Preoperative N stage evaluation in advanced gastric cancer patients using multidetector CT: can the sum of the diameters of metastatic LNs be used for N stage evaluation?

AIM: To compare the diagnostic performance of total counts of metastatic lymph nodes (LN-sum) and conventional multidetector (MD) computed tomography (CT) staging in the nodal evaluation of advanced gastric cancer (AGC) patients. MATERIALS AND METHODS: In total, 127 consecutive patients who underwent preoperative MDCT and gastrectomy for AGC were identified. Metastatic LNs on MDCT were defined as LNs with a short axis ≥8 mm, marked or heterogeneous enhancement, and morphological features (central necrosis, round shape, clustering). The sensitivity, specificity, accuracy, positive predictive value (PPV), and negative predictive value (NPV) of the N-stage using LN-sum and conventional MDCT staging were generated and compared. In addition, metastatic LN counts between the MDCT and the histopathological examinations and correlation between LN-sum and histopathological nodal status were analysed. RESULTS: The total counts of metastatic LNs on MDCT was significantly smaller than those detected in histopathological assessments (p<0.0001). LN-sum showed significant correlation with the pathological N stage and the number of metastatic LNs (rho=0.69, 0.73, p<0.0001). The areas under the receiver operating characteristic curve were 0.896, and 0.835, for N stage ≥N2 and N3, with cut-off values of 12.5 and 23.5 mm, respectively. LN-sum provided better diagnostic performance than conventional MDCT staging for discriminating N0-2 versus N3; sensitivity, accuracy, PPV and NPV of LN-sum were significantly higher (80.4 versus 52.2%, 81.1 versus 68.5%, 71.2 versus 57.1%, and 88 versus 74.1%). CONCLUSION: LN-sum may be sufficiently useful in assessing the N3 stage of AGC and may help to plan appropriate therapy for AGC patients.

Characterization of cottage cheese using Weissella cibaria D30: Physicochemical, antioxidant, and antilisterial properties.

This study aimed to evaluate the potential of Weissella cibaria D30 as an adjunct culture in cottage cheese, including an assessment of antioxidant, antilisterial, and compositional parameters. Cottage cheese samples were manufactured using a commercial starter culture and probiotic strains Lactobacillus rhamnosus GG (GG) or W. cibaria D30 (W) and without probiotic (control). Samples were stored at 4 ± 1°C for 28 d. Bacterial cell counts (log cfu/g) of control, GG, and W samples were counted at 0, 7, 14, 21, and 28 d. Counts of W. cibaria D30 in the W samples remained at 6.85 log cfu/g after 28 d. Total solids, fat, protein, ash, and pH were measured and no significant differences were observed in compositional parameters or pH after 28 d of storage in all cheeses except those inoculated to Listeria monocytogenes. To measure the antilisterial effect, Listeria monocytogenes was inoculated into the cottage cheese samples and bacterial cell counts were obtained at 0, 6, 12, 24, 48, 72, 96, 120, and 144 h. Listeria monocytogenes counts were less than the analytical limit of detection (<10 cfu/g) in the inoculated GG and W samples, whereas the counts of L. monocytogenes in the inoculated control sample remained at 3.0 log cfu/g after 144 h. We used the DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS [2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)] radical scavenging activity assays to assess antioxidant activity: GG and W samples exhibited significant increases in antioxidant activity compared with the control sample. These results indicate that W. cibaria D30 has potential as an adjunct culture in the dairy industry.

Identification of Novel Macropinocytosing Human Antibodies by Phage Display and High-Content Analysis.

Internalizing antibodies have great potential for the development of targeted therapeutics. Antibodies that internalize via the macropinocytosis pathway are particularly promising since macropinocytosis is capable of mediating rapid, bulk uptake and is selectively upregulated in many cancers. We hereby describe a method for identifying antibodies that internalize via macropinocytosis by screening phage-displayed single-chain antibody selection outputs with an automated fluorescent microscopy-based high-content analysis platform. Furthermore, this method can be similarly applied to other endocytic pathways if other fluorescent, pathway-specific, soluble markers are available.

Mixed endometrial stromal and smooth muscle tumor of the uterus in a postmenopausal woman: morphologic and immunohistochemical features.

Mixed endometrial stromal and smooth muscle tumor of the uterus is a rare occurrence, and it is truly challenging to diagnose or dif- ferentiate mesenchymal tumors of the uterine corpus, due to their many overlapping features. In most cases, the gross pathology of mixed endometrial stromal and smooth muscle tumor differs from that of pure endometrial stromal and pure smooth muscle tumors. A 59-year-old postmenopausal woman presented with vaginal spotting, low abdominal pain, and an uterine mass. Subsequent pelvic magnetic resonance imaging revealed a 4.0x3.8x3.4-cm sized uterine mass with enhancement. The mass showed restricted diffusion on diffusion-weighted images, and thus, was suspected to be uterine sarcoma rather than degenerative leiomyoma. Levels of tumor markers, CA 125, CA 19-9, and SCC, were within their normal ranges. The patient underwent exploratory laparotomy. Morphological and immunohistochemical evaluations were performed, and a final diagnosis of mixed endometrial stromal and smooth muscle tumor of the uterus was rendered. Her postoperative course was uneventful, and aromatase inhibitor adjuvant therapy was administered.

Short communication: Physicochemical and antioxidant properties of Cheddar-type cheese fortified with Inula britannica extract.

Cheddar-type cheese was fortified with the antioxidant Inula britannica flower extract (IBE). Cheddar-type cheeses manufactured with varying concentrations of IBE (0, 0.25, 0.5, 0.75, and 1% wt/vol) were analyzed during storage at 4°C, 0, 1, 2, and 3 wk after production. Higher IBE concentrations resulted in higher protein and ash contents, with a concomitant decrease in pH, total solid, and fat content relative to the unfortified control cheese. The total phenolic content also increased with IBE concentration, but decreased over longer storage periods. The antioxidant activities of the cheeses, determined as 2,2-diphenyl-1-picrylhydrazyl (DPPH) free-radical scavenging activity and ferric thiocyanate assay results, increased proportionally to the total phenolic content. The highest antioxidant effect was observed in the 1% IBE-fortified cheese, showing 79 and 86% antioxidant effects in the DPPH and ferric thiocyanate assays, respectively. At the 1-wk time point, the 5 cheese preparations underwent sensory evaluation for odor, taste, texture, color, and overall quality, determined using a descriptive analysis by a trained panel (n=20). The addition of IBE resulted in some increases in extract odor and taste. Overall, IBE showed good potential as an antioxidant supplement for dairy products.

Short communication: Physicochemical and antioxidant properties of milk supplemented with red ginseng extract.

This study investigated the effect of red ginseng extract (RGE) on the physicochemical properties, sensory test, and antioxidant activity of milk. The milk samples with RGE added at 0.5, 1, 1.5, and 2% were analyzed during storage at 4°C. The physicochemical properties included composition of milk, pH, titratable acidity, and color. The antioxidant activity of milk samples was determined using the 2,2-diphenyl-1-picrylhydrazyl method, ß-carotene bleaching assay, and ferric thiocyanate assay. An increase in the amount of RGE in milk resulted in an increase of lactose and total solids content, titratable acidity, and a* and b* values, whereas fat and protein contents remained unchanged. Also, pH and L* value decreased. The antioxidant activity of milk samples supplemented with RGE was higher than that of the control sample. Sensory evaluation was performed using a quantitative descriptive analysis. Two types of samples were used: (1) sterilized milk fortified with RGE (0.5, 1, 1.5, and 2%) and (2) 2% RGE, 2% RGE with oligosaccharide, and 2% RGE with oligosaccharide and cyclodextrin. The addition of oligosaccharide and cyclodextrin could effect an increase of sweetness, a decrease of bitterness and flavor of RGE, and aftertaste. Therefore, milk supplemented with RGE could be useful as a functional food.

The effectiveness of gadolinium MRI to improve target delineation for radiotherapy in hepatocellular carcinoma: a comparative study of rigid image registration techniques.

To achieve consistent target delineation in radiotherapy for hepatocellular carcinoma (HCC), image registration between simulation CT and diagnostic MRI was explored. Twenty patients with advanced HCC were included. The median interval between MRI and CT was 11 days. CT was obtained with shallow free breathing and MRI at exhale phase. On each CT and MRI, the liver and the gross target volume (GTV) were drawn. A rigid image registration was taken according to point information of vascular bifurcation (Method[A]) and pixel information of volume of interest only including the periphery of the liver (Method[B]) and manually drawn liver (Method[C]). In nine cases with an indefinite GTV on CT, a virtual sphere was generated at the epicenter of the GTV. The GTV from CT (VGTV[CT]) and MRI (VGTV[MR]) and the expanded GTV from MRI (V+GTV[MR]) considering geometrical registration error were defined. The underestimation (uncovered V[CT] by V[MR]) and the overestimation (excessive V[MR] by V[CT]) were calculated. Through a paired T-test, the difference between image registration techniques was analyzed. For method[A], the underestimation rates of VGTV[MR] and V+GTV[MR] were 16.4 ± 8.9% and 3.2 ± 3.7%, and the overestimation rates were 16.6 ± 8.7% and 28.4 ± 10.3%, respectively. For VGTV[MR] and V+GTV[MR], the underestimation rates and overestimation rates of method[A] were better than method[C]. The underestimation rates and overestimation rates of the VGTV[MR] were better in method[B] than method[C]. By image registration and additional margin, about 97% of HCC could be covered. Method[A] or method[B] could be recommended according to physician preference.

Is elective nodal irradiation beneficial in patients with pathologically negative lymph nodes after neoadjuvant chemotherapy and breast-conserving surgery for clinical stage II-III breast cancer? A multicentre retrospective study (KROG 12-05).

BACKGROUND: To evaluate the effects of elective nodal irradiation (ENI) in clinical stage II-III breast cancer patients with pathologically negative lymph nodes (LNs) (ypN0) after neoadjuvant chemotherapy (NAC) followed by breast-conserving surgery (BCS) and radiotherapy (RT). METHODS: We retrospectively analysed 260 patients with ypN0 who received NAC followed by BCS and RT. Elective nodal irradiation was delivered to 136 (52.3%) patients. The effects of ENI on survival outcomes were evaluated. RESULTS: After a median follow-up period of 66.2 months (range, 15.6-127.4 months), 26 patients (10.0%) developed disease recurrence. The 5-year locoregional recurrence-free survival and disease-free survival (DFS) for all patients were 95.5% and 90.5%, respectively. Pathologic T classification (0-is vs 1 vs 2-4) and the number of LNs sampled (<13 vs ≥13) were associated with DFS (P=0.0086 and 0.0012, respectively). There was no significant difference in survival outcomes according to ENI. Elective nodal irradiation also did not affect survival outcomes in any of the subgroups according to pathologic T classification or the number of LNs sampled. CONCLUSIONS: ENI may be omitted in patients with ypN0 breast cancer after NAC and BCS. But until the results of the randomised trials are available, patients should be put on these trials.

Diffusion-weighted MRI for differentiation of benign from malignant lesions in the gallbladder.

AIM: To investigate the value of diffusion-weighted imaging (DWI) for differentiating benign from malignant gallbladder lesions. MATERIALS AND METHODS: One hundred and twenty-six patients who had undergone magnetic resonance imaging (MRI) with DWI, in whom the histopathological diagnosis of their gallbladder lesions was confirmed by biopsy or surgery were retrospectively analysed. Thirty-six malignant and 90 benign lesions were included. Two radiologists categorized gallbladder lesions into seven types on two imaging sets [T2-weighted imaging (WI) alone and combined T2WI and DWI (b = 800 s/mm(2))] according to the presence of wall thickening, layered patterns, morphology of the mass, and diffusion restriction. Disagreements were resolved in consensus. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of each imaging set for diagnosing gallbladder carcinoma were calculated. The diagnostic performance of each imaging set was calculated using receiver operating characteristic (ROC) curve analysis. Additionally, ADC values of malignant and benign gallbladder lesions were compared separately for 1.5 and 3 T MRI. RESULTS: The sensitivity, specificity, PPV, and NPV of diagnosis at T2WI were 97.2%, 86.7%, 74.5%, and 98.7%, respectively. The sensitivity, specificity, PPV, and NPV using combined T2WI and DWI were 97.2%, 92.2%, 83.3%, and 98.8%, respectively. Diagnostic accuracy for gallbladder carcinoma slightly improved after adding DWI, from 0.92 to 0.95 (p < 0.05). ADC values for gallbladder carcinoma were significantly lower than those for benign lesions. Mean ADC values of malignant and benign lesions were 0.97 ± 0.25 × 10(-3) and 1.72 ± 0.56 × 10(-3) mm(2)/s, respectively, at 1.5 T (p < 0.001), and 1.04 ± 0.38 × 10(-3) and 2.2 ± 0.72 × 10(-3) mm(2)/s, respectively, at 3 T (p < 0.001). CONCLUSION: DWI can improve diagnostic accuracy for differentiating benign from malignant gallbladder lesions.

Interfractional variability in intensity-modulated radiotherapy of prostate cancer with or without thermoplastic pelvic immobilization.

PURPOSE: To determine the variability of patient positioning errors associated with intensity-modulated radiotherapy (IMRT) for prostate cancer and to assess the impact of thermoplastic pelvic immobilization on these errors using kilovoltage (kV) cone-beam computed tomography (CBCT). MATERIALS AND METHODS: From February 2012 to June 2012, the records of 314 IMRT sessions in 19 patients with prostate cancer, performed with or without immobilization at two different facilities in the Korea University Hospital were analyzed. The kV CBCT images were matched to simulation computed tomography (CT) images to determine the simulation-to-treatment variability. The shifts along the x (lateral)-, y (longitudinal)- and z (vertical)-axes were measured, as was the shift in the three dimensional (3D) vector. RESULTS: The measured systematic errors in the immobilized group during treatment were 0.46 ± 1.75 mm along the x-axis, - 0.35 ± 3.83 mm along the y-axis, 0.20 ± 2.75 mm along the z-axis and 4.05 ± 3.02 mm in the 3D vector. Those of nonimmobilized group were - 1.45 ± 7.50 mm along the x-axis, 1.89 ± 5.07 mm along the y-axis, 0.28 ± 3.81 mm along the z-axis and 8.90 ± 4.79 mm in the 3D vector. The group immobilized with pelvic thermoplastics showed reduced interfractional variability along the x- and y-axes and in the 3D vector compared to the nonimmobilized group (p < 0.05). CONCLUSION: IMRT with thermoplastic pelvic immobilization in patients with prostate cancer appears to be useful in stabilizing interfractional variability during the planned treatment course.

MDCT features in the differentiation of T4a gastric cancer from less-advanced gastric cancer: significance of the hyperattenuating serosa sign.

OBJECTIVE: The purpose of our study was to evaluate CT findings to differentiate between T4a and less advanced gastric cancers. METHODS: The institutional review board approved this study and waived informed consent. This study included 228 retrospectively identified patients with surgically confirmed gastric cancer (138 T1, 25 T2, 24 T3 and 41 T4a) and who had also undergone pre-operative CT scan. Transverse and multiplanar reconstruction scans were reviewed in consensus by two other blinded radiologists. The following CT findings that differentiate T4a from less advanced cancers were evaluated: nodular or an irregular outer layer of the gastric wall, haziness of the perigastric fat and a hyperattenuating serosa sign. The CT features of T4a and less advanced gastric cancers were compared by means of univariate and multivariate analyses. RESULTS: In univariate analysis, nodular or an irregular outer layer of the gastric wall, haziness of the perigastric fat and the hyperattenuating serosa sign were significant in differentiation between T4a and less advanced gastric cancers. In addition, nodular or an irregular outer layer of the gastric wall and the hyperattenuating serosa sign were significant in differentiation between T3 and T4a. In multivariate logistic analysis, the hyperattenuating serosa sign was the most significant finding in differentiation between T3 and T4a (odds ratio, 4.210; 95% confidence intervals, 1.581-11.214; p=0.004). CONCLUSION: The hyperattenuating serosa sign may be a useful CT finding in differentiation between T4a and less-advanced gastric cancers. ADVANCES IN KNOWLEDGE: The hyperattenuating serosa sign is associated with gastric cancer with invading the serosa and can facilitate planning of the optimal pre-operative evaluation and treatment.

The relationship between mast cell density and tumour grade in transitional cell carcinoma of the bladder.

This study investigated the relationship between mast cell numbers and the grade of bladder transitional cell carcinoma (TCC). Bladder TCC biopsies were obtained via transurethral resection and 45 stage T1 specimens were included in the study. Specimens were sorted into two groups, low grade (grade I) and high grade (grades II and III). Samples were stained using haematoxylin and eosin, toluidine blue and immunohistochemical staining for tryptase. Mast cells were examined by light microscopy and cell density was recorded. Mast cell density was significantly higher in high-grade TCC than low-grade TCC. There was also a significant relationship between the number of mast cells identified using toluidine blue staining or immunohistochemical staining for tryptase. Detailed studies of mast cell function will enable the development of more effective antitumour therapies via mast cell manipulation.

CT differentiation of pyogenic liver abscesses caused by Klebsiella pneumoniae vs non-Klebsiella pneumoniae.

OBJECTIVE: Klebsiella pneumoniae is one of the organisms most commonly isolated from pyogenic liver abscesses in Asian populations. We compared CT findings in liver abscesses caused by K. pneumoniae with those caused by other bacterial pathogens. METHODS: Of 214 patients with liver abscesses examined over a 5 year period, 129 patients with positive blood or aspirate cultures were enrolled. The patients were divided into two groups: the K. pneumoniae monomicrobial liver abscess (KLA) group (n = 59) and the non-K. pneumoniae monomicrobial or polymicrobial liver abscess (non-KLA) group (n = 70). Two radiologists blinded to the culture results evaluated the CT images, recording the number, size, location and configuration of abscesses, the thickness of the abscess wall, the pattern of rim enhancement, septal enhancement, the double target sign, internal necrotic debris, internal gas bubbles and underlying biliary disease. The presence of diabetes and metastatic infection was also compared between groups. Statistical analyses were performed using univariate (Student's t-test and χ(2) test) and multivariate analyses. RESULTS: Multivariate analysis showed that a thin wall, necrotic debris, metastatic infection and the absence of underlying biliary disease were the most significant predictors of KLA. When three of the four criteria were used in combination, a specificity of 98.6% was achieved for the diagnosis of KLA. CONCLUSION: A thin-walled abscess, internal necrotic debris, the presence of metastatic infection and the absence of underlying biliary disease may be useful CT findings in the early diagnosis of K. pneumoniae liver abscesses.

Adherence and anticarcinogenic effects of Bacillus polyfermenticus SCD in the large intestine.

AIMS: To investigate the probiotic properties of Bacillus polyfermenticus SCD such as their adherence to Caco-2 cells and anticarcinogenic effects on human colon cancer cells and rat colon cancer carcinogenesis. METHODS AND RESULTS: The cell surface hydrophobicity and cell agglutination of B. polyfermenticus SCD was 64.04 +/- 1.25% and 297.3 +/- 8.7 mg ml(-1), respectively. This strain was strongly adherent to Caco-2 cells. Bacillus polyfermenticus SCD was also found to inhibit the growth of colon cancer cells in a dose-dependent manner as detected by the MTT assay. After 10 weeks of B. polyfermenticus SCD supplementation with 3 x 10(6) CFU day(-1) in F344 male rats, dimethylhydrazine-induced aberrant crypts and preneoplastic lesions decreased by 40% compared with the control rats. CONCLUSIONS: Bacillus polyfermenticus SCD has strong adherent properties in the colon, and anticarcinogenic effects in vitro and in vivo. SIGNIFICANCE AND IMPACT OF THE STUDY: These new findings on the characteristics of B. polyfermenticus SCD will be valuable in the evaluation of this commercial probiotic. Also, B. polyfermenticus SCD can be useful for the inhibition of colon cancer cells as an ingredient of medicinal foods or new drugs.

Urinary ascites and anuria caused by bilateral fungal balls in a premature infant.

A case is reported of anuria and urinary ascites secondary to bilateral ureteropelvic obstruction by fungal balls. Management consisted of bilateral nephrostomy drainage with local irrigation with amphotericin B, and systemic antifungal treatment without surgery. Aspiration by paracentesis was performed for the urinary ascites and continuous drainage through an 8 Fr pig tail catheter for the urinoma. The literature on renal fungus balls in neonates and infants is reviewed.

Two cases of perineal ectopic testis.

PURPOSE: Perineal ectopic testis is seen very rarely and here we report on two patients with perineal ectopic testis. METHODS/RESULTS: We experienced two cases of perineal ectopic testis on which orchiopexies to the corresponding hemiscrotums were performed. CONCLUSIONS: On surgical exploration, the gubernaculum testes were found to lie fixed to the perineum.

Evidence for basolateral but not apical membrane localization of outwardly rectifying depolarization-induced Cl(-) channel in airway epithelia.

The rat primary cultured-airway monolayer had been an excellent model for deciphering the ion channel after nystatin permeabilization of its basolateral or apical membrane (Hwang et al., 1996). After apical membrane permeabilization of rat primary cultured-airway monolayer, 4,4'-diisothiocyanatostilbene-2, 2'-disulfonic acid (DIDS)-sensitive outwardly rectifying depolarization-induced Cl(-) (BORDIC) currents were observed across the basolateral membrane in symmetrical NMG-Cl solution in this study. No significant Cl(-) current induced by the application of voltage clamping was observed across the apical membrane in symmetrical NMG-Cl solution after basolateral membrane permeabilization. The halide permeability sequence for BORDIC current was Br(-) = I(-) > Cl(-). BORDIC current was not affected by basolaterally applied bumetanide (0.5 mm). Basolateral DIDS (0.2 mm) but not apical DIDS inhibited CFTR mediated short-circuit current (I(sc)) in an intact monolayer of rat airway epithelia, a T84 human colonal epithelial cell line, and a Calu-3 human airway epithelial cell line. This is the first report showing that depolarization induced Cl(-) current is present on the basolateral membrane of airway epithelia.

Hazardous alcohol use: its delineation as a subthreshold disorder, and approaches to its diagnosis and management.

The last 20 years have seen a significant paradigm shift in how we view alcohol misuse. The dichotomous model of "alcoholism" and "normal drinking" has now been replaced by the concept of a spectrum of disorders. In this new framework, "hazardous alcohol use" is defined as a repeated pattern of drinking that confers the risk of harmful consequences. It is a typical example of a subthreshold disorder. Where actual physical or psychological harm or social problems have occurred, the terms "harmful alcohol use" and "alcohol abuse," respectively, are applied. These conditions would typically be considered to be above the clinical threshold. The most severe disorder, alcohol dependence, is a psychobiological syndrome with often severe physical, psychological, and social sequelae. This article describes how the concept of hazardous alcohol use originated, and reviews the intervention techniques that have been developed to induce and assist hazardous drinkers to reduce their consumption to nonhazardous levels. The findings from a series of World Health Organization (WHO) collaborative studies on brief interventions for hazardous alcohol use are described. This work has resulted in the development of the Alcohol Use Disorders Identification Test (AUDIT) screening instrument, which can detect over 90% of hazardous drinkers in a range of settings, and the demonstration that 5 minutes' structured advice can reduce hazardous consumption by 30%. The later phases of this program of work have examined strategies to promote the dissemination of brief interventions for hazardous alcohol use throughout primary health care, and the nationwide, systematic, and sustained utilization of these interventions.

Alternatamides A-D: new bromotryptamine peptide antibiotics from the Atlantic marine bryozoan Amathia alternata.

Four new bromotryptamine peptides, alternatamides A-D (1-4), have been isolated from the Atlantic bryozoan Amathia alternata. The structures of the alternatamides were assigned primarily on the basis of 2D NMR data. The absolute stereochemistry of the N-methylleucine amino acid was shown to be L (2'S) by hydrolysis and comparison with standards. The alternatamides show modest antibacterial activities against several Gram-positive bacteria.

Frequent allelic loss at chromosome arm 3p is distinct from genetic alterations of the Von-Hippel Lindau tumor suppressor gene in head and neck cancer.

Previous molecular genetic studies revealed that allelic loss of chromosome arm 3p is a frequent event in upper aerodigestive tract squamous cell carcinoma (UADT SCC). Recently, the Von-Hippel Lindau (VHL) tumor suppressor gene was identified at chromosome band 3p25-26. To determine if the VHL locus is altered in these tumors, a paired series of 26 tumors and blood from patients with UADT SCC that were previously shown to exhibit allelic loss of 3p were tested for LOH surrounding the VHL locus using four different polymorphic markers. All of the samples (100%) exhibited LOH for at least 1 marker. However, no LOH was detected using a polymorphism within exon 1 of the VHL gene which was informative for 18 of the 26 cases. Furthermore, mutations of the VHL gene could not be identified by single-strand conformation polymorphism, dideoxyfingerprint or direct DNA sequence analysis. In addition, the VHL gene was not inactivated by hypermethylation in any of the 26 tumor samples studied. These findings demonstrate that allelic loss of chromosome arm 3p in UADT SCC involves regions surrounding the VHL locus but does not include the VHL gene. The VHL gene, therefore, does not appear to be involved in the pathogenesis of UADT SCC.