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At least 235 records · Page 13Linked to original sources
PubMed2005

Assessment of HOPE fixation in vitrectomy specimens in patients with chronic bilateral uveitis (masquerade syndrome).

BACKGROUND: Cytological examination of rapidly transported, unfixed vitreous specimens is considered the gold standard in exclusion of primary intraocular lymphoma (PIOL) in patients with idiopathic steroid resistant chronic uveitis. These specimens are difficult to interpret, and reports of "false negatives" or "false positives" are common. Fixation using HOPE solution (Herpes-glutamic acid buffer mediated Organic solvent Protection Effect) has been successfully applied in the investigation of cytospin preparations using immunocytology, in situ hybridisation and polymerase chain reaction (PCR). The purpose of this study was to compare the cytologic features of vitrectomy specimens from patients with clinical reactive vitritis and PIOL prepared following HOPE fixation with those in conventional cytospins and to identify any diagnostic pitfalls. METHODS: Pars plana vitrectomy was performed in 15 cases of patients with chronic uveitis. The vitreous samples were halved: one half was fixed in HOPE solution; the other half remained unfixed. All samples were subject to conventional staining, immunocytology and clonality analysis using polymerase chain reaction (IgH-PCR) and GeneScan. The specimens were assessed following by two pathologists who were masked to the cytological preparation method. The specimens were evaluated for cellularity, cellular appearance, cytoplasmic and nuclear features as well as quality of the immunostains. RESULTS: Twelve of the fifteen vitreous samples were diagnosed as reactive vitritis; in three specimens, a primary intraocular lymphoma of B-cell type was diagnosed. Compared to the unfixed vitreal specimens, the quality of the cytomorphology and immunocytology improved in the HOPE-fixed specimens. IgH-PCR and GeneScan analysis demonstrated polyclonal amplification products in the reactive cases, and monoclonal B-cell populations in the B-PIOL. CONCLUSION: Our results demonstrate that cytomorphology and immunoreactivity of vitreous specimens are well preserved following HOPE fixation. DNA of sufficient quality could be extracted from HOPE-fixed vitreous biopsies, in order to perform clonality analyses. HOPE fixation appears to be promising in simplifying the transportation of these specimens, and may improve the diagnostic reliability of vitreous specimens in patients with masquerade syndrome.

Adult↗
PubMed2005

Species of Pseudorhabdosynochus Yamaguti, 1958 (Monogenea: Diplectanidae) from Epinephelus fasciatus and E. merra (Perciformes: Serranidae) off New Caledonia and other parts of the Indo-Pacific Ocean, with a comparison of measurements of specimens prepared using different methods, and a description of P. caledonicus n. sp.

Species of Pseudorhabdosynochus were studied from fresh specimens collected from Epinephelus fasciatus and E. merra off New Caledonia, South Pacific, and specimens deposited in Museums. Experiments on two species demonstrated that the sclerotised hollow organs, such as the quadriloculate male copulatory organ and the vagina, may show differences in measurements of up to 50% when flattened. P. caledonicus n. sp. is described from E. fasciatus in New Caledonia, on which it is relatively rare; it is distinguished on the basis of the quadriloculate organ, which has a very thin anterior wall, the sclerotised parts of the vagina in form of a straight tube with a star-shaped lateral structure, and the squamodiscs composed of 11 open rows of rodlets. P. cupatus (Young, 1969) is redescribed from abundant material from E. fasciatus off New Caledonia (new geographical record) and compared with paratype specimens from Australia (from E. fasciatus and E. merra) and specimens from E. fasciatus in the Red Sea (both herein redescribed and figured); a specimen was also found on a slide from E. merra off Vanuatu. P. melanesiensis (Laird, 1958) is redescribed from material from E. merra off New Caledonia (new geographical record) and compared with type-specimens (herein redescribed and figured) from the same host off Vanuatu. The structure of the sclerotised vagina in P. cupatus and P. melanesiensis is very similar, with a thin-walled tube and a heavily sclerotised structure with three loculi. P. vagampullum (Young, 1969) is redescribed from the paratypes from E. merra from Australia, but was not found in New Caledonia; specimens included among its paratypes (from E. merra in Australia), but different, are herein attributed to Pseudorhabdosynochus sp. 3. P. lantauensis (Beverley-Burton & Suriano, 1981) is redescribed from the paratype specimens from E. longispinis off Hong-Kong. A specimen found among the paratypes of P. cupatus belongs to a different species, herein designated as Pseudorhabdosynochus sp. 1. Specimens from E. longispinis off Hong-Kong, previously attributed to P. cupatus, are attributed to another species, Pseudorhabdosynochus sp. 2. The three species P. cupatus, Pseudorhabdosynochus sp. 1 and Pseudorhabdosynochus sp. 2 have in common a 'lamellosquamodisc' composed of central telescopic lamellae and peripheral rows of rodlets; they can be distinguished by the shape of the sclerotised vagina and measurements of the haptoral hard-parts. Specimens from E. longispinis off Hong-Kong, previously attributed to P. vagampullum, probably belong to a different species. Consequently, after these modified determinations, P. cupatus parasitises only E. fasciatus and E. merra, and P. melanesiensis and P. vagampullum parasitise only E. merra. With their wide geographical distribution and different species of Pseudorhabdosynochus in different localities, E. fasciatus and E. merra appear to represent excellent models for investigating monogenean biogeography in the Indo-Pacific Ocean.

Animals↗
PubMed1982

Biomechanical studies of rabbit abdominal wall. Part II.--The mechanical properties of specimens in relation to length, width, and fibre orientation.

In a biomechanical investigation of tractus iliotibialis and abdominal wall tissue from rabbits, an analysis was made of the influence that the fibre orientation in specimens as well as the length and width of specimens has on the measured parameters. The mechanical testing was performed with a materials testing machine (Alvetron). The investigation showed that the mechanical characteristics of specimens, when the width was kept constant, depended upon the fibre orientation in the specimens and the length of the specimens. The investigation further demonstrated that in intact abdominal wall no proportionality exists between, on the one side, the width of specimens (2, 4, 6 and 8 mm) and, on the other, the breaking strength, the energy absorption, and the elastic stiffness, respectively. For the 6 day old abdominal wall wounds a proportionality, however, could be demonstrated between the 4 and 8 mm broad specimens on the one side and the breaking strength, the energy absorption, and the elastic stiffness on the other. A possible explanation of the difference between the mechanical characteristics of short and long specimens can be given by the theoretical possibility that the short specimens contain relatively more fibres running all the way through the specimen from clamp to clamp than do the long specimens. The consequences on biomechanical investigations rendered by the result of the analyses are discussed.

Abdominal Muscles↗
PubMed1988

Effects of sonication and centrifugation of clinical specimens on the recovery of herpes simplex virus.

We have analyzed frozen and fresh specimens for herpes simplex virus (HSV) to determine the effect of sonication and centrifugation on virus recovery. After sonication, titers of 24/27 specimens increased from 1.3-30.8-fold with a mean increase of 6.8-fold. One hundred seventy-four fresh specimens were inoculated before and after sonication into CV-1 tube cultures. There was not a statistically significant difference in time to positivity between the sonicated and unsonicated portions. Thus, although sonication of specimens can sharply increase the viral titer of positive specimens, sonication of fresh specimens does not significantly enhance the isolation of HSV. To determine the effect of centrifugation of clinical specimens on recovery of HSV, thirty-one culture positive frozen specimens were centrifuged, and the supernatants and resuspended sonicated pellets were titered. Twenty-five specimens had sufficient recoverable virus to titer; in 21/25 (87.5%) specimens the titers were reduced in the supernatants after centrifugation (mean reduction 38%; range of 4-92%). Two of 31 (7%) supernatants were negative in culture while the sonicated pellets were positive. Thus, centrifugation of specimens prior to cell inoculation may compromise recovery of HSV.

Animals↗
PubMed2005

Kappa statistics to measure interrater and intrarater agreement for 1790 cervical biopsy specimens among twelve pathologists: qualitative histopathologic analysis and methodologic issues.

BACKGROUND: As part of a program project to evaluate emerging optical technologies for cervical neoplasia, we performed fluorescence and reflectance spectroscopic examinations of patients with abnormal Papanicolaou smears. Biopsy specimens were taken from each area and measured optically, and study pathologists performed qualitative histopathologic readings. Several methodologic issues arose in this analysis: (1) the interpathologist and intrapathologist agreement between institutions for the 1790 biopsy specimens; (2) the interinstitutional agreement among the two institutions conducting the trials on 117 randomly chosen biopsy specimens; (3) the interinstitutional agreement among the two institutions and a third expert gynecologic pathologist to ensure the expert readings were comparable to those outside both institutions on 117 randomly chosen biopsy specimens; and (4) an additional three reviews of the 106 difficult biopsy specimens by all three institutions. METHODS: All 1790 specimens from 850 patients were reviewed three times at each institution in blinded fashion; those for which the first and second reviews were identical were not reviewed a third time. A randomly selected sample of 117 specimens was randomly ordered and read by study pathologists at The University of Texas M. D. Anderson Cancer Center, British Columbia Cancer Agency (BCCA), and Brigham and Women's Hospital (BWH). The 106 difficult cases were treated in the same manner as the randomized and random-ordered cases. Generalized, unweighted, and weighted kappas and their 95% confidence intervals were used to assess agreement. Binary comparisons were used to compare diagnostic categories. FINDINGS: The kappas for the three readings of the overall data set using eight-category World Health Organization (WHO) criteria were as follows: 0.66 for the generalized, 0.72 for weighted, and ranged from 0.59 to 0.94 unweighted binary categories; those read using four-category Bethesda criteria: 0.70 for generalized, 0.69 for weighted, and 0.56-0.94 for unweighted binary categories. For the pool versus the study pathologist readings, the eight-category kappa was 0.51 for generalized, 0.72 for weighted, and 0.56-0.82 for unweighted binary categories; for those read using Bethesda criteria: 0.70 for generalized, 0.70 for weighted, and 0.59-0.82 for the unweighted binary categories. The interpathologist and intrapathologist readings were fair by Landis standards at the low end of the diagnostic scale (atypia, human papillomavirus, and CIN1) and substantial to almost perfect at the high end (CIN2, CIN3, and CIS). The randomly selected and randomly ordered sample of 117 specimens read with the WHO system yielded a generalized kappa of 0.45; among the three institutions (M. D. Anderson Cancer Center vs. BCCA, M. D. Anderson vs. BWH, and BCCA vs. BWH), the unweighted kappas were 0.46, 0.41, and 0.49 and the weighted were 0.65, 0.66, and 0.68, respectively; for the Bethesda, a generalized kappa of 0.65, unweighted kappas of 0.66, 0.65, and 0.47, and weighted of 0.74, 0.72, and 0.74. The difficult specimens read with the WHO system yielded a generalized kappa of 0.23; among the three institutions the unweighted kappas were 0.20, 0.30, and 0.37, and the weighted were 0.17, 0.34, and 0.31; for the Bethesda, a generalized kappa of 0.25; among the three institutions, the unweighted kappas were 0.21, 0.32, and 0.37, and the weighted were: 0.07, 0.21, and 0.37, respectively. INTERPRETATION: Kappas in this expert group of pathologists were in the moderate, substantial, and almost perfect ranges for the overall and randomized samples. The randomized sample was representative of the larger sample. The kappa of the specimens for which disagreements arose was, predictably, in the slight range. Our findings will aid both the correlations with optical measurements using fluorescence and reflectance spectroscopy and the quantitative histopathologic analysis of these study specimens.

Biopsy↗
PubMed1997

Loss of the CDKN2A/p16 locus detected in bladder irrigation specimens by fluorescence in situ hybridization.

PURPOSE: We measured the CDKN2A/p16 tumor suppressor gene locus in bladder irrigation specimens and correlated the measurement with the clinical status of patients with bladder cancer. MATERIALS AND METHODS: Irrigation specimens were obtained at cystoscopy from 10 normal controls, 21 patients with bladder cancer in whom no concurrent bladder tumor was seen and 23 patients with bladder tumors. Deoxyribonucleic cytometry and fluorescence in situ hybridization measurements were made. One fluorescence in situ hybridization probe was specific to the chromosome 9 centromere and the other, COSp16, targeted the CDKN2A/p16 region on chromosome 9p21. Three rates were calculated, including the hyperdiploid fraction from deoxyribonucleic acid cytometry, disomic fraction from the 9 centromere count and COSp16F, the frequency of COSp16 in association with 9 centromere. Specimens were classified as positive or negative for each of these rates using cutoff points based on previous studies and the distribution of values obtained for the normal control specimens. RESULTS: Hyperdiploid fraction values were positive (greater than 8%) in 1 normal and 1 nontumor specimen. Ten specimens from patients with tumor showed elevated hyperdiploid fraction values. In 4 nontumor and 13 tumor irrigation specimens the chromosome 9 disomic fraction values were positive (less than 80%). COSp16F was positive (less than 83%) for 18 nontumor irrigation specimens and 18 tumor irrigation specimens. One normal, and 39 of 44 nontumor and tumor irrigation specimens were positive by at least 1 test. CONCLUSIONS: COSp16 loss is measurable in irrigation specimens and it correlates with clinical status. This assay may prove useful in screening for and managing bladder cancer.

DNA, Neoplasm↗
PubMed2003

Incidental healed postinfectious glomerulonephritis: a study of 1012 renal biopsy specimens examined by electron microscopy.

Glomerulonephritis (GN) characterized by immune complex deposits typical of postinfectious GN but with a paucity or absence of overt clinical symptoms and/or urinary abnormalities may occur after a group A streptococcus infection. The overall incidence of this type of subclinical GN is not known. To address this question, electron microscopy findings in 1012 consecutive renal biopsy specimens (952 native kidney, 60 transplant) examined by a single renal pathologist from August 1999 to April 2002 were retrospectively reviewed for the presence of distinct subepithelial and intramembranous deposits indicative of postinfectious GN. Such deposits were noted in 83 biopsy specimens, including 26 with a primary diagnosis of postinfectious GN (acute, persistent, or latent) and 57 in which these deposits were an incidental finding. In each of the latter 57 cases, some or all of the deposits showed partial or extensive loss of electron density typical of partially or largely resorbed deposits. A diagnosis of incidental postinfectious GN was not made in any biopsy specimen exhibiting another immune complex-related glomerular disease that could possibly account for the deposits, composing 443 of the 1012 biopsy specimens examined. Thirty of the 57 biopsy specimens with incidental postinfectious GN showed mesangial hypercellularity, although this was focal and segmental in all but 3 cases and was not accompanied by the endocapillary hypercellularity typical of acute postinfectious lesions. Immunofluorescence microscopy revealed glomerular deposits of C3 in >90% of these biopsy specimens and IgM deposits in 66%, but only rare IgG, IgA, and Cq deposits. Twenty-three (40%) of these 57 biopsy specimens exhibited diabetic nephropathy, either alone or in combination with another lesion; for perspective, only 128 (13%) of the 1012 biopsy specimens examined showed evidence of diabetic nephropathy. In summary, incidental evidence of resolving or largely healed postinfectious GN was noted in up to 10.5% of renal biopsy specimens (57 of 543, not including specimens with a primary diagnosis of an immune complex-related glomerular disease). The recognition of such lesions is potentially important in the interpretation of certain renal biopsy specimens.

Adolescent↗
PubMed2003

[Sonographical breast biopsy: how many core biopsy specimens are needed?].

PURPOSE: To determine the optimum number of specimens of a sonographically guided core biopsy of the breast. MATERIALS AND METHODS: From January 2001 to April 2001, sonographically guided core biopsies (coaxial needle: 11 G; core needle: 12 G) were performed on 106 patients with 115 BI-RADS trade mark 4-5 lesions that had corresponding sonographic findings. Five specimens were obtained in anteroposterior direction parallel to the chest wall and each specimen examined histologically. RESULTS: A total of 575 core biopsies produced 545 specimens (94.8 %), with no material obtained in 30 biopsies (5.2 %). The first and second specimen confirmed the diagnosis in 99 of 115 lesions (86 %), with subsequent specimens confirming the diagnosis in 87 % (3 rd specimen), 88 % (4 th specimen) and 82 % (5 th specimen). The additive accuracy of the five core breast specimens was 86 %, 96 %, 98 %, 99 % and 100 %. The possible volume of 78.9 mm 3 as determined by the sample notch of the core needle only furnished an average volume of 38.1 mm 3 (41.1 mm 3 for the 1 st, 40.1 mm 3 for the 2 nd, 37.5 mm 3 for the 3 rd, 36.7 mm 3 for the 4 th and 35.0 mm 3 for 5 th core specimen). CONCLUSION: Sonographically guided core breast biopsy (coaxial needle: 11 G; core needle: 12 G) may require five passes to achieve a diagnostic accuracy of more than 99 %.

Adult↗
PubMed2006

Validation of the quality of histological images obtained of fresh and formalin-fixed specimens of esophageal and gastric mucosa by laser-scanning confocal microscopy.

BACKGROUND AND STUDY AIMS: We have previously reported the success of a method of virtual histology using laser-scanning confocal microscopy (LCM) in vitro on untreated fresh specimens obtained from the gastrointestinal mucosa. In the present study, we aimed to apply LCM to both fresh and formalin-fixed specimens, without additional treatment, in order to validate and compare the quality of the images obtained. METHODS: We obtained 18 specimens from 11 patients, either by endoscopic biopsy or following surgical resection. First, we observed the fresh, saline-immersed specimen with LCM using the Fluroview microscope (Olympus Co. Ltd., Tokyo, Japan). We then fixed the specimen with formalin and obtained further LCM images 1 hour, 3 hours, and 24 hours after fixation. Three independent observers observed the images and were asked to assess the origin of the samples, the treatment of the samples, the time after formalin fixation, and whether they showed benign or malignant lesions. We used kappa statistics to compare the agreement among the three observers in each of these four areas of interest. RESULTS: Between January and March 2003, we obtained 191 LCM images from 18 specimens. Thirty images were randomly selected for observation. The overall accuracy for differentiating between esophagus and stomach specimens was 96.6 %. The accuracy of differentiating normal from cancerous lesions was 92.2 %. The differentiation between saline-immersed and formalin-fixed specimens was 59.7 % accurate and the assessment of the time interval after formalin fixation was only 37.3 % accurate. The kappa statistics showed that there was strong interobserver agreement on the differentiation of specimen origin and of cancerous from benign lesions. However, there was no agreement among the observers on the method of specimen preparation or on the estimated time interval after formalin fixation. CONCLUSIONS: We concluded that images obtained from fresh specimens using LCM were of a quality good enough to make an accurate diagnosis of upper gastrointestinal carcinoma.

Biopsy↗
PubMedDate not supplied

Cervical, urine and vaginal specimens for detection of Chlamydia trachomatis by ligase chain reaction in women: a comparison.

Screening for Chlamydia trachomatis in women is generally done using only one specimen from each patient in order to minimize costs. In this study the aim was to compare the performances of vaginal, cervical and urinary specimens in a population of young women with sparse symptoms. During 1998, specimens from 1,001 women at the Departments of Venereology and Youth Health Care at the University Hospital of Uppsala, Sweden were examined by both ligase chain reaction and cell culture for detection of C. trachomatis. The samples from the cervix, vagina and urine were tested by ligase chain reaction, while specimens for cell culture were collected from the cervix and urethra. The prevalence of genital C. trachomatis infections was 5.1%. A single urine specimen had a sensitivity of 80.0%, while the sensitivity of a single vaginal specimen was 96.0%. The specificity was 100% for the urine specimens and 99.4% for the vaginal specimens. The sensitivity and specificity of a single cervical specimen was 92.0% and 99.6%, respectively. Although the urine ligase chain reaction seemed to have the lowest sensitivity of the compared specimens for testing of C. trachomatis infections in this population, the differences in sensitivity between urine, cervical and vaginal specimens were not statistically significant.

Adolescent↗
PubMed2004

The use of specimens from various genitourinary sites in men, to detect Trichomonas vaginalis infection.

Variations in estimates of prevalence of trichomoniasis in men may reflect true differences in the burden of disease but are also affected by the performance of diagnostic methods and the type of specimen tested. In this study, men were evaluated at baseline and at follow-up, to evaluate syndromic management of urethritis and the effects of human immunodeficiency virus and trichomoniasis, in Lilongwe, Malawi. First-void urine specimens and urethral swabs were obtained at enrollment, for Trichomonas vaginalis culture; semen specimens were also obtained at follow-up. The sensitivities of testing methods using urine specimens and urethral swabs were equal; 67% of cases were identified by use of either specimen, and, in 47% of cases, both specimens tested positive. When semen specimens were included, all 3 specimens tested positive in only 19% of cases. Semen was the most sensitive single specimen, and, in 25.6% of cases, only semen specimens tested positive. Thus, prevalence of T. vaginalis infection in men is underestimated if only 1 specimen is tested.

Adolescent↗
PubMed1987

Rapid diagnosis of invasive cytomegalovirus infection by examination of tissue specimens in centrifugation culture.

Two hundred eight tissue specimens from 81 patients were examined by centrifugation culture for the rapid diagnosis of invasive cytomegalovirus (CMV) infection. Results were compared with those obtained by both conventional viral cultures and histologic examination. CMV was identified by centrifugation culture in 52 (25%) specimens at 16 hours after inoculation, including 39 lung specimens and 12 gastrointestinal specimens. One additional esophageal specimen was positive (53 of 208 specimens; 25.5%) at 36 hours. In contrast, CMV was recovered from only 50 of these 53 specimens by conventional cell culture at a mean of 11 days after inoculation. Forty-three of the 208 specimens (20.5%) were positive for CMV when examined by histologic methods, including light microscopy and tissue immunofluorescence. All 43 were positive by centrifugation culture. Specimens that were positive by centrifugation culture but negative by histologic methods included three lung specimens and seven gastrointestinal specimens. The technic of centrifugation culture is more sensitive for diagnosis of invasive CMV infection by the examination of affected tissue than either conventional viral culture or histologic technics, including tissue immunofluorescence, and more rapid than conventional culture or routine histologic examination.

Antibodies, Monoclonal↗
PubMed2000

Comparison of direct examination of three types of bronchoscopy specimens used to diagnose nosocomial pneumonia.

OBJECTIVE: To compare direct examination of bronchial aspirate and plugged telescopic catheter specimens (PTC) with infected cell counts in bronchoalveolar lavage (BAL) specimens for the diagnosis of nosocomial pneumonia. DESIGN: Prospective study of critically ill patients. SETTING: Intensive care unit in a university hospital. PATIENTS: A total of 64 patients hospitalized for >48 hrs with suspected nosocomial pneumonia. INTERVENTIONS: Fiberoptic bronchoscopy with bronchial aspirate and quantitative protected specimen brush, PTC, and BAL cultures. PTC and bronchial aspirate specimens were Gram-stained. BAL specimens for infected cell counts were examined as described previously in the literature. MEASUREMENTS AND MAIN RESULTS: Nosocomial pneumonia was diagnosed by the medical staff based on all available clinical, radiologic, laboratory test, and microbiological data and on the course before and after appropriate therapy. A total of 71% of patients were ventilated, and 70.1% were receiving antibiotics. Nosocomial pneumonia was diagnosed in 54% of the cases. On direct examination, sensitivity (Se) and specificity (Sp) of bronchial aspirate specimens were Se, 82% and Sp, 60%; of BAL with 5% infected cells, Se, 56% and Sp, 100%; of BAL with 3% infected cells, Se, 74% and Sp, 96%; of PTC specimens, Se, 65% and Sp, 76%; and of PTC specimens plus BAL with 3% infected cells, Se, 83% and Sp, 78%. BAL with 3% infected cells was significantly better for predicting nosocomial pneumonia than direct examination of bronchial aspirate or PTC specimens (p = .0012). When the BAL showed 3% infected cells, neither direct examination of bronchial aspirate nor direct examination of PTC specimens was useful (p = .24 and p = .38, respectively). Combined use of direct examination of PTC specimens plus BAL with 3% infected cells markedly improved sensitivity. The total cost of each procedure was taken into account for the final evaluation. CONCLUSIONS: Our data suggest that BAL with 3% infected cells is currently the only test whose predictive value for nosocomial pneumonia is sufficiently high to be of use for guiding the initial choice of antimicrobial class while waiting for quantitative culture results.

Adult↗
PubMed2000

Gastric metaplasia: a frequently overlooked feature of duodenal biopsy specimens in untreated celiac disease.

BACKGROUND: Duodenal gastric metaplasia is rarely reported in untreated celiac disease, although it is seen in 60% to 100% of duodenal biopsy specimens in nonceliac patients with histologic duodenitis. The low incidence could represent underreporting, a decreased incidence in pediatric patients generally, or the more distal sampling site that is customary for most biopsy specimens that are obtained to diagnose celiac disease. It could also be a unique feature of the inflammatory reaction that characterizes this disease. The purpose of this study was to examine the incidence of gastric metaplasia in duodenal specimens from children with untreated celiac disease with special reference to patient age and biopsy site. METHOD: Formalin-fixed paraffin-embedded specimens of duodenal mucosa were selected from the pathology department's archival material. Sections were either stained histochemically or by immunochemical methods, according to an antigen-retrieval protocol. Forty-four duodenal specimens from untreated patients with celiac disease (n = 22) and control subjects of similar age with normal histology (n = 22) were examined. Ten of each were obtained during upper endoscopy from the proximal duodenum (proximal site) and 12 of each by Crosby capsule near the ligament of Treitz (distal site). RESULTS: All specimens from patients with celiac disease exhibited marked villous atrophy. None had been noted to have gastric metaplasia during routine examination of sections stained by hematoxylin and eosin. Fifteen (68%) of 22 of the celiac specimens and 2 of 22 (9%) control specimens contained gastric metaplasia, identified as patches of gastric-type cells containing MUC5AC (gastric mucin), pS2 (gastric trefoil factor) and neutral (periodic acid-Schiff-positive) mucin. Five of the seven celiac specimens that had no metaplasia showed increased numbers of goblet cells expressing gastric markers. The incidence of gastric metaplasia was not different for endoscopic (70%) or capsule (67%) specimens. Sixty-eight percent (7/11) of patients aged less than 3 years had gastric metaplasia. CONCLUSION: The presence of gastric metaplasia has been previously underreported in celiac disease specimens. Detection would be improved by the routine use of period acid-Schiff/ alcian blue staining. The incidence of gastric metaplasia in celiac disease is not significantly influenced by biopsy site or age at time of the biopsy.

Age Factors↗
PubMed1998

Comparative studies of the detection rates of Leishmania parasites from formalin, ethanol-fixed, frozen human skin specimens by polymerase chain reaction and Southern blotting.

In this study, detection rates of Leishmania parasites from human skin were compared among three different types of specimens, formalin-fixed, ethanol-fixed, and frozen, by polymerase chain reaction (PCR) and Southern blotting. For this purpose, we used biopsy specimens collected from 19 leishmaniasis patients and performed PCR and Southern hybridization with the probe specific for Leishmania (Viannia) braziliensis complex. Among these 19, 16 specimens were from cutaneous leishmaniasis (CL), one, diffuse cutaneous leishmaniasis (DCL) and 2, mucocutaneous leishmaniasis (MCL) and were formalin-fixed and paraffin-embedded. The causative agents for one case of CL and one case of DCL were already identified as L. (Leishmania) complex. Six specimens of CL were preserved in 100% ethanol. Two specimens of MCL were frozen tissues. PCR using the formalin-fixed and paraffin-embedded specimens revealed positive bands at 70 bp in 9 (47.4%) out of 19 specimens of CL, MCL and DCL. Southern blotting detected the signals in 12 (63.2%) out of the 19. PCR using the 100% ethanol-fixed specimens revealed positive bands in 4 (66.7%) out of 6, and Southern blotting also detected the signals in 4 (66.7%) out of the 6. PCR and Southern blotting using 2 frozen specimens of MCL were always positive (100%). Although we failed to detect significant differences by Chi-square test between the results from the formalin-fixed, paraffin-embedded specimens and those from 100% ethanol-fixed ones, we concluded that ethanol-fixed specimens, convenient for transportation and storage, would be more useful for diagnosis of leishmaniasis by PCR in a developing country.

Adolescent↗
PubMed1981

Evaluation of transport and storage techniques for isolation of Campylobacter fetus subsp. jejuni from turkey cecal specimens.

Immediate culturing of fecal specimens is not always possible, and appropriate methods for transport and storage of Campylobacter fetus subsp. jejuni specimens have not been fully evaluated. Using nine techniques, we studied the survival of C. fetus subsp. jejuni in cecal specimens from infected turkeys. The organisms survived in specimens held without transport medium for 3 to 15 days (median, 9 days) at 4 degrees C, and 2 to 9 days (median, 4 days) at 25 degrees C. Only 20% of specimens frozen for 24 h at either -20 or -70 degrees C yielded C. fetus subsp. jejuni. Specimens dried on filter paper strips were negative for C. fetus subsp. jejuni within 1.5 h. Cary-Blair medium with decreased agar was the best of the six transport media tested, it enabled recovery of the organism from 100% (3 days) and 71% (7 days) of cecal samples held at 4 degrees C and 94% (3 days) and 85% (7 days) of cecal specimens held at 25 degrees C. In contrast, more than half of all cecal specimens held at 4 or 25 degrees C in Culturettes or buffered glycerol saline were negative by 3 days, and all were negative at 7 days. Results with the other three media studied (Campy-thio, thioglycolate medium, and alkaline peptone water) were intermediate. Overnight incubation of specimens in alkaline peptone water at 37 or 42 degrees C did not enhance recovery of C. fetus subsp. jejuni. Therefore, refrigeration without a transport medium is satisfactory for up to 3 days for recovery of C. fetus subsp. jejuni from specimens, however, we recommend the use of Cary-Blair medium with decreased agar for specimens that must be transported or stored for longer than 3 days and for rectal swabs, to prevent drying.

Animals↗
PubMed1997

Influence of endocervical specimen adequacy on PCR and direct fluorescent-antibody staining for detection of Chlamydia trachomatis infections.

The cellular quality of the endocervical swab specimen used for the detection of Chlamydia trachomatis may dramatically impact the sensitivity of the diagnostic assay used. An evaluation of the adequacy of 319 endocervical swab specimens from women attending two inner-city sexually transmitted disease and family planning clinics, as well as five high school-based family planning clinics, was performed, and the resulting data were compared with the diagnostic results obtained by both Amplicor PCR and Microtrak direct fluorescent-antibody (DFA) staining. The swab from each patient was rolled across the open circular area of a DFA slide and then used to inoculate a transport tube for PCR (Roche), after which the swab was discarded. The slides were stained and examined by epifluorescence microscopy for the presence of C. trachomatis elementary bodies and for the presence and number of cell types to determine specimen adequacy. Cellular adequacy for a cervical swab specimen was defined as the presence of one or more columnar epithelial or metaplastic epithelial cells or the presence of more than 100 erythrocytes per high-power microscopic field. Of the 319 specimens read by DFA, 204 (63.9%) were determined to be adequate. There were 34 (10.7%) positive specimens by DFA and/or PCR. Twenty-nine (9.1%) specimens were positive by PCR, 20 (6.3%) specimens were DFA positive, and 15 (4.7%) were concordantly positive by both tests. The prevalence of chlamydia among adequate specimens was 14.2% (29/204), compared to 4.3% (5/115) for inadequate specimens (P < 0.0001). Variations in specimen quality and the sensitivity of the diagnostic assay used have a significant impact on determining the prevalence of C. trachomatis in a population.

Cervix Uteri↗
PubMed1997

Evaluation of the MB/BacT system and comparison to the BACTEC 460 system and solid media for isolation of mycobacteria from clinical specimens.

The MB/BacT automated system is designed for the isolation of mycobacteria from clinical specimens. It utilizes a colorimetric sensor and reflected light to continuously monitor the CO2 concentration in the culture medium. We compared its performance to that of the BACTEC 12B media for the radiometric BACTEC 460 instrument and that of solid culture media. Respiratory specimens and urine samples were decontaminated with 4% NaOH. The vials of the two instruments were inoculated with 500 microl of sample and two solid egg-based media at 200 microl each. All vials were incubated at 37 degrees C for 6 weeks. A total of 1,078 specimens (633 respiratory specimens, 78 cerebrospinal fluid specimens, 177 other body fluid specimens, 87 urine specimens, and 103 other types of specimens) were cultured in parallel. Mycobacteria could be identified from 73 (6.8%) specimens: 67 M. tuberculosis, 3 M. kansasii, 1 M. xenopi, 1 M. terrae, and 1 mixed M. avium with M. scrofulaceum. Of these, 63 (86.3%) specimens were positive with the MB/BacT system, 67 (91.8%) were positive with the BACTEC 460 instrument, and 58 (79.5%) were positive with the two egg-based media. MB/BacT cultures were positive on average after 17.5 (+/-6.4) days, BACTEC cultures with a growth index of >20 (mean, 200) were positive after 14.3 (+/-8.2) days, and egg-based media were positive after 24.2 (+/-7.5) days. Microorganisms other than mycobacteria contaminated 46 (4.3%) MB/BacT cultures and 31 (2.9%) BACTEC cultures, which had to be discarded. The MB/BacT system is a well-automated system for the detection of M. tuberculosis in clinical specimens without using radioactive reagents. Further trials are required to determine whether it is suitable for the culture of nontuberculous mycobacteria.

Bacterial Typing Techniques↗