Pre-embedding immunohistochemistry as an approach to high resolution antigen localization at the light microscopic level

Summary Pre-embedding immunohistochemistry with subsequent embedding in hydroxypropyl methacrylate enables one to obtain high resolution staining of antigens in 1 tissue sections. A routine method using formaldehyde fixation, methanol permeation, and an indirect method with fluoresceinlabeled second antibody is described. This method is compared with other pre-embedding staining procedures. To illustrate the method the mouse small intestine was chosen as a model and stained with antibodies to tubulin, actin, and fibronectin. Some anticipated and some unusual staining patterns were found.     

Isolation and biochemical characterization of delta-aminolevulinic acid dehydratase from Streptomyces yokosukanensis ATCC 25520

In this study, delta-aminolevulinic acid dehydratase from Streptomyces yokosukanensis ATCC 25520, producer of an unusual purine riboside antibiotic called nebularine, was purified and characterized. Purification procedures involved with ammonium sulphate precipitation and gel filtration techniques by use of Sephacryl S-200. After gel filtration a 90.76-fold purification was obtained. The maximum enzymic activity was observed in the supernatant after 100% precipitation. According to the data obtained from investigation, the enzyme was found to be a single polypeptide having molecular mass around 34.8 kDa. This was determined by SDS-PAGE. Its optimal temperature around 45 degrees C, and optimal pH was found to be 8.0. Some heavy metals, Pb2+, Zn2+, Fe3+, Co2+, Mn2+, Mg2+ inhibited its activity between 20-51%, Ni2+ increased its activity up to 15%.     

Unusual structural and chemical characteristics of the Dufour gland in the ant Meranoplus diversus

Abstract The Dufour gland of both queens and workers of Meranoplus diversus is very large. The reservoir is lined with flat epithelial cells, which usually contain smooth endoplasmic reticulum but also show the unusual accumulation of electron‐dense inclusions. Chemically, the gland contains an unusual mixture of long‐chain fatty acids and their derivatives, including hydrocarbons, ketones and amines with long‐chain substituents, hydroquinone, and sugars. Some of the substances identified are already known as insect defensive compounds against other arthropods.     

Viral immunodiagnosis.

This review discusses the commonly employed methods for viral immunodiagnosis, mentions unusual or novel procedures, and briefly refers to the use of monoclonal antibody.     

Isolation and properties of the glycolytic enzymes from Zymomonas mobilis. The five enzymes from glyceraldehyde-3-phosphate dehydrogenase through to pyruvate kinase.

The five glycolytic enzymes glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase, phosphoglycerate mutase, enolase and pyruvate kinase were each purified from extracts of Zymomonas mobilis cells, by using dye-ligand chromatography as the principal step. Two procedures, producing three and two of the enzymes respectively, are described in detail. Z. mobilis glyceraldehyde-phosphate dehydrogenase was found to be similar in most respects to the enzyme from other sources, except for having a slightly larger subunit size. Phosphoglycerate kinase has properties typical for this enzyme; however, it did not show the sulphate activation effects characteristic of this enzyme from most other sources. Phosphoglycerate mutase is a dimer, partially independent of 2,3-bisphosphoglycerate, and has a high specific activity. Enolase was found to be octameric; otherwise its properties were very similar to those of the yeast enzyme. Pyruvate kinase is unusual in being dimeric, and not requiring K+ for activity. It is not allosterically activated by sugar phosphates, having a high activity in the absence of any effectors. Some quantitative differences in the relative amounts of these enzymes, compared with eukaryotic species, are ascribed to the fact that Z. mobilis utilizes the Entner-Doudoroff pathway rather than the more common Embden-Meyerhoff glycolytic route.     

Rotifers from Turkey

Seventy-eight rotifer species are reported from 19 localities in Turkey. About half are first records for that country. Some interesting species are illustrated and the composition of the regional fauna is discussed. Wide-ranging taxa are dominant, with notbale additions of tropical, subtropical, afrotropical and circumboreal species. Many are restricted to saline environments, where some show an unusual vertical distribution. Endemism is suspected in a few taxa.     

An unusual heparan sulfate isolated from lobsters (Homarus americanus)

An unusual heparan sulfate was isolated from lobsters (Homarus americanus). The polysaccharide has a composition and properties intermediate to heparin and the more common heparan sulfates. The sulfate and D-glucuronic acid content is high, while anticoagulant activity is low. The major repeating unit appears to consist of D-glucuronic acid and D-glucosamine N-O-disulfate. Some N-acetyl groups are present, the content of L-iduronic acid O-sulfate is low, and monosulfated or nonsulfated disaccharide-repeating units (very common in heparan sulfates) appear to be very rare. The data obtained again emphasize the heterogeneity of heparan sulfates and the need for adequate characterization when dealing with unusual or unexplored sources.     

Additional Observations on Gametogenesis and Fertilization in Trichonympha

SUMMARY. Some of the unusual types of sexual behavior in Trichonympha are considered: cytoplasmic incompatibility of gametes as shown in the fertilization process, pronuclear incompatibility, independence of pronuclear and cytoplasmic incompatibilities, loss of extranuclear organelles of male gamete, the formation of gametes with no chromosomes, those with the diploid number, and those with numbers between these extremes.     

Analysis of the protein kinome of Entamoeba histolytica

Protein kinases play important roles in almost all major signaling and regulatory pathways of eukaryotic organisms. Members in the family of protein kinases make up a substantial fraction of eukaryotic proteome. Analysis of the protein kinase repertoire (kinome) would help in the better understanding of the regulatory processes. In this article, we report the identification and analysis of the repertoire of protein kinases in the intracellular parasite Entamoeba histolytica. Using a combination of various sensitive sequence search methods and manual analysis, we have identified a set of 307 protein kinases in E. histolytica genome. We have classified these protein kinases into different subfamilies originally defined by Hanks and Hunter and studied these kinases further in the context of noncatalytic domains that are tethered to catalytic kinase domain. Compared to other eukaryotic organisms, protein kinases from E. histolytica vary in terms of their domain organization and displays features that may have a bearing in the unusual biology of this organism. Some of the parasitic kinases show high sequence similarity in the catalytic domain region with calmodulin/calcium dependent protein kinase subfamily. However, they are unlikely to act like typical calcium/calmodulin dependent kinases as they lack noncatalytic domains characteristic of such kinases in other organisms. Such kinases form the largest subfamily of kinases in E. histolytica. Interestingly, a PKA/PKG-like subfamily member is tethered to pleckstrin homology domain. Although potential cyclins and cyclin-dependent kinases could be identified in the genome the likely absence of other cell cycle proteins suggests unusual nature of cell cycle in E. histolytica. Some of the unusual features recognized in our analysis include the absence of MEK as a part of the Mitogen Activated Kinase signaling pathway and identification of transmembrane region containing Src kinase-like kinases. Sequences which could not be classified into known subfamilies of protein kinases have unusual domain architectures. Many such unclassified protein kinases are tethered to domains which are Cysteine-rich and to domains known to be involved in protein-protein interactions. Our kinome analysis of E. histolytica suggests that the organism possesses a complex protein phosphorylation network that involves many unusual kinases.     

Widespread pyocyanin over-production among isolates of a cystic fibrosis epidemic strain

Background  

Some isolates of the Liverpool cystic fibrosis epidemic strain of Pseudomonas aeruginosa exhibit an unusual virulence-related phenotype, characterized by over-production of quorum sensing-regulated exoproducts such as pyocyanin and LasA protease. Our aim was to determine the prevalence of this unusual phenotype amongst isolates of the epidemic strain, and to study other intraclonal phenotypic and genotypic variations.     

Unusual structural features of the 5S ribosomal RNA from Streptococcus cremoris

The nucleotide sequence of the 5S ribosomal RNA of Streptococcus cremoris has been determined. The sequence is 5' (sequence in text) 3'. Comparison of the S. cremoris 5S RNA sequence to an updated prokaryotic generalized 5S RNA structural model shows that this 5S RNA contains some unusual structural features. These features result largely from uncommon base substitutions in helices I, II and IV. Some of these unusual structural features are shared by several of the known 5S RNA sequences from mycoplasmas. However, the characteristic bloc of deletions found in helix V of these mycoplasma 5S RNAs is not present in the 5S RNA of S. cremoris.     

Maximizing the use of the abdominoplasty incision

Candidates for abdominoplasty often request multiple procedures at the time of consultation. Some of these patients have the potential opportunity to have ancillary procedures performed through the abdominoplasty incision, such as breast augmentation or suction-assisted lipectomy. Access via the abdominoplasty incision can also limit the need for distant donor sites, for instance, when autologous fillers or rib graft are necessary. The techniques described are straightforward and are based on standard principles that should be considered when ancillary procedures are performed in conjunction with abdominal contouring procedures. In a review of 70 consecutive patients undergoing abdominoplasty, 91 ancillary procedures were performed in conjunction with the abdominoplasty. Of the total number of patients undergoing abdominoplasty, 29 patients underwent 30 procedures facilitated through their abdominoplasty incision, including 23 suction-assisted lipectomies of the flanks, six breast augmentations, and one rib cartilage harvesting for rhinoplasty. A review of the author's experience and discussion for potential options afforded by this exposure are presented.     

Should we expand the indications for analysis of fetal chromosomes?

The usual indications of amniocentesis for prenatal diagnosis are briefly reviewed. Some situations which are note included in these usual indications are discussed: mothers of 34 years, young aunts of a trisomic child, patients with a previous aborted conceptus diagnosed as trisomic by histological examination of the placenta or tissue culture. It is anticipated that, if accepted, these "unusual" criteria would not overload a given laboratory with an excess of analysis, and would certainly relief anxiety of motivated couples.     

Identification of ORD, a Drosophila protein essential for sister chromatid cohesion.

Attachment between the sister chromatids is required for proper chromosome segregation in meiosis and mitosis, but its molecular basis is not understood. Mutations in the Drosophila ord gene result in premature sister chromatid separation in meiosis, indicating that the product of this gene is necessary for sister chromatid cohesion. We isolated the ord gene and found that it encodes a novel 55 kDa protein. Some of the ord mutations exhibit unusual complementation properties, termed negative complementation, in which particular alleles poison the activity of another allele. Negative complementation predicts that protein-protein interactions are critical for ORD function. The position and nature of these unusual ord mutations demonstrate that the C-terminal half of ORD is essential for sister chromatid cohesion and suggest that it mediates protein binding.     

Detection of pulmonary tuberculosis; comparative value of routine radiologic examinations and routine laboratory procedures

The abnormalities disclosed by the usual laboratory diagnostic procedures (blood cell count and hemoglobin determination, sedimentation rate determination, urinalysis, and serologic test for syphilis) are compared with the number revealed by stereophotoroentgen chest examination in a series of 951 patients. The number of significant abnormalities uncovered by the routine stereophotoroentgen examination of the chest was comparable to the number disclosed by the usual laboratory procedures used in patient evaluation. Some form of radiologic examination of the chest should be included in the routine laboratory procedures required for patient evaluation.     

DETECTION OF PULMONARY TUBERCULOSIS—Comparative Value of Routine Radiologic Examinations and Routine Laboratory Procedures

The abnormalities disclosed by the usual laboratory diagnostic procedures (blood cell count and hemoglobin determination, sedimentation rate determination, urinalysis, and serologic test for syphilis) are compared with the number revealed by stereophotoroentgen chest examination in a series of 951 patients.The number of significant abnormalities uncovered by the routine stereophotoroentgen examination of the chest was comparable to the number disclosed by the usual laboratory procedures used in patient evaluation.Some form of radiologic examination of the chest should be included in the routine laboratory procedures required for patient evaluation.     

STRUCTURALLY COLOURED PATTERN MARKS ON THE INNER WEBS OF FLIGHT FEATHERS.

Some Psittacidae and Coraciidae show the unusual feature of blue and green pattern marks restricted to reverse surfaces of flight feathers.
The optical causes of the colours of these marks are identical with, and the morphological configurations similar to, those which typically cause these colours on obverse surfaces of feathers in the same families. The atypical marks may, however, have been evolved independently from the typically located colour.
Colour patterns on flight feathers are discussed in relation to the mechanics of flight and to optical effect.