Cervical Cytology Specimen Stability in Surepath Preservative and Analytical Sensitivity for HPV Testing with the cobas and Hybrid Capture 2 Tests.

None of the commercial HPV tests are U.S. FDA-approved for testing of cervical cytology specimens in SurePath preservative. Still, ~30% of HPV testing is performed on specimens in this formalin-containing preservative. Formalin-induced DNA fragmentation and cross-linking may interfere with HPV detection. We evaluated analytical sensitivity and specimen stability of the cobas 4800 HPV (Roche) and Hybrid Capture 2 HPV (HC2, Qiagen) tests with residual cervical cytology samples in SurePath preservative available within 1 week of collection. Cobas testing was performed with and without heating samples at 120°C for 20 min diluted 1:1 in an alkaline environment (pretreatment) to revert DNA crosslinking. Stability was tested after 2 weeks of storage at ambient temperature followed by ≤10 weeks at 4°C. Analytical sensitivity and positivity rates (HC2, 18%; cobas pretreated, 46%; cobas untreated, 47%) were greater for cobas than HC2 (n = 682). After 6 weeks of storage, mean HC2 ratios were lower (mean 0.9, SD 6.3) but high variability limited statistical power to detect trends. Cobas threshold cycles (Ct's) increased in untreated (mean 2.1) but not pretreated samples (mean 0.3; n = 110; p≤0.0001). Overall, cobas had greater analytical sensitivity for samples in SurePath preservative. Although pretreatment introduced a manual sample transfer step and 30 min of incubation times, it improved stability without negatively affecting analytical sensitivity. While awaiting results of large trials to evaluate the clinical performance of cobas, the addition of the pretreatment step may improve the detection of HPV, especially after prolonged sample storage.

Optimization of PCR for quantification of simian immunodeficiency virus genomic RNA in plasma of rhesus macaques (Macaca mulatta) using armored RNA.

INTRODUCTION: Quantification of plasma viral load (PVL) is used to monitor disease progression in SIV-infected macaques. This study was aimed at optimizing of performance characteristics of the quantitative PCR (qPCR) PVL assay. METHODS: The PVL quantification procedure was optimized by inclusion of an exogenous control hepatitis C virus armored RNA (aRNA), a plasma concentration step, extended digestion with proteinase K, and a second RNA elution step. Efficiency of viral RNA (vRNA) extraction was compared using several commercial vRNA extraction kits. Various parameters of qPCR targeting the gag region of SIVmac239, SIVsmE660, and the LTR region of SIVagmSAB were also optimized. RESULTS: Modifications of the SIV PVL qPCR procedure increased vRNA recovery, reduced inhibition and improved analytical sensitivity. The PVL values determined by this SIV PVL qPCR correlated with quantification results of SIV RNA in the same samples using the 'industry standard' method of branched-DNA (bDNA) signal amplification. CONCLUSIONS: Quantification of SIV genomic RNA in plasma of rhesus macaques using this optimized SIV PVL qPCR is equivalent to the bDNA signal amplification method, less costly and more versatile. Use of heterologous aRNA as an internal control is useful for optimizing performance characteristics of PVL qPCRs.

Student evaluations of the portfolio process.

OBJECTIVE: To evaluate pharmacy students' perceived benefits of the portfolio process and to gather suggestions for improving the process. METHODS: A questionnaire was designed and administered to 250 first-, second-, and third-year pharmacy students at the University of Arizona College of Pharmacy. RESULTS: Although the objectives of the portfolio process were for students to understand the expected outcomes, understand the impact of extracurricular activities on attaining competencies, identify what should be learned, identify their strengths and weaknesses, and modify their approach to learning, overall students perceived the portfolio process as having less than moderate benefit. First-year students wanted more examples of portfolios while second- and third-year students suggested that more time with their advisor would be beneficial. CONCLUSIONS: The portfolio process will continue to be refined and efforts made to improve students' perceptions of the process as it is intended to develop the self-assessments skills they will need to improve their knowledge and professional skills throughout their pharmacy careers.

Use of an interrupted time-series design to evaluate a cancer screening program.

An alternative approach to intervention-control designs to evaluate community health education studies is to use a quasi-experimental design in which the outcomes of interest are examined over time in the intervention unit. The Forsyth County Cancer Screening Project (FoCaS) was a comprehensive clinic- and community-based education program to increase screening for cervical cancer and breast cancer among low-income women. This paper reports the use of piecewise regression accounting for potential effects of auto-correlation in the data to evaluate the effectiveness of the project in increasing mammography screening. Data for the evaluation of trends in screening consisted of all mammograms performed during the period of May 1992 through June 1995 at the Reynolds Health Center in Forsyth County, North Carolina. The results suggested that the FoCaS project was effective in increasing mammography screening among women age 40 or older in the study population. Analysis of the trends by age indicated that the program had differential effects on women age 40-49 and 50 or older. The results demonstrate that analyses of the type presented here can either complement or serve as an alternative to more traditional intervention-control analyses.

Community-based interventions to improve breast and cervical cancer screening: results of the Forsyth County Cancer Screening (FoCaS) Project.

The FoCaS (Forsyth County Cancer Screening) Project was one of six projects funded by the National Cancer Institute "Public Health Approaches to Breast and Cervical Cancer" initiative. The goal of this project was to improve the use of breast and cervical cancer screening among low-income, predominately African-American, women age 40 and older. Strategies implemented in the intervention city included public health clinic in-reach strategies (chart reminders, exam room prompts, in-service meetings, and patient-directed literature) and community outreach strategies (educational sessions, literature distribution, community events, media, and church programs). Baseline and follow-up data from independent cross-sectional samples in both the intervention and comparison cities were used to evaluate the effects of the intervention program. A total of 248 women were surveyed at baseline, and 302 women were surveyed 3 years later at follow-up. The proportion of women reporting regular use of mammography increased (31 to 56%; P < 0.001) in the intervention city. In the comparison city, a nonsignificant (ns) increase in mammography utilization was observed (33 to 40%; P = ns). Pap smear screening rates also improved in the intervention city (73 to 87%; P = 0.003) but declined in the comparison city (67 to 60%; P = ns). These relationships hold in multivariate models. The results suggest that a multifaceted intervention can improve screening rates in low-income populations. These results have important implications for community-based research and efforts in underserved populations.

Conservative management of an 11-week cervical pregnancy. A case report.

BACKGROUND: Cervical pregnancy is a rare form of ectopic pregnancy. This condition is usually treated with hysterectomy. CASE: A woman with an 11.3-week cervical pregnancy was treated with methotrexate, angiographic embolization of the anterior division of the hypogastric artery, dilatation and curettage, and minimal rollerball ablation of bleeding cervical vessels only. The patient was discharged on postoperative day 1. Complications included a malodorous discharge, which was treated with oral antibiotics, and claudication of the thighs for two weeks. CONCLUSION: Advanced cervical pregnancies can be treated conservatively.

Clinic-based interventions to promote breast and cervical cancer screening.

BACKGROUND: Breast and cervical cancer continue to claim the lives of women. Early detection modalities for these cancers are available; however, utilization rates are far from optimal. Studies have documented the motivating effect that physician recommendations have on compliance with preventive health behaviors. The goal of this study was to develop and implement strategies to improve the use of cervical and breast cancer screening among African-American women age 40 and older who resided in low-income housing communities. METHODS: Baseline surveys among clinic providers and a random sample of women in the target population indicated areas to be included in intervention material. Community health center-based strategies included educational interventions for providers and patients, follow-up interventions for abnormal screening tests, and the implementation of a computer tracking system. Pap smear and mammogram utilization rates at the health center were tracked throughout the project period to assess the effect of the clinic-based interventions. RESULTS: Both Pap smear and mammography rates increased over time. Fifteen cases of breast cancer and 1 case of invasive cervical cancer have been detected. Compliance rates for follow-up for cervical dysplasia have increased from 50 to 90%. CONCLUSIONS: These results suggest that clinic-based interventions can improve the use of breast and cervical cancer screening and follow-up among low-income women.

Development and implementation of outreach strategies for breast and cervical cancer prevention among African American women. FoCaS Project. Forsyth County Cancer Screening.

BACKGROUND: The purpose of the FoCaS (Forsyth County Cancer Screening) Project was to develop and implement strategies that would improve the beliefs, attitudes, and preventive health habits of populations typically considered hard to reach. Conventional health education methods have not produced substantial results; thus, innovative and unusual strategies are needed. METHODS: The FoCaS project implemented specific methods to reach the targeted population of African American women aged 40 and older that resides in public housing communities. Five outreach strategies were used: 1) educational classes (group setting and one-on-one sessions) on various topics that relate not to breast and cervical cancer but to women's issues in general; 2) media campaigns strategically scheduled throughout the year; 3) the inclusion of religion in educational classes and community outreach; 4) the use of information centers to distribute materials; and 5) a community-wide cancer-awareness event. RESULTS: These strategies reached women in nonthreatening environments that permitted heavy involvement and easy understanding of the importance of breast and cervical cancer screening. CONCLUSION: The effects of these strategies on promoting screening will be evaluated using data from the follow-up survey conducted during the spring of 1996.

Cancer screening behaviors of low-income women: the impact of race.

Cancer mortality rates are greater for African Americans than for whites. Reasons for this are due in part to the disproportionate number of the poor who are African American. Of particular concern are breast, cervical, and colorectal cancer, as screening exams, when used regularly, can reduce mortality. As part of an National Cancer Institute-funded study to improve breast and cervical cancer screening among low-income, predominately African American women, a survey was done to collect data on knowledge, attitudes, and practices related to breast, cervical, and colorectal cancer. A total of 300 women, African American and white residents of low-income housing communities, completed the survey. More African American women than white women had a mammogram within guidelines (52% vs. 40%), a clinical breast exam within the last year (60% vs. 56%), a Pap smear within the last 3 years (80% vs. 59%), and a Fecal Occult Blood Test within the last year (21% vs. 17%). Slightly more white women had a flexible sigmoidoscopy (FS) exam within the last 5 years (31% vs. 24%). When adjusted for age differences in the two populations, the differences in receiving regular screening exams were not statistically significant. Variables related to receiving these tests for all women included receiving regular check-ups (breast cancer); beliefs (breast and colorectal cancer screening), and knowledge (cervical cancer). Among African American women, barriers to screening were important for breast screening and regular checkups were related to Pap smear screening (odds ratio [OR] = 13.9, p < .01). High perceived risk of colorectal cancer was related to recent FS only for white women (OR = 47.9, p = .012). Women in this homogenous income group had similar rates of screening and had similar barriers to receiving recommended screening tests; thus, interventions should address beliefs and knowledge of risk targeted to all low-income women.

Management of malignant biliary stricture with self-expanding metallic stent.

BACKGROUND: Self-expanding metallic mesh stents are designed to remain patent longer than polyethylene (PE) stents, which generally clog in 3 to 4 months. Though more expensive, metal stents may therefore be a better choice for malignant strictures. METHODS: From January 1991 to October 1995, we performed ERCP in 212 patients with malignant or benign strictures, and 34 ultimately had insertion of a metallic stent. These stents were placed by the percutaneous transhepatic route in 17 patients and endoscopically in 17. RESULTS: Metallic stent insertion was successful in each case and relieved the preoperative jaundice and cholangitis. There were no procedure-related deaths; complications were pancreatitis (one) and hemorrhage (one). Overall stent patency was 6.2 months. Three of 34 stents occluded due to tumor ingrowth at 3, 4.5, and 8 months and were treated by placing a new PE stent through the blocked metal stent. The remaining 31 stents remained patent until patient death (n = 15, mean survival = 4.9 months) or are still open (n = 16, mean patency = 12.2 months). CONCLUSIONS: Self-expanding metal stents provide effective palliation of malignant biliary strictures and should be considered an alternative to open surgery. Metal stents remain patent much longer than PE stents and usually a single session of metal stenting can palliate biliary obstruction for life.

The recruitment of African-Americans to cancer prevention and control studies.

BACKGROUND: African-Americans have the highest overall age-adjusted cancer incidence and mortality rates of any population group in the United States. Despite this fact, this group remains underrepresented in cancer prevention and control research studies, primarily because most recruitment strategies result in limited access to African-American populations. METHODS: As part of three large-scale cancer prevention and control studies, effective strategies for recruiting African-American participants were developed and implemented. RESULTS: Eight strategies have been identified as successful recruitment strategies for involving African-Americans in cancer prevention and control studies. Utilizing these strategies resulted in recruiting a representative number of African-American participants, in relation to the local population, into the three studies. CONCLUSIONS: African-Americans can be recruited to participate in cancer control and prevention studies if plans include special strategies targeted to addressing unique barriers, beliefs, and concerns.

Validation of self-reported breast and cervical cancer screening tests among low-income minority women.

The objective of the Forsyth County Cancer Screening Project is to assess barriers to breast and cervical cancer screening among low-income women and to develop an educational program to address these barriers. To properly assess the barriers, it was first necessary to determine if self-reported rates of breast and cervical cancer screening were accurate. All women who participated in the baseline survey (n = 555) were asked to provide information regarding if, where, and when they had obtained mammograms and Pap smears. Identified health care facilities were then contacted to verify this information. Approximately 80% of responses were verified for at least one of the exams with the information provided. For mammography, 77% of self-reports were correct, whereas 67% of self-reports of Pap smear screening were correct (kappa = 0.54 and 0.15, respectively). For both tests, women thought they had received them more recently than they actually had, by an average of 3 months for mammography and 23 months for Pap smears. Using validated reports of screening did not substantially change identified predictors of screening for mammography. For Pap smear screening, however, most of the identified predictors of screening became nonsignificant when medical chart reports were used instead of self-reports, suggesting that caution should be used in relying on self-reports to design programs to improve cervical cancer screening practices.

Use of a photoessay to teach low-income African American women about mammography.

BACKGROUND: Although incidence rates of breast cancer are lower among African American women than white women, mortality rates among African American women are higher, especially for women of lower socioeconomic levels. Reasons for this situation include the lesser use of breast cancer screening examinations by low-income, primarily African American women, late stage of diagnosis, and delays in treatment. METHODS: As part of community outreach and public health clinic inreach programs for the Forsyth County Cancer Screening Project, approximately 908 African American women who reside in low-income housing communities were targeted for educational efforts related to breast cancer screening. Early in the project, it was discovered that many of the women were unfamiliar with mammography and had very little awareness of how the entire examination was conducted. This low level of knowledge was particularly important because it was a barrier for obtaining regular mammography. A photoessay depicting the process of getting a mammogram was developed and used in community outreach efforts in three formats: in educational classes, as a display in the housing communities, and in physicians' offices. The acceptability of this photoessay to communicate knowledge of and reduce fears about mammography was assessed through evaluation surveys in interviews with a sample of 47 women from the target population. RESULTS: Overall, these women liked the photoessay and felt that it provided knowledge about mammography and reduced fears associated with anticipating mammography. CONCLUSIONS: Strategies such as this may be ideal to communicate important information about cancer prevention and control in low-literacy populations.

Effect of multilevel sequential stenosis on lower extremity arterial duplex scanning.

BACKGROUND: The sensitivity of lower extremity arterial duplex scanning in detecting a > 50% stenosis may be decreased in extremities with multilevel sequential stenosis. PATIENTS AND METHODS: The results of lower extremity arterial duplex scanning of the common femoral, upper and lower superficial femoral, above- and below-knee popliteal arteries, and tibioperoneal trunk in 80 extremities of 44 patients were compared to those of arteriography. Thirty-one arterial segments in 27 extremities had a > 50% stenosis. The stenoses were categorized as first order (first or only stenotic segment in the extremity, n = 23) and second order (stenosis occurring distal to a > 50% stenosis, n = 4, or occlusion, n = 4). RESULTS: The sensitivity, specificity, and positive predictive values of duplex scan detection of a > 50% arterial stenosis or occlusion were 86%, 96%, and 67% for the common femoral artery; 95%, 98%, and 95% for the upper superficial femoral artery; 97%, 90%, and 88%, for the lower superficial femoral artery; 84%, 90%, and 87% for the above-knee popliteal artery; 47%, 98%, and 90% for the below-knee popliteal artery; and 25%, 100%, and 100% for the tibioperoneal trunk. Duplex scanning detected 18 (78%) of the 23 first-order stenoses compared to only 1 (13%) of the 8 second-order stenoses in limbs with multilevel sequential disease (P < 0.01). The peak systolic velocity at the stenotic site was significantly higher for first-order (mean +/- SD 168 +/- 54 cm/s) compared to second-order (38 +/- 13 cm/s) stenoses (P < 0.00002). CONCLUSIONS: Duplex scanning was highly sensitive in detecting lower extremity first-order stenoses. Low peak systolic velocities at second-order stenoses of limbs with multilevel sequential disease significantly decreased the sensitivity of duplex scanning.

Sinus augmentation. Rationale, development, long-term results.

Sinus grafting is a proven procedure with predictable results. However, practitioners need to be aware of and adhere to the details that dictate the success of the procedure.

Urokinase infusion: feasibility of monitoring for complications in a non-intensive care setting.

In an effort to more effectively use critical care facilities and to reduce costs, during a 2 1/2-year period, the condition of 32 patients who received 37 local intraarterial urokinase (UK) infusions was monitored in a non-intensive care unit (ICU) setting. Techniques of infusion, mean total dose of lytic agent used (1.7 million IU), and mean duration of infusion (22 hours) were similar to those reported previously in series of patients monitored in the ICU. Complete lysis (no angiographically detectable residual clot within the treated segment) was achieved in 28 of 37 infusions (76%). Major complications occurred during two infusions (5.4%). In 33 of the 37 cases, systemic heparin was administered during UK infusion. No cases of pericatheter thrombosis were encountered. At the authors' institution, patients can be safely monitored during local UK infusion in a non-ICU setting without compromising effectiveness of therapy. This approach has resulted in enhanced cost-effectiveness of thrombolytic therapy and more effective use of critical care facilities.

Tube tamponade: potential pitfall in angiography of arterial hemorrhage associated with percutaneous drainage catheters.

Diagnostic angiography performed to search for a source of hemorrhage in three patients with percutaneous transhepatic biliary catheters and one patient with a percutaneous nephrostomy catheter was initially unrewarding when performed with the drainage catheter in place. In each patient, removal of the drainage catheter resulted in severe pulsatile hemorrhage from the parenchymal tract and allowed angiographic localization of the bleeding site. Temporary control of the hemorrhage was then obtained by inflating an angioplasty balloon within the tract. Transcatheter embolotherapy provided definitive control of bleeding in three patients. When initial angiographic evaluation for bleeding in patients with percutaneous biliary and nephrostomy catheters fails to depict a source, the study should be repeated immediately after removal of the drainage catheter. Because hemorrhage can be severe once tamponade is relieved, the drainage catheter should be withdrawn over a guide wire so that a tamponading catheter can be rapidly reinserted to control hemorrhage until more definitive therapy is undertaken.

Stereochemistry of methylene transfer involving 5,10-methylenetetrahydrofolate.

The stereochemistry of the transfer catalyzed by rabbit liver serine transhydroxymethylase (EC 2.1.2.1) of the prochiral hydroxymethyl group from serine to tetrahydrofolate to form 5,10-methylenetetrahydrofolate was studied. Initial kinetic studies on labeled 5,10-methylenetetrahydrofolate showed that nonenzymatic racemization of the prochiral methylene center was buffer dependent and was slow under the conditions employed. Specifically tritiated (3R)- and (3S)-serines were employed to study the transfer reaction. Reactions were carried out under various conditions and the stereochemistry of the methylene carbon of the 5,10-methylenetetrahydrofolate produced was determined. The enzyme was shown to be partially stereospecific for this transfer reaction, proceeding with a loss of about 50% of the stereochemical purity of the transferred carbon center. Possible mechanistic interpretations of this finding are discussed.

Serine transhydroxymethylase: evidence for a sequential random mechanism.

Initial velocity patterns in the presence of product and dead-end inhibitors suggest that in reaction 1 the addition of substrates and release of products occur by a sequential random mechanism: L-serine + tetrahydrofolate in equilibrium glycine + 5,10-methylenetetrahydrofolate. This interpretation is supported by equilibrium isotope-exchange studies. The relative maximum rates of exchange of L-serine in equilibrium glycine and L-serine in equilibrium 5,10-methylenetetrahydrofolate in reaction 1 were not inhibited by high levels of substrates. The relative rates of these two exchange reactions were similar but were not identical. These results suggest that the catalytic interconversion and dissociation of substrates are of the same order of magnitude. Reaction 1 represents the transfer of a one-carbon group from the third carbon of L-serine to tetrahydrofolate. Inhibition studies showed that abortive enzyme ternary complexes are formed with L-serine and tetrahydrofolate compounds, which also contain a one-carbon group, e.g., 5-methyltetrahydrofolate and 5,10-methylenetetrahydrofolate. This suggests that the one-carbon binding site can accomodate two one-carbon groups simultaneously without serious steric hindrance.