Stability of PreservCyt for Hybrid Capture (HC II) HPV test.

The Food and Drug Administration (FDA) has approved the Hybrid Capture II (HC II) assay to test for the presence of high-risk types of human papilloma virus (HPV) DNA using specimens in PreservCyt fixative for up to 21 days after collection. The ability of HC II to determine the presence of HPV DNA in actual patient samples after longer periods of storage has not been shown. To determine if specimens older than 21 days can yield useful results, 207 patient specimens that had been tested for HPV DNA by HC II (primary test) were tested again after a significant period of storage ranging from approximately 2.5 to 13.5 mo (retest). The results of the primary test and the retest agreed in 86% of the cases. The high level of agreement in the results suggests that the presence of high-risk types of HPV DNA can be determined from actual cervical cytology material in PreservCyt with the HC II assay for at least 3 mo after specimen collection.

Feasibility of laparoscopic partial nephrectomy using pledgeted compression sutures for hemostasis.

PURPOSE: To develop a technique for laparoscopic partial nephrectomy (LPN) without the use of hilar occlusion that allows large renal resection and excellent hemostasis. MATERIALS AND METHODS: Five female domestic pigs underwent right laparoscopic transperitoneal lower-pole partial nephrectomy after placement of pledgeted parenchymal compression sutures tied intracorporeally to induce regional renal hypoperfusion. Postoperatively, serial serum creatinine measurements were obtained to monitor renal function. The pigs were allowed to recover and 2 weeks later underwent an identical procedure on the left side. The animals were sacrificed after the second procedure, and both renal units were removed for ex vivo retrograde urograms and histologic analysis. RESULTS: The median operative time was 154.5 minutes (range 110-305 minutes), and the median blood loss was 137.5 mL (range 100-300 mL). On average, 35% (range 31%-36.8%) of the kidney was resected. All cases required use of adjunctive hemostatic clips to control bleeding from central vessels. All animals survived 2 weeks and had no evidence of urinary extravasation clinically or on ex vivo retrograde urograms. CONCLUSIONS: In the porcine model, LPN with placement of pledgeted sutures allows resection of large renal segments, although technical refinements are required to improve hemostasis. Currently, the need for adjunctive hemostatic measures limits the initial clinical application of this technique to small, exophytic tumors.

Clinical relevance of benign endometrial cells in postmenopausal women.

Our objective was to determine if the finding of benign endometrial cells on a Papanicolaou (Pap) smear of a postmenopausal woman is associated with endometrial/uterine pathology, independent of symptomatology and hormone replacement therapy (HRT) status. The medical records of 146 postmenopausal patients who had a Pap smear showing normal-appearing endometrial cells between January 9, 1997 and January 12, 2000 were reviewed. Uterine pathology for each patient was determined by reviewing the results of endometrial sampling (endometrial biopsy or dilatation and curettage), hysterectomy, or pelvic sonogram, which were performed within 24 mo of the cytologic smear. The results were then correlated with clinical symptomatology and HRT status of each patient at the time the cytologic smear was obtained. Of the 146 Pap smears coded with "endometrial cells in a postmenopausal woman," 50 were excluded due to prior hysterectomy, perimenopausal status, and absence of further follow-up. Of the remaining 96 women, 27 (28%) had benign pathologic findings including polyps, leiomyomata, and simple hyperplasia without atypia, whereas 11 (12%) had significant pathologic findings including hyperplasia with atypia, adenocarcinoma, mixed Mullerian tumor, and leiomyosarcoma. Of the 11 patients with significant pathology, only one patient did not have abnormal vaginal bleeding but instead had a 30-wk-size irregular uterus on examination, and only 2 patients received hormone replacement therapy. In conclusion, Reporting endometrial cells on Pap smears in postmenopausal women did not lead to the diagnosis of any cases of significant pathology that would have gone unsuspected clinically. Moreover, HRT status did not affect the incidence of normal endometrial cells on Pap smears in postmenopausal women, nor did it aid in distinguishing which postmenopausal women had endometrial/uterine pathology. This calls into question the usefulness of the current Bethesda guideline to report "benign endometrial cells in a postmenopausal woman."

Cholestatic presentation of chronic hepatitis C: a clinical and histological study with a review of the literature.

Bile duct lesions are observed in the livers of chronic hepatitis C patients, but are inconstant and rarely associated with other features of chronic cholestasis and progressive bile duct injury or loss. We aimed to identify the clinical and biochemical characteristics of patients with chronic hepatitis C from our patient database presenting with prominent cholestatic features to determine if there is a correlation between histological evidence of bile duct injury and clinical or biochemical features observed in these patients. We retrospectively reviewed a hepatitis C database including 620 patients to identify those who presented with either alkaline phosphatase (AP) > or = 400 units/liter (normal 30-126 units/liter) or AP > or = 250 units/liter with pruritus. All patients were negative for anti-mitochondrial antibody (AMA). Appropriate exclusion criteria were used to exclude patients with other confounding factors. Histological features were compared with age- and sex-matched controls selected randomly from our hepatitis C database. Thirty-two patients were identified as meeting the above criteria. Twenty-four were excluded for the presence of other confounding factors and two for lack of liver biopsy. There were two men and four women. The mean age was 47 +/- 9 years. Four of the six presented with pruritus, which was severe in three. Liver biopsy showed evidence of moderate to severe fibrosis in all but one patient. Evidence of bile duct injury was seen in all patients and tended to be more severe than in controls. Bile ductular proliferation and mild ductopenia were the most commonly observed findings. A subset of patients with chronic hepatitis C may present with prominent cholestatic features. The majority of these patients present with pruritus and have histological evidence of bile duct injury, which may be progressive.

Assessment of HER-2/neu status in breast cancer. Automated Cellular Imaging System (ACIS)-assisted quantitation of immunohistochemical assay achieves high accuracy in comparison with fluorescence in situ hybridization assay as the standard.

This retrospective study of formalin-fixed infiltrating breast cancer specimens compared manual immunohistochemical assay with a new image analyzer-assisted immunohistochemical quantitation method, using fluorescence in situ hybridization assay (FISH) as the standard. Following the manual immunohistochemical assay, 189 cases, including most manual immunohistochemically positive and some random negative cases, were analyzed by FISH assay for Her-2/neu gene amplification and by the Automated Cellular Imaging System (ACIS) for immunohistochemical staining. Using the FISH standard, the ACIS immunohistochemical assay attained a higher concordance rate and sensitivity than the manual immunohistochemical assay (91.0% and 88% vs 85.7% and 71%, respectively), with only a slight decrease in specificity (93% vs 96%, respectively). In particular, the ACIS immunohistochemical assay resulted in a higher correlation with the FISH assay in the manual immunohistochemical assay 2+ cases. The ACIS immunohistochemical assay achieved higher accuracy than the manual method according to receiver operating characteristic curve analysis. The ACIS method represents a substantial improvement over the manual method for objective evaluation of the HER-2/neu status.

Fine-needle aspiration biopsy features in a case of giant cell fibroblastoma of the chest wall.

Giant cell fibroblastoma is an unusual tumor of childhood, primarily occurring in the superficial soft tissues. We describe the fine-needle aspiration biopsy features of a case of giant cell fibroblastoma of the chest wall in a 3-year-old child. The aspirates comprised bland spindle to oval cells entrapped in a metachromatic matrix, accompanied by rare multinucleated giant cells with wreathlike nuclei. Although we were unable to render a definitive diagnosis on fine-needle aspiration biopsy, surgical resection of the mass established the diagnosis of giant cell fibroblastoma. We review the distinctive cytologic features of some common soft tissue tumors arising in this age group that may give rise to a diagnostic conundrum on fine-needle aspiration biopsy.

The use of fine needle aspiration biopsy in the evaluation of lymphadenopathy.

Fine needle aspiration biopsy is an important diagnostic tool in the evaluation and triage of patients with lymphadenopathy. It offers a simple and inexpensive test for diagnosis of reactive hyperplasia, infections, granulomatous lymphadenopathies, and metastatic diseases. Although previously regarded as limited in its use for diagnosing primary lymphoid malignancies, fine needle aspiration in combination with immunophenotypic and genotype studies is gaining respect in providing accurate diagnosis of lymphoma for primary treatment in selected patients.

The utility of transbronchial (Wang) fine needle aspiration in lung cancer diagnosis.

We evaluated our experience with transbronchial fine needle aspiration (TBNA) in cancer diagnosis over a period of 1 year. A total of 51 aspirates were performed by specialist chest physicians in the presence of a cytopathologist who made on spot evaluation of Diff-Quik smears for adequacy and guided the aspirator for additional sampling if necessary. Two clusters of at least 10 malignant cells were required on the Diff-Quik smears to render an on the spot positive diagnosis of malignancy. Aspirates showing atypical cells or few malignant cells not fulfilling the above criteria were placed in a suspicious category and additional material was requested. The TBNA results were correlated with the transbronchial biopsy when available.

Chronic graft-versus-host disease after hematopoietic cell transplantation presenting as an acute hepatitis.

A variety of illnesses involving the gut and liver follow hematopoietic cell transplantation (HCT). A 20 yr-old white male developed severe acute hepatitis 36 wk (day 252) after matched, unrelated, allogeneic HCT for chronic myelogenous leukemia (CML). Mild skin graft-versus-host disease (GVHD) had occurred at about 20 wk (day 140) after transplant. Liver biopsy showed bile duct injury and a diffuse lobular injury pattern most consistent with a GVHD variant and not reminiscent of drug-induced or viral hepatitis. No findings suggestive of herpesvirus, adenovirus, or varicella-zoster virus were found. High-dose steroids resulted in marked improvement of his liver enzyme levels. We report this patient as representing the acute hepatitic presentation of chronic GVHD of the liver.

Creatine supplementation increases renal disease progression in Han:SPRD-cy rats.

The growing use of creatine as a potential ergogenic aid among active individuals has raised concern regarding its effects on the kidney, particularly among those individuals with compromised renal function. The object of this study is to investigate the effects of oral creatine supplementation in an accepted animal model of renal cystic disease. Han:Sprague-Dawley (SPRD)-cy rats with cystic kidney disease were administered a creatine supplement at a loading dose of 2.0 g/kg of diet for 1 week, followed by 5 weeks during which the dose was one fifth this amount, mimicking typical human consumption on a body-weight basis. Cystic kidney disease progression was assessed by measuring kidney size and fluid content and determining cyst scores. Renal function was assessed by measuring serum urea and creatinine concentrations and creatinine clearance. Creatine supplementation resulted in greater cyst growth and worsened renal function in the Han:SPRD-cy rat, evidenced by greater kidney weights (2.87 +/- 0.08 versus 2.61 +/- 0.09 g/100 g of body weight; P: = 0.0365), renal fluid contents (89.22 +/- 0.41 versus 87.38 +/- 0.48 g/100 g of kidney weight; P: = 0.0057), cyst scores (0.49 +/- 0.02 versus 0.40 +/- 0.03; P: = 0.0167) and serum urea concentrations (23.96 +/- 0.92 versus 20.65 +/- 1.06 mmol/L; P: = 0.0230), and lower creatinine clearances (0.125 +/- 0.098 versus 0.162 +/- 0.011 mL/min/100 g of body weight; P: = 0.0159). These results indicate that creatine supplements may exacerbate disease progression in an animal model of cystic renal disease. Although systematic research of the effects of creatine supplementation in humans with compromised renal function is awaited, it follows that creatine should be used with particular caution in individuals with or at risk for renal disease.

Hyalinizing clear-cell carcinoma of salivary glands in fine-needle aspiration.

Hyalinizing clear-cell carcinoma (HCCC) is a recently described distinctive salivary gland neoplasm. Because of its cytoplasmic clearing and the bland nuclear features, HCCC resembles other tumors. The authors describe the cytomorphologic features of four cases of HCCC in fine-needle aspirates (FNA) and discuss the differential diagnosis. Fine-needle aspirates from 4 patients with primary HCCC of minor salivary glands were reviewed. Smears were stained with Diff-Quik and Papanicolaou stains. The cytologic features of the epithelial and the stromal components were analyzed. Cell blocks were prepared, and findings were correlated with prior or subsequent surgical specimens in each case. The smears contained numerous cohesive small and large epithelial cell groups and sheets which had sharp outlines and showed focal nuclear overlapping. The cells had uniform round to ovoid nuclei, granular chromatin, and small nucleoli. The abundant, well-defined cytoplasm was clear in many cells but denser in others. No myoepithelial cells or hyaline globules were identified. HCCC seems to have characteristic cytomorphologic findings on FNA smears. Because these cytologic features are not specific, and overlap with those of a number of salivary gland neoplasms that contain clear cells, a high level of suspicion, clinico-pathologic correlation, and examination of cell blocks are necessary to suggest the diagnosis. A diagnosis of HCCC by FNA was suspected in 3 of the 4 cases reported here.

High dietary fat intake increases renal cyst disease progression in Han:SPRD-cy rats.

The effect of a high level of dietary fat on renal cyst disease was examined in the Han:SPRD-cy rat model of polycystic kidney disease. Control and diseased rats at 4 wk of age were fed either a low fat or high fat diet (5 or 20 g/100 g diet) for 6 wk. In rats with kidney disease fed the high fat rather than the low fat diet, kidneys were 17% larger, renal fluid content was 19% higher and cyst scores were 30% higher, indicating greater disease progression. In diseased rats fed the high fat diet, serum urea was 25% higher, indicating worsened renal function. Serum creatinine was 49% higher only in males. To examine whether high dietary fat worsened renal cyst disease by altering sex hormone concentrations, serum testosterone and estrogen concentrations were determined. In normal compared with diseased male rats, serum testosterone concentrations were one to three times higher. Serum testosterone concentrations were higher in normal male rats fed the high compared with the low fat diet, but were not affected by diet in diseased rats. Serum estrogen concentrations were unaffected by dietary fat levels or by disease state. Although it remains to be elucidated how dietary fat influences sex hormone concentrations in this disease, the current study demonstrates that a high dietary fat intake increases kidney disease progression in Han:SPRD-cy rats.

The role of fine-needle aspiration biopsy in the primary diagnosis of mesenchymal lesions: a community hospital-based experience.

BACKGROUND: The objective of this study was to determine the utility of fine-needle aspiration biopsy (FNAB) in the primary diagnosis of mesenchymal lesions. A total of 162 cases with a diagnosis of benign or malignant mesenchymal lesion (excluding lipoma) on FNAB were retrieved from the cytopathology archives for the years 1990-1997. METHODS: Patients selected for inclusion in this study underwent FNAB as the primary diagnostic modality without a previous tissue diagnosis and had a subsequent surgical procedure for definitive histologic correlation. Seventy-two patients were selected on the basis of the above criteria. RESULTS: Cytologic diagnoses were categorized as benign, malignant, or suspicious for malignancy. Among the 72 cases selected, 42 (58%) benign, 18 (25%) malignant, and 12 (16%) suspicious diagnoses were rendered. Of the patients with benign FNAB diagnoses, 39 of 42 (93%) had a benign lesion on histologic follow-up, and 3 of 42 (7%) had a malignancy. Of the patients with malignant FNAB diagnoses, 17 of 18 (94%) had a malignant lesion and 1 of 17 (6%) proved to be benign. In the subset of suspicious lesions, subsequent histology was benign in 5 of 12 (42%) and malignant in 7 of 12 (58%). CONCLUSIONS: Based on our study, FNAB has excellent accuracy (88%), sensitivity (89%), and specificity (87%) for classifying a mesenchymal tumor as benign or malignant. FNAB can be a rapid and effective tool for the primary categorization of mesenchymal lesions and provide reliable information to the clinician for triage of patients.

Laboratory assessment of the status of Her-2/neu protein and oncogene in breast cancer specimens: comparison of immunohistochemistry assay with fluorescence in situ hybridisation assays.

AIM: To evaluate the clinical usefulness of three commercially available assays for Her-2/neu oncogene and protein measurements. The Her-2/neu protein is overexpressed, mostly as a result of gene amplification, in 20-30% of human breast cancers, and has been shown to have prognostic and predictive value for treatment with chemotherapy or the new monoclonal antibody, Herceptin. METHODS: An immunohistochemistry (IHC) assay using the Dako polyclonal antibody A0485, which measures the Her-2/neu protein, was compared with two new Food and Drug Administration (FDA) approved fluorescence in situ hybridisation (FISH) assays--INFORM and PathVysion, in a cohort of 52 formalin fixed, paraffin wax embedded breast tissues. These tissues were selected randomly from 84 consecutive infiltrating breast cancer specimens, which were first stratified according to the Her-2/neu protein levels as measured by IHC. RESULTS: The two FISH assays achieved a 98% concordance rate: 14 specimens (27%) showed Her-2/neu gene amplification and 37 specimens (71%) showed no Her-2/neu gene amplification. The PathVysion assay had certain advantages over the INFORM assay. In contrast, the IHC assay detected Her-2/neu overexpression in a high percentage of cases, including 13 high positive specimens (25%) and 13 medium positive specimens (25%). Although 10 of these 13 IHC high positive specimens showed gene amplification by FISH, nine of 13 IHC medium positive specimens showed no gene amplification. Statistical analyses showed that the differences between IHC and FISH assays were primarily in the specimens with medium positive IHC, but negative FISH results. CONCLUSIONS: Because of the increasing importance of the Her-2/neu oncogene and oncoprotein in the clinical management of patients with breast cancer, the accurate and consistent evaluation of Her-2/neu status is crucial. This study suggests that the best approach is to combine both IHC and FISH assays; that is, to use the IHC assay as a triage step, followed by the PathVysion FISH assay to analyse the IHC medium and high positive cases.

Expression of the human mismatch repair gene hMSH2: a potential marker for urothelial malignancy.

BACKGROUND: The human mismatch repair (MMR) gene hMSH2 (human mutS homolog-2) is a DNA repair gene that has been reported to be mutated in 40% of hereditary nonpolyposis colon cancer (HNPCC) kindreds and a small percentage of sporadic tumors. HNPCC is a cancer predisposition syndrome with an increased risk of carcinoma of the colon, endometrium, stomach, small intestine, ovary, ureter, and renal pelvis. Immunohistochemical analysis demonstrated increased hMSH2 expression in sporadic colon carcinoma and in the replicative compartment of normal epithelium. A recent immunohistochemical analysis of hMSH2 in bladder tumors correlated reduced hMSH2 expression with recurrence and higher tumor grade. In the current study, we examined hMSH2 expression in urothelial malignancy using immunohistochemical analysis and developed a molecular assay for the detection of hMSH2 expression in bladder washes. METHODS: Immunohistochemical analysis of 17 tumors from the genitourinary tract and reverse transcription coupled with polymerase chain reaction (RT-PCR) of 40 bladder washes were used to investigate hMSH2 expression in noninvasive and invasive urothelial malignancies. RESULTS: Increased expression of hMSH2 was detected in all tumors examined using immunohistochemical analysis independent of grade or stage. Reverse transcription-PCR of hMSH2 mRNA from bladder washes detected 17 of 21 patients with primary or recurrent urothelial neoplasms or tumors involving the urothelial system. Four patients with urothelial malignancies without detectable hMSH2 expression from their bladder washes had high grade lesions. Ten of 13 patients without pathologic or cystoscopic evidence of bladder tumors were negative for hMSH2 expression in bladder washes. Two patients with bladder tumors and bladder washes that were positive for hMSH2 subsequently were found to be negative for hMSH2 after treatment of their tumors and at last follow-up had remained recurrence free for at least 1 year. CONCLUSIONS: The results of the current study suggest that hMSH2 expression is increased in low and high grade urothelial neoplasms, similar to the expression pattern in sporadic colon carcinoma. However, a fraction of high grade lesions may not express hMSH2 as detected by RT-PCR from bladder washes. The ability to detect hMSH2 expression in bladder washes may allow the use of hMSH2 expression as a marker for urothelial malignancy. In addition, the ability to define hMSH2 deficient tumors using bladder washes may have prognostic significance in the treatment of patients with urothelial carcinoma.