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[Melanocyte tumors. Classical and new diagnostic possibilities].

Melanocytic tumors are one of the major problems in diagnostic dermatopathology as they comprise benign nevi, malignant melanomas and borderline cases. Apart from a proportion of congenital lesions, most benign nevi are acquired tumors that arise during early adulthood and eventually may undergo regressive change. Histologically, they present as so-called common nevi, so-called dysplastic nevi, Spitz's nevi, blue nevi and their variants, and combined nevi. In typical cases, the distinction from a malignant melanoma is not difficult. However, benign simulators of malignancy exist as much as deceptively bland appearing melanomas and in some cases the diagnosis remains dubious despite careful weighting of criteria. Indeed, morphological features may not always suffice to disclose the nature of a melanocytic tumor. Ancillary techniques including immunohistochemistry and measurement of telomerase activity may be of assistance in this respect. One should nevertheless be aware that the biological behavior of certain borderline cases cannot be predicted with certainty.

Diagnosis, Differential↗

Vector for IS element entrapment and functional characterization based on turning on expression of distal promoterless genes.

We constructed and characterized a novel trap vector for rapid isolation of insertion sequences. The strategy used for the isolation of IS elements is based on the ability of many IS elements to turn on the expression of otherwise silent genes distal to some sites of insertion. The simple transposition of an IS element can sometimes cause the constitutive expression of promoterless antibiotic resistance genes resulting in selectable phenotypes. The trap vector pAW1326 is based on a pBR322 replicon, it carries ampicillin and streptomycin resistance genes, and also silenced genes that confer chloramphenicol and kanamycin resistance once activated. The trap vector pAW1326 proved to be efficient and 85 percent of all isolated mutations were insertions. The majority of IS elements resident in the studied Escherichia coli strains tested became trapped, namely IS2, IS3, IS5, IS150, IS186 and Tn1000. We also encountered an insertion sequence, called IS10L/R-2, which is a hybrid of the two IS variants IS10L and IS10R. IS10L/R-2 is absent from most E. coli strains, but it is detectable in some strains such as JM109 which had been submitted to Tn10 mutagenesis. The distribution of the insertion sequences within the trap region was not random. Rather, the integration of chromosomal mobile genetic elements into the offered target sequence occurred in element-specific clusters. This is explained both by the target specificity and by the specific requirements for the activation of gene transcription by the DNA rearrangement. The employed trap vector pAW1326 proved to be useful for the isolation of mobile genetic elements, for a demonstration of their transposition activity as well as for the further characterization of some of the functional parameters of transposition.

DNA Transposable Elements↗

Prion diseases in man.

Human spongiform encephalopathies (HSE) are uncommon neurodegenerative diseases of unknown etiology. Whilst numerous observations provide arguments for an infectious disorder mediated by an unconventional agent of the so-called prion-type, the recent discovery of predisposing allelic variants suggests that HSE might be considered as genetically linked diseases.

Animals↗

Sarcomatoid salivary duct carcinoma.

The so-called sarcomatoid salivary duct carcinoma (SSDC) is one of the variants of salivary duct carcinoma (SDC). This neoplasm is characterized by the presence of both a carcinomatous and a sarcomatoid tumor component. The histology and nomenclature of such neoplasms has been a matter of debate for many years. The histologic, immunohistochemical, and electron microscopic findings including those of 4 previously described cases of SSDC are defined and the different attitudes concerning their etiology will be discussed. In addition, the fine-needle aspiration biopsy of such a case is presented for the first time. In analogy to typical SDC there seems to be a predilection for elderly men and a location in major salivary glands. The resected SSDC tumors measured between 1.5 and 3.5 cm. Histologically, each case was a composite of SDC and sarcomatoid carcinoma. Immunohistochemical positivity for epithelial membrane antigen (EMA) and cytokeratins (AE1/AE3, CAM 5.2) was shown in the sarcomatoid tumor component. The important cytomorphologic feature of SSDC is the presence of cohesive clusters and flat sheets of cells with a cribriform pattern, in combination with an atypical spindle cell component. The use of the term SSDC seems more appropriate than the term carcinosarcoma , as the immunohistochemical, electron microscopic, and recent molecular findings in this and other biphasic neoplasms imply a monoclonal origin.

Adult↗

[Maxillary sinus hypoplasia].

Maxillary sinus hypoplasia is rare, with an estimated prevalence of 1-5%. Out of the CT scans performed in sinusal patients between March 1998 and June 1999, we report on 4 isolated maxillary sinus hypoplasia, 4 maxillary sinus hypoplasia associated to concha bullosa, and 10 isolated conchae bullosas. All cases were evaluated by nasosinusal endoscopy and CT scan. Size, location and uni/bilateral presentation of concha bullosa is correlated to maxillary sinus hypoplasia presence, specially with regards to uncinate process presence, medial or lateral retraction. The pathogenesis of maxillary sinus hypoplasia is reviewed, and its relation to concha bullosa, evaluating how this could explain some cases of the so called chronic maxillary sinus atelectasia, as an acquired and progressive variant of maxillary sinus hypoplasia in adults.

Adolescent↗

Synthesis of highly diversified carrier ampholytes. Evaluation of the resolving power of isoelectric focusing in the Pi system (alpha-1-antitrypsin genetic polymorphism).

The use of condensing reagents such as epoxypropanol, diepoxyoctane, acrylamide and N,N'-methylenebisacrylamide in the synthesis of carrier ampholytes increased the diversity of amphoteric components. The quality of these synthetic carrier ampholytes has been tested in the separation of variants of alpha-1-anti-trypsin, a genetic polymorphism called the Pi system. A resolving power of the order of 0.005 pH unit was obtained.

Ampholyte Mixtures↗

Mutations in the adiponectin gene in lean and obese subjects from the Swedish obese subjects cohort.

Adiponectin (also called AdipoQ, gelatin-binding protein 28, Acrp30) DNA sequence variants were determined in 96 unrelated female subjects with severe obesity (mean body mass index [BMI], 42.3 kg/m2) and in 96 non-obese female controls (mean BMI, 23.0 kg/m2) from the Swedish Obese Subjects (SOS) cohort. A single base substitution (T45G) at codon 15 of exon 2 resulting in no change in amino acid (Gly15Gly) was found in equal frequencies among obese and control subjects. However, this polymorphism was associated with serum cholesterol and waist circumference (P=.023 and.043, respectively) in the obese group. A IVS2 + G62T sequence variation was also identified, but had similar prevalence rates in obese and control subjects. Blood glucose was highest in the obese female subjects who were homozygotes for the G allele (GG) of the IVS2 + G62T polymorphism (N=56; P=.033) and all the diabetics (n=6) in this sample were in this group. IVS2 + G62T polymorphism was also associated with BMI (P=.014), diastolic blood pressure (P=.009), and sagittal diameter (P=.032). A missense point mutation at codon 111 (Tyr111His) was not associated with any obesity-related phenotypes. In conclusion, adiponectin DNA sequence variations might play a role in the complications of morbid obesity and should be further investigated.

Adiponectin↗

Studies on proteins in post-ecdysial nymphal cuticle of locust, Locusta migratoria, and cockroach, Blaberus craniifer.

Proteins were extracted from the cuticle of mid-instar nymphs of locusts, Locusta migratoria, and cockroaches, Blaberus craniifer. Seven proteins were purified from the locust extract and five from the cockroach extract, and their amino acid sequences were determined. Polyacrylamide gel electrophoresis indicates that the proteins are present only in the post-ecdysially deposited layer of the nymphal cuticles. One of the locust and one of the cockroach nymphal proteins contain a 68-residue motif, the RR-2 sequence, which has been reported for several proteins from the solid cuticles of other insect species. Two of the cockroach proteins contain a 75-residue motif, which is also present in a protein from the larval/pupal cuticle of a beetle, Tenebrio molitor, and in proteins from the exoskeletons of a lobster, Homarus americanus, and a spider, Araneus diadematus. The motif contains a variant of the Rebers-Riddiford consensus sequence, and is called the RR-3 motif. One of the locust and three of the cockroach post-ecdysial proteins contain one or more copies of an 18-residue motif, previously reported in a protein from Bombyx mori pupal cuticle. The nymphal post-ecdysial proteins from both species have features in common with pre-ecdysial proteins (pharate proteins) in cuticles destined to be sclerotised; they show little similarity to the post-ecdysial cuticular proteins from adult locusts or to proteins from soft, pliable cuticles. Possible roles for post-ecdysial cuticular proteins are discussed in relation to the reported structures.

Amino Acid Sequence↗

The role of Glu39 in MnII binding and oxidation by manganese peroxidase from Phanerochaete chrysoporium.

Manganese peroxidase (MnP) is a heme-containing enzyme produced by white-rot fungi and is part of the extracellular lignin degrading system in these organisms. MnP is unique among Mn binding enzymes in its ability to bind and oxidize Mn(II) and efficiently release Mn(III). Initial site-directed mutagenesis studies identified the residues E35, E39, and D179 as the Mn binding ligands. However, an E39D variant was recently reported to display wild-type Mn binding and rate of oxidation, calling into question the role of E39 as an Mn ligand. To investigate this hypothesis, we performed computer modeling studies which indicated metal-ligand bond distances in the E39D variant and in an E35D--E39D--D179E triple variant which might allow Mn binding and oxidation. To test the model, we reconstructed the E35D and E39D variants used in the previous study, as well as an E39A single variant and the E35D--E39D--D179E triple variant of MnP isozyme 1 from Phanerochaete chrysosporium. We find that all of the variant proteins are impaired for Mn(II) binding (K(m) increases 20--30-fold) and Mn(II) oxidation (k(cat) decreases 50--400-fold) in both the steady state and the transient state. In particular, mutation of the E39 residue in MnP decreases both Mn binding and oxidation. The catalytic efficiency of the E39A variants decreased approximately 10(4)-fold, while that of the E39D variant decreased approximately 10(3)-fold. Contrary to initial modeling results, the triple variant performed only as well as any of the single Mn ligand variants. Interestingly, the catalytic efficiency of the triple variant decreased only 10(4)-fold, which is approximately 10(2)-fold better than that reported for the E35Q--D179N double variant. These combined studies indicate that precise geometry of the Mn ligands within the Mn binding site of MnP is essential for the efficient binding, oxidation, and release of Mn by this enzyme. The results clearly indicate that E39 is a Mn ligand and that mutation of this ligand decreases both Mn binding and the rate of Mn oxidation.

Amino Acid Substitution↗

The cGMP-gated channel and related glutamic acid-rich proteins interact with peripherin-2 at the rim region of rod photoreceptor disc membranes.

The rod cGMP-gated channel is localized in the plasma membrane of rod photoreceptor outer segments, where it plays a central role in phototransduction. It consists of alpha- and beta-subunits that assemble into a heterotetrameric protein. Each subunit contains structural features characteristic of nucleotide-gated channels, including a cGMP-binding domain, multiple membrane-spanning segments, and a pore region. In addition, the beta-subunit has a large glutamic acid- and proline-rich region called GARP that is also expressed as two soluble protein variants. Using monoclonal antibodies in conjunction with immunoprecipitation, cross-linking, and electrophoretic techniques, we show that the cGMP-gated channel associates with the Na/Ca-K exchanger in the rod outer segment plasma membrane. This complex and soluble GARP proteins also interact with peripherin-2 oligomers in the rim region of outer segment disc membranes. These results suggest that channel/peripherin protein interactions mediated by the GARP part of the channel beta-subunit play a role in connecting the rim region of discs to the plasma membrane and in anchoring the channel.exchanger complex in the rod outer segment plasma membrane.

Amino Acid Sequence↗

An unusual Neisseria isolated from conjunctival cultures in rural Egypt.

Seven isolates of an unusual Neisseria sp. were obtained from eye cultures of children in two rural Egyptian villages. These Neisseria utilized only glucose, they exhibited a positive reaction when tested with antisera to crude antigen from Neisseria meningitidis and N. gonorrhoeae, and they did not react with the fluorescent antibody tests for N. gonorrhoeae or with the monoclonal antibodies used to serotype gonococci. The Egyptian isolates had colony morphology more typical of meningococci than gonococci and showed opaque and transparent colony variants. On SDS-PAGE, the major outer-membrane proteins had different patterns than those noted for comparable proteins of meningococci and gonococci; heat-modifiable outer-membrane proteins were present. Four of the six isolates examined had cryptic plasmids of 2.8 megadaltons, which were slightly larger than the cryptic plasmid of N. gonorrhoeae. These plasmids were homologous to the gonococcal cryptic plasmid, but had different restriction enzyme fragment patterns. The DNA from the Egyptian isolates, like DNA from N. meningitidis but unlike DNA from N. gonorrhoeae, could be cut with the restriction enzyme HaeIII. The frequency of transformation into a temperature-sensitive mutant of N. gonorrhoeae was 0.2 for the Egyptian isolates and 0.1 for N. meningitidis, a frequency that was 5-10-fold lower than that for the N. gonorrhoeae control isolates. Whole-cell DNA from the Egyptian isolates showed 68%-73% homology with N. gonorrhoeae and 57%-63% with N. meningitidis. On the basis of our observations, the Egyptian isolates are distinct from N. meningitidis and may represent a variant of N. gonorrhoeae. We suggest that the isolates be called Neisseria gonorrhoeae ssp. kochii.

Bacterial Proteins↗

Malignant chondroid syringoma: immunohistopathology.

Malignant chondroid syringoma, also called malignant mixed tumor of the skin, is a rare variant of a malignant tumor derived from sweat gland cells. Histologically the tumor is composed of an epithelial and a mesenchymal component. The epithelial structures show glandular differentiation and carcinomatous features. They are embedded in a mucinous stroma with spindle mesenchymal cells and areas of chondroid differentiation. The epithelial cells express cytokeratins. The luminal epithelial cells show binding to the lectin Ulex europaeus; intraluminal cells are carcinoembryonic antigen positive. The stromal cells are cytokeratin negative and sporadically positive for vimentin. Chondroid areas are S-100 protein and vimentin positive. No labeling for actin is seen.

Adenoma, Pleomorphic↗

Pathologic and clinical features of 77 Hodgkin's lymphoma patients treated in a lymphoma protocol (LNH87): a GELA study.

Between 1987 and 1993, 77 of 2855 lymphomas included in the LNH87 protocol of the GELA as non-Hodgkin lymphoma (NHL) and reviewed by a panel of pathologists had a diagnosis changed to Hodgkin lymphoma (HL). Some of these lymphomas had been initially interpreted as anaplastic large-cell lymphoma Hodgkin-like (ALCL-HL subtype). The purpose of this study was to analyze the histologic pitfalls initially encountered, to define more clearly the diagnostic criteria of lymphomas placed in the gray zone around HL, and to follow the survival of these 77 patients affected with HL and initially treated with NHL regimens. The 77 cases of HL were reviewed by three hematopathologists and immunostained with a large panel of antibodies, including CD30, CD15, CD3, CD20, CD45, CD43, LMP-1, EMA, BNH-9, TiA1, and ALK1. Each case was classified according to the Lukes-Rye system and the British National Lymphoma Investigation (BNLI) grading. The initial clinical presentation of patients was analyzed, and the overall and event-free survival rates of the 77 patients were estimated. Among the 77 HLs, 46 were misinterpreted as NHL by primary individual pathologists (12 as ALCL, 8 as ALCL-HL, 12 as peripheral T-cell lymphoma (PTCL), 6 as B-cell lymphoma, and 8 as unclassifiable NHL). The other 31 cases had been first considered by the panel as consistent with ALCL-HL (n = 18) or with PTCL (n = 13) and were changed later in view of an immunophenotype concordant with HL. Fifty-five percent of the patients completed the full NHL treatment. The 5-year event-free and overall survival rates were 54% and 77%, respectively. The current results indicate that lymphomas initially called ALCL-HL should not be regarded as a variant of ALCL, but as HL. The clinical consequences of misdiagnoses seem to be a lower event-free survival rate compared with that of classical HL, probably because of more relapses of initially inappropriately treated HL.

Adolescent↗

Comparison between MAP and postprocessed ML for image reconstruction in emission tomography when anatomical knowledge is available.

Previously, the noise characteristics obtained with penalized-likelihood reconstruction [or maximum a posteriori (MAP)] have been compared to those obtained with postsmoothed maximum-likelihood (ML) reconstruction, for emission tomography applications requiring uniform resolution. It was found that penalized-likelihood reconstruction was not superior to postsmoothed ML. In this paper, a similar comparison is made, but now for applications where the noise suppression is tuned with anatomical information. It is assumed that limited but exact anatomical information is available. Two methods were compared. In the first method, the anatomical information is incorporated in the prior of a MAP-algorithm and is, therefore, imposed during MAP-reconstruction. The second method starts from an unconstrained ML-reconstruction, and imposes the anatomical information in a postprocessing step. The theoretical analysis was verified with simulations: small lesions were inserted in two different objects, and noisy PET data were produced and reconstructed with both methods. The resulting images were analyzed with bias-noise curves, and by computing the detection performance of the nonprewhitening observer and a channelized Hotelling observer. Our analysis and simulations indicate that the postprocessing method is inferior, unless the noise correlations between neighboring pixels are taken into account. This can be done by applying a so-called prewhitening filter. However, because the prewhitening filter is shift variant and object dependent, it seems that MAP reconstruction is the more efficient method.

Algorithms↗

Endocrine differentiation in mucoid carcinoma of the breast.

Mucoid carcinoma of the breast is not a single homogeneous entity. It comprises two main variants separable on structural and cytological grounds and a smaller transitional type. The salient pathological differences are detailed. One variant is usually arygrophilic and contains dense core granules of the type seen in endocrine tumours. Both endocrine and amphicrine cells have been identified in this sub-type. The relationship of this endocrine variant of mucoid carcinoma to other argryophil carcinomas (so-called 'carcinoids') of the breast is discussed.

Adenocarcinoma, Mucinous↗

Identification of a novel HLA-B allele (B*4202) in a Saudi Arabian family with Behçet's disease.

A new HLA-B antigen, tentatively called HLA-B42AND, was identified as a B42 serologic variant in a Saudi Arabian family. DNA sequencing analysis of the second and third exon of this new B allele revealed that B42AND was identical to B*4201 except for a single T to C substitution at position 97 of exon 2. This substitution results in histidine (CAC) at codon 9 in B42AND instead of tyrosine (TAC) in B*4201. The antigen frequency of B42AND in a Saudi Arabian population was around 10%. This novel B42AND has officially been named HLA-B*4202.

Alleles↗

The incidence and significance of birthmarks in a cohort of 4,641 newborns.

An unselected cohort of 4,641 newborns was ascertained prospectively for the purpose of detecting any cutaneous lesion. These were catalogued into pigmented lesions, vascular lesions, and miscellaneous lesions. Several important findings were elucidated: congenital nevocellular nevi are speckled at their borders; no white newborn in the study had more than one café au lait mark; a hypopigmented tuft of hair was seen in one infant with tuberous sclerosis but is found more commonly in normal individuals; a previously undescribed lesion called zosteriform melanocytic nevus was seen as a normal pigmentary variant in blacks; and hypopigmented macules seen at birth are seen primarily in normal infants. It is hoped that these findings will allow the pediatrician and dermatologist to offer more meaningful prognostic information to their patients.

Alopecia↗

Isolation of a lysogenic bacteriophage carrying the stx(1(OX3)) gene, which is closely associated with Shiga toxin-producing Escherichia coli strains from sheep and humans.

A specific PCR for the detection of a variant of the gene encoding Shiga toxin 1 (stx(1)) called stx(1(OX3)) (GenBank accession no. Z36901) was developed. The PCR was used to investigate 148 Stx(1)-producing Escherichia coli strains from human patients (n = 72), cattle (n = 27), sheep (n = 48), and a goat (n = 1) for the presence of the stx(1(OX3)) gene. The stx(1(OX3)) gene was present in 38 Shiga toxin-producing E. coli (STEC) strains from sheep belonging to serogroups O5, O125, O128, O146, and OX3 but was absent from Stx(1)-positive ovine STEC O91 strains. The stx(1(OX3)) gene was also detected in 22 STEC strains from humans with nonbloody diarrhea and from asymptomatic excreters. Serotypes O146:H21 and O128:H2 were most frequently associated with stx(1(OX3))-carrying STEC from sheep and humans. In contrast, Stx(1)-producing STEC strains from cattle and goats and 50 STEC strains from humans were all negative for the stx(1(OX3)) gene. The stx(1(OX3))-negative strains belonged to 13 serotypes which were different from those of the stx(1(OX3))-positive STEC strains. Moreover, the stx(1(OX3)) gene was not associated with STEC belonging to enterohemorrhagic E. coli (EHEC) serogroups O26, O103, O111, O118, O145, and O157. A bacteriophage carrying the stx(1(OX3)) gene (phage 6220) was isolated from a human STEC O146:H21 strain. The phage was able to lysogenize laboratory E. coli K-12 strain C600. Phage 6220 shared a similar morphology and a high degree of DNA homology with Stx(2)-encoding phage 933W, which originates from EHEC O157. In contrast, few similarities were found between phage 6220 and Stx(1)-encoding bacteriophage H-19B from EHEC O26.

Animals↗