Prevalence of synchronous bilateral tonsil squamous cell carcinoma: A retrospective study.

OBJECTIVES: The prevalence of synchronous bilateral tonsil cancer remains unexplored. To date, only 38 cases have been described in the literature. With an aim to substantiate the Danish recommendation of performing bilateral tonsillectomy in patients with suspected or proven tonsil cancer and in patients with cervical carcinoma metastasis from an unknown primary tumour, this study was undertaken to determine the prevalence of synchronous bilateral tonsil cancer. DESIGN: A retrospective review of all patients diagnosed with tonsil cancer in the period 2000-2015, Aarhus University Hospital, Denmark, was performed. RESULTS: Seven of 211 (3.3%) consecutive patients with tonsil cancer, who had undergone bilateral tonsillectomy (n = 180) or unilateral tonsillectomy (clinically normal side) combined with contralateral tonsil biopsy (side with suspected cancer) (n = 31), had synchronous bilateral tonsil cancer. Furthermore, dysplasia was found in the contralateral tonsil in two patients with unilateral tonsil cancer. Four of 171 (2.3%) patients with suspected unilateral tonsillar cancer had additional contralateral tonsil cancer. Three of 34 (8.8%) patients without clinical signs of tonsillar malignancy on any side (32 patients with carcinoma of unknown primary) had synchronous bilateral tonsil cancer. In none of the patients were bilateral tonsil cancer suspected. Patients with unilateral vs synchronous bilateral tonsil cancer had similar clinical and tumour characteristics. CONCLUSION: Knowledge on additional contralateral synchronous tonsil cancer is crucial for avoidance of early recurrence of oropharyngeal cancer in patients with tonsil cancer. Based on our findings, we recommend bilateral tonsillectomy in all patients with suspected or proven tonsil cancer and carcinoma of unknown primary.

A systematic review of Fusobacterium necrophorum-positive acute tonsillitis: prevalence, methods of detection, patient characteristics, and the usefulness of the Centor score.

The purpose of this study was to review the current evidence for an association between Fusobacterium necrophorum (FN) and acute tonsillitis (AT), to assess the prevalence of FN in AT, to identify the better FN detection method, and to characterize the clinical characteristics of FN-positive patients with uncomplicated AT. A systematic literature search was conducted in the PubMed, EMBASE, and SweMed+ databases for studies reporting on the recovery rates of FN in patients with AT. A total of 498 unique records were identified. Eleven studies were included in a qualitative synthesis and six studies were included in a meta-analysis. Considerable heterogeneity (I2 = 60 %) and risk of bias and confounders was found among the studies, though a subset of studies (prospective) had lower heterogeneity and higher study quality. FN was recovered significantly more frequently from patients (21.2 %) compared to healthy controls (7.6 %) (p < 0.001). FN recovery rates were similar between culture-based studies (20.3 %) and studies using polymerase chain reaction (PCR) methodology (22.2 %) (p = 0.462). The typical FN-positive AT patient was a smoking young (15-25 years) male presenting with tonsillar exudates and a Centor score of 2 or higher. A clear association between FN and AT was found. FN is likely to be a significant and prevalent pathogen in AT, especially in teenagers and young adults. However, no evidence for causality between FN and AT has been established and it is unexplored if timely antibiotic therapy directed against FN accelerates the resolution of symptoms and decreases the risk of complications.

Prevalence of tonsillar human papillomavirus infections in Denmark.

The incidence of tonsillar carcinomas associated with Human Papillomavirus (HPV) infection has increased dramatically over the last three decades. In fact, currently in Scandinavia, HPV-associated cases account for over 80 % of tonsillar carcinoma cases. Yet, the epidemiology and natural history of tonsillar HPV infections remains poorly characterized. Our aim was to characterize such infections in the Danish population in tumor-free tonsillar tissue. Unlike previous studies, we considered both palatine tonsils. We examined both tonsils from 80 patients with peritonsillar abscess (n = 25) or chronic tonsillar disease (n = 55). HPV was detected by nested PCR with PGMY 09/11 and GP5+/GP6+L1 consensus primers, and typed by sequencing. Samples were also analyzed using a higher-throughput method, the CLART HPV 2 Clinical Array Assay. The overall prevalence of HPV tonsillar infection was 1.25 % (1/80, 95 % CI 0.03-6.77 %) by nested PCR, and 0 % by CLART HPV2 Clinical Array. The HPV-positive patient was a 16-year-old female with recurrent tonsillitis and tonsillar hypertrophy. The type detected was HPV6. HPV was not detected in the contralateral tonsil of this patient. Compared to cervical HPV infections in Denmark, tonsillar HPV infections are 10- to 15-fold less frequent. In the HPV-positive patient in this study, HPV was detected in only one of the tonsils. This raises the possibility that prior studies may underestimate the prevalence of HPV infections, as they do not consider both palatine tonsils.

Incidence and microbiology of peritonsillar abscess: the influence of season, age, and gender.

To explore the correlations among the incidence, microbiology, season, gender, and age in patients with peritonsillar abscess (PTA) in order to identify risk factors for PTA development. All patients with PTA treated at private ENT practices in Aarhus County and in the Ear-Nose-Throat Departments at Aarhus University Hospital and Randers Hospital from January 2001 to December 2006 were included in the study. Age- and gender-stratified population data for Aarhus County for the same 6 years were obtained. The incidence rate of PTA increased from childhood to peak in teenage life and declined afterward gradually until old age. Girls predominated over boys until the age of 14 years. Subsequently, men were more frequently affected than women. Fusobacterium necrophorum (FN) was significantly more prevalent than group A Streptococcus (GAS) among patients aged 15-24 years (P < 0.001). In contrast, GAS was significantly more frequently recovered among children aged 0-9 years and adults aged 30-39 years compared with FN (P < 0.001 and P = 0.017 respectively). The seasonal variation of PTA was statistically insignificant (P = 0.437). However, GAS was significantly more frequently recovered in the winter and spring than in the summer (P = 0.002 and P = 0.036 respectively). There was a trend toward a higher incidence of FN infection during the summer than the winter (P = 0.165). Although the collected PTA incidence was stable throughout the year, the microbiology fluctuated with seasons. Patients aged 15-24 years are at an increased risk of PTA due to FN, which may have clinical implications for the diagnostic work-up and treatment of patients with acute tonsillitis.

Antibody development to Fusobacterium necrophorum in patients with peritonsillar abscess.

A polymicrobial mixture of aerobic and anaerobic bacteria is commonly recovered from peritonsillar abscess (PTA) aspirates. Previous studies have suggested a role for Fusobacterium necrophorum (FN) in the development of PTA. The purpose of the current study was to explore whether anti-FN antibodies were produced in patients with PTA. We developed a novel immunofluorescence-based method to measure anti-FN antibody levels in acute and convalescent sera from 15 patients with PTA and 47 patients with chronic tonsillar conditions (controls) undergoing acute or elective tonsillectomy, respectively. Bacterial cultures were performed on tonsillar cores and surfaces, pus aspirates, and blood. An increase in anti-FN antibody levels (of at least doubling of the previous level) was observed in 8 of 11 (73 %) PTA patients with FN-positive pus aspirate cultures (FN-positive patients). In contrast, the four FN-negative PTA patients did not have an increase in anti-FN antibody levels (p = 0.026). The change in anti-FN antibody levels in FN-positive PTA patients was also significantly greater than that for FN-positive electively tonsillectomized patients (p = 0.0014) and all electively tonsillectomized patients (p < 0.001). Our results validate FN as a significant and prevalent pathogen in PTA. This finding has implications for the diagnostic work-up of PTA and may also have implications for the treatment of acute tonsillitis.

A Retrospective Study of 291 Patients With Head and Neck Sarcomas: Treatment, Outcomes, and Prognostic Factors.

AIMS: Sarcomas constitute a group of rare malignant neoplasms, commonly subcategorized into soft tissue sarcomas (STS) and bone sarcomas. This study aims to describe the treatment modalities and outcome of head and neck sarcoma (HNS) patients in western Denmark and to identify prognostic factors for overall survival and recurrence in HNS patients. MATERIALS AND METHODS: The Aarhus sarcoma registry, The National Danish Sarcoma Database, and the Danish National Pathology Registry were used to identify HNS adult patients diagnosed between 1979 and 2022. RESULTS: Altogether, 291 patients were included in this study. The prevalent histological subtypes were undifferentiated pleomorphic sarcoma (16%; 48/291) and leiomyosarcoma (15%; 44/291) for STS patients (n = 230) and chondrosarcoma (10%; 28/291) and osteosarcoma (7%; 19/291) for bone sarcoma patients (n = 61). Surgery with curative intent was performed in 71% (164/230) and 70% (43/61) of STS and bone sarcoma patients, respectively. Clear resection was achieved in 59% (97/164) of STS patients and 44% (19/43) of bone sarcoma patients. Eighty-nine patients relapsed (STS n = 66, bone sarcoma n = 23) after a median time of 2.7/5.5 years for STS/bone sarcoma patients. The five-year overall survival rates were 45% for STS patients and 66% for bone sarcoma patients. The following factors were significantly, negatively associated with overall survival in STS patients: Age (hazard ratio (HR)) = 1.02, p < 0.001), tumour size ≥5 cm (HR = 1.75, p = 0.003), metastatic disease (HR = 3.17, p < 0.001), high grade tumour (HR = 2.24, p = 0.004), previous cancer (HR = 2.84, p < 0.001), and high Aarhus composite biomarker score (ACBS) (HR = 4.56, p = 0.001). For relapse in STS patients, higher tumour grade (HR = 3.19, p = 0.014), intralesional margins (HR = 2.84, p < 0.001), ≥2 previous cancers (HR = 3.00, p = 0.004), and high ACBS (HR = 3.29, p = 0.047), were negatively associated. For bone sarcomas only higher age (HR = 1.02, p = 0.049) and intralesional margins (HR = 2.91, p = 0.042) were significant negative factors for overall survival. CONCLUSION: Multiple prognostic factors for overall survival and relapse were identified, especially for STS patients.

The role of viruses in the pathogenesis of peritonsillar abscess.

Peritonsillar abscess (PTA) is the most frequent complication of acute tonsillitis and a prevalent cause for acute admission to otorhinolaryngology departments. Our aim was to examine the role of viruses in the pathogenesis of PTA, as this has not previously been considered. We examined both palatine tonsils from 25 patients undergoing acute tonsillectomy for PTA, using PCR-based assays for herpes simplex virus-1 and -2 (HSV-1 and -2), adenovirus, Epstein-Barr virus (EBV), influenza A and B, and respiratory syncytial virus (RSV) A and B. We similarly examined tonsils from 55 patients undergoing elective tonsillectomy due to chronic tonsillar conditions. These patients served as a control group, as they did not have a clinically apparent infection at the time of surgery. Only HSV-1 (5/80, 6.3%), adenovirus (11/80, 13.8%), and EBV (71/80, 88.8%) were detected in our study population. There was no statistically significant difference in the frequency of these viruses across different diagnostic groups. Quantification of EBV load demonstrated no differences between the PTA and the elective tonsillectomy group, nor between the abscessed and non-abscessed tonsil of PTA patients. In summary, our data do not support a significant role for the examined viruses in the pathogenesis of PTA.

Significant pathogens in peritonsillar abscesses.

Peritonsillar abscesses (PTA) are polymicrobial infections, with a diverse aerobic and anaerobic flora. The aim of the present study is to compare bacteriologic culture results from patients with PTA to those from patients undergoing elective tonsillectomy (clinically non-infected tonsils), to better elucidate the pathogenic significance of various isolates. A prospective study was conducted on 36 PTA patients undergoing acute tonsillectomy and on 80 electively tonsillectomised patients. Fusobacterium necrophorum (FN) and Streptococcus group A (GAS) were isolated significantly more frequently from the tonsillar cores of PTA patients, from both the abscessed (p = 0.001 and p = 0.046, respectively) and non-abscessed sides (p < 0.001 and p = 0.046, respectively), than from the tonsillar cores of electively tonsillectomised patients. Our findings indicate that FN and GAS are the prominent pathogens in PTA. In patients with PTA, the incidence of FN and GAS isolated from the abscessed tonsil was the same as from the non-abscessed contralateral side, and the growth was comparable by a semi-quantitative approach. Our findings suggest that FN is also of pathogenic importance in acute tonsillitis, and that FN growth is not a subsequent phenomenon once an abscess has formed. Our findings further suggest that other factors influence the development of PTA.

An overview of the microbiology of acute ear, nose and throat infections requiring hospitalisation.

This study is the first to provide an extensive overview of the microbiology of acute ear, nose and throat infections requiring hospitalisation. All 2,028 cases of acute infections admitted between 1 January 2001 and 31 December 2006 were reviewed to assess the use of pre-admission antibiotics, microbiological results, antibiotic and surgical management and length of hospitalisation. Infections of the oropharynx accounted for the vast majority of admissions, followed by ear infections, and cutaneous neck abscesses. Peritonsillar abscess was the most frequent diagnosis, accounting for over one third of admissions (39.8%, 808 out of 2,028). Complete microbiological data were available for 1,430 cultures, and were analysed for trends with respect to diagnosis, age, gender and use of pre-admission antibiotics. Forty-six percent (657 out of 1,430) of cultures yielded no growth or normal flora. This value increased to 77.0% (298 out of 387) in patients with pre-admission antibiotics. The distribution of microbiological isolates varied significantly between patients with and without pre-admission antibiotics, as well as with respect to age and gender. The most frequently isolated bacteria were Group A Streptococcus (13.7%, 196 out of 1,430), Fusobacterium necrophorum (13.6%, 195 out of 1,428) and Staphylococcus aureus (8.0%, 114 out of 1,430). Fusobacterium necrophorum was primarily isolated from cases of peritonsillar abscess (90.8%, 177 out of 195). This study suggests that F. necrophorum is a far more widespread pathogen in otorhinolaryngology than previously reported, and questions the value of routine culturing, as results rarely altered initial treatment. Further research on the microbiological variations with age and gender is recommended to better target culturing and treatment, and enhance our understanding of the pathogenesis of ear, nose and throat infections.

CA 19-9 and CA 125 levels in the sera of normal blood donors in relation to smoking history.

CA 19-9 and CA 125 serum levels were evaluated among smoking and nonsmoking healthy blood donors. Smoking did not elevate mean levels of either CA 19-9 or CA 125 in the sera of 496 of these blood donors from Philadelphia, PA. Mean CA 19-9 levels were slightly higher among females than among males. Among smokers there was a trend toward slightly increasing CA 19-9 serum levels with increased age, which was significant among the male donors. Trends toward slightly decreased mean serum levels of CA 125 among smokers were of borderline significance. Serum CA 19-9 and CA 125 levels in none of these donor subpopulations was elevated compared to levels reported by others for gastrointestinal or ovarian carcinoma patients, respectively. Therefore, smoking status should not interfere with use of either the CA 19-9 or CA 125 assays for diagnostic or monitoring applications.

Purification and composition of a novel gastrointestinal tumor-associated glycoprotein expressing sialylated lacto-N-fucopentaose II (CA 19-9).

Monoclonal antibody 1116NS 19-9 (Mab 19-9) exhibits selective reactivity with human gastrointestinal carcinomas and recognizes a carbohydrate determinant (CA 19-9) defined as a sialylated lacto-N-fucopentaose II. A scheme was devised for the purification of a human gastrointestinal tumor-associated glycoprotein antigen expressing CA 19-9 from colorectal carcinoma cell line SW1116 culture media in high yield. The key steps in the purification were immunoaffinity column chromatography with Mab 19-9 followed by reduction and alkylation of the specifically bound proteins in the presence of 6 M guanidine hydrochloride and a second Mab 19-9 immunoaffinity fractionation. The purified CA 19-9 containing glycoprotein ran as a single band on sodium dodecyl sulfate-polyacrylamide gradient gels with an apparent molecular mass of 210 kilodaltons. In the absence of detergents, this purified glycoprotein apparently reassociated to form aggregates of 600-2000 kilodaltons molecular mass as determined by size-exclusion chromatography. Amino acid analysis of CA 19-9 containing glycoprotein revealed that serine, threonine, and proline together accounted for greater than 35% of the amino acid residues, consistent with a mucin-like structure for the protein. Carbohydrate compositional analysis, however, was in contrast to a typical mucin with a fucose:mannose:galactose:N-acetylgalactosamine: N-acetylglucosamine:N-acetylneuraminic acid molar ratio of 4:1:12:2.5:5:5. The presence of both N-acetylgalactosamine and mannose suggested that both O- and N-linked oligosaccharides may exist on CA 19-9 containing glycoprotein. Protein and carbohydrate analyses indicated that this novel tumor-associated glycoprotein was 85% carbohydrate by weight. This purification procedure may be applicable to the isolation of other epithelial tumor-associated antigens.

Characterization of the CA 125 antigen associated with human epithelial ovarian carcinomas.

The murine monoclonal antibody OC125 reacts with an antigenic determinant (CA 125) found on a high-molecular-weight glycoprotein complex present in the serum of greater than 80% of women with epithelial ovarian cancer. The antigen expressing CA 125 (CA 125 antigen) isolated from the sera of ovarian carcinoma patients was shown by gel electrophoresis, molecular size exclusion chromatography, and buoyant density ultracentrifugation to have similar immunological and physical characteristics to antigen isolated from an ovarian cancer cell line (OVCA 433) and human milk. A composite sodium dodecyl sulfate: polyacrylamide:1.0% agarose gel resolved the CA 125 activity from the three sources of antigen into disperse bands of similar electrophoretic mobilities with apparent masses of 200,000 to 1 million daltons. The buoyant densities of the CA 125 antigen complexes from human serum, OVCA 433 cells, and human milk were in the range of 1.36 to 1.46 g/ml. Isolation of CA 125 antigen of higher purity from OVCA 433 supernatant was achieved by a series of steps including OC125 immunoaffinity chromatography. Subsequent resolution of this purified CA 125 antigen complex by sodium dodecyl sulfate:polyacrylamide gel electrophoresis gave rise to a band at approximately 200,000 daltons. Treatment of the CA 125 antigen from OVCA 433 cells with 10 mM periodic acid resulted in no loss of activity. Reduction and alkylation in 6 M guanidine-HCl or treatment at 100 degrees C for 20 min resulted in complete loss of activity. Exoglycosidase treatments did not result in loss of activity, whereas protease digestion eradicated all activity. These data strongly suggest that the CA 125 antigenic determinant is composed of, at least in part, conformationally dependent peptide.

Monoclonal antibody immunoradiometric assay for an antigenic determinant (CA 125) associated with human epithelial ovarian carcinomas.

CA 125 is an antigenic determinant expressed by greater than 80% of nonmucinous epithelial ovarian carcinomas. An immunoradiometric assay has been developed using a murine monoclonal antibody (OC125) to quantitate CA 125 in human serum. This immunoradiometric assay was optimized for specificity, sensitivity, and performance characteristics. Using a simultaneous immunoradiometric assay, the mean CA 125 concentration in 56 sera from healthy individuals was 11.2 +/- 5.4 (S.D.) units/ml, with 9.7 +/- 3.2 units/ml for 30 males and 13.1 +/- 6.8 units/ml for 26 females. A reference value of 35 units/ml included all 56 normals and excluded 86 of 105 (82%) ovarian carcinoma patients. This reference value also excluded 9 of 142 patients (6%) with benign diseases, but if the upper limit of normal was set at 65 units/ml, only 3 of 142 (2%) patients with benign diseases had elevated serum CA 125 levels, whereas 77 of 105 (73%) ovarian carcinoma patient sera remained positive. The ability of researchers, with this assay, to discriminate between CA 125 values in sera of patients with ovarian carcinoma and those of healthy individuals and patients with benign disease suggests that the assay deserves continued evaluation for monitoring and early diagnosis of ovarian cancer.

Coexpression of different antigenic markers on moieties that bear CA 125 determinants.

CA 125 has been extensively evaluated as a serum marker for monitoring patients with epithelial ovarian carcinoma. Recently, consideration has been given to the use of CA 125 as one component in a strategy for early detection of this disease. A number of benign conditions can, however, increase CA 125 in serum, limiting the utility of a single antigen determination for identifying ovarian cancer patients. Coexpression of different epitopes on the high molecular weight complexes that express CA 125 determinants might provide a more specific test for malignant disease, provided that adequate sensitivity were maintained. To determine how frequently determinants are coexpressed, macromolecular moieties containing CA 125 determinants have been isolated from ascites fluid of ovarian cancer patients by immunoaffinity chromatography. CA 125+ moieties have been probed on Western transfers with several murine monoclonal antibodies that recognize distinct tumor-associated epitopes. Marked heterogeneity was observed between patients with regard to antigenic determinants that could be coexpressed with CA 125. A fraction of ascites fluids from different ovarian cancer patients contained moieties which bound to OC 125 on a solid phase immmunoadsorbent and which also bound 125I-labeled monoclonal antibodies NS 19-9, B72.3, DF3, or the novel murine monoclonal antibody OC 3632 in a double determinant immunoradiometric assay. Serum samples were evaluated from patients with ovarian cancer and from apparently healthy individuals. Coexpression of TAG 72 and CA 125 was observed most frequently. When the double determinant assay for coexpression of TAG 72 and CA 125 was compared to assays for the individual antigens, the assay for coexpression was substantially less sensitive than those for the individual markers.

Tumorigenicity in athymic mice of the human colon carcinoma cell line SW1116 expressing the tumor-associated antigenic determinant CA 19-9.

The human colorectal carcinoma cell line SW1116 under optimal growth conditions synthesized and shed antigens bearing the monoclonal antibody-defined carbohydrate determinant CA 19-9. Antigen expressing CA 19-9 in cell culture supernatant was quantitated by an immunoradiometric assay for CA 19-9. Injection of SW1116 cells s.c. into athymic BALB/c mice resulted in the growth of moderately differentiated tumors possessing a distinct morphological resemblance to a typical adenocarcinoma of the colon. Intervals to tumor appearance were dependent on inoculum dose, but 95% of mice at both 5 X 10(6) and 10(7) cells/mouse developed tumors within 14 to 21 days. CA 19-9 antigen was detected in the sera of all nude mice with SW1116 tumors, and antigen concentration correlated (r = 0.77) with tumor volume throughout the 9-week study. The half-life of this antigen in serum following tumor excision from nude mice was 6.5 +/- 1.5 (S.D.) hr. Carcinoembryonic antigen was also detected in serum from mice bearing SW1116 tumors by an immunoradiometric assay for carcinoembryonic antigen, but its concentration correlated (r = 0.86) with tumor volume for only the first 4 weeks of tumor growth. Significant levels of endogenous immunoglobulin G1 and immunoglobulin G3 antibodies to CA 19-9 antigen were found in the serum of nude mice with SW1116 tumors by radioimmunodiffusion, but no apparent relationship between antibody titer and tumor growth or CA 19-9 antigen level in serum was evident. This tumor model may be useful in devising radioimmunodetection and immunotherapeutic strategies for primary and metastatic human colon carcinomas.

[The influence of transport on organ cultured corneas. An experimental study of porcine corneal endothelium]. / Einfluss des Transportes auf organkultivierte Hornhäute. Experimentelle Studie über die Schädigung des Schweinehornhautendothels.

BACKGROUND: To investigate the influence of transport on the quality of corneas in organ culture, based on the vitality of the corneal endothelium. METHODS: Transport was simulated for 222 porcine corneal disks. These were placed in standard transportation containers filled with organ culture medium II, kept in an incubator and then shaken on a laboratory shaker. RESULTS: Agitated corneas at all acceleration rates always showed less endothelial cell damage than corneas kept motionless. The best condition was found after maximum accelerations of 0.10 g and 0.72 g. Damage consisted predominantly of disseminated cell loss and circular cell damage. A storage temperature of 37 degrees C caused most harm to the endothelium. CONCLUSION: The reduction in endothelial damage found in corneas after agitation can be attributed to a better distribution of nutrients in the transportation container. Assuming transferability, a better quality of human grafts can be achieved by inducing slight motions of corneas in organ culture.

The oxidative metabolism of estradiol conditions postmenopausal bone density and bone loss.

Because lifelong exposure to estrogen is a strong determinant of bone mass, we asked whether metabolic conversion of estrogen to either inactive or active metabolites would reflect postmenopausal bone mineral density (BMD) and rate of bone loss. Biochemical markers of inactive estrogen metabolites, urinary 2-hydroxyestrogen (2OHE1) and 2-methoxyestrogen (2MeOE1), and active metabolites, urinary 16alpha-hydroxyestrone (16alphaOHE1), estradiol (E2), and estriol (E3), were determined in 71 untreated, healthy postmenopausal women (age, 47-59 years) followed prospectively for 1 year. Urinary 2MeOE1 was correlated negatively with baseline vertebral (anteroposterior [AP] projection, r = -0.23 andp < 0.05; lateral view, r = -0.27 and p < 0.05) and proximal femur bone density measured by dual-energy X-ray absorptiometry (DXA; total, r = -0.38 and p < 0.01; neck, r = -0.28 and p = 0.02; trochanter, r = -0.44 and p < 0.01). BMDs of women in the lowest quartile of urinary 2MeOE1 (< 15 ng/g) were significantly higher than those in the highest quartile at all skeletal sites (p < 0.05). Likewise, women in the lowest quartile of urinary 2OHE1/16alphaOHE1 ratio (< 1.6) did not experience bone loss after 1 year, in contrast to women in the higher quartiles. We propose that the rate of inactivation of estrogens through 2-hydroxylation may contribute to postmenopausal osteoporosis.

Application of an improved ELISA assay to the analysis of urinary estrogen metabolites.

Work from Strang and other laboratories has established that the 2-/16 alpha-hydroxyestrone ratio is inversely correlated with the risk for breast and cervical cancer. In order to measure these metabolites in urine samples, it is essential to have an assay for these compounds that is both sensitive and reproducible. The present paper describes such an ELISA assay, which overcomes problems that existed in prior approaches to measuring these compounds. The new ELISA procedure supplies greater sensitivity and reproducibility than earlier assay procedures. The ELISA assay has also been found to correlate well with the GC-MS procedure of Adlercreutz.

Monoclonal antibody-based enzyme immunoassay for simultaneous quantitation of 2- and 16 alpha-hydroxyestrone in urine.

Alterations in the metabolism of estrogen have been implicated as an important factor in the etiology of diseases such as gynecological cancers and lupus erythematosus. The major metabolites of estradiol are hydroxylated at the C-2 or C-16 alpha position yielding products with estrogen antagonist and agonist activities, respectively. A sensitive and specific immunodiagnostic assay to determine the balance between these competing pathways might serve as a routine biomarker for management of estrogen-related diseases. We describe here the generation of high affinity, specific murine monoclonal antibodies to 2-hydroxyesterone and 16 alpha-hydroxyestrone by high efficiency fusion protocols. With these antibodies, we have developed a rapid and simple enzyme immunoassay (EIA) kit for the simultaneous quantitation of 2- and 16 alpha-hydroxyestrone in unextracted urine. Initial validation studies established that urinary metabolite 2- and 16 alpha-hydroxyestrone concentrations found by the EIA correlate well with values found by gas chromatography-mass spectroscopy. Preliminary studies with the EIA kit found total recovery of metabolites from spiked urine samples. The EIA inter- and intra-assay coefficients of variation for 2-hydroxyestrone and 16 alpha-hydroxyestrone and the ratio of 2-hydroxyesterone to 16 alpha-hydroxyestrone with the current EIA kit were consistently less than 9%. This kit, designated ESTRAMET 2/16 may provide an important new tool for research in estrogen-related diseases.