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Whole mounted radical prostatectomy specimens do not increase detection of adverse pathological features.

PURPOSE: The optimal method to process radical prostatectomy specimens to maximize the detection of adverse pathological features is unclear and accurate staging is critical. We compare the ability of whole mounted sections to detect these features compared to partially submitted radical prostatectomy specimens. MATERIALS AND METHODS: A total of 93 consecutive radical prostatectomy specimens were processed as whole mounts. Tissue sections were analyzed and the pathological outcomes measured included Gleason score, surgical margin status, and presence or absence of extraprostatic tumor extension and/or seminal vesicle invasion. The pathological outcomes of the preceding cohort were compared to those of a similar cohort consisting of 554 men whose radical prostatectomy specimens were processed as partially submitted glands. RESULTS: A multivariate logistic regression analysis was performed to determine the effect of the method of tissue processing on the pathological outcomes. When considered alone or adjusted for various preoperative patient characteristics (prostate specific antigen, biopsy Gleason score and clinical stage), there were no significant differences in the ability of whole mounted specimens to detect the various outcomes compared to partially submitted specimens (all p >0.4). CONCLUSIONS: Whole mounted sampling of the radical prostatectomy specimen does not improve detection of adverse pathological features.

Histocytological Preparation Techniques↗

[Comparison of monolayer specimens and conventional smears].

INTRODUCTION: Monolayer specimens were compared with conventional smears in a split sample study. The quality of the specimens, sensitivity, time consumption, and costs were elucidated. METHOD: Conventional smears and fixated residual material from 1701 women were available. RESULTS: The number of inadequate/less adequate specimens was reduced significantly (p < 0.001). Diagnostic agreement was found in 1531 (90%) of the 1701 cases (kappa = 0.52, SE (kappa) = 0.026). Dysplasia (NOS) was diagnosed in seven monolayer specimens (0.4%) versus 33 smears (1.9%) and ratio atypia/low grade was reduced by 33%, which indicates greater accuracy in diagnosis in monolayers. Histological follow-up showed sensitivities of 95% (monolayers) and 94% (smears). The time consumed (laboratory work, screening) on a smear versus a monolayer specimen was 11.5 minutes versus 9.3 minutes. Utensil costs of a smear are kr. 10.69. compared to kr. 26.50 for a monolayer specimen. CONCLUSION: The higher costs should be set against the saving in significantly improved specimen quality, improved diagnostic accuracy, and shorter time consumption. The use of the monolayer technique (direct to vial) is recommended to replace the conventional smear.

Cervix Uteri↗

Comparison of blood specimens from plain and gel vacuum blood collection tubes.

This study was set in the Division of Laboratory Medicine, Chulalongkorn Hospital. All 2,000 blood specimens were randomly collected using evacuated blood collection by plain or gel vacuum tubes. After collection, each specimen was considered and judged using criteria of specimen rejection to determine how proper the specimen presentations were. All data were reviewed, collected and interpreted. It revealed that there were only 20 (1%) improper specimens and all were improper in quality. There was no significant difference between the ratio of improper specimens in both groups (P > 0.30). From this study, it revealed that efficacy of both types of vacuum tubes was not different. The new gel vacuum tube seems to be an effective tool in the evacuated blood collection system due to its advantage in reduction of time in specimen processing.

Blood Specimen Collection↗

Vaginal extraction of the intact specimen following laparoscopic radical nephrectomy.

PURPOSE: We report the detailed technique and results of transvaginal extraction of the intact laparoscopic radical nephrectomy specimen. MATERIALS AND METHODS: Since June 2000, 10 select female patients with a median age of 67 years underwent transvaginal extraction of the intact specimen after laparoscopic radical nephrectomy. In 5 patients open surgery had previously been performed on the uterus, including transabdominal hysterectomy in 2 and cesarean section in 3. Laparoscopic nephrectomy was performed via the transperitoneal and retroperitoneal approach in 5 cases each. After completion of the primary laparoscopic procedure a sponge stick was externally inserted into the sterile prepared vagina and tautly positioned in the posterior fornix. Laparoscopically a transverse posterior colpotomy was created at the apex of the tented up posterior fornix and the drawstring of the entrapped specimen was delivered into the vagina. After laparoscopic exit was completed the patient was placed in the supine lithotomy position. The specimen was extracted intact via the vagina and the posterior colpotomy incision was repaired transvaginally. Patients were mailed a linear scale analog questionnaire to assess various aspects of recovery with responses graded from 0--no pain and/or change to 10--severe pain and/or change. RESULTS: Vaginal extraction was successful in all 10 patients. Median operative time for the vaginal extraction procedure was 35 minutes. Blood loss was minimal. Median tumor size was 3.6 cm. (range 2.4 to 7.4) and median specimen weight was 327 gm. (range 152 to 484). No intraoperative complications occurred. Postoperatively blood spotting via the vagina in 1 patient resolved spontaneously. Postoperative questionnaires revealed excellent patient satisfaction and convalescence. CONCLUSIONS: Vaginal extraction is an efficacious and minimally morbid technique for removing the intact entrapped specimen after laparoscopic radical nephrectomy. It has now become our preferred technique of intact specimen extraction in appropriate female patients.

Adult↗

The use of retained patient specimens for haematology quality control.

Patient blood specimens constitute ideal quality control material in many respects. Although stability is a problem, patient specimens are sufficiently stable to allow their use in the control of short-term systematic error. The principal challenges involve the design of a system which combines excellent performance characteristics (probability of error detection and probability of false rejection) with a minimum of extra work. In the past, guidelines have been presented for an optimized quality control program using retained patient specimens in haematology. These guidelines call for the use of three retained specimens initially analysed only once, with subsequent analyses judged 'out of control' if they deviate from the initial result by a prescribed multiple of the long-term standard deviation. In practice, this system may result in a relatively high probability of false rejection of data (Pfr) due to inadequately established control ranges. Also, the maintenance of three retained patient specimens may be an excessive burden on small laboratories. We present data on the optimization of a quality control program using one retained specimen that is appropriate for use by smaller laboratories. The control rules and the number of initial analyses are adjusted to yield the highest possible probability of error detection (Ped while maintaining a low Pfr and a low additional workload. In addition, we present data concerning the control of satellite instruments by sharing patient specimens between the satellite instrument and a 'reference' instrument.

Computer Simulation↗

Prevalence of antibody to human immunodeficiency virus and hepatitis B surface antigen in blood samples submitted to a hospital laboratory. Implications for handling specimens.

The prevalence of hepatitis B surface antigen (HBsAg) and antibody to human immunodeficiency virus (HIV) was determined in serum or plasma specimens of 506 patients submitted to the clinical chemistry laboratory of an urban teaching hospital, and the results were correlated with "biohazard" warning labels on the specimens. Hepatitis B surface antigen, HIV antibody, or either of these were present in 32 (6.3%), 15 (3.0%), and 44 specimens (8.7%), respectively. Ten (67%) of 15 specimens with HIV antibody and nine (28%) of 32 with HBsAg bore biohazard labels. Among 473 unlabeled specimens, HIV antibody was present in five (1.1%), HBsAg was present in 23 (4.9%), and 27 (5.7%) contained either or both of these markers. All clinical and laboratory personnel should be vaccinated against hepatitis B and should handle all blood specimens as if they were infected, regardless of biohazard labeling. By fostering complacency in handling unlabeled specimens, the use of biohazard labels may paradoxically increase the risk that health care workers will be exposed to HIV and hepatitis B virus.

Chemistry, Clinical↗

Problems in quantitative x-ray microanalysis of biological specimens.

Use of quantitative methods in X-ray microanalysis of biological specimens has expanded the scope of this technique. Reliable quantitation in biological microanalysis is, however, not simple, and a number of difficulties are typical for biological specimens. The purpose of this tutorial is to signal problems in quantitative X-ray microanalysis of biological specimens and to discuss possible solutions. First of all, the instrumental configuration has to be optimized and the contribution of extraneous sources to the spectrum minimized. Contamination and mass loss during irradiation have to be reduced. After the spectrum is obtained, background subtraction and deconvolution routines are applied. The contribution of extraneous sources to the spectrum has to be accurately determined and subtracted from the initial spectrum. Standards have to be prepared that contain the elements of interest in a known amount and are homogeneous. The intensities in specimen and standard are then compared and corrections are applied to correct for differences in composition between specimen and standard that are relevant to the resulting X-ray spectrum. These corrections are relatively simple for thin specimens but may be more intricate for bulk specimens. It is then important to know under which conditions the quantitative procedure can be simplified.

Biology↗

Evaluation of evacuated blood-collection tubes: effects of three types of polymeric separators on therapeutic drug-monitoring specimens.

The potential of three types of separator materials found in conventional blood-collection tubes for interference in therapeutic drug measurements was assessed. None of the separators (based on acrylic, silicone, or polyester polymers) had any significant effect on the concentrations of seven drugs (theophylline, digoxin, phenytoin, phenobarbital, gentamicin, ethanol, and cyclosporine) in blood specimens that were processed and analyzed promptly. Storage of specimens for 24 h resulted in an average 2.4% increase in theophylline values in specimens collected in tubes with the acrylic separator (P = 0.024); an average 8.1% decrease in phenytoin in specimens collected in tubes with the polyester-based separator (P < 0.001); and an average 4.2% decrease in phenobarbital in specimens collected in tubes with the polyester-based separator (P = 0.02). All other drug concentrations were not significantly affected. A small decrease in phenytoin (7.9%; P < 0.01) was seen when the specimen volume in 7-mL tubes containing polyester-based separator was reduced to 1.0 mL; all other drug concentrations were unaffected by partial filling of tubes. Paired blood specimens from pediatric patients, when collected in plain tubes and tubes containing acrylic separator, yielded no significant differences for theophylline, digoxin, tobramycin, phenytoin, or phenobarbital concentrations. The three commercially available separators had only small effects on therapeutic drug concentrations, and a newly developed separator based on an acrylic resin was suitably inert.

Acrylic Resins↗

[Media for transportation of specimens from gynecologic infections].

Specimens from the urogenital tract of 458 out-patients of the gynaecological clinic were examined using two collection kits including Amies and C.A.T. Swab medium. In the examined group, trichomonads were found in 165 cases (30.6% of all specimens), yeasts in 131 specimens (38.6%) and in 49 specimens (10.7%) both agents were detected. In 48 cases (10.5%), yeasts were detected only microscopically from the C.A.T. Swab medium. Examination of the wet mounts from C.A.T. Swab medium after 48 hours' incubation proved best for the diagnosis of trichomonads. Anaerobic bacteria were found in 378 specimens (82.5%), mycoplasmas and ureaplasmas in 281 specimens (61.4%). The simultaneous use of both media facilities detection of yeasts, trichomonads, aerobic and anaerobic bacteria and of mycoplasmas in specimens from the female urogenital tract.

Culture Media↗

Feasibility and quality of biological banking of human normal and tumor tissue specimens as sources of DNA for the Malmö Diet and Cancer Study.

Human tumor and normal tissue specimens, which were collected from autopsy material 1-6 days postmortem, were compared with similar tissue specimens collected within 2 h after surgical resection and transport to the pathology department. The end point criteria used to evaluate the quality of the specimens for biological banking purposes were the extractability and yield of high molecular weight DNA and UV absorption ratios at 260:280 after collection and immediate storage of the specimens at -80 degrees C. The data demonstrated that autopsy material was a quality source of DNA, although of not such high quality as surgical biopsy specimens <2 h after resection. The advantages of using autopsy material to supplement surgical specimen collection sent to pathology, as opposed to using specimen collection at surgery wards or formalin-fixed material, as sources of DNA are: (a) large amounts of tumor and normal tissues from a variety of organ sites can be obtained without regard to the patient's health status; (b) a higher percentage of retrieval of incident cases of cancer in prospective designed trials is more likely to be achieved; and (c) the extractable DNA is of sufficiently high enough quality to permit direct analyses by molecular hybridization and sequence methodologies.

Autopsy↗

Smith-Lemli-Opitz syndrome diagnosed in a 130-year-old anatomical specimen.

The Museum Vrolik collection of human anatomy comprises 360 recently re-described specimens with congenital anomalies. The external findings in one of these specimens, originally described by Willem Vrolik (1801-1863) 130 years ago, were suggestive of Smith-Lemli-Opitz (SLO) syndrome. Cholesterol synthesis was analyzed in skin biopsies, obtained from the suspected SLO specimen and a control specimen. The cholesterol levels in the SLO specimen and in the control specimen were in the proportion of 1 to 45. This confirms the diagnosis in this specimen which, to our knowledge, represents the oldest known case of SLO syndrome.

Cadaver↗

The diagnosis and reporting of adenocarcinoma of the prostate in core needle biopsy specimens.

BACKGROUND: There has been relatively little written on the diagnosis and reporting of adenocarcinoma of the prostate diagnosed in core needle biopsy specimens. METHODS: This article reviews issues concerning diagnosing, grading, and quantification of prostate carcinoma in core needle biopsy specimens. RESULTS: The diagnosis of prostate carcinoma in core needle biopsy specimens is discussed, including the relative frequency and utility of various architecture, cytologic, and ancillary features. Grading of prostate carcinoma in core needle biopsy specimens is evaluated along with the relationship of core needle biopsy grade to corresponding radical prostatectomy grade. The relationship between the extent of carcinoma in core needle biopsy specimens to extent of tumor in the radical prostatectomy is summarized. Finally, this article summarizes articles supporting the use of high molecular weight cytokeratin in the diagnosis of adenocarcinoma of the prostate in core needle biopsy specimens. CONCLUSIONS: Pathologists are not only called upon to diagnose limited cancer in core needle biopsy specimens, but also to quantify and grade these cancers accurately. Issues relating to this pathologic evaluation are critical for physicians treating men with adenocarcinoma of the prostate.

Adenocarcinoma↗

Image cytometry as a discriminatory tool for cytologic specimens obtained by endoscopic retrograde cholangiopancreatography.

BACKGROUND: Routine brush cytology is relatively insensitive for the diagnosis of biliary and pancreatic malignancy. Sensitivity can be improved by measuring DNA and proliferation. The goal of this study was to assess the discriminatory capacity of image cytometry using pancreaticobiliary brush cytology specimens obtained during endoscopic retrograde cholangiopancreatography (ERCP). Analysis included morphometry, DNA quantification, and characterization of nuclear chromatin distribution and condensation. METHODS: Brush cytology specimens were obtained during ERCP from 22 chronic pancreatitis specimens, 11 pancreatic adenocarcinoma specimens, 13 primary sclerosing cholangitis specimens, and 11 cholangiocarcinoma specimens and contrasted with 25 normal epithelia specimens. A SAMBA 2005 image processor was used to analyze Feulgen stained chromatin density and distribution. Discriminant analysis of 37 morphonuclear variables was performed to characterize differences between: 1) chronic pancreatitis and pancreatic adenocarcinoma, and 2) primary sclerosing cholangitis and cholangiocarcinoma. RESULTS: Chronic pancreatitis was distinguished from pancreatic adenocarcinoma (P < or = 0.001); sensitivity and specificity were both estimated to be 82%. Primary sclerosing cholangitis was distinguished from cholangiocarcinoma (P < or = 0.01); sensitivity and specificity were estimated to be 82% and 85%, respectively. CONCLUSIONS: Multiparameter image cytometry has potential as an adjuvant diagnostic technique in patients with pancreaticobiliary malignancy.

Adenocarcinoma↗

Diagnosing Helicobacter pylori by imprint cytology: can the same biopsy specimen be used for histology?

Imprint cytology of the gastric mucosa has been found to be very simple, inexpensive, and rapid for diagnosing Helicobacter pylori infection. However, there is a fear that preparing imprint smears may damage the biopsy specimen for subsequent histologic examination. This study was planned to investigate whether this damage happens. Four antral biopsy specimens were obtained from each of the 100 patients undergoing upper gastrointestinal endoscopy. Imprint smears were made from two biopsy specimens, which were then fixed in 10% formal saline and sent for histologic study. The third and fourth biopsy specimens were directed fixed in 10% formal saline for histologic examination. Two pathologists examined the imprint smears. Agreement between the two observers was observed in 97% of cases. Beyond-chance agreement was good with a kappa index of 0.90. H. pylori organisms were seen in 82% of biopsy specimens from which imprint smears were prepared and in the same percentage of biopsy specimens that were processed directly. The pathologists could not identify the histologic sections form which imprints were made. It is concluded that imprint cytology is an excellent method of diagnosing H. pylori infection and that preparing imprint smears does not alter the quality of the tissue. The same biopsy specimen can be used for histologic studies.

Cytodiagnosis↗

Laboratory implementation of a rapid three-stain technique for detection of microorganisms from lower respiratory specimens.

A rapid, cost-effective method for the evaluation of lower respiratory specimen has become increasingly important in the diagnosis of pulmonary diseases in immunocompromised patients. In the past, the technically demanding, time-consuming, and expensive Gomori-methenamine-silver (GMS) stain was the principal means for the evaluation of these specimens. In this study, we compared the GMS stain with a new rapid, three-stain protocol for the evaluation of lower respiratory specimens. Lower respiratory specimens were obtained by bronchoalveolar lavage (BAL). Conventional Wright/Giemsa and Gram stains were utilized, as well as a contemporary strain, calcofluor white (CW). A cell count was performed on the BAL specimens, and cytospins were stained by the three stains. The calcofluor white-stained slides were examined with an epi-fluorescent microscope, whereas the other stains were evaluated with a conventional light microscope. Gomorimethenamine-silver (GMS), acid-fast bacillus (AFB), and Papanicolaou (PAP) stains were performed as controls. Thirty-two BAL procedures were performed in 20 (63%) male patients and 12 (37%) female patients. The clinical diagnosis was pneumonia in 31% of the patients, malignant hematologic disease in 28%, acute respiratory distress syndrome (ARDS) in 9%, and acquired immunodeficiency syndrome (AIDS) in 28%. Of these specimens, 78% were adequate for interpretation and 22% were inadequate. Bacteria were found in 50% (16/32) of all BALs, fungi were found in 9% (3/32), and Pneumocystis carinii was found in 9% (3/32). Gram-positive bacteria were most frequently found in patients with pneumonia (80%, 4/5), whereas P. carinii was identified in patients with AIDS. There were no false-positive results. One CW stain was equivocal for P. carinii due to high fluorescent background. Laboratory implementation of the rapid, three-staining technique was accomplished without difficulty in microbiology and hematology laboratory sections. Specimen evaluation with the rapid staining protocol was technically easy to perform; however, experience in ultraviolet fluorescent microscopy was crucial for interpretation of CW stain. All results were available in 2 hr, cost was reduced by 30%, and the assays were available 7 days/week. Further studies are ongoing to substantiate the sensitivity, specificity, and predictive value of this technique, as well as clinical guidelines for its optimal utilization.

Adult↗

The mucosa of the gastric remnant harboring malignancy. Histologic findings in the biopsy specimens of 504 asymptomatic patients 15 to 46 years after partial gastrectomy with emphasis on nonmalignant lesions.

Endoscopic bioptic screening of 504 asymptomatic postgastrectomy patients, 15 to 46 years after initial surgery, revealed ten gastric stump cancers of which six turned out to be early cancers; three of the early carcinomas were found during follow-up after prior severe dysplasia. At first endoscopy mild dysplasia was found in 58, moderate dysplasia in 11, and severe dysplasia in none of the patients. Follow-up biopsies in 177 patients showed mild dysplasia in 30, moderate dysplasia in six, and severe dysplasia in six patients. Regression of severe dysplasia was not observed. Both progression and regression of mild and moderate dysplasia occurred. At the first endoscopy the frequency of atrophy, intestinal metaplasia, cystic dilatation of glands, and foveolar hyperplasia in the stomal biopsy specimens was 45%, 35%, 54%, and 47%, respectively; during follow-up, 47%, 48%, 56%, and 56% respectively. These four lesions were significantly more frequent in the biopsy specimens of patients with gastric carcinoma or dysplasia than in the other patients and they were present in the environment of the tumor in all the surgical specimens of the six early cancers detected by the screening. Preoperatively a combination of these four lesions could be demonstrated in only those early gastric cancer patients, in whom more than eight stomal biopsy specimens were taken. Of 34 patients with severe atrophy in three or more stomal biopsy specimens taken at the same time, two manifested early stump cancer during follow-up. Severe dysplasia is a marker of malignancy and demands close follow-up; the value of mild and moderate dysplasia is less clear. The combination of atrophy, especially severe atrophy, intestinal metaplasia, cystic dilatation, and foveolar hyperplasia in the biopsy specimens of a single patient may also point to increased cancer risk. It is advisable to obtain multiple biopsy specimens of the anastomosis, also because early gastric cancer may occur without a suspect macroscopic appearance.

Adolescent↗

Clinical sensitivity of p53 mutation detection in matched bladder tumor, bladder wash, and voided urine specimens.

BACKGROUND: Mutations in the p53 tumor suppressor gene may correlate with an increased risk of recurrence and disease progression in patients with bladder carcinoma. The ability to accurately and sensitively detect p53 mutations in cytology specimens may be of benefit in the treatment of bladder carcinoma patients with superficial, minimally invasive disease. METHODS: Genomic DNA was isolated from 49 cases, each of which was comprised of matched bladder tumor tissue, bladder wash, and voided urine specimens obtained concurrently at a single institution. The genomic DNA was analyzed for mutations in the p53 tumor suppressor gene using a p53 mutation detection assay. Automated dideoxy sequencing of mutant specimens also was performed. RESULTS: Of the 49 cases, 29 (59%) showed no evidence of p53 mutations in the tumor, bladder wash, or voided urine specimens. Of the remaining 20 cases, 19 showed evidence of mutations in the tumor. Of these 19 p53 mutant bladder tumors, 16 (84%) were detected in the matched bladder wash and 16 (84%) were detected in the matched voided urine specimens. One case resulted in the detection of mutant p53 in the voided urine and the bladder wash, but not in the tumor. Analysis of the results between tumor tissue and bladder wash or tumor and voided urine showed 84.2% sensitivity, 96.8% specificity, and 91.8% accuracy. Sequence analysis of the mutant cases showed that the mutations detected in the tumor tissue were the same mutations detected in the bladder wash and the voided urine specimens. CONCLUSIONS: Both voided urine and bladder wash specimens from patients with bladder carcinoma were found to provide a high rate of clinical accuracy for the determination of the p53 gene status in patients with bladder tumors.

Aged↗

Variant estrogen receptor alpha mRNAs in human breast cancer specimens.

A panel of human breast cancer specimens was examined for single base change mutations by DNA sequencing and for larger deletions using a PCR-based assay. In the cancer specimens examined, no sequencing variants were detected other than a previously characterized polymorphism. Although most of the specimens contained estrogen receptor (ER) variants at a low level, 2 of 118 specimens exhibited variants which, after amplification, constituted most of the amplified ER cDNA. One specimen contained a single variant form, and there was little evidence of the wild-type ER mRNA by PCR, Northern blotting or immunocytochemistry. The second specimen, despite the presence of a normal-sized mRNA by Northern blotting and normal immunocytochemical staining for ER, contained at least 5 different variant forms as well as the wild-type ER. All but 1 of the variant forms were processing variants, and 3 of these processing variants have not been described before. One variant, although lacking exons 2-4, has break points in exons 1 and 5 that do not correspond to intron-exon boundaries. This variant might reflect more widespread damage to the genome in this breast cancer specimen. The low level of occurrence of variants suggests that ER variant forms, at least in the coding region, do not contribute generally to the progression of breast cancer.

Blotting, Northern↗