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PubMed2006

Near-infrared spectroscopic determination of salinity and internal pressure of fluid inclusions in minerals.

A near-infrared (NIR) spectroscopic method is proposed to achieve the simultaneous determination of salinity and internal pressure of fluid inclusions in natural minerals. A combination band between the anti-symmetric stretching and bending vibrations of molecular water at approximately 5180 cm-1 was observed for standard salt solutions and natural minerals containing fluid inclusions with known salinities. A curve-fitting procedure was used to analyze the change in the band shape of the combination. Justification of the calibration was confirmed by observation of fluid inclusions in natural minerals whose salinities had already been determined using microthermometry. The detection limit of the present method is 1 NaCl-eq wt. %. The minimum size of fluid inclusions that produced well-resolved spectra was approximately 30 microm. This method was applied to assess micro fluid inclusions in a natural diamond with cubic growth habit (cuboid). The salinity and residual pressure of those fluid inclusions were estimated respectively as 4.4 wt. % NaCl-eq and 0.6-0.8 GPa. The present method is complementary to Raman microscopy and microthermometry for the determination of salinity in fluid inclusions of geological samples.

Journal Article↗
PubMed2004

Orientation and interaction of oblique cylindrical inclusions embedded in a lipid monolayer: a theoretical model for viral fusion peptides.

We consider the elastic behavior of flat lipid monolayer embedding cylindrical inclusions oriented obliquely with respect to the monolayer plane. An oblique inclusion models a fusion peptide, a part of a specialized protein capable of inducing merger of biological membranes in the course of fundamental cellular processes. Although the crucial importance of the fusion peptides for membrane merger is well established, the molecular mechanism of their action remains unknown. This analysis is aimed at revealing mechanical deformations and stresses of lipid monolayers induced by the fusion peptides, which, potentially, can destabilize the monolayer structure and enhance membrane fusion. We calculate the deformation of a monolayer embedding a single oblique inclusion and subject to a lateral tension. We analyze the membrane-mediated interactions between two inclusions, taking into account bending of the monolayer and tilt of the hydrocarbon chains with respect to the surface normal. In contrast to a straightforward prediction that the oblique inclusions should induce tilt of the lipid chains, our analysis shows that the monolayer accommodates the oblique inclusion solely by bending. We find that the interaction between two inclusions varies nonmonotonically with the interinclusion distance and decays at large separations as square of the distance, similar to the electrostatic interaction between two electric dipoles in two dimensions. This long-range interaction is predicted to dominate the other interactions previously considered in the literature.

Computer Simulation↗
PubMed2002

[Application of beta-cyclodextrin inclusion technique in new dosage form of angelica sinensis essential oil].

OBJECTIVE: To make a solid form of Angelica Sinensis essential oil by preparing its inclusion compound with beta-cyclodextrin and evaluate the inclusion rate, dissolution rate and liberating rate of this complex. METHOD: GC-MS, X-ray diffraction and thin-layer chromatography were applied to characterize the essential oil and the inclusion compound. RESULT: The inclusion compound was shown to form a new phase by X-ray diffraction analysis. GC-MS and TLC check-up corroborated the composition of the free oil and the inclusion oil remaining the same. CONCLUSION: The results showed Angelica Sinensis essential oil was more stable with beta-cyclodextrin inclusion. The preparation of this inclusion compound has advantages in limited amount, long time effect, easily preserved and convenient administration as a new dosage form.

Angelica sinensis↗
PubMed2004

[Comparing students in inclusive education to those in special schools: the view of parents of children with learning disabilities].

The paper presents the results of a survey of 755 parents of learning disabled children with certified special needs who either attended classes within regular education or special schools. All parents were involved in the decision on the school placement of their children. The experiences of 547 parents of learning disabled students in inclusive classes were contrasted with those of 207 parents of children in special schools. Besides a rather high satisfaction with previous school experiences of their children a number of differences between the two groups of parents could be observed. Parents of students in special schools viewed their children as rather little challenged by their educational requirements whereas those in inclusive education found their children to be overtaxed. The social development of the students in inclusive education was judged as more positive and, generally, a higher rate of parents of learning disabled students in inclusive classes were satisfied with their choice of the educational setting. Although the requirements for parental support concerning studying were higher in inclusive classes this cannot solely explain the differences of experiences with school. In a second step, satisfied parents were compared to dissatisfied parents. It could be found that the group of dissatisfied parents had to make their choice on the educational setting of their children under less favourable conditions and many could not accept that their child had been classified as having special needs. This applied to parents of students in inclusive education as well as to parents of children in special schools. Additionally, parents of students with German as a second language reported to be discontented more frequently. No significant discrepancies could be found between different grades or federal states with different quotas of inclusive education.

Attitude↗
PubMed2006

[UV spectroscopic characterization of the inclusion interaction of beta-cyclodextrin and sym-diphenyl-thiourea and sym-diphenyl-urea].

The inclusion interaction of beta-cyclodextrin and sym-diphenyl-thiourea and sym-diphenyl-urea was studied by UV spectra. The stoichiometry ratio for the formation of the inclusion complexes was determined by Hildebrand-Benesi equation linear analysis and molar ratio method. The standard molar Gibbs energies, enthalpies, and entropies were derived for the inclusion process by Ks at different temperatrues. The result showed that the host:guest ratio of inclusion complex between the two diphenyl compounds and beta-CD is 2 : 1, the stability constant (Ks) of 2 : 1 inclusion complexes was higher than that of 1 : 1 inclusion complexes due to cooperative binding in the close two hydrophobic cyclodextrin cavities, and the association of the guest molecule with beta-CD was favored by enthalpy changes, proving that the Van der Waals interaction and the dipole-dipole interaction were main binding forces of cyclodextrin inclusion complex.

Algorithms↗
PubMed1991

[HSP 70 is associated with abnormal cytoplasmic inclusions characteristic of neurodegenerative diseases].

Several degenerative diseases of the central nervous system are characterized by the presence of neuronal inclusions. One of these inclusions, neurofibrillary tangles in Alzheimer's disease, has been shown to contain ubiquitin that belongs to a group of proteins known as heat shock proteins. Subsequent studies revealed that ubiquitin is also associated with various neuronal inclusions including Lewy bodies, Pick bodies and hyaline inclusions. Very recently, ubiquitin has been found also to be associated with glial inclusions that are unique to multiple system atrophy. The close association of ubiquitin with varying cellular inclusions, together with its function in the proteolytic process, raised the hypothesis that ubiquitin may be involved in the degradation of abnormal proteins appearing in the damaged neurons and glial cells. In central nervous system, a group of heat shock proteins collectively known as HSP 70 is also present which is constitutive and/or inducible. Since HSP 70 has been suspected to play a crucial role degradation and repair of abnormal intracellular proteins, we hypothesized that HSP 70 may be associated with those inclusions, as the case with ubiquitin. To test this we performed immunohistochemical studies on brain tissues from patients with various neurodegenerative conditions by using specific polyclonal antibody to HSP 70. Brain tissues were obtained at autopsy from each three patients with Alzheimer's disease, Pick's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS) and multiple system atrophy. Tissues were fixed in buffered formalin and embedded in paraffin. Immunostaining was performed by the standard ABC method using diaminobenzidine as a chromogen. Sections were lightly stained with hematoxylin. Polyclonal antibodies were raised in rabbits against mouse HSP 70.(ABSTRACT TRUNCATED AT 250 WORDS)

Alzheimer Disease↗
PubMed1995

[Clinical importance of inclusion cysts of nevi in bulbar conjunctiva].

We investigated 30 cases of nevi in bulbar conjunctiva histopathologically from 1980 to 1992. The patients' ages were between 5 and 85 years old (mean, 21 years). Investigated parameters of the tumors were the location, growth pattern, and the presence of inclusion cysts, melanin granules, goblet cells, and feeding vessels. Of 30 cases, 26 cases (87%) had inclusion cysts which were characteristic findings of conjunctival nevi. The mucous secretion from goblet cells may play an important role in the synthesis of inclusion cysts, but goblet cells were not found in 9 cases. This finding suggests that other secretory cells may be present. Four cases without inclusion cysts consisted of flat tumors. From this finding, the presence of inclusion cysts may be related to the growth of tumors. We conclude that the growth of the pigmented conjunctival tumors with inclusion cysts suggests a benign growth pattern. A periodical follow-up study or biopsy should be performed, if inclusion cysts are not found.

Adolescent↗
PubMed1997

Occurrence of birefringent retinal inclusions in cynomolgus monkeys after high doses of canthaxanthin.

PURPOSE: To reproduce and investigate in a primate animal model the phenomenon of the red carotenoid canthaxanthin (beta, beta-carotene-4'4'-dione) to induce crystal-like retinal deposits as they have been observed in the ocular fundus of humans after high canthaxanthin intake (i.e., more than 30 mg/day). METHODS: Groups of four cynomolgus monkeys (Macaca fascicularis) per gender and dose were administered 5.4, 16.2, or 48.6 mg canthaxanthin/kg body weight daily by oral gavage for 2.5 years. Eight control animals received placebo. In vivo ophthalmoscopy was performed at intervals of 3 months along with electroretinography after 12 and 24 months and retinal biomicroscopy just before the monkeys were killed. Retinal wholemounts or frozen sections were investigated postmortem by polarization, bright field, and differential interference contrast microscopy. Retinal and preterminal plasma canthaxanthin concentrations were determined by high-performance liquid chromatography (HPLC). RESULTS: By ophthalmoscopy and retinal biomicroscopy in vivo, no crystals or other light-reflecting particles were observed in the central paramacular retina. However, postmortem polarization microscopy of all 24 canthaxanthin-treated animals showed a circular zone in the peripheral retina containing birefringent, polymorphous red, orange, or white inclusions. The density of these inclusions was diminished within 1 to 8 mm posterior to the ora serrata. These inclusions were located mainly in the inner retinal layers, that is the nerve fiber layer and ganglion cell layer, inner plexiform layer, and inner nuclear layer. Twelve of the 24 canthaxanthin-treated animals showed yellow, golden birefringent inclusions in the macula. Retinas of placebo-treated monkeys were free of birefringent, crystal-like inclusions. The HPLC confirmed the presence of all-trans canthaxanthin, and 4-OH-echinenone and isozeaxanthin as well, in the retinas of all canthaxanthin-treated animals. Neither electroretinography nor histopathology indicated any adverse effects of the canthaxanthin-induced retinal inclusions seen in this study. CONCLUSIONS: A high intake of canthaxanthin for 2.5 years led to the deposition of crystal-like birefringent inclusions in the inner layers of the peripheral retina and, to some extent, the central retina of cynomolgus monkeys. The presence of these deposits did not interfere with morphology nor with retinal function.

Animals↗
PubMed1997

Unique crystalline inclusions in neonatal pig small intestine epithelial cells.

In this study we describe the presence of novel crystalline inclusions in the jejunal and ileal epithelium of newborn pigs. Hampshire/Yorkshire piglets (0-3 days) were anesthetized with pentobarbital and tissues were recovered following laparotomy. Sections of jejunum and ileum were prepared with a variety of fixatives for paraffin and methacrylate embedment for the light and electron microscopy. The characteristics of these crystalline structures have been demonstrated by cytochemical staining, immunohistochemical staining, and ultrastructural studies. Using light microscopy, it was determined that 10.8% of villus epithelial cells in the jejunum contained inclusions, whereas inclusions were present in 26.6% of the villus epithelial cells in the ileum. Inclusions were absent in crypt epithelial cells of both areas of the intestine, and the percentage of cells with inclusions increased towards the villus tip. Preliminary histochemical analysis has been unrevealing. The inclusions are periodic acid/Schiff's (PAS) reagent-negative, and do not stain for the presence of concentrated proteins. Using electron microscopy, the inclusions are found to be clustered in the supra-nuclear region of the cells and are generally oriented with the long axis of the cell. The crystals vary in size and have a dense core region surrounded by a regular less dense periphery.

Animals↗
PubMed1993

Adenovirus and intranuclear inclusions in appendices in intussusception.

AIMS: To examine the light and electron microscopic features of appendices removed at the time of surgical reduction of intussusception in children; and to confirm that the viral inclusions seen in some of them are due to adenovirus. METHODS: A series of 39 appendices from cases of intussusception and 15 control appendices were reviewed. Light microscopic examination of haematoxylin and eosin stained sections was performed on all of them and one appendix with large numbers of inclusions was examined by electron microscopy. Non-isotopic in situ hybridisation using a biotinylated DNA probe was carried out on sections of appendix from 30 of the cases of intussusception and from the 15 controls. RESULTS: Light microscopic examination showed viral inclusions in 19 of the appendices from the cases of intussusception and in none of the controls. Electron microscopic examination showed viral particles with the typical features of adenovirus. Most of the appendices with viral inclusions in the haematoxylin and eosin stained sections also contained adenovirus DNA as shown by in situ hybridisation. CONCLUSIONS: Viral inclusions seen in appendices from cases of intussusception are caused by adenovirus. Adenovirus DNA was not demonstrable in appendices from cases of intussusception without viral inclusions and the aetiological factors involved in intussusception in these children remain unknown.

Adenoviridae↗
PubMed1999

Chemical treatment of Escherichia coli: 3. Selective extraction of a recombinant protein from cytoplasmic inclusion bodies in intact cells.

In previous parts of this study we developed procedures for the high-efficiency chemical extraction of soluble and insoluble protein from intact Escherichia coli cells. Although high yields were obtained, extraction of recombinant protein directly from cytoplasmic inclusion bodies led to low product purity due to coextraction of soluble contaminants. In this work, a two-stage procedure for the selective extraction of recombinant protein at high efficiency and high purity is reported. In the first stage, inclusion-body stability is promoted by the addition of 15 mM 2-hydroxyethyldisulfide (2-HEDS), also known as oxidized beta-mercaptoethanol, to the permeabilization buffer (6 M urea + 3 mM ethylenediaminetetraacetate [EDTA]). 2-HEDS is an oxidizing agent believed to promote disulfide bond formation, rendering the inclusion body resistant to solubilization in 6 M urea. Contaminating proteins are separated from the inclusion-body fraction by centrifugation. In the second stage, disulfide bonds are readily eliminated by including reducing agent (20 mM dithiothreitol [DTT]) into the permeabilization buffer. Extraction using this selective two-stage process yielded an 81% (w/w) recovery of the recombinant protein Long-R3-IGF-I from inclusion bodies located in the cytoplasm of intact E. coli, at a purity of 46% (w/w). This was comparable to that achieved by conventional extraction (mechanical disruption followed by centrifugation and solubilization). A pilot-scale procedure was also demonstrated using a stirred reactor and diafiltration. This is the first reported study that achieves both high extraction efficiency and selectivity by the chemical treatment of cytoplasmic inclusion bodies in intact bacterial cells.

Bioreactors↗
PubMed1980

Unusual intracytoplasmic inclusions in acute myeloblastic leukemia.

Unusual intracytoplasmic inclusions within early granulocyte precursor cells from a patient with acute myeloblastic leukemia (AML) are described. Based upon their staining characteristics and electron- and light-microscopic appearance, the inclusions are distinctly different from any previously described. The inclusions display a variety of shapes, including rectangles, squares, circles, ovals, and irregular, globular forms. Most of the inclusions are refractile and crystal-like. The possible composition of these inclusions is discussed. They are compared with inclusions previously described within leukemic and granulocytic cells.

Aged↗
PubMed1995

Common variable immunodeficiency and inclusion body myositis: a distinct myopathy mediated by natural killer cells.

Inclusion body myositis developed in two men, 36 and 48 years old with long-standing common variable immunodeficiency. Immunophenotypic analysis of the endomysial cells showed an increased number of natural killer (NK) cells (defined as CD57+, CD56+, CD3-, CD8-, CD68-) accounting for 8.5 to 9.5% of the total cells, compared with a mean of 1% in sporadic inclusion body myositis. The remaining cells were CD8+, macrophages, and CD4+ T cells. NK cells were positive for intercellular cell adhesion molecule-1 and invaded muscle fibers negative for major histocompatibility complex (MHC) class I. In contrast to ubiquitous endomysial expression of MHC class I antigen in sporadic inclusion body myositis, the MHC class I in common variable immunodeficiency and inclusion body myositis was absent or weakly expressed in only some of the muscle fibers surrounded by CD8+ cells. Enteroviral or retroviral RNA sequences were not amplified. Treatment with intravenous immunoglobulin improved strength in 1 patient whose repeated muscle biopsy specimen showed normal NK cells. We conclude that inclusion body myositis can develop in patients with common variable immunodeficiency. Common variable immunodeficiency with inclusion body myositis is an immune myopathy mediated by NK cells in a non-MHC class I-restricted cytotoxicity, and by CD8+ cells in an MHC class I-restricted process. This is the first description of an inflammatory myopathy in which NK cells participate in the myocytotoxic process.

Adult↗
PubMed1996

Multiple mitochondrial DNA deletions in sporadic inclusion body myositis: a study of 56 patients.

Inclusion body myositis, a chronic inflammatory disorder, is the most common cause of myopathy in adults over the age of 50. Diagnosis is based on clinical features and distinctive morphological findings by both light and electron microscopy. The causes of inclusion body myositis are still unknown. Ultrastructural mitochondrial changes and ragged-red fibers are common in patients with sporadic inclusion body myositis, and multiple [correction of mutiple] mitochondrial DNA (mtDNA) deletions have been reported in 3 such patients, suggesting that mtDNA mutations may have a pathogenetic role. We studied 56 patients with sporadic inclusion body myositis, using a combination of clinical, morphological, biochemical, and molecular genetic analyses to determine the frequency and the distribution of mtDNA deletions. Using the polymerase chain reaction, we found multiple mtDNA deletions in 73% of patients, compared to 40% of normal age-matched control subjects and 47% of disease control subjects. The presence of deletions correlated with morphological evidence of ragged-red, cytochrome c oxidase-negative fibers, and with defects of complexes I and IV of the electron transport chain. Although aging may account for a proportion of mtDNA deletions in patients with sporadic inclusion body myositis and control subjects, mtDNA alterations may be accelerated in sporadic inclusion body myositis.

Adult↗
PubMed1993

Inclusion body myositis and renal cell carcinoma. Report of two cases and review of the literature.

OBJECTIVE: To describe the clinical course of 2 patients with concurrent inclusion body myositis and renal cell carcinoma and review published reports of inclusion body myositis associated with malignancy. METHODS: Prospective followup of 2 patients. Review of published case reports and series of patients with inclusion body myositis. RESULTS: Our 2 patients with inclusion body myositis and renal cell carcinoma had no improvement of strength following nephrectomy. Seven previously reported cases of inclusion body myositis and malignancy were identified and are discussed. CONCLUSION: Findings in our 2 patients suggest that there is no etiopathologic relationship between inclusion body myositis and malignancy.

Carcinoma, Renal Cell↗
PubMed2007

High and compact formation of baculoviral polyhedrin-induced inclusion body by co-expression of baculoviral FP25 in Escherichia coli.

Previously, we found that baculoviral polyhedrin (Polh) can successfully be used in Escherichia coli as a fusion partner for the expression of special foreign proteins as inclusion bodies, and the resulting, easily isolatable Polh-induced fusion inclusion bodies had almost the same characteristics as the native Polh. Here, we investigated the effects of co-expression of baculoviral FP25 protein on Polh-induced inclusion-body production in an E. coli expression system, as FP25 is known to be involved specifically in polyhedra formation. Using several analytical tools, including SDS-PAGE, pronase proteolysis, solubilization under alkaline conditions, and electron microscopy, we found that co-expressed FP25 was associated with Polh-induced inclusion bodies and that its co-expression led to formation of compact inclusion bodies as well as high production levels. We confirmed that FP25 co-expression induced higher production levels of other heterologous protein, antimicrobial peptide Hal18, fused with aggregation-prone Polh. Therefore, co-expression of baculoviral FP25 can be promisingly used to increase the levels of baculoviral Polh-fused foreign proteins, especially harmful proteins, expressed as inclusion bodies in an E. coli expression system.

Baculoviridae↗
PubMed1995

Mitochondrial constituents of corpora amylacea and autofluorescent astrocytic inclusions in senescent human brain.

Corpora amylacea (CA) are cytoplasmic inclusions that accumulate in human brain in the course of normal aging, and to an even greater extent, in Alzheimer's disease and other neurodegenerative conditions. In senescent and Alzheimer-diseased human brains, astrocytes in limbic and periventricular regions exhibit red autofluorescent inclusions, homologous to Gomori-positive astrocyte granules previously described in the brains of aging rodents and other vertebrates. We have shown that Gomori inclusions in situ and in culture are derived from autophagocytosed mitochondria exhibiting iron-mediated peroxidase activity. In the human brain, the autofluorescent inclusions share many properties with CA. Both types of inclusion progressively accumulate in periventricular regions with advancing age, are largely astrocytic in origin, and contain various heat shock proteins and ubiquitin. Using histochemistry in conjunction with cofocal microscopy, we demonstrated that both CA and the red autofluorescent granules exhibit non-enzymatic peroxidase activity and an affinity for CAH and PAS. The only major divergent histochemical feature between the Gomori-positive astrocyte granules and CA is the presence of orange-red autofluorescence in the former and the absence of endogenous fluorescence in the latter. On the basis of numerous shared topographic and histochemical features, we hypothesized that CA are largely derived from autofluorescent (Gomori-positive) astrocyte granules which reside in periventricular regions of the senescent CNS. Immunofluorescent labeling and laser scanning confocal microscopy demonstrated consistent colocalization of the mitochondrial proteins, sulfite oxidase, and heat shock protein 60, to both CA and the autofluorescent astroglial inclusions. In addition, both CA and the autofluorescent astrocyte granules exhibit staining for DNA which colocalizes to mitochondrial antigens and therefore likely represents mitochondrial nucleic acid in dual-labeled preparations. These observations suggest that a) Gomori-positive astrocyte granules in human brain are homologous to those described in rodents, b) Gomori-positive granules may be structural precursors of CA in senescent human brain, and c) in the aging human brain, degenerate mitochondria within periventricular astrocytes give rise to autofluorescent cytoplasmic granules and corpora amylacea.

Aging↗
PubMed1992

alpha B crystallin expression in non-lenticular tissues and selective presence in ubiquitinated inclusion bodies in human disease.

alpha B crystallin is a lens protein which has homology with the small heat-shock proteins and is also expressed in non-lenticular tissues. Polyclonal antibodies have been raised to a synthetic peptide corresponding to residues 1-10 of alpha B crystallin. The antiserum detects a 20 kDa polypeptide on nitrocellulose replicas after polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate of extracts of heart muscle known to be rich in alpha B crystallin. Staining of normal human tissues reveals immunoreactivity of lens capsular epithelium, skeletal muscle, cardiac muscle, smooth muscle, renal tubular epithelium, Schwann cells, and glial cells, as has been described by other workers. In addition, positive staining of normal thyroid epithelium, colonic epithelium, and stratified squamous epithelium was seen. Tissues known to contain ubiquitinated inclusion bodies were immunostained with the anti-alpha B-crystallin antiserum. Staining of cortical Lewy bodies, astrocytic Rosenthal fibres, and hepatic Mallory bodies was seen, but only a proportion of inclusions were positive. Neurones containing the ubiquitinated inclusions of Alzheimer's disease were only very rarely immunostained and the ubiquitinated inclusions of motor neurone disease were not detected by the antiserum. Reactive astrocytes in cerebral tissues were strongly immunostained. The results suggest that alpha B crystallin is involved in the formation of ubiquitinated inclusion bodies that have associated intermediate filaments and support previous observations on the localization of a brain-specific ubiquitin carboxy-terminal hydrolase which similarly divides ubiquitinated filamentous inclusions in the central nervous system into two main groups.

Alzheimer Disease↗