKC activity in RC cells. 1,25-(OH)2D3 and 3a decreased PKC in matrix vesicles from GC cultures, whereas plasma membrane PKC activity was increased, with 1,25-(OH)2D3 having a greater effect. 24,25-(OH)2D3 caused a significant decrease in PKC activity in matrix vesicles from RC cultures; 24,25-(OH)2D3, 3a, and 3b increased PKC activity in the plasma membrane fraction, however. Thus, with little or no binding to calf thymus VDR, 3a and 3b can affect cell proliferation, proteoglycan production, and PKC activity. The direct membrane effect is analogue-specific and cell maturation-dependent. By studying analogues with greatly reduced affinity for the VDR, we have provided further evidence for the existence of a membrane receptor(s) involved in mediating nongenomic effects of vitamin D metabolites.

Rapid and long-term effects of PTH(1-34) on growth plate chondrocytes are mediated through two different pathways in a cell-maturation-dependent manner

The aims of this study were to clarify the role of cell maturation stage on chondrocyte response to parathyroid hormone (PTH) by examining the effect of PTH(1-34) on alkaline-phosphatase-specific activity (ALPase) of chondrocyte cultures at two distinct stages of maturation, and to determine the signaling pathways used by the cells to mediate this effect. Confluent, fourth passage rat costochondral resting zone (RC) and growth zone (GC) chondrocytes were used. ALPase was measured in the cell layer, as well as in matrix vesicles (MV) and plasma membranes (PM), after the addition of 10 −7 to 10 −11 mol/L bovine PTH(1-34), the active peptide, or bovine PTH(3-34), the inactive peptide, to the cultures. PTH(1-34) increased ALPase in the GC cultures at two separate times: between 5 and 180 min, with maximal stimulation at 10 min, and 36 to 48 h. In contrast, PTH(3-34) had no effect. At 10 min and 48 h, PTH(1-34) produced a dose-dependent increase in ALPase of both MV and PM isolated from GC cultures. Addition of forskolin and IBMX to increase cAMP increased ALPase in GC cultures to a level similar to that seen after addition of PTH(1-34). In contrast, the addition of PTH(1-34) to RC cells only increased ALPase between 5 and 60 min, with peak activity at 10 min. As with GC, PTH increased ALPase in both MV and PM. Moreover, the addition of PTH(3-34) or forskolin and IBMX had no effect on ALPase in RC. PTH(1-34) had no effect on GC protein kinase C (PKC) activity; however, the addition of PTH(1-34) to RC caused a dose-dependent increase in PKC activity. H8, an inhibitor of PKA, had no effect on PTH-stimulated ALPase in RC cells, but inhibited the PTH-dependent response in GC cells. In contrast, chelerythrine, an inhibitor of PKC activity, inhibited PTH-stimulated ALPase in RC cells, but had no effect on PTH-stimulated ALPase in GC cells. This study shows that the effect of PTH(1-34) on RC and GC cells is maturation dependent in terms of time course and mechanism. Whereas both cell types exhibit a rapid response to PTH, only GC cells show a long-term response. In GC, the effects of PTH are associated with changes in cAMP and may also involve at least one other pathway, whereas, in RC, the PTH effects appear to be associated with changes in PKC. (Bone 21:249–259; 1997)

Vitamin D regulation of metalloproteinase activity in matrix vesicles.

Matrix vesicles (MVs) are enriched in matrix metalloproteinases (MMPs) capable of degrading proteoglycans. The aim of the present study was to identify which MMPs are present in MVs and determine whether these MMPs are regulated by 1,25-(OH)2D3 [1,25] and 24,25-(OH)2D3 [24,25]. To do this, growth zone (GC) and resting zone (RC) chondrocytes were isolated from rate costochondral cartilage and placed into culture. At confluence, GCs were treated with 1,25 and RCs with 24,25 for 24 hours. MVs, plasma membranes (PMs), and conditioned media were then collected from the cultures. RTPCR demonstrated the presence of mRNA for stromelysin-1 and 72 kDa gelatinase in both RCs and GCs, Casein zymography revealed activity at M(r) 48 and 28 kDa in MV, but not PM or conditioned media; Western analysis confirmed that this activity was stromelysin-1. Gelatinolytic activity, at low levels, was also found in MVs, but not PMs or conditioned media. When enzyme activity was measured using a proteoglycan bead assay, it was found that both GCs and RCs produced MVs and PMs containing neutral metalloproteinase. Both cells also produced MVs and PMs containing plasminogen activator. The addition of 1,25 to GCs caused a significant 4- to 5-fold increase in metalloproteinase activity in MVs, but not PMs. In contrast, MVs from cultures of RCs treated with 24,25 contained decreased metalloproteinase activity; enzyme activity in PMs was unaffected by 24,25. Plasminogen activator in MVs from RC was increased by treatment with 24,25, while MV enzyme activity was decreased after treatment of GC cultures with 1,25. This study shows that both RCs and GCs produce stromelysin-1 and 72 kDa gelatinase and that these enzymes are preferentially localized in MVs. Further, MMP and plasminogen activator activities in MVs and PMs are regulated by vitamin D metabolites.

Effectiveness of Repeated Gonorrhea Cultures in the Third Trimester

Objective: With the high cost of health care today, the universal prophylactic measures recommended, and the availability of effective treatment should infection occur, the practice of routinely repeating the endocervical gonorrhea (GC) culture in the third trimester of pregnancy may be unwarranted. Methods: To test this hypothesis, we reviewed charts from patients who had received routine prenatal care during a 2-year period at the Lafayette and Opelousas parish health units. Those charts, which had documented results of both the initial and repeat GC cultures, were then used for retrospective review. The results ofthe initial GC culture were compared with that taken in the third trimester. Other data recorded included age, gravidity, race, and history of gonorrhea, syphilis, or multiple sexual partners. Results: Two hundred fifty charts were available for extraction; 130 of these had documentation of both GC cultures. Of the 130 cultures obtained during the initial prenatal visit, only 6 (4.6%) were positive. Of the repeat cultures taken during the third trimester, none were positive. Thirteen patients (10.0%) had a documented history of GC infection; none of them had positive cultures during the study period. Conclusions: Screening for GC during pregnancy is important and appropriate. This is commonly accomplished by taking a GC culture during the initial prenatal visit. Based upon the present study, we found that repeating this culture in the third trimester, even in a relatively high-risk population, seems unnecessary, whether the initial culture is negative or not.

Sexually transmitted diseases and human immunodeficiency virus screening in a population of sexually abused girls

Study Objective: We wanted to determine the prevalence of sexually transmitted diseases (STD) and human immunodeficiency virus (HIV) seropositivity in pediatric and adolescent female victims of sexual abuse. Participants and Setting: Over a three and one-half year period, 288 girls (ages 3 months to 18 years) were referred to the Pediatric Sexual Abuse Clinic of a major medical center in Hartford, Connecticut. Design: Examination of these patients included genital and anal colposcopy, and triple site cultures for Neisseria gonorrhoeae (GC) and Chlamydia trachomatis (CT). Anti-HIV enzyme-linked immunosorbent assay and syphilis (RPR) testing were offered in all cases and done each time informed consent was obtained. Main Outcome Measurements and Results: Medical findings were “consistent with sexual abuse” in 90 cases. The medical findings in the remaining cases were called “inconclusive of sexual abuse.” There were 5 positive GC cultures, 19 positive CT cultures, 1 positive genital herpes simplex culture, 11 cases of condyloma acuminatum and 3 cases of Trichomonas vaginalis. Spermatozoa were recovered from three patients and three pregnancies were diagnosed. There were 126 anti-HIV tests done. Of these patients tested, 80 alleged sexual abuse of at least 6 months duration. All results were nonreactive. Of 204 patients tested for syphilis, none was found to be positive. Conclusion: HIV infection and syphilis were not identified in our population of sexually abused pediatric and adolescent female patients for whom informed consent was granted.

Clinical findings and legal resolution in sexual assault

Female sexual assault victims who received medical evaluation at Detroit Receiving Hospital from July to December 1980 and were seen by the Rape Counseling Center (RCC) were evaluated. Data were abstracted from the emergency department record, RCC file, and gynecology clinic; 372 patients were evaluated. One hundred twenty-seven patients (34%) were given DES, including 17 women who had been raped at least once before. Data regarding DES dosage were available only on 34 patients, with 27 stating they took the entire course; 5, an incomplete course; and 2 refusing to answer. DES is still approved for post-coital contraception in rape. However, if there is one major concern about its administration, it is that close follow-up be provided to anticipate or eliminate complications, especially those associated with inadvertent pregnancy. The extremely poor medical follow-up, the surprising lack of an "appropriate chief complaint" when follow-up does occur, the large number of sexual assault victim repeaters in an inner city population, and the socioeconomic and demographic characteristics of an urban, inner-city sexual assault victim population which make it impossible to determine the effectiveness of DES are strong arguments for the severe restriction of use of DES in the urban, inner city setting. There is no need for pretreatment GC cultures or VDRL assays. Patients should be treated in conformity with current CDC recommendations. Where a rape counseling center is available, it is suggested that the unit assume close and direct responsibility of directing patients for medical follow-up, regardless of emergency department treatment.

Retrograde labeling, enrichment, and characterization of retinal ganglion cells from the neonatal rat

We have developed a method for labeling retinal ganglion cells in neonatal rats by retrograde transport of the fluorescent dye, True Blue (TB), injected into the optic chiasm. Following proteolytic dissociation of labeled retinas into single cells, the labeled cells could be enriched 50- to 100-fold by centrifugation in a 5%/10% metrizamide gradient. When plated in Ham's F-10 medium in the presence of fetal calf serum and chick optic tectum-conditioned medium, the labeled cells could be maintained in vitro up to 48 hr. In these cultures, the ganglion cells (GCS) constituted 50 to 70% of the total cell population. When GC-rich fractions or GC cultures were stained with a monoclonal antibody to Thy-1 antigen, greater than 90% of the TB-labeled cells were reactive. In order to localize voltage-sensitive sodium channels, GC-rich cultures were reacted with 125I-scorpion toxin. Analysis of the autoradiograms showed that the density of silver grains was about 10-fold higher on TB-labeled cells than on nonfluorescent cells, or in controls which contained excess of unlabeled toxin. When GC cultures were incubated with micromolar concentrations of putative GC transmitters, aspartate and glutamate, the amino acids were accumulated by 15 to 20% of labeled cells. Several lectin receptors were also localized on TB-labeled cells in situ. Whereas the lectins wheat germ agglutinin, concanavalin A, peanut agglutinin, Dolichos biflorus agglutinin, and Limulus polyphemus agglutinin bound to TB-labeled cells, others such as Ricinus communis agglutinin I, Ulex, and Lotus lectins showed no binding. The lectin binding was specific since preincubation with the appropriate hapten sugar blocked lectin binding.