Complexity of 12q13–22 amplicon in liposarcoma: Microsatellite repeat analysis

The 12q13-22 amplicon from four liposacroma specimens evaluated by comparative genomic hybridization was studied analyzing 55 microsatellite markers by PCR. All four specimens were informative in at least 34 loci; an amplification or allelic imbalance was identified with four to 17 markers. The amplicons were discontinuous; there were non-amplified marker loci between the amplified marker loci. These findings indicate the presence of separate amplicons in the 12q13–22 region. Evidence of the concomitant gain of one allele and loss of the other allele was found with several markers in one tumor and with one marker in two tumor specimens. Southern blotting showed amplification of CDK4 and MDM2 in all four specimens. Genes Chromosom. Cancer 18:66–70, 1997. © 1997 Wiley-Liss, Inc.     

RT-PCR based diagnosis and molecular characterisation of mumps viruses derived from clinical specimens collected during the 1996 mumps outbreak in Portugal

Clinical specimens collected during an outbreak of mumps were characterised by RT-PCR, nested PCR, and nucleotide sequencing. Mumps virus was positively identified in 12/21(57%) saliva, 9/21(43%), throat and 1/33(3%) urine specimens and further sequence comparison revealed that at least six strains of viruses, which differed from 0–9.43% at the nucleotide levels, were co-circulating during the epidemic. However, phylogenetic analysis showed that these viruses grouped with two previously identified lineages which were mostly composed of other European mumps virus isolates. J. Med. Virol. 52:349–353, 1997. © 1997 Wiley-Liss, Inc.     

Mixed genogroup SRSV infections among a party of canoeists exposed to contaminated recreational water

Samples of faeces collected from a party of canoeists involved in a gastroenteritis outbreak were examined by electron microscopy and RT-PCR for evidence of infection with SRSVs. A broadly reactive primer pair was used to detect SRSVs followed by application of genogroup-specific primers to SRSV-positive specimens. Exposure data were collected by means of a questionnaire. SRSVs were detected in 1/4 specimens examined by EM and 3/4 by RT-PCR. Genogrouping, and sequencing of PCR products revealed two distinct strains: a genogroup I strain, related to the Desert Shield virus, and a genogroup II strain, related to the Lordsdale virus to be associated with the outbreak. Exposure data indicated that capsising and eating food before getting changed were associated with an increased risk of gastroenteritis and was consistent with infection following the consumption of contaminated water. This study confirms the greater sensitivity of RT-PCR for the diagnosis of SRSV infections and its utility, when incorporating genogroup-specific primers, in establishing more complex epidemiological data. J. Med. Virol. 52:425–429, 1997. © 1997 Wiley-Liss, Inc.     

Trypanosoma rangeli : increase in virulence with inocula of different origins in the experimental infection in mice

We compared two murine models of Trypanosoma rangeli infection. The same inoculum dose and age-matched hosts were used in both cases. One group was infected with trypomastigotes obtained from passages in mice and the other, with trypomastigotes obtained from cell culture after a passage in mice. We observed that trypomastigotes obtained from the in vitro cellular infection showed increased virulence in␣experimental animals, with a 70% rate of death being noted in experimental mice instead of the lack of mortality seen when in vivo-derived parasites were used. The greatest levels of parasitemia and tissual lesions in the presence of the parasite also occurred when in vitro-derived parasites were used. Received: 10 March 1997 / Accepted: 10 April 1997     

Helicobacter pylori antigen in the glomeruli of patients with membranous nephropathy

 Renal biopsy specimens from patients with membranous nephropathy (MN) were studied using immunohistochemical labelling to clarify the aetiological significance of Helicobacter pylori antigen in this disease. Sixteen specimens were examined, from 7 male and 9 female MN patients. Renal specimens from patients with diabetic nephropathy and IgA nephropathy, and from autopsied patients without renal diseases were obtained as controls. Immunohistochemical labelling was performed using one polyclonal antibody and three monoclonal antibodies against H. pylori. Specimens from 11 of the MN patients revealed granular deposits along the glomerular capillary walls, which reacted positively with polyclonal antibody after trypsin pretreatment. None of the control specimens revealed positive labelling. The MN specimens showed no positive reaction with the primary antibody, which had been treated for immunoabsorption testing using sonicated H. pylori.We also determined H. pylori status in these MN patients histologically and/or serologically. Of the 11 patients whose glomeruli were positive for anti-H. pylori antibody, 7 were suitable for analysis, and all were regarded as positive for H. pylori infection. These results suggest that the presence of a specific antigen in the glomeruli of patients with MN and H. pylori infection may be involved in the pathogenesis of MN. Received: 18 October 1996 / Accepted: 3 March 1997     

Cytologic specimens from the eye: A clinicopathologic study of 33 patients

Cytologic specimens from the eye (CSE) (fine-needle aspirates and intraocular [vitrectomy] washings) are uncommon in most cytology practices. We reviewed our experience and diagnostic accuracy in lesions from this site over a 13-yr period. Thirty-three patients (12 males, 21 females, age 7–85 yr; mean 56 yr) were identified. Eighteen specimens were submitted to rule out malignancy. Of these, four CSE were positive, including melanoma (n = 1), large-cell lymphoma (n = 2), and carcinoma (n = 1). None of the 14 patients with negative cytology showed clinical evidence of malignancy over a 1–28-mo follow-up period (mean, 8.4 mo). Fourteen specimens were submitted to rule out infectious agents. Cytology material contained culture-proven organisms in one case each of Candida andAspergillus. Of serology-confirmed specimens, four CSE failed to reveal toxoplasma, and three lacked herpes viral inclusions. One case each of culture-positiveP. acnes andC. albicans lacked the organisms in the CSE. Twelve specimens were submitted to rule out inflammatory lesions. CSE showed chronic inflammation (n = 5), acute and chronic inflammation (n = 1), macrophages (n = 5), and nonnecrotizing granulomas (n = 1). Of 3 patients with sarcoidosis, CSE contained mature lymphocytes in all 3 cases, but no granulomas. Although the numbers are small, there is greater sensitivity in the detection of malignancy than of infectious agents. Diagn. Cytopathol. 1997;17:262–266. © 1997 Wiley-Liss, Inc.     

Identification and typing of molluscum contagiosum virus in clinical specimens by polymerase chain reaction

A polymerase chain reaction (PCR) which enables the detection of molluscum contagiosum virus (MCV) genomes in either fresh or formalin-fixed clinical specimens is described. The primers used were designed to amplify a 167 bp region of the 3.8kbp HindIII fragment K of the MCV 1 genome. The ability of this PCR to detect three common MCV types (1, 1v and 2) in clini-cal specimens was confirmed using frozen extracts from 75 molluscum lesions, and digests of single sections of 11 formalin-fixed, paraf-fin-embedded lesions; all of which had been previously typed by Southern hybridisation. In addition, 2 specimens previously negative by hybridisation were shown to be positive for MCV DNA by PCR. Confirmation of the identity of the PCR products and distinction between the two major MCV types (MCV 1/1v versus MCV 2) was achieved by comparison of the results of cleavage with the restriction endonucleases HhaI and SacI. Sequencing of the PCR products revealed complete homology between MCV 1 and 1v, but minor nucleotide variations between MCV 1/1v and MCV 2 were identified. As well as providing a highly sensitive means of diagnosis, the technique may also prove useful for investigations into the pathogenesis, epidemiology and natural history of molluscum contagiosum infection. J. Med. Virol. 53:205–211, 1997. © 1997 Wiley-Liss, Inc.     

Microhabitat distribution and coexistence of Microcotylidae (Monogenea) on the gills of the striped mullet Mugil cephalus : chance or competition?

The two monogenean species Metamicrocotyla cephalus and Microcotyle mugilis have specific microhabitats on the gills of Mugil cephalus on which they may or may not coexist. M. cephalus is found in sector 1 of the posterior hemibranch of arch I. M. mugilis is found on the filaments of sector 5 of the posterior hemibranch of arch I. The coexistence of these two monogenean species on the same fish does not induce a change in their respective distribution, except for a marked preference of M. cephalus for the left side and of M. mugilis for the right. This 15-month-long study shows that when the two species of monogeneans coexist the infection prevalence and intensity are higher than in cases of monospecific infection. The specific character of the microhabitat, the apparent absence of interspecific competition, and the existence of positive species interactions are discussed. Received: 2 July 1997 / Accepted: 28 August 1997     

Significant increase in antigastric autoantibodies in a long-term follow-up study of H. pylori gastritis

In 30% of H. pylori-infected patients a certain type of antigastric autoantibodies, reacting against canalicular structures within human parietal cells, is detectable. Furthermore, it has been shown that these autoantibodies are correlated with atrophy of the mucosa in the corpus. The aim of this study was to analyse the prevalence of these anticanalicular autoantibodies (ACAB) and their significance for development of gastric mucosa atrophy in a 12-year follow-up period. Gastric biopsy specimens from 62 persons in Saaremaa Island, Estonia, were collected in 1997 and assessed independently by two pathologists in accordance with the updated Sydney system. The sera of these persons were immunohistochemically screened for ACAB and for classic parietal cell antibodies (PCA). In addition, for 37 of the 62 persons, gastric biopsies and sera collected 12 years earlier (1985) were investigated in an analogous manner. ACAB increased significantly, from 8 out of 37 in 1985 to 17 out of 37 in 1997 (P=0.004; McNemar test). In 1997 a significant correlation existed between the presence of ACAB and corpus mucosa atrophy (19 out of 30 versus 10 out of 32 without atrophy; P=0.01; odds ratio (OR)=3.8, 95% CI 1.4–10.6). However, no correlation was found between ACAB and development of atrophy in the period from 1985 to 1997. All 37 persons were PCA negative in 1985, whereas in 1997, 2 turned out to be PCA positive. ACAB increased significantly with duration of H. pylori gastritis. The correlation between ACAB and presence of gastric corpus atrophy was confirmed. However, it is possible that ACAB are the consequence of and not a causative factor in gastric mucosa atrophy, insofar as the association of ACAB with progression of corpus atrophy was not significant. Received: 20 October 1999 / Accepted: 13 January 2000     

Comparison between vaginal tampon and cervicovaginal lavage specimen collection for detection of human papillomavirus DNA by the polymerase chain reaction

The aim of the study was to compare the accuracy of self-administered vaginal tampon (VT) specimens for the detection of human papillomaviruses (HPVs) with that of cervicovaginal lavage specimens (CVL). Two hundred seventy-four paired VT and CVL specimens were collected prospectively from women at risk of sexually transmitted diseases. Specimens were treated and amplified with the polymerase chain reaction (PCR). Each woman served as her own control. One hundred and forty-four of 272 (52.9%) CVLs and 159 of 271 (58.7%) VTs contained HPV DNA sequences (correlation of 88%). The sensitivity and specificity of vaginal tampons reached 93.9% (138/147) and 80.5% (99/123), respectively. HPV typing results were concordant for 99 negative paired samples and 114 paired samples positive for the same type(s) (correlation of 78.9%). It is concluded that these sampling methods collect cells from different areas of the genital epithelium, highlighting the importance of further assessment of the comparative predictive value of HPV detection in each sample. J Med Virol 51: 42–47, 1997. © 1997 Wiley-Liss, Inc.     

Diagnostic pitfalls in fine-needle aspiration biopsy of the mediastinum

A retrospective review of 189 fine-needle aspiration (FNA) biopsies of the mediastinum from four university medical centers was performed. Review of Diff-Quik- and Papanicolaou-stained direct smears was performed from a series of 189 FNA biopsies along with surgical pathology correlation obtained in 42% of the cases. There were 28 (14.8%) nondiagnostic or unsatisfactory for diagnosis cases. Of the satisfactory FNA specimens with histologic correlation, 12 cases (6%) were discordant. These errors primarily involve subclassification of small-cell malignancies involving the mediastinum, including a misdiagnosis of small-cell carcinoma for lymphoma. Large-cell lesions that were problematic included the accurate diagnosis of Hodgkin's lymphoma including the separation from non-Hodgkin's lymphoma. Large-cell lymphoma with sclerosis was misinterpreted in two cases due to distortion of cells by the mesenchymal tissue and sparsely cellular smears. In two cases classification of primary germ-cell tumors and separation from metastatic carcinoma was a problem. In general, FNA of the mediastinum is an accurate procedure, but can be challenging in a minority of cases due to sparse cellularity of the lesions and accurate classification of a variety of neoplasms that occur in this region. These 12 discordant cases serve as the basis of our report. Diagn. Cytopathol. 17:121–126, 1997. © 1997 Wiley-Liss, Inc.     

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

The Museum Vrolik collection of human anatomy comprises 360 recently redescribed 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. Am. J. Med. Genet. 68:257–259, 1997. © 1997 Wiley-Liss, Inc.     

An outbreak of febrile gastroenteritis associated with corn contaminated by Listeria monocytogenes

BACKGROUND: On May 21, 1997, numerous cases of febrile gastrointestinal illness were reported among the students and staff of two primary schools in northern Italy, all of whom had eaten at cafeterias served by the same caterer. METHODS: We interviewed people who ate at the cafeterias about symptoms and foods consumed on May 20. There were no samples of foods left at the cafeterias, but we tested routine samples taken on May 20 by the caterer and environmental specimens at the catering plant. The hospitalized patients were tested for common enteropathogens and toxins. RESULTS: Of the 2189 persons interviewed (82 percent of those exposed), 1566 (72 percent) reported symptoms; of these, 292 (19 percent) were hospitalized. Among samples obtained from hospitalized patients, all but two of the stool specimens and all blood specimens were negative for common enteropathogens. Listeria monocytogenes was isolated from one blood specimen and from 123 of the 141 stool specimens. Consumption of a cold salad of corn and tuna was associated with the development of symptoms (relative risk, 6.19; 95 percent confidence interval, 4.81 to 7.98; P<0.001). L. monocytogenes was isolated from the caterer's sample of the salad and from environmental specimens collected from the catering plant. All listeria isolates were serotype 4b and were found to be identical on DNA analysis. Experimental contamination of sterile samples of the implicated foods showed that L. monocytogenes grew on corn when kept for at least 10 hours at 25 degrees C. CONCLUSIONS: Food-borne infection with L. monocytogenes can cause febrile illness with gastroenteritis in immunocompetent persons.     

Decalcified Bone: Twenty Years of Successful Specimen Management

Abstract

Producing quality slides of paraffin embedded, decalcified bone depends on the interaction of several independent mechanisms that all must work in concert. Grossing, fixation, decalcification, processing, embedding, sectioning, and staining each comprise a unique procedure when examining the quality of a slide. A system that integrates all these concepts in a proven workable formula is essential for specimen management and necessary for greater reproducibility. A thorough understanding of each of the component concepts is necessary to tailor the system to a variety of hard tissue types and specimens. (The J Histotechnol 20:267, 1997)     

Antibodies against enteroviruses in intravenous Ig preparations: Great variation in titres and poor correlation with the incidence of circulating serotypes

Antibody titres in immunoglobulin preparations for intravenous use were tested against 24 different enterovirus serotypes and varied between 1 : 100 and 1 : 10,000 within a single batch. Differences up to 8-fold were found for homologous titres between two different batches that were prepared 6 years apart. The lowest titre obtained was 1 : 40. The observed differences within and between the two batches could not be explained by different incidence of serotypes of enteroviruses circulating at the time blood was collected. Differences in titres of up to 18-fold were observed when several strains of the same serotype were tested suggesting that intratypic variation influences antibody titres. It is concluded that immunoglobulin preparations contain antibodies against many enteroviruses, but that titres can be low and cannot be predicted from the incidence of any particular serotype circulating in the community. Because of intratypic variation, selection of a batch for specific treatment should be based on results obtained with the patient's own isolate, and not with a reference strain. J. Med. Virol. 53:273–276, 1997. © 1997 Wiley-Liss, Inc.     

Evaluation of bone marrow and blood cultures for the recovery of mycobacteria in the diagnosis of disseminated mycobacterial infections

This study evaluated the validity of bone marrow (BM) and blood specimens for the diagnosis of disseminated mycobacterial infections (DMIs). From 1990 to February 1997, all specimens were processed with the lysis-centrifugation procedure; thereafter (until December 2001), they were processed with the BACTEC Myco/F Lytic system. Twenty-three paired BM-blood specimens with mycobacteria in at least one specimen were studied from 23 patients. The strains isolated were 14 Mycobacterium avium complex (MAC) and nine M. tuberculosis complex (MTBC). Blood specimens had a statistically significant greater sensitivity for the isolation of MAC than BM (100% vs. 57.1%, respectively), whereas sensitivity for the isolation of MTBC was equal for the two specimen types (66.7%). Although not statistically significant, the times required to detect mycobacteria from blood specimens were lower than those from BM in the MycoF/Lytic system. Overall, blood cultures represented a more sensitive and less invasive alternative to BM cultures for the diagnosis of disseminated mycobacteriosis caused by MAC, especially when the MycoF/Lytic system was used, but provided no advantage for the diagnosis of DMI caused by MTBC.     

Features of the Femoral Proximocaudal Joint Capsule Insertion Among Canids

This observational study was conducted to evaluate the anatomic relationship between the proximocaudal femoral joint capsule insertion and the femoral caudolateral curvilinear osteophyte (CCO), across ancient and modern domestic and non-domestic canids. Museum specimens of proximal femora were screened for presence of remnant enthesophytes of the caudal joint capsule insertion (first inclusion criterion) and then for the CCO (second inclusion criterion). The initially screened population included 267 dry bone specimens: Six Canis species, hybrid coyote × domestic dog, and five vulpines (three Vulpes species, one Urocyon, and one Nyctereutes). Proximocaudal joint capsule insertion remnant enthesophytes were limiting at n = 19 specimens: Seven ancient domestic dogs, four modern coyotes, two ancient coyotes, two modern hybrid coyote × dog, two modern red foxes, and two modern raccoon dogs. The joint capsule enthesophytes are associated with inflammation, but are observed far less frequently than the CCO. The CCO is seen radiographically but is visible more frequently by direct inspection. The primary inclusion criterion necessarily was a visible caudal joint capsule insertion; spatial relationships of the CCO can be assigned with confidence only when a capsule insertion line can be recognized clearly. We demonstrate that the anatomic CCO associates with the joint capsule insertion being nonspecific and species-independent. A joint capsule insertion-CCO spatial relationship across species is an important new observation, strongly indicating that both are pathological features. Our data indicate need for new research to characterize the canid coxofemoral joint and its overt and incipient pathology in a phylogenetic context. Anat Rec, 302:2164–2170, 2019. © 2019 American Association for Anatomy     

Bronchoalveolar lavage: Comparison of nucleopore filters and direct smear preparations

To determine whether cytopreparation affects the diagnostic yield of bronchoalveolar lavage specimens, we compared 50 lavage samples by using two methods. One nucleopore filter preparation and four direct slides were prepared on each sample submitted. All slides were stained by the Papanicolaou method. Assessment of cellularity, cellular preservation, and an independent diagnosis were rendered on each sample for both preparatory methods. Statistical analysis showed no difference in cellularity between the two methods (P = .06). The degree of nuclear and cytoplasmic preservation was higher using the direct method, although this did not appear to affect diagnosis in this study. One major discrepancy in diagnosis was observed between the two methods. By prospectively comparing nucleopore filter and direct preparation of bronchoalveolar lavages, we found that there was minimal affect on either cellularity or diagnosis. We conclude that either method delivers reliable cytologic results. Diagn. Cytopathol. 17:160–164, 1997. © 1997 Wiley-Liss, Inc.