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PubMed1997

Comparison of manual Amplicor PCR, Cobas Amplicor PCR, and LCx assays for detection of Chlamydia trachomatis infection in women by using urine specimens.

We compared the Roche Amplicor PCR, Roche Cobas Amplicor PCR, and Abbott LCx assays by using urine specimens for the detection of Chlamydia trachomatis infections in a female population. First-catch urine and endocervical swab specimens were collected from a total of 442 patients. Urine specimens were tested by the manual Roche Amplicor PCR, the automatic Roche Cobas Amplicor PCR, and the Abbott LCx assays as instructed by the manufacturers. For the Cobas Amplicor PCR, the internal control protocol was used for every specimen to reveal the presence of polymerase inhibitors. Cell culture of cervical specimens was used as a reference method. Of 442 patients, 50 (11.3%) were confirmed to have chlamydial infection. The diagnostic sensitivity and specificity of cell culture with cervical swab specimens were 88 and 100%, respectively. With urine specimens the sensitivity and specificity for the manual Amplicor PCR assay were 100 and 99.7%, respectively; those for the automatic Cobas Amplicor PCR assay were 94 and 99.2%, respectively; and those for the LCx assay were 94 and 100%, respectively. Thus, all amplification methods with urine specimens proved to be highly sensitive and specific for the detection of C. trachomatis infection in women. No statistically significant differences in the test performances could be demonstrated for specimens from this population. All three amplification techniques with urine specimens proved to be superior to cell culture with cervical swab specimens in diagnosing C. trachomatis infections in women.

Chlamydia Infections↗
PubMed1998

Rapid shell vial culture technique for detection of enteroviruses and adenoviruses in fecal specimens: comparison with conventional virus isolation method.

Detection of enteroviruses and adenoviruses mainly in fecal specimens by rapid culture with inoculation onto cell monolayers in flat-bottom tubes by centrifugation and immunofluorescence staining with genus-specific monoclonal antibodies was compared with that by the conventional virus isolation procedure. For both conventional culture and shell vial culture human lung fibroblast cells and tertiary monkey kidney cells were used. For enterovirus detection, 979 clinical specimens (916 stool specimens, 56 cerebrospinal fluid specimens, and 7 nasopharyngeal swabs) were used. Conventional culture detected 74 enterovirus isolates. A cytopathic effect compatible with the presence of an enterovirus after 3 days of incubation occurred in 25 of the 74 (34%) specimens that eventually became positive. The detection rate for enteroviruses by rapid cell culture after 2 to 3 days of incubation was 42 of 74 (57%). The genus-specific enterovirus monoclonal antibody did not react with strains of echovirus types 22 and 23 or enterovirus type 71. Rapid cell culture for the detection of adenoviruses was performed with 567 clinical specimens (536 stool specimens, 25 cerebrospinal fluid specimens, and 6 miscellaneous specimens), in which 42 adenoviruses were found by conventional culture. Nine of the 42 (21%) adenovirus isolates were detected by conventional culture within 3 days after inoculation, whereas 21 (50%) were found by rapid cell culture within 2 to 3 days. Only two of the nine specimens found to be positive for the enteric adenovirus type 41 by conventional culture as well by a type-specific enzyme-linked immunosorbent assay (ELISA) tested positive by rapid cell culture. In conclusion, the rapid shell vial assay allows the early detection and identification of enteroviruses and adenoviruses in clinical specimens but is markedly less sensitive than the conventional isolation procedure according to the eventual results of the conventional isolation procedure. Conventional cell culture remains a prerequisite for serotyping of enteroviral isolates. On the basis of the results for adenovirus type 41, the rapid detection of adenoviruses was not considered to be useful for the detection of clinically relevant adenoviruses in fecal samples.

Adenoviruses, Human↗
PubMed1999

Utility of multiple-stool-specimen ova and parasite examinations in a high-prevalence setting.

A retrospective analysis of the results of 2,704 ova and parasite (O & P) examinations performed on stool specimens collected from 1,374 patients between October 1996 and September 1997 was performed to evaluate the utility of performing O & P examinations on multiple, independently collected stool specimens in a high-prevalence setting. A total of 995 specimens (36.8%) examined during the study contained parasites; 546 (20.2%) contained pathogenic organisms. The positivity rate (54.5%) for the patients from whom three specimens were examined was significantly higher than for the patients from whom either two specimens (33.3%) or a single specimen (19.8%) was submitted for examination. For the group of patients from whom at least 3 specimens were submitted for O & P examination, 373 independent opportunities for diagnosing infection with intestinal parasites could be analyzed. The first stool specimen collected proved to be adequate in only 75.9% (283 of 373) of evaluated cases; however, examination of two specimens increased the sensitivity of O & P detection to 92% (343 of 373). The third specimen collected provided additional information on only 30 of 373 occasions (8%). These data indicate that in populations with a high prevalence of intestinal parasitic infections, two independently collected stool specimens should be subjected to O & P examination to ensure adequate diagnostic sensitivity.

Feces↗
PubMed2001

Direct detection of Legionella species from bronchoalveolar lavage and open lung biopsy specimens: comparison of LightCycler PCR, in situ hybridization, direct fluorescence antigen detection, and culture.

We developed a rapid thermocycling, real-time detection (also known as real-time PCR) method for the detection of Legionella species directly from clinical specimens. This method uses the LightCycler (Roche Molecular Biochemicals, Indianapolis, Ind.) and requires approximately 1 to 2 h to perform. Both a Legionella genus PCR assay and Legionella pneumophila species-specific PCR assay were designed. A total of 43 archived specimens from 35 patients were evaluated, including 19 bronchoalveolar lavage (BAL) specimens and 24 formalin-fixed, paraffin-embedded open lung biopsy specimens. Twenty-five of the specimens were culture-positive for Legionella (9 BAL specimens and 16 tissue specimens). BAL specimens were tested by LightCycler PCR (LC-PCR) methods and by a direct fluorescent antibody (DFA) assay, which detects L. pneumophila serogroups 1 to 6 and several other Legionella species. Tissue sections were tested by the two LC-PCR methods, by DFA, by an in situ hybridization (ISH) assay, specifically designed to detect L. pneumophila, and by Warthin-Starry (WS) staining. The results were compared to the "gold standard" method of bacterial culture. With BAL specimens the following assays yielded the indicated sensitivities and specificities, respectively: Legionella genus detection by Legionella genus LC-PCR, 100 and 100%; Legionella genus detection by DFA assay, 33 and 100%; and L. pneumophila detection by L. pneumophila species-specific LC-PCR, 100 and 100%. With open lung biopsy specimens the following assays yielded the indicated sensitivities and specificities, respectively: Legionella genus detection by LC-PCR 68.8 and 100%; Legionella genus detection by DFA assay, 44 and 100%; Legionella genus detection by WS staining, 63 and 100%; L. pneumophila species-specific detection by LC-PCR, 17 and 100%; and L. pneumophila species-specific detection by ISH, 100 and 100%. The analytical sensitivity of both LC-PCR assays was <10 CFU/reaction. LC-PCR is a reliable method for the direct detection of Legionella species from BAL specimens. The Legionella genus LC-PCR assay could be performed initially; if positive, L. pneumophila species-specific LC-PCR could then be performed (if species differentiation is desired). The speed with which the LC-PCR procedure can be performed offers significant advantages over both culture-based methods and conventional PCR techniques. In contrast, for the methods evaluated, culture was the best for detecting multiple Legionella species in lung tissue. WS staining, Legionella genus LC-PCR, and L. pneumophila species-specific ISH were useful as rapid tests with lung tissue.

Antigens, Bacterial↗
PubMed2004

Suicide PCR on skin biopsy specimens for diagnosis of rickettsioses.

As rickettsioses may be severe diseases and Rickettsia prowazekii is a potential agent of bioterrorism, highly efficient diagnostic techniques are required to detect rickettsiae in patients. We developed a nested PCR assay using single-use primers targeting single-use gene fragments present in the genomes of both Rickettsia conorii and R. prowazekii. We used this "suicide" PCR with DNA from 103 skin biopsy specimens from patients who definitely had a rickettiosis, 109 skin biopsy specimens from patients who possibly had a rickettsiosis, and 50 skin biopsy specimens from patients with nonrickettsial diseases. The suicide PCR detected "R. conorii conorii" in 38 biopsy specimens, R. africae in 28 biopsy specimens, R. slovaca in 12 biopsy specimens, "R. sibirica mongolotimonae" in 5 biopsy specimens, R. aeschlimannii in 2 biopsy specimens, and "R. conorii caspia" and "R. sibirica sibirica" in 1 biopsy specimen each. The technique had a specificity of 100% and a sensitivity of 68%. It was 2.2 times more sensitive than culture (P < 10(-2)) and 1.5 times more sensitive than regular PCR (P < 10(-2)). The efficacy of the suicide PCR was reduced by antibiotic therapy prior to biopsy (P < 10(-2)) and was increased when it was performed with eschar biopsy specimens (P = 0.03). We propose the use of the suicide PCR as a sensitive, specific, and versatile technique for improving the diagnosis of rickettsioses, especially when it is used on eschar biopsy specimens taken prior to antibiotic therapy.

Base Sequence↗
PubMed2005

Comparison of the COBAS AMPLICOR MTB and BDProbeTec ET assays for detection of Mycobacterium tuberculosis in respiratory specimens.

The performances of the BDProbeTec ET (Becton Dickinson) and COBAS AMPLICOR MTB (Roche) were retrospectively evaluated for detecting Mycobacterium tuberculosis complex in various respiratory specimens. The BACTEC and MGIT liquid culture system (Becton Dickinson) was used as a reference method. A total of 824 respiratory specimens, comprised of sputa, bronchoalveolar lavage fluid, and bronchial and tracheal aspirates from 580 patients, were evaluated. Out of 824 clinical specimens, 109 specimens from 43 patients were culture positive for M. tuberculosis. Of these 109 specimens, 67 were smear positive, 85 were positive by the COBAS AMPLICOR MTB test, and 94 were positive by the BDProbeTec ET. Of the 715 culture-negative specimens, 17 were positive by the auramine staining, 11 were positive by the COBAS AMPLICOR MTB test, and 12 were positive by the BDProbeTec ET. After discrepancy analysis and review of the patients' clinical data, 130 specimens from 50 patients were considered "true-positive" specimens. This resulted in the following sensitivities: microscopy, 61.5%; COBAS AMPLICOR MTB test, 78.0%; and BDProbeTec ET, 86.2%. The specificities of each system, based on the clinical diagnosis, were 99.7% for microscopy, 99.9% for the COBAS AMPLICOR MTB test, and 99.9% for the BDProbeTec ET. The data presented represent a considerable number of specimens evaluated with a considerable number of culture- and auramine-positive and culture-positive and auramine-negative results and therefore give a realistic view of how the data should be interpreted in a daily routine situation. Specifically, the data with regard to the culture-positive and auramine-negative specimens are useful, because in a routine situation, auramine-negative specimens are sometimes accepted, on clinical indications, to be analyzed by an amplification method.

Benzophenoneidum↗
PubMed1992

Complications associated with percutaneous needle biopsy of the liver when one, two or three specimens are taken.

The diagnostic yield of blind percutaneous liver biopsy is improved when two or three specimens are taken during the procedure by redirecting the needle through a single entry-site, without exposing the patient to a greater risk of complications provided that standard precautions are taken. This study was designed to obtain further data on the safety of this procedure at King Edward VIII Hospital, Durban. During the period 1984-1990 (inclusive) a total of 2,646 biopsies were carried out: a single specimen was obtained in 834 patients, two specimens in 983 patients and three in 829 patients. Complications directly attributable to the procedure occurred in 24 patients who had one specimen, 20 who had two, and 19 who had three specimens taken during the biopsy. A single specimen had been obtained from three of the eight patients who had died, two specimens had been taken from another patient, and three specimens were obtained from the other four patients, i.e. patients in whom two or three specimens were taken did not have a higher incidence of pain, symptomatic hypotension, biliary peritonitis or death than those in whom one specimen was taken. Accordingly, when blind percutaneous needle biopsy of the liver is carried out, two specimens should be obtained by redirecting the needle through a single entry site as this improves the diagnostic yield without increasing complications. The morbidity and mortality associated with liver biopsy in this hospital is, however, high. Good technique, careful monitoring of patients after biopsy and prompt and aggressive resuscitation are essential if the mortality rate is to be reduced.

Biopsy, Needle↗
PubMed1998

Detection of Chlamydia trachomatis in vaginal specimens from female commercial sex workers using a new improved enzyme immunoassay.

OBJECTIVE: To evaluate the performance of a new improved enzyme immunoassay (EIA) kit for the detection of Chlamydia trachomatis in vaginal swab and endocervical swab specimens from female commercial sex workers, in comparison with a conventional EIA test and a polymerase chain reaction (PCR) assay. METHODS: A high risk group of 163 female commercial sex workers who visited a sexually transmitted disease (STD) clinic in order to undergo screening for major STDs, including chlamydial infection, were enrolled. A total of four swab specimens, including two vaginal and two endocervical specimens, were collected from each woman by a clinician. To identify C trachomatis, a new improved EIA kit (IDEIA PCE), a conventional EIA kit (IDEIA), and PCR assay (Amplicor) were used. Discrepancies in the results were resolved using supplementary PCR assay. A female patient was considered to be infected with C trachomatis if the IDEIA PCE test and PCR test for both sample sites (endocervical and vaginal) gave positive results. Following resolution of these discrepancies, relative sensitivity and specificity, confidence intervals, and predictive values for each type of specimen by each assay were calculated. RESULTS: Of the 163 women tested, 35 (21.5%) were shown to be infected with C trachomatis. The relative sensitivities in vaginal swab specimens were 88.8%, 68.6%, and 91.4% using IDEIA PCE, IDEIA, and PCR, respectively. The relative specificities in vaginal swab specimens were 99.2%, 99.2%, and 100%, respectively. The relative sensitivities in endocervical swab specimens were 85.7%, 77.1%, and 91.4% with IDEIA PCE, IDEIA, and PCR, respectively. The relative specificities in endocervical swab specimens were all 100%. CONCLUSIONS: The results obtained in this study suggest that the sensitivity and specificity of IDEIA PCE test on vaginal swab and endocervical swab specimens were similar to those of PCR assay on the two types of specimen. It is concluded that IDEIA PCE test on vaginal swab specimens is an acceptable, sensitive, and less invasive approach for the detection of C trachomatis in commercial sex workers with a high prevalence of C trachomatis infection.

Adult↗
PubMed1997

Quantitative measurement of volume changes induced by oral plastic surgery: validation of an optical method using different geometrically-formed specimens.

The purpose of this research was to study the validity and variability of a projection Moiré system, measuring volume differences of geometrically different formed specimens mimicking localized alveolar ridge defects. Nine pairs of specimens were fabricated, each of which simulated a preoperative ridge defect and a corresponding surgically-corrected postoperative ridge defect. All specimen pairs had a mathematically defined form which allowed the accurate assessment of their volume differences by a mechanical 3-D coordinate measuring machine or by a software-controlled milling machine. Measurements achieved with these methods were used as the references for comparison. Six specimen pairs, A1 to A6, possessed a simple rectangular geometrical form which facilitated their fabrication. Three specimen pairs, B1 to B3, were milled and consisted of geometrically more complex 3-D sculptured surfaces, which came closest to a true imitation of a localized ridge defect. An optical measurement system in the form of the projection Moiré was utilized, applying a 4-phase shift technique, and results obtained with this device were regarded as test volumes. The absolute variability of the test volume measurements differed between 0.397 mm3 to 15.872 mm3, corresponding to a relative variability of 0.83% to 2.83%. The mean of the relative variability was within 1.68% for the "A" specimens and 2.15% for the "B" specimens. However, the difference was not significant, probably due to the limited number of "B" specimens. The systematic error of the Moiré measurements in relation to the reference methods was surprisingly low, ranging from -0.12 mm3 to 7.67 mm3. The relative systematic error, expressed as a percentage of reference volume, ranged between 0.06% and -2.23%. The mean of the relative error for the more complex "B" specimens was 1.37%, which was less accurate in comparison to the more simply formed "A" specimens with a relative systematic error of 0.35%. Therefore, in this in vitro model it was possible to measure volume differences of geometrically different formed specimens, mimicking localized alveolar ridge defects, with a validity within 2.2% and with a variability of less than 2.8%.

Algorithms↗
PubMedDate not supplied

Comparative ultrastructure of needle aspiration biopsy and surgical resection specimens of lung tumors.

Ultrastructural examination affords conclusive evidence for classification of lung tumors. Tissue properly fixed for electron microscopy is not available in many cases, however. Ultrastructural diagnosis of resected specimens obviously follows, rather than directs, the surgical treatment. Fine-needle aspiration (FNA) of lung masses is recommended as a means to obtain lung tumor tissue for electron microscopy. Nevertheless, no comparison has been made between ultrastructural information gained from aspiration specimens and resected specimens. Electron microscopy was performed on transthoracic FNA specimens of 10 lung tumors for which surgical resection was subsequently performed. Glutaraldehyde-fixed specimens from FNA and surgical resection were prepared for electron microscopy according to routine procedures. Fixation of the FNA specimens was equivalent or superior to that of the resected specimens in 9 of the cases. Three of the FNA specimens contained necrotic as well as viable tissue. Features essential for diagnosis such as desmosomes, junctions, neurosecretory granules, intermediate filaments, glycogen, lipid, mucin, and microvilli were identifiable in both FNA and resected specimens. FNA specimens therefore yield a representative sample of the ultrastructural features of lung tumors when adequate cellular material is obtained. Use of a coaxial needle sampling technique with immediate microscopic assessment reduces the likelihood of retrieving only blood or necrotic tissue in the electron microscopy specimens.

Adult↗
PubMed1990

Standard specimens for stain calibration: application to Romanowsky-Giemsa staining.

Standardized specimens with reproducible staining properties were fabricated from extracts of biological objects (bovine liver, nucleoprotamine and defatted muscle). The standard specimens were stained with two formulations of the Romanowsky-Giemsa stain (RG), using the same azure B and eosin Y. One formulation used methanol and Sorensen's buffer and the other DMSO and Hepes buffer as solvents. The standard specimens were stained either in the composite stain or in the individual dyes dissolved in the same solvents and at the same concentration as the composite stain. Solution spectroscopy demonstrated different spectra for the two formulations with some wavelength regions varying by more than an order of magnitude. The RG spectra were also very different from those of the individual dyes dissolved at the RG concentration in the respective solvents. The stained standard specimens were analyzed by microspectrophotometry and were found to have spectra similar to those of cell smears. Furthermore, the standard specimens were shown to be a repeatable substrate for stain uptake. The transmitted light intensity from random fields of the same standardized specimen varied +/- 5%. When specimens were stained at the same time, the specimen-to-specimen variation depended on preparation conditions and the measurement wavelength, but was as good as +/- 5% for some conditions. The quantitative stain performance of both formulations was studied and compared. The standardized specimens provide a tool for the quantitative study of staining processes and specimen preparation procedures and for stain calibration.

Animals↗
PubMed1999

Cytologic examination of specimens obtained by means of tracheal washes performed before and after high-speed treadmill exercise in horses with a history of poor performance.

OBJECTIVE: To evaluate results of cytologic examination of specimens obtained by means of tracheal washes (TW) in 42 horses with a history of poor performance. DESIGN: Cross-sectional case series. ANIMALS: 42 horses with a history of poor performance. PROCEDURE: A TW was performed via endoscopy before and after horses exercised on a high-speed treadmill, and specimens were evaluated microscopically and graded. RESULTS: Ten (24%) horses were considered to be clinically normal before and after exercise. Pulmonary hemorrhage was diagnosed in 8 (19%) horses. One horse had evidence of exercise-induced pulmonary hemorrhage (EIPH) before exercise and pulmonary hemorrhage and allergic or inflammatory airway disease (IAD) after exercise. Five (12%) horses had IAD, and 1 had IAD and pulmonary hemorrhage after exercise. Seven (17%) horses had evidence of EIPH and IAD in both specimens. Four (10%) horses with EIPH had an increase in the proportion of hemosiderophages in the specimen obtained after exercise. Specimens obtained before exercise in 6 (14%) horses were not representative of the respiratory tract and could not be compared with specimens obtained after exercise. CLINICAL IMPLICATIONS: Interpretation of TW specimens obtained before and after exercise differed for only 5 of 36 (14%) horses. Specimens obtained after exercise were more likely to reveal airway disease. All specimens obtained after exercise adequately represented the respiratory tract, whereas 6 specimens obtained before exercise did not. Specimens obtained after exercise contained more airway secretions and had less cytologic evidence of pharyngeal contamination. Therefore, we recommend that TW samples be obtained after exercise in horses.

Animals↗
PubMed1999

Immunohistochemical Analysis of Radical Prostatectomy Specimens After 8 Months of Neoadjuvant Hormonal Therapy.

Neoadjuvant hormone therapy (NHT) prior to radical prostatectomy (RP) results in residual foci of atrophic glands, which can be difficult to identify with hematoxylin-eosin staining, raising the possibility that the low positive-margin rates are an artifact of pathologic understaging. The purpose of this study was to evaluate changes in prostate specific antigen (PSA) and prostatic acid phosphatase (PAP), as well as proliferation marker Ki-67 and Bcl-2 oncoprotein, by immunostaining after 8 months of NHT. Twenty-nine men with clinically confined prostate cancer who received 8 months of NHT and had both pretreatment biopsy and RP specimens obtained at Vancouver Hospital constituted the treatment group. The control group consisted of 23 RP specimens from patients not receiving NHT. Sections were stained with antibodies against PSA, PAP, proliferation marker Ki-67, and the antiapoptosis protein Bcl-2. The PSA and PAP immunostaining were graded for percentage of positive tumor cells and intensity of staining, while Ki-67 and Bcl-2 staining was graded according to the percentage of positive tumor cells. Absent or low percentage-positive PSA and PAP staining was observed in 40% to 50% of the NHT-treated RP specimens but none of the needle biopsy or untreated control RP specimens. Low-intensity PSA and PAP staining was detected only in RP specimens after NHT treatment, and then in only 20% of cases. Low or absent Ki-67 staining was noted in 78% of the NHT- treated RP specimens, compared with only 13% of the matched pre-NHT needle biopsies and 26% of untreated RP specimens. The percentage of specimens with high (>5%) Ki-67 staining decreased from 37% in the pre-NHT needle biopsies to 8% in the NHT-treated RP specimens. Bcl-2 staining increased after treatment with NHT, with 20% of the needle biopsies having high (>5%) Bcl-2 staining compared with 53% of the NHT-treated RP specimens. The frequency of low Bcl-2 staining (<1%) decreased from 53% in the pre-NHT needle biopsies to 27% in the NHT-treated RP specimens. Although PAP and PSA staining decreased after NHT, both markers remain sufficiently positive to be used as epithelial markers to help detect residual foci of prostate cancer that are difficult to identify on H&E-stained slides after NHT. Increased Bcl-2 after NHT, even in early-stage disease, is consistent with its role in the prevention of apoptosis and progression to androgen independence. Low levels of Ki-67 staining indicates a low probability of proliferation and outgrowth of androgen-independent clones after 8 months of NHT.

Journal Article↗
PubMedDate not supplied

Plastination of whole brain specimen and brain slices.

BACKGROUND: The human dead body specimens are plastinated for teaching purposes in medical institutions, using silicone. The silicone impregnated whole brain specimens and brain slices do not give satisfactory results. METHODS: In the present study the brain specimens were plastinated with another polymer known as Polyester-Copolymer. The brain specimens were first preserved and then fixed with 5% formalin. The specimens were then dehydrated and degreased in a volatile solvent acetone. The specimens were placed in Polyester-Copolymer solution which penetrated the brain tissue both intracellulary and intercelluraly. The specimens were then cured by gas method. RESULTS: The whole brain specimens and brain slices plastinated with Polyester-Copolymer were dry, odorless, handy and durable. It also gives a clear visual contrast between grey and white matter in brain slices whereas the brain specimens plastinated with silicone are flexible and sticky. There was no color contrast between grey and white matter. CONCLUSION: The polyester impregnated brain specimens and slices are non-toxic and ideal for teaching purposes and examinations. They require minimal aftercare. The whole organ serial sections of plastinated brain specimens will help 3-dimensional study of the normal brain and will improve the assessment of brain pathology.

Brain↗
PubMed1994

Investigation of oxygen-derived free radical generation in cancellous bone specimens obtained from dogs.

Generation of free radicals and the ability of various antioxidants to attenuate radical production in freshly procured cancellous bone specimens was investigated, using spin-trapping and electron spin resonance (ESR) techniques. Seven core cancellous bone specimens, 10 mm long and 7.9 mm in diameter, were obtained using aseptic technique, from the proximal portion of the humerus of 9 adult mixed-breed dogs. One core cancellous bone specimen from each dog was incubated in spin trap alpha-phenyl-N-tert-butylnitrone in Eagle's minimum essential medium and served as a control. The other 6 specimens from each dog were incubated in alpha-phenyl-N-tert-butylnitrone/Eagle's minimum essential medium plus 1 of the following antioxidants: superoxide dismutase, catalase, superoxide dismutase/catalase, indomethacin, allopurinol, or deferoxamine mesylate. All specimens were incubated at 26 C for 90 minutes, then frozen at -20 C until they were prepared for analysis by ESR spectroscopy. Each specimen was thawed, homogenized, and extracted in a low-dielectric organic solvent prior to obtaining an ESR spectrum which was analyzed for hyperfine splitting constants to identify radicals. Each first-derivative spectrum was digitally double-integrated to obtain an area: these areas were used to compare intensities of the spin. For each treatment group, the areas from the treated specimens were compared with the areas from the control specimens, using a paired t-test. Significance was accepted at P < or = 0.05. Spin adducts were detected in all cancellous bone specimens. Specimens incubated in deferoxamine (P = 0.0017) and superoxide dismutase/catalase (P = 0.0452) had significantly smaller areas than did control specimens.(ABSTRACT TRUNCATED AT 250 WORDS)

Allopurinol↗
PubMed2003

Potential margin distortion in breast tissue by specimen mammography.

HYPOTHESIS: There is a potential for margin distortion associated with the use of a compression plate in specimen mammography (SM) devices. DESIGN: Prospective observational study. SETTING: Two tertiary care teaching hospitals. PATIENTS: Breast biopsy tissue dimensions were recorded in 15 patients undergoing 18 operations before and after SM (group 1). After review of these data, we supervised the positioning of tissue for SM and obtained measurements for 12 breast biopsy specimens from 12 additional patients before and after SM (group 2). MAIN OUTCOME MEASURES: Breast biopsy specimen dimensions recorded before and after SM, including length, width, and depth (the specimen depth along the axis of compression); data on age, menopausal status, hormone therapy, and the presence or absence of calcifications or tumor; and a national telephone survey on technique and understanding of SM at 21 institutions. RESULTS: The specimen depth in group 1 was markedly decreased (P<.001) along the axis of compression. The mean length and width of the specimens were marginally increased by the effect of the SM device. Specimen depth in group 2 was unchanged when minimal pressure was used on the compression plate. Tissue was adequately immobilized in both groups, and all lesions were identified by SM. Our survey revealed that 71% of hospitals reported using firm compression in SM, and diverse techniques were employed. CONCLUSIONS: This study demonstrates the potential for specimen margin distortion with compression SM. Undue specimen compression is common in routine SM. Surgeon oversight in the method of securing the specimen within the SM container will minimize or avoid the potential for margin distortion.

Biopsy↗
PubMed1990

Amorphisation of MgO single crystal specimens prepared by ion milling for transmission electron microscopy studies.

Single crystal MgO specimens having low load Vickers indentations were thinned in an ion milling machine employing a single ion gun, and their characteristics were investigated with optical microscopy and high voltage electron microscopy (HVEM). It was found that the state of cleanliness of the specimen chamber of the ion milling machine had a very marked influence on the quality of the thinned specimens. If the specimen chamber was not well cleaned before ion milling a fresh specimen, the latter tended to show amorphisation due to the deposition on the specimen of the debris left in the chamber from the previously ion-milled specimens. Such observations were made from MgO specimens ion milled in several different types of commercial ion milling machine employing a single gun. It is proposed that to obtain good-quality ion milled TEM specimens, it is important to clean the specimen chamber thoroughly prior to milling.

Magnesium Oxide↗
PubMed2003

Four-year experience of use of the Cobas Amplicor system for rapid detection of Mycobacterium tuberculosis complex in respiratory and nonrespiratory specimens in Greece.

To evaluate the experience of a clinical microbiology laboratory with a DNA amplification assay for routine detection of Mycobacterium tuberculosis, the Cobas Amplicor Mycobacterium tuberculosis (MTB) polymerase chain reaction (PCR) assay (Roche Diagnostics Systems, USA) was performed on 7,722 respiratory and 1,451 nonrespiratory specimens collected from 3,321 patients. The results were compared with those of culture in conventional Lowenstein-Jensen medium, culture in the MB/BacT system (Organon Teknika, France), and clinical investigations. A total of 240 of the 254 respiratory specimens culture positive for Mycobacterium tuberculosis were also positive in the PCR assay. Of the 7,300 culture-negative specimens, 45 (0.6%) were positive in the PCR. After detailed interpretation, the overall sensitivity, specificity, and positive and negative predictive values of the PCR assay were 84.5, 99.8, 94.1, and 99.4%, respectively, for respiratory specimens. The PCR assay was more sensitive for smear-positive respiratory specimens (97.1%) than for smear-negative respiratory specimens (48.6%). Of the 18 culture-positive (smear-negative) nonrespiratory specimens, 9 were positive in the PCR. None of the 1,384 culture-negative nonrespiratory specimens were positive in the PCR. The inhibition rates detected by the internal control of the test were 2.2% for respiratory specimens and 3.4% for nonrespiratory specimens. After resolving the discrepancies, the overall sensitivity, specificity, and positive and negative predictive values of the PCR assay were 82.5, 99.8, 94.3, and 99.4%, respectively, when compared to the results of diagnostic culture. In conclusion, the use of the Cobas Amplicor MTB-PCR assay might enable clinical microbiology laboratories with considerable previous experience in molecular biology testing to perform PCR and confirm tuberculosis infection immediately, leading to improved patient management.

Bacteriological Techniques↗