Metered Cryospray™: a novel uniform, controlled, and consistent in vivo application of liquid nitrogen cryogenic spray.

In this article, a novel cryotherapy approach using a uniform, controlled, and consistent in vivo application of liquid nitrogen (LN2) spray as a Metered Cryospray™ (MCS) process is described. Although MCS may be used for many potential clinical applications, this paper focuses on the development that led to the controlled and consistent delivery of radial LN2 cryogen spray in order to generate a uniform circumferential effect and how the amount of MCS can be adapted to specifically ablate targeted diseases within a patient's lumen such as an airway or esophagus.

N2 gas egress from patients' airways during LN2 spray cryotherapy.

Spray cryotherapy using liquid nitrogen (LN2) is a general surgical tool used to ablate benign or malignant lesions. Adequate egress of the gaseous nitrogen (N2) generated during this process must be provided for safe use when LN2 is used within the body rather than topically. When delivered to either the gastrointestinal tract (requiring active venting via a suction tube) or body cavities open to room barometric pressure (such as lung airways) allowing for passive venting, the N2 gas generated from the boiling process must be evacuated. This work will examine the egress of N2 during procedures requiring passive venting from human airways undergoing liquid nitrogen spray cryotherapy. Venting characteristics for safe N2 egress will be presented and discussed based on analytical modeling using fluid mechanics simulations and experimental studies of N2 venting with laboratory and porcine models.

Immunotherapy of human cervical high-grade cervical intraepithelial neoplasia with microparticle-delivered human papillomavirus 16 E7 plasmid DNA.

OBJECTIVE: The purpose of this study was to evaluate the safety of the administration of a bacterial expression plasmid encoding a 13 amino acid sequence that is highly homologous with human papillomavirus E7 within poly (lactide-co-glycolide) microparticles (ZYC101) in women with HLA A2+ antigen and persistent cervical intraepithelial neoplasia grade 2/3 and human papillomavirus 16. STUDY DESIGN: Fifteen women entered an institutional review board-approved dose-escalating phase I study with the use of three levels of blood monitoring and urine studies, Papanicolaou tests, and colposcopy. Escalation required no serious adverse events. Immunologic responses were evaluated in peripheral blood with the use of human papillomavirus peptide-stimulated interferon gamma enzyme-linked immunosorbent assay for T-cell reactivity. In cervical secretions, immunoglobulin A anti-human papillomavirus 16 E2 concentrations were measured. Three doses every 3 weeks were followed 4 weeks later by surgical excision. RESULTS: No serious adverse events occurred. Five women had complete histologic responses; 11 women had human papillomavirus-specific T-cell responses. Four of five complete histologic responses developed immunoglobulin A anti-E2-specific antibody. CONCLUSION: ZYC101 warrants further investigation because of a 33% complete histologic responses, a 73% immunologic response, and no serious adverse events.

Use of loop electrosurgical excision procedure specimens to provide tissue fractions for immunologic analyses.

OBJECTIVE: To determine if dividing loop electrosurgical excision procedure (LEEP) specimens to provide tissue for research increases rates of LEEP specimen misdiagnosis and recurrent cervical intraepithelial neoplasia. MATERIALS AND METHODS: In this chart review, 42 women with biopsy-confirmed cervical intraepithelial neoplasia (CIN) 2,3 had up to 20% of their LEEP specimens sectioned and used for immunologic analysis. The remainder of each specimen was assessed routinely. Follow-up cytologic analyses and cervical biopsies also were assessed. This cohort was compared with a control cohort of 80 patients with biopsy confirmed CIN 2,3 whose LEEP specimens were not divided. Statistical significance was defined as a p value of < .05. RESULTS: There were no statistically significant differences between the groups with regard to histologic assessment of LEEP specimens or follow-up outcomes. CONCLUSIONS: Use of up to 20% of LEEP specimens for research purposes neither adversely affects histologic evaluation of LEEP specimens nor leads to poorer follow-up outcomes.

Endocervical curettage at conization to predict residual cervical adenocarcinoma in situ.

OBJECTIVE: To determine if performing an endocervical curettage (ECC) at the time of conization is a useful diagnostic tool for predicting residual cervical adenocarcinoma in situ (AIS) among women who might wish to preserve their fertility. METHODS: All patients diagnosed with AIS from 1995 to 2000 at four institutions were identified. Data were retrospectively extracted from clinical records. Women included in the statistical analysis were (1) younger than 40 years, (2) had an ECC performed at the time of the initial cone biopsy, (3) had a clearly demarcated surgical margin pathologically, and (4) underwent a second surgical procedure. RESULTS: Twenty-nine (24%) of 123 AIS patients met criteria for inclusion. The median age was 33 years (range, 17 to 39) and 13 (46%) were nulliparous. Initial surgery was a cold-knife conization (n = 17) or loop electrosurgical excision procedure (n = 12). Twelve (41%) ECCs and 15 (52%) cone margins were histologically positive. Sixteen patients underwent a repeat conization; 13 underwent hysterectomy. Thirteen (45%) patients had residual AIS at the time of their second surgical procedure. ECC had a superior positive predictive value (100% vs 47%; P < 0.01) and negative predictive value (94% vs 57%; P = 0.01) compared to cone margin in predicting residual AIS. None of the women undergoing fertility-sparing surgery developed recurrent AIS or adenocarcinoma. CONCLUSION: ECC performed at the time of conization may be a useful tool for predicting residual AIS in women considering fertility preservation.

Cytologic and biopsy findings leading to conization in adenocarcinoma in situ of the cervix.

OBJECTIVE: To investigate the utility of currently available screening tests in preoperatively detecting adenocarcinoma in situ of the cervix. METHODS: Patients with a cone biopsy diagnosis of adenocarcinoma in situ from 1987 to 2000 at our institution were identified. Results from Papanicolaou smears, cervical biopsies, and endocervical curettages preceding the diagnostic cone biopsy were collected from medical records and referring providers. Fisher exact test (two-tail) was used for statistical analysis. RESULTS: The preoperative screening results preceding a cone biopsy containing adenocarcinoma in situ were available in 118 patients. Among 94 Papanicolaou smears, 65 (69%) glandular lesions and 29 (31%) squamous or unspecified lesions were reported. Biopsy and/or endocervical curettage after the 29 squamous or unspecified lesions on Papanicolaou smear detected 15 additional glandular lesions, totaling 80 (85%) of 94 cases of glandular disease detected before conization. Among all 118 cases with some form of preoperative data available, glandular disease was predicted in 100 cases (85%). In cases of suspected glandular disease, 86% were treated with cold knife cone compared with 22% in cases of suspected squamous abnormalities (P <.001). CONCLUSION: The sensitivity of detecting a glandular abnormality before a cone biopsy containing adenocarcinoma in situ is 69% with the Papanicolaou smear and 85% with the addition of biopsy and endocervical curettage. This underscores the importance of using preoperative assessment to appropriately plan treatment for a suspected glandular lesion.

Trimodal spectroscopy for the detection and characterization of cervical precancers in vivo.

OBJECTIVE: The objective of this study was to assess the potential of 3 spectroscopic techniques (intrinsic fluorescence, diffuse reflectance, and light scattering) individually and in combination (trimodal spectroscopy) for the detection of cervical squamous intraepithelial lesions. STUDY DESIGN: The study was conducted with 44 patients who underwent colposcopy for the evaluation of an abnormal Papanicolaou smear. Fluorescence and reflectance spectra were collected from colposcopically normal and abnormal sites and analyzed to extract quantitative information about tissue biochemistry and morphologic condition. This information was compared with histopathologic classification, and diagnostic algorithms were developed and validated with the use of logistic regression and cross-validation. RESULTS: Diagnostically significant differences exist in the composition of fluorescing biochemicals, the scattering properties, and the epithelial cell nuclear morphology of cervical squamous intraepithelial lesions and non-squamous intraepithelial lesions. Trimodal spectroscopy is a superior tool for the detection of cervical squamous intraepithelial lesions than any 1 of the techniques alone. CONCLUSION: Trimodal spectroscopy has the potential to improve the in vivo detection of precancerous cervical changes.

Managing atypical squamous cells of undetermined significance (ASCUS): human papillomavirus testing, ASCUS subtyping,or follow-up cytology?

OBJECTIVE: This study related morphologic subtype, human papillomavirus status, and a second cytologic examination to the follow-up biopsy-proven high-grade squamous intraepithelial lesion (HSIL; grade II or III cervical intraepithelial neoplasia) after a cytologic diagnosis of atypical squamous cells of undetermined significance (ASCUS). STUDY DESIGN: Seven hundred four liquid-based cervical cytology specimens were classified as normal, "ASCUS, favor reactive" (AFR), "ASCUS, not otherwise specified," "ASCUS, favor low-grade squamous intraepithelial lesion," "ASCUS, favor HSIL" (AFHS), low-grade squamous intraepithelial lesion, and HSIL. Human papillomavirus typing used polymerase chain reaction-restriction fragment length polymorphism analysis. A longitudinal review of the cytologic and histologic records of ASCUS cases with > or =1 follow-up test or biopsy ascertained the frequency of a follow-up diagnosis of biopsy-proven HSIL (grade II or III cervical intraepithelial neoplasia). RESULTS: Three hundred eighty-six cases (208 ASCUS, 68 normal, 86 with low-grade squamous intraepithelial lesions, and 24 with HSIL) were evaluated. High-risk human papillomavirus (HRHPV positive) was lowest with normal cytology (13%), highest with HSIL (71%), and was present in 29.8% of ASCUS cases, ranging from 22.2% (AFR) to 75% (AFHS). Most ASCUS tests (64%) were followed by a negative cytologic or histologic examination. Overall, 3.8% and 11% of ASCUS and HRHPV-positive ASCUS had histologic outcomes of HSIL. AFHS had the highest (25%) and AFR had the lowest (1.1%) proportion of HSIL outcomes. Sensitivity, specificity, and positive predictive values of human papillomavirus testing for biopsy-proven HSIL were 87.5%, 72.5%, and 11.3%, respectively. CONCLUSION: HSIL and AFHS are distinguished by the highest frequency of HRHPV types and higher rates of HSIL outcome. The remaining categories of ASCUS are heterogeneous with respect to human papillomavirus type and HSIL risk, and the value of subclassification of these entities is dependent on the practice. A human papillomavirus detection system based on polymerase chain reaction-restriction fragment length polymorphism identifies a smaller percentage of high-risk human papillomaviruses than mixed probe-based methods, probably because of the more precise exclusion of cross-reacting low-risk human papillomavirus. Negative HRHPV findings by either system show a markedly reduced risk of an HSIL outcome. However, the relative advantage of human papillomavirus testing over follow-up cytology will be influenced by the frequency of negative follow-up cytologic examination and sensitivity of liquid-based preparations in a given practice.

NAD(P)H and collagen as in vivo quantitative fluorescent biomarkers of epithelial precancerous changes.

During the development of neoplasia, epithelial tissues undergo biochemical and structural changes that can manifest in tissue fluorescence. There have been several reports on different in vivo fluorescence characteristics between normal and precancerous (dysplastic) tissues. However, it has been difficult to identify and quantify the origins of these changes, mainly because of distortions introduced in measured tissue fluorescence spectra by tissue scattering and absorption. Such distortions can be removed by combining information in simultaneously measured fluorescence and reflectance spectra. Thus, we can recover the intrinsic (undistorted) tissue fluorescence. In this report, we show that extraction of the intrinsic fluorescence allows us: (a) to determine the fluorescence spectra of NAD(P)H and collagen in an in vivo environment, and (b) to use these NAD(P)H and collagen spectra to describe, quantitatively, diagnostically significant biochemical changes between normal and dysplastic tissues. Specifically, by analyzing intrinsic fluorescence of human epithelial tissue as it becomes deoxygenated in vivo, we can resolve the fluorescence spectra of NAD(P)H and collagen, two of the major tissue fluorophores. This is important because fluorescence depends on the local environment of the chromophore. Then, we extract the intrinsic fluorescence spectra of sites from 35 patients with suspected cervical lesions and 7 patients with Barrett's esophagus and describe them accurately as a linear combination of NAD(P)H and collagen contributions. In both tissue cases, we find that low collagen and high NAD(P)H fluorescence characterizes the high-grade dysplastic lesions when compared with nondysplastic tissues. These data present evidence for the presence of detectable levels of NAD(P)H fluorescence in human epithelial tissues in an in vivo setting and demonstrate that NAD(P)H and collagen may be used as quantitative fluorescence biomarkers for in vivo detection of dysplasia in the cervix and the esophagus.

Atypical glandular cells of undetermined significance in conventional cervical/vaginal smears and thin-layer preparations.

BACKGROUND: Biopsy follow-up of a cervical/vaginal smear interpretation of atypical glandular cells of undetermined significance (AGUS) most often reveals either a benign reactive process or a squamous cervical intraepithelial neoplasia (CIN) rather than a glandular one. The ThinPrep Papanicolaou test (TP) has been shown to increase diagnostic sensitivity for CIN. To the authors' knowledge there are few studies examining its effectiveness in diagnosing uterine glandular lesions, either endocervical or endometrial. The authors compared outcomes after AGUS interpreted in TP specimens and conventional smear preparations (CPs). METHODS: Follow-up was sought in all cases that were interpreted as AGUS during a 3-year period in which practice groups converted from CPs to TPs. A tissue diagnosis of adenocarcinoma in situ, CIN of Grade 2 or 3, or invasive carcinoma was considered to be a positive follow-up result. Either a benign biopsy finding with a subsequent benign cytologic result or two consecutive benign cytologic results were considered to be negative follow-up results. RESULTS: AGUS was reported in 116 of 75,002 TPs (0.15%) and 151 of 79,322 CPs (0.19%). Follow-up information was available in 91 (78%) and 130 (86%) cases, respectively. The predictive value of AGUS for a positive follow-up result was 22% in the TP group and 15% in the CP group. An AGUS interpretation with a positive follow-up result was found in 0.027% of all cases using TPs and in 0.025% of all cases using CPs. CONCLUSIONS: The use of TPs may increase the diagnostic specificity of AGUS for a high-grade precursor lesion or carcinoma.

Management of adenocarcinoma in situ, micro-invasive, and early stage adenocarcinoma of the cervix.

The incidence of glandular neoplasms of the uterine cervix has been steadily increasing over the past two decades. Given the fact that these lesions are more difficult to diagnosis and are relatively infrequent, less is known about them compared with their squamous counterparts. In addition, because these lesions tend to arise in women of childbearing age, there is a particular need to understand whether the in-situ and early invasive forms of adenocarcinoma are amenable to conservative treatment measures that spare fertility like their squamous counterparts. Recent publications have addressed the underlying causes of the increasing incidence, the pros and cons of conservative management, and the difficulties in definitively identifying each glandular subtype. The most difficult diagnostic lesions are adenocarcinoma in situ cases and early invasive adenocarcinomas.

Early-stage cervical adenocarcinoma treated by surgical intent: the role of para-aortic lymph node dissection.

OBJECTIVE: Previous reports suggest that cervical adenocarcinomas have a unique pattern of spread and are more apt to metastasize to para-aortic lymph nodes. The purpose of this study was to further define the node of para-aortic lymph node dissection in early-stage cervical adenocarcinoma treated by surgical intent. METHODS: Institutional review board approval was obtained to perform a computerized search of the data of all women diagnosed with cervical adenocarcinoma between 1982 and 2000. Hospital charts were retrospectively reviewed. Follow-up was obtained from the tumor registry, medical records, and correspondence with health care providers. RESULTS: Three hundred (87%) of 345 early-stage (FIGO IA(1)-IIA) cervical adenocarcinoma patients were primarily treated by surgical intent. Two hundred seventy-six underwent pelvic and para-aortic node dissection (n = 69) or pelvic node dissection only (n = 207); 24 had no lymph node dissection. The median number of lymph nodes removed was 13 pelvic (range, 1-58) and 3 para-aortic (range, 1-17). Three (4%) of 69 patients had para-aortic nodal metastases. Each had either grossly evident para-aortic adenopathy (n = 2) or an adnexal metastasis. Thirty-six of 40 women developing recurrent disease had at least some component of pelvic recurrence; 4 had only extrapelvic disease. Three patients undergoing para-aortic node dissection developed an isolated extrapelvic recurrence despite originally negative para-aortic nodes (n = 2) or treatment by extended-field radiation for para-aortic metastases. One woman undergoing only pelvic node dissection had an isolated extrapelvic recurrence despite originally negative nodes. CONCLUSIONS: Early-stage cervical adenocarcinoma primarily treated by surgical intent has a very low risk of para-aortic metastases. These were detected only when there was gross evidence of nodal or adnexal disease.

Can Monsel's Transmit Nosocomial Infections? A Microbiological Investigation.

OBJECTIVE.: Our objective was to test whether common nosocomial pathogenic bacteria can be cultured in Monsel's paste, 20% ferric subsulfate, an astringent solution with styptic qualities that causes hemostasis when applied topically to small cuts. MATERIALS AND METHODS.: First, samples of Monsel's paste were collected and plated onto 4 media. Bacterial growth was evaluated at 48 hours. Second, thioglycollate medium was inoculated with a combination of Monsel's paste and actively growing stock bacteria. After 2 days, the samples were subcultured. They were evaluated for growth at 5 days. Next, samples of Monsel's were inoculated with 10Pseudomonas aeruginosa or 10Clostridium perfringens. The samples of inoculated paste were examined at 5-minute intervals thereafter to evaluate bacterial survival. RESULTS.: Neither the open samples of Monsel's solution nor the inoculated samples of Monsel's solution grew bacteria. When the samples inoculated with 10 organisms were examined at timed intervals after inoculation, no bacteria were noted after 5 minutes. CONCLUSIONS.: It is unlikely that nosocomial bacteria could survive exposure to Monsel's solution. Thus, potential contamination of the solution is unlikely, and subsequent transmission of infection is improbable.

Stage IIB-IVB cervical adenocarcinoma: prognostic factors and survival.

OBJECTIVE: The incidence of cervical adenocarcinoma is increasing relative to squamous cell carcinoma and all cervical cancers. Few reports have described the outcome of patients with advanced cervical adenocarcinoma. The purpose of this study was to determine the prognostic factors and survival for patients with stage IIB-IVB disease. METHODS: Institutional Review Board approval was obtained to perform a computerized search of all women diagnosed with cervical adenocarcinoma at our three institutions between 1982 and 2000. Medical records were retrospectively reviewed. Clinical follow-up was obtained from the SGO database and tumor registry and via correspondence with health care providers. Statistical analysis was performed using logistic regression for clinical variables and the log-rank test to compare Kaplan-Meier survival estimates. RESULTS: Eighty-three women with FIGO stage IIB-IVB cervical adenocarcinoma were identified. The median patient age was 53 years (range, 22-88). The median follow-up of 17 (20%) surviving patients was 33 months (range, 6-147); 66 (80%) died during the study interval. Stage IIB disease, young patient age, and grade 1 histology were independent variables having a favorable impact on survival (each P < 0.02). Stage IIB patients (n = 41) were more likely to be alive at 2 (64% vs 8%) and 5 years (30% vs 0%) than women with stage IIIA-IVB disease (n = 42; P < 0.01). CONCLUSIONS: Women diagnosed with advanced stage cervical adenocarcinoma have a poor prognosis. Prospective, multicenter trials of platinum-based chemoradiation or other novel therapies are urgently needed in the treatment of this highly lethal disease.