Proof of concept: predicting the onset of meat spoilage by an integrated oxygen sensor spot in MAP packages.

During storage of modified atmosphere packaged (MAP) meat, the initial microbiota grows to high cell numbers, resulting in perceptible spoilage after exceeding a specific threshold level. This study analyses, whether elevated oxygen consumption in the headspace of MA-packages would enable a prediction method for meat spoilage. We monitored the growth of single spoiling species inoculated on high-oxygen MAP beef and poultry, performed sensorial analysis and determined oxygen concentrations of the headspace via a non-invasive sensor spot technology. We detected microbial headspace oxygen consumption occurring prior to perceptible meat spoilage for certain species inoculated on beef steaks. However, headspace oxygen consumption and cell counts at the onset of spoilage were highly species-dependent, which resulted in a strong (Brochothrix thermosphacta) and moderate (Leuconostoc gelidum subspecies) decrease of the headspace oxygen content. No linear decrease of the headspace oxygen could be observed for Carnobacterium divergens and Carnobacterium maltaromaticum inoculated on poultry meat. We demonstrate the applicability of an incorporated oxygen sensor spot technology in MAP meat packages for detection of spoilage in individual packages prior to its perceptible onset. This enables individual package evaluation and sorting within retail, and consequently reduces meat disposal as waste.

Diversity and anaerobic growth of Pseudomonas spp. isolated from modified atmosphere packaged minced beef.

AIMS: This study aimed to monitor development of spoilage-associated microbiota on high-oxygen modified atmosphere packaged (MAP) minced beef, assess diversity of Pseudomonas sp. therein employing a polyphasic approach and probe their ability to grow anaerobically in the presence of carbon dioxide. METHODS AND RESULTS: Headspace atmosphere and total viable count of MAP minced beef were monitored, and spoilage-associated microbiota was identified using matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS). Pseudomonas spp. represented a major part of the spoilage-associated microbiota throughout the spoilage process and were characterized with a polyphasic approach including MALDI-TOF, randomly amplified polymorphic DNA biotyping, 16S rDNA and rpoD sequence analysis, and carA multiplex polymerase chain reaction. Pseudomonas isolates displayed high diversity and varying assertiveness under conditions employed in MAP minced beef with P. fragi, P. lundensis and P. weihenstephanensis as dominant species. CONCLUSIONS: The polyphasic approach enabled high-throughput characterization of Pseudomonas sp. Their adapted capability to grow anaerobically and resistance to high levels of CO2 is suggested to be a general feature within the genus, which is hitherto underexplored. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows that diverse Pseudomonas generally regarded as strict aerobes and CO2 -sensitive appear well adapted to grow under MAP conditions, leading to high cell counts in minced beef and ultimately contribute to spoilage of the product.

Monitoring of spoilage-associated microbiota on modified atmosphere packaged beef and differentiation of psychrophilic and psychrotrophic strains.

AIMS: This study aimed to explore the discriminatory power of MALDI-TOF MS as a high-throughput method to monitor growth dynamics of meat-spoiling bacteria on modified atmosphere packaged (MAP) beef and to differentiate psychrophilic and psychrotrophic spoilage-associated strains. METHODS AND RESULTS: MAP beef steaks were incubated for 21 days at constant 4 and 10°C. Development of headspace gas composition, pH, CFU and spoilage-associated microbial composition were monitored. MALDI-TOF MS exhibited high discriminatory power for reliable, high-throughput identification of spoilage-associated, psychrotrophic microbiota. Microbiota development was highly dependent on initial abundance of specific species. Organoleptic onset of spoilage was concomitant with an alteration of headspace atmosphere and pH. Screening for psychrophiles at 4°C on beef revealed the abundance of Leuconostoc gelidum subsp. gelidum TMW2·1998 with characteristic psychrophilic growth behaviour. CONCLUSIONS: We suggest that control of initial contaminants is crucial to predict the onset of spoilage and that headspace atmosphere and pH are important parameters with spoilage-indicative potential. SIGNIFICANCE AND IMPACT OF THE STUDY: MALDI-TOF MS proved suitable for high-resolution monitoring of psychrotrophic and psychrophilic spoilage-associated microbiota and enables improved insights in the spoilage progress. The presence of psychrophilic strains on beef is the likely causative for unexplained spoilage events.

Assertiveness of meat-borne Lactococcus piscium strains and their potential for competitive exclusion of spoilage bacteria in situ and in vitro.

AIMS: This study aimed to investigate intraspecies assertiveness of meat-borne Lactococcus piscium isolates, inhibitory effects on unwanted and harmful meat spoilers, and the prevalence on beef deliberately inoculated with Lc. piscium. METHODS AND RESULTS: Co-inoculation of Lc. piscium isolates and spoilers (Brochothrix thermosphacta, Leuconostoc gelidum subsp. gasicomitatum, Carnobacterium divergens, Pseudomonas weihenstephanensis, Serratia liquefaciens, Hafnia alvei) were conducted in sterile meat simulation medium. Differentiation of Lc. piscium strains was carried out with colony-based RAPD-PCR. Selective cultivation was used to differentiate spoilers from Lc. piscium. Intraspecies assertiveness revealed Lc. piscium TMW2.1614 as most assertive strain. Co-inoculation of selected Lc. piscium strains caused substantial growth reduction of spoilers while the extent was strain- and spoiler dependent. Monitoring the microbiota on beef steaks deliberately inoculated with Lc. piscium revealed prevalence over the endogenous microbiota while maintaining a ripened sensory impression without undesired alterations. CONCLUSIONS: This study reveals Lc. piscium strains TMW2.1612/2.1614/2.1615 as highly competitive against spoilers in vitro while beef deliberately inoculated with these strains maintained acceptable organoleptics. SIGNIFICANCE AND IMPACT OF THE STUDY: Selected Lc. piscium strains exhibit high potential for application as bioprotective cultures for competitive exclusion on beef in order to extend minimum shelf life and enhance product safety of meat.

An adapted isolation procedure reveals Photobacterium spp. as common spoilers on modified atmosphere packaged meats.

The genus Photobacterium comprises species of marine bacteria, commonly found in open-ocean and deep-sea environments. Some species (e.g. Photobacterium phosphoreum) are associated with fish spoilage. Recently, culture-independent studies have drawn attention to the presence of photobacteria on meat. This study employed a comparative isolation approach of Photobacterium spp. and aimed to develop an adapted isolation procedure for recovery from food samples, as demonstrated for different meats: Marine broth is used for resuspending and dilution of food samples, followed by aerobic cultivation on marine broth agar supplemented with meat extract and vancomycin at 15°C for 72 h. Identification of spoilage-associated microbiota was carried out via Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry using a database supplemented with additional mass spectrometry profiles of Photobacterium spp. This study provides evidence for the common abundance of multiple Photobacterium species in relevant quantities on various modified atmosphere packaged meats. Photobacterium carnosum was predominant on beef and chicken, while Photobacterium iliopiscarium represented the major species on pork and Photobacterium phosphoreum on salmon, respectively. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates highly frequent isolation of multiple photobacteria (Photobacterium carnosum, Photobacterium phosphoreum, and Photobacterium iliopiscarium) from different modified-atmosphere packaged spoiled and unspoiled meats using an adapted isolation procedure. The abundance of photobacteria in high numbers provides evidence for the hitherto neglected importance and relevance of Photobacterium spp. to meat spoilage.

Comparative genome analysis, predicted lifestyle and antimicrobial strategies of Lactococcus carnosus and Lactococcus paracarnosus isolated from meat.

Strains of the psychrotrophic bacterium Lactococcus piscium have gained increasing attention as potentially bioprotective cultures due to their assertiveness against fish and meat spoilage bacteria as well as pathogenic bacteria. Recently, we have described two novel species within the genus Lactococcus (Lc.) namely Lc. carnosus (TMW 2.1612T) and Lc. paracarnosus (TMW 2.1615T) isolated from modified atmosphere packaged meat. Within this study, we compared the genomes of two Lc. carnosus strains, two Lc. paracarnosus strains and 16 Lc. piscium strains from our laboratory and five publicly available genomes previously affiliated to the species Lc. piscium. Our phylogenetic analysis supports reclassification of 20 of the strains to either Lc. carnosus or Lc. paracarnosus, so far limiting the Lc. piscium type strain (DSM 6634T) as sole representative of this species. Comparative genomics approach was conducted to predict underlying mechanisms involved in interspecies competition strategies of Lc. carnosus and Lc. paracarnosus against meat spoilers and predict their lifestyle in meat environments. In general, strains of the three species were highly similar regarding metabolic pathways for most of the relevant meat-derived substrates. In silico analyses enabled prediction of homolactic hexose fermentation by Lc. carnosus, Lc. paracarnosus and Lc. piscium. Further, genes required for the heterofermentative metabolism of hexoses and pentoses were only found in the Lc. pisicum type strain (DSM 6634T). We predict a low spoilage potential for Lc. carnosus and Lc. paracarnosus strains. No genes for decarboxylation of amino acids yielding biogenic amines were found in the genomes. Regarding their antimicrobial mechanisms against spoilers, we found a strain-specific putative polymorphic toxin system predictively delivered by the type VIIb secretion system, enabling cell-to-cell contact-dependent growth inhibition. Furthermore, we found additional genes predictively involved to the suppression of spoilers within the food microbiome (prophages, lytic domains, bacteriocins, metabolites).

Induction of apoptosis by proteasome inhibitors in B-CLL cells is associated with downregulation of CD23 and inactivation of Notch2.

Recently, proteasome inhibitors (PI) have attracted interest as novel anticancer agents in B-cell chronic lymphocytic leukemia (B-CLL). A prominent feature of B-CLL cells is the high expression of CD23, which is closely related to cell survival and is regulated by Notch2. Since several components of the Notch signaling cascade are tightly regulated by proteasomal degradation, we studied the effect of PI on Notch2 activity and CD23 expression. Exposure of B-CLL cells to PI led to induction of apoptosis, a time- and dose-dependent downregulation of CD23 expression and a decline in DNA binding of transcriptionally active Notch2. In contrast, the transcription factor NF-AT and its putative target gene CD5, which is highly expressed in B-CLL cells, were unaffected. When the late phase of PI-induced apoptosis was arrested by inhibition of caspase 3, the reduction of Notch2 activity was still observed, indicating that reduction of active Notch2 took place already during an earlier phase of apoptosis. Enforced expression of constitutively active Notch2 decreased PI-mediated apoptosis in a human B-cell line. These data indicate that downregulation of CD23 and loss of Notch2 activity are early steps in PI-induced apoptosis of B-CLL lymphocytes and may be part of the full apoptotic response.

Reconstitution of endogenous interferon a by recombinant interferon in hairy cell leukemia.

Recombinant human IFN alpha (rhIFN-alpha) plays an important role in the treatment of hairy cell leukemia (HCL). However, the mechanisms leading to its beneficial effect are not completely clarified, and there is no information on IFN-alpha gene expression in this disease. Therefore, we investigated the pattern of IFN-alpha gene expression and protein production in HCL and their potential regulation by rhIFN-alpha. Blood samples from 10 patients with HCL and 8 healthy donors (HD) were investigated. Expression of IFN-alpha mRNA was assessed by reverse transcription-PCR analysis in peripheral blood mononuclear cells (PBMCs) under basal conditions and on induction with rhIFN-alpha and polyionosinic-polycytidylic acid [poly(I.C)]. IFN-alpha concentrations in plasma and culture supernatants were measured by immunoassays, and intracellular IFN-alpha was evaluated by fluorescence-activated cell sorting analysis. Results showed that, in contrast to blood samples from HDs, freshly isolated PBMCs from un treated HCL patients did not express IFN-alpha mRNA, whereas IFN-alpha transcripts were found in patients who were under rhIFN-alpha therapy Plasma of untreated patients contained no, or extremely low levels of IFN-alpha as compared with plasma of treated patients and HDs. Ex vivo treatment of PBMCs with rhIFN-alpha or poly(I.C) resulted in a remarkable up-regulation of IFN-alpha at the mRNA and protein level. In HCL, however the amounts of IFN-alpha protein remained less than in HD. Inhibition of IFN-alpha transcription was found after exposure of PBMCs to serum fron untreated patients. Finally, a reduced capacity to produce IFN-alpha was found within B- cell, T-cell, and monocyte compartments in HCL patients which could be enhanced by rhIFN-alpha. The results demonstrate the ability, of rhIFN-alpha to up-regulate the expression of IFN-alpha gene and protein production and suggest that priming the production of endogenous IFN-alpha is a critical step in the mechanism of action of rhIFN-alpha in HCL.

Inadequate production of hematopoietic growth factors in hairy cell leukemia: up-regulation of interleukin 6 by recombinant IFN-alpha in vitro.

The course of hairy cell leukemia (HCL) is characterized by progressive pancytopenia. The pathogenesis of this phenomenon is still not fully understood. To study if the decrease in hematopoiesis in HCL is accompanied by abnormal concentrations of growth factors, we investigated the production of granulocyte colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, interleukin 3 (IL-3), interleukin 6 (IL-6), and tumor necrosis factor alpha by peripheral blood mononuclear cells (PBMCs) of eight patients with HCL. The results point to a severe deficiency of production of all cytokines tested as compared to healthy donors. However, enrichment of autologous monocytes by counterflow centrifugation resulted in a marked increase of the levels of granulocyte colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, IL-6, and tumor necrosis factor alpha. The most pronounced effects were seen with IL-6. Reverse transcription-PCR analysis indicated that pokeweed mitogen, IFN-alpha, and poly(I:C) are capable of inducing the expression of IL-6-specific mRNA in HCL cells. These findings are substantiated on the protein level by immunofluorescence analysis. Incubation of PBMCs with IFN-alpha resulted in a significant increase of intracellular IL-6 in HCL but not in healthy donors. This increase was also seen in hairy cells positive for CD19 and CDllc. Furthermore, IFN-alpha induced the secretion of IL-6 from PBMCs of HCL patients but not healthy donors. In conclusion, our studies with PBMCs from patients with HCL revealed an inadequate supply of hematopoietic growth factors that might, in part, be due to the monocytopenia characteristic for this disease. The findings also indicate that IFN-alpha is capable of inducing the production of IL-6 in the patients' PBMCs as well as in their hairy cells. These data from our in vitro studies support the clinical observation that treatment with IFN-alpha leads to reconstitution of hematopoiesis.

Soluble CD23 reliably reflects disease activity in B-cell chronic lymphocytic leukemia.

PURPOSE: This study was initiated to evaluate whether soluble CD23 (sCD23) reflects disease activity and tumor load in B-cell chronic lymphocytic leukemia (B-CLL) and to determine its prognostic potential for this disease. PATIENTS AND METHODS: The concentration of sCD23 was measured in the serum of 45 B-CLL patients, 50 patients with other lymphoproliferative disorders, and 41 healthy donors (HD). sCD23 serum levels from B-CLL patients were correlated with parameters of disease activity and total tumor mass (TTM) score. In selected cases, sCD23 was measured repeatedly over a 24-month period. Expression, density, and calculated total amount of membrane CD23 on peripheral-blood B-CLL cells, as well as its correlation to sCD23 levels in serum and supernatants, were determined. RESULTS: sCD23 in B-CLL patients serum was highly elevated as compared with other lymphoproliferative disorders, with the exception of immunocytoma (IC). Both advanced Rai stages and active forms of B-CLL were associated with higher levels of sCD23. There was a highly significant reciprocal relationship between sCD23 and lymphocyte count doubling time (LCDT). Serum sCD23 correlated positively with serum deoxythymidine kinase activity and TTM score, but not with absolute lymphocyte counts. The repetitive measurement of serum sCD23 showed the usefulness of this marker in monitoring disease progression in B-CLL. The total amount of membrane CD23 on in vitro-cultured B-CLL cells correlated significantly with sCD23 levels in the supernatant, whereas correlation between serum sCD23 and membrane CD23 on freshly isolated B-CLL cells was absent. CONCLUSION: Our results indicate that sCD23 is a highly sensitive and specific parameter with prognostic potential for B-CLL, which may be used as a tumor marker and may help to assess disease activity.

The diagnosis of endometrial carcinoma by means of jet wash and DNA flow-through cytophotometry.

Flow-through cytophotometric determination of nuclear DNA content on jet wash material from the endometrium was employed in the diagnosis of endometrial carcinoma. One hundred and thirty cases were studied. In comparison to results from cytodiagnosis, flow-through photometry yielded a false negative rate of 31.6% and a false positive rate of 42.2%. The false negative findings resulted in part from the small relative frequency of atypical cells in a mixed population of normal and atypical cells in certain cases. Besides this, we often found carcinomas with a diploid DNA stem line, which could not be distinguished cytophotometrically from normal corpus endometrium (Sandritter, 1952; Atkin et al., 1959; Hustin, 1976). The flow-through photometrically false positive findings may have resulted either from cell aggregates or from a nuclear DNA content elevated over the diploid value in proliferating cells (D. Wagner et al., 1968; D. Wagner and Richard, 1968). The observed false negative and false positive rates demonstrate that flow-through photometric determination of nuclear DNA content is unsuitable for the diagnosis of endometrial carcinoma.

The colposcopic screening.

The objective of this paper is a valuation of colposcopy as a screening tool. As database, 392 patients with histologically confirmed intraepithelial neoplasia were used. Colposcopic and cytologic findings were compared with the final histologic diagnosis. The following results were obtained: (1)The colposcopic findings correlated with the histologic diagnosis to a significantly higher degree than the cytologic findings. Depending upon the rate of dysplasia, the colposcopic findings predicted the diagnosis in 84-97%. (2) The false-negative rate of cytology in condylomatous lesions and mild dysplasia was high (39 and 26%, respectively), in particular in comparison with the false-negative rate of colposcopy of 5% for both lesions. Thus, a negative smear does not exclude consistently a dysplasia of the cervix. (3) The false-negative rate of cytology for the high grade lesions (CIN II and CIN III) was 13 and 1% respectively and, thus, lower than in the low grade lesions. There were, however, considerable discrepancies in comparison with the histologic rating of the lesion. In CIN III cytology correlated with histology in only 61%, colposcopy, however, 85% (P < 0.001). Our results demonstrate that colposcopy is an excellent tool for detecting HPV caused lesions especially subclinical lesions and CIN I. Colposcopy is also a corrective for the false-negative cyto-smear rate (about 20-40%). Thus, colposcopy may be used as an effective quality assurance method and an excellent screening method in that colposcopy is superior in grading dysplastic lesions of the cervix. The application of the European terminology was advantageous.

Human papillomavirus, type 16, DNA in multicentric anogenital neoplasia associated with idiopathic panmyelopathy. A case report.

A 27-year-old woman suffering from panmyelopathy for six years presented with a cervical low grade squamous intraepithelial lesion (SIL), vulvar high grade SIL and perianal squamous cell carcinoma with an inguinal metastasis. Southern blot hybridization with 32P-labeled human papillomavirus (HPV) DNA revealed HPV 16 DNA in varying copy numbers in material from the four locations. HPV 16 genomes persisting after surgery on the perianal tumor area were no longer detectable after betatron radiotherapy.

Human papillomavirus standardization and DNA cytophotometry in cervical intraepithelial neoplasia.

Human papillomavirus (HPV) standardization and DNA cytophotometry were carried out in 29 cases of cervical intraepithelial neoplasia (CIN) grades I-III. A prognostically unfavorable DNA distribution pattern with an aneuploid stemline was found in 14 of the 16 dysplasias with HPV types 16 and 18, while in 11 of 13 dysplasias with HPV types 6 and 11 there was a favorable DNA distribution with a euploid-polyploid stemline. Among 178 colposcopically, cytologically and histologically confirmed cervical lesions, there was a statistically significant incidence of HPV 16/18 infections in severe dysplasias and carcinomas, while HPV 6/11 was found predominantly in mild cervical lesions. It seems that CIN can be divided into high- and low-risk lesions not only by the degree of severity and the DNA distribution pattern but also by HPV typing.

DNA-cytometric diagnosis of prospective malignancy in borderline lesions of the uterine cervix.

Interactive DNA cytometry was used for the diagnosis of prospective malignancy in 48 smears with borderline lesions (mild and moderate dysplasias) of the uterine cervix. In addition, 183 smears with benign squamous epithelia, 38 with carcinoma in situ and 7 with invasive squamous carcinoma were also measured. Nuclear Feulgen-DNA measurements were performed using various methods, and the resulting data were analyzed by an algorithm for a DNA-cytophotometric diagnosis of malignancy. The results were compared with the data on follow-up and subsequent histologic studies in these cases. There was no false-positive diagnosis in the 183 benign smears and only 1 false-negative diagnosis in the 76 histologically proven squamous-cell carcinomas, which yields a specificity of 100% and a sensitivity of 98.6%. The sensitivity for the detection of subsequent histologically proven malignancy in cases with cytologically mild or moderate dysplasia amounted to 97%. In 13 borderline cases, there was a mean interval of 21 months between the taking of the cytologic smear on which the DNA diagnosis of malignancy was made and the date on which the histologic confirmation of malignancy was made. In 17% of the cytologically dysplastic cases, the DNA diagnosis of malignancy was not verified by subsequent histologic investigation. These results indicate that interactive DNA cytometry is able to detect prospective malignancy in smears from borderline lesions of the uterine cervix with a high sensitivity.

Lymphocyte subpopulations in malignant ascites of serous papillary ovarian adenocarcinoma. An immunocytochemical study.

The aim of this study was to investigate lymphocyte subpopulations in 17 patients with malignant ascites due to serous papillary adenocarcinoma of the ovary. Eight patients had not been treated prior to the study whereas nine patients had been treated by surgery and chemotherapy. A panel of monoclonal antibodies against surface markers that correlate with the immune functions of the lymphocytes was used. The lymphocyte subpopulations were identified by the immunoperoxidase adhesive slide assay, and the results in treated and untreated patients were compared. Both groups of patients showed lymphocytosis (41 +/- 25% and 33 +/- 14% of the total cells, respectively). The untreated patients had a significantly higher proportion of B cells (14 +/- 4% of lymphocytes) than did treated patients (7 +/- 2%). No differences were found between both groups regarding the helper-inducer/suppressor-cytotoxic T lymphocyte ratio. The proportion of lymphocytes expressing interleukin-2-receptors was higher in treated patients (6 +/- 2%) than in untreated patients (1.2 +/- 1%). Both groups showed a high percentage of natural killer/cytotoxic cells (17 +/- 7% and 18 +/- 5%, respectively). In the only chylous effusion in this study, there was an increase in helper-inducer and activated T lymphocytes. Future studies are required to document whether surface marker analysis of lymphocytes in malignant effusions may be useful for assessment of the prognosis and the results of treatment.

Malignant ascites of serous papillary ovarian adenocarcinoma. An immunocytochemical study of the tumor cells.

In 17 malignant peritoneal effusions due to papillary serous adenocarcinoma of the ovary, the reaction patterns of the tumor cells to monoclonal antibodies (MAbs) against surface antigens were studied and compared with the reaction patterns of mesothelial cells in the same effusions. The following surface markers were used with the adhesive slide method: epithelial membrane antigen (EMA), human epithelium-specific cell surface antigen (HEA-125), human endothelial antigen (BMA-120), carcinoembryonic antigen (CEA 3-13), an antibody against natural killer cells and cytotoxic cells (BMA-070), granulocyte antigen (Leu M1) and leukocyte antigen of class I (HLA-1). In all cases, from 30% to 95% of the tumor cells reacted with EMA and HEA-125. Tumor cells showed a positive staining with CEA 3-13 in only five cases. In all cases, from 75% to 95% of the tumor cells reacted positively with BMA-120. The reactivity of a few mesothelial cells with EMA and of all mesothelial cells with BMA-120 did not interfere with the identification of positive tumor cells since the reaction patterns were different. Interestingly, our study demonstrated that BMA-070, an MAb identifying natural killer cells and cytotoxic cells, is also a most useful tumor marker. The same was found to be true for Leu M1, an MAb originally thought to react only with granulocytes. The tumor cells showed a partial or total loss of the expression of HLA-1 reactivity. Since all cases were immunocytochemically positive for tumor cells while conventional cytology was positive in only 13 of the cases, the immunocytochemical analysis of malignant peritoneal effusions due to papillary serous adenocarcinoma of the ovary seems able to improve the cytologic diagnosis of the fluids.

Colposcopy, cervicography, speculoscopy and endoscopy. International Academy of Cytology Task Force summary. Diagnostic Cytology Towards the 21st Century: An International Expert Conference and Tutorial.

ISSUES: The colposcope was developed in 1925 and is well established in clinical gynecologic practice for defining and delineating cytologically detected lesions mainly of the cervix but also the vagina and vulva. Additionally, various endoscopic procedures in gastroenterology, pulmonary and urologic lesions enhance the cytologic detection and histologic verification of precancerous and cancerous lesions. The cost-effectiveness of all these devices and their applicability, particularly in countries with a limited health budget, is a major issue. This task force considered aspects of the present state of the art and the challenges in the 21st century. CONSENSUS POSITION: Automated cytology can interface with colposcopic examination in a number of significant ways. Automated cytologic analysis of conventional cervical smears can potentially direct colposcopic examination by predicting the nature of a lesion, assist in determining which patients should receive colposcopy and, in some settings, thereby reduce the number of colposcopies. Potentially, various combinations of automated cytology and colposcopy may be used to generate screening protocols that might result in more effective and inexpensive screening. The role of cervicography, or high-resolution cervical photography, as a screening device remains to be defined. Sensitivity for high grade lesions is generally no greater than that in cytology, and specificity appears lower. The interpretation of cervical photographs in triage of mildly abnormal cytology may prove to be useful in countries with established cytology programs. In areas of the world where cytology screening programs are not in place, the interpretation of cervical photographs may have its most dramatic effect. Cost-effectiveness analyses are needed. There are, at present, insufficient data for the evaluation of speculoscopy, a procedure using chemiluminescent illumination of the cervix for visualization of acetowhite areas. Basic training in colposcopy should be integrated into the residency programs of obstetrics and gynecology. Criteria for the adequate training of colposcopists should be developed. Continuing education programs in colposcopy should be developed when they are not already in existence. The cost-effectiveness of integrating colposcopy as a primary screening technique should be evaluated. Following a high-grade squamous intraepithelial lesion (HSIL) cytology result, colposcopically directed punch biopsy should be taken with or without endocervical curettage. This generally should precede the loop electrosurgical excision procedure (LEEP); however, in certain circumstances direct LEEP may be indicated. LEEP under colposcopic vision is an efficient way to treat an HSIL lesion of the cervix because the histologic extent and margins can be determined, unlike with laser surgery or cryosurgery. It is also more cost-effective than cold knife conization because general anesthesia and an operating room are unnecessary. Following LEEP, the endocervical canal should be examined colposcopically for any evidence of involvement. Lesions in the endocervix can then be removed with a different-shaped loop. Further research into Raman spectroscopy as a diagnostic aid in cervical pathology is needed, as is the use of micrococolpohysteroscopy for in vivo cytologic analyses, especially of the endocervical canal and transformation zone. Hysteroscopy is the most direct method for the diagnosis and treatment of intrauterine diseases. Hysteroscopic endometrial biopsy is more accurate than conventional biopsy methods. Cervical invasion of endometrial cancer can be detected by hysteroscopy. The depth of invasion, however, is more accurately determined by magnetic resonance imaging or computed tomography. ONGOING ISSUES: Many topics for ongoing research and/or implementation are mentioned under "Consensus Position," above. (ABSTRACT TRUNCATED)