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PubMed2000

Dynamics of the development of Chlamydophila psittaci inclusions in epithelial and fibroblast host cells.

The development of Chlamydophila psittaci (formerly Chlamydia psittaci, avian strains) inclusions in fibroblast L-929 and epithelial BGM cell lines was studied along the bacterial growth cycle using a BGM cell-adapted strain in the presence or absence of cycloheximide and cycloheximide + polyethylene glycol. Evolution of the inclusions was determined in terms of their number and size at 24, 30, 36, 48 and 54 h after infection. Significant differences in the chlamydial growth were found between both host cells, throughout the study. Higher numbers of inclusions (P < 0.05) were observed in L cells while larger inclusions (P < 0.01) were found in BGM cells. In both fibroblast and epithelial cells, inclusions showed a significant (P < 0.001) increase in size at the later times studied. Free extracellular chlamydial particles were noticed at 48 and 54 h post-infection (p.i.) in infected L cells, and at 54 h p.i. in BGM cells. Addition of cycloheximide or cycloheximide + polyethylene glycol had no significant effect on the number of inclusions or their size. The results suggest that host cell characteristics and innate compatibility between Chlamydophila strain and host cell are more important than host cell adaptation for the development of the microorganism.

Animals↗
PubMed2002

Golgi fragmentation occurs in the cells with prefibrillar alpha-synuclein aggregates and precedes the formation of fibrillar inclusion.

Amyloid-like fibrillar aggregates of intracellular proteins are common pathological features of human neurodegenerative diseases. However, the nature of pathogenic aggregates and the biological consequences of their formation remain elusive. Here, we describe (i) a model cellular system in which prefibrillar alpha-synuclein aggregates and fibrillar inclusions are naturally formed in the cytoplasm with distinctive kinetics and (ii) a tight correlation between the presence of prefibrillar aggregates and the Golgi fragmentation. Consistent with the structural abnormality of Golgi apparatus, trafficking and maturation of dopamine transporter through the biosynthetic pathway were impaired in the presence of alpha-synuclein aggregates. Reduction in cell viability was also observed in the prefibrillar aggregate-forming condition and before the inclusion formation. The fibrillar inclusions, on the other hand, showed no correlation with Golgi fragmentation and were preceded by these events. Furthermore, at the early stage of inclusion formation, active lysosomes and mitochondria were enriched in the juxtanuclear area and co-aggregate into a compact inclusion body, suggesting that the fibrillar inclusions might be the consequence of an attempt of the cell to remove abnormal protein aggregates and damaged organelles. These results support the hypothesis that prefibrillar alpha-synuclein aggregates are the pathogenic species and suggest that Golgi fragmentation and subsequent trafficking impairment are the specific consequence of alpha-synuclein aggregation.

Animals↗
PubMed2003

Casein kinase II-mediated phosphorylation regulates alpha-synuclein/synphilin-1 interaction and inclusion body formation.

Alpha-synuclein is a phosphoprotein that accumulates as a major component of Lewy bodies in the brains of patients with Parkinson disease. Synphilin-1, which is also present in Lewy bodies, binds with alpha-synuclein and forms cytoplasmic inclusions in transfected cells. Yet the molecular determinants of this protein-protein interaction are unknown. Here we report that casein kinase II (CKII) phosphorylates synphilin-1 and that the beta subunit of this enzyme complex binds to synphilin-1. Additionally, both CKII alpha and beta subunits are present within cytoplasmic inclusions in cells that overexpress synphilin-1. Notably, the interaction between synphilin-1 and alpha-synuclein is markedly dependent on phosphorylation. Inhibition of CKII activity by 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole blocks the binding between these two proteins and significantly reduces the percentage of cells that contain eosinophilic cytoplasmic inclusions. Mutation of the major CKII phosphorylation site in alpha-synuclein (S129A) has no significant impact on the binding between alpha-synuclein and synphilin-1 or on the formation of synphilin-1/alpha-synuclein-positive inclusions. These data suggest that the CKII-mediated phosphorylation of synphilin-1 rather than that of alpha-synuclein is critical in modulating their tendency to aggregate into inclusions. These observations collectively indicate that a ubiquitous post-translational modification such as phosphorylation can regulate inclusion body formation in the context of alpha-synuclein and synphilin-1 interaction.

Animals↗
PubMedDate not supplied

Microvillous inclusion disease: report of a case with atypical features.

Microvillous inclusion disease is a rare lethal disorder characterized by intractable, severe, watery diarrhea beginning in early infancy. The underlying defect is thought to be an autosomal recessive genetic abnormality resulting in defective brush-border assembly and differentiation. Normally, this diagnosis is easily established through the electron microscopic demonstration of characteristic microvilli-lined inclusions lying within the apical cytoplasm of surface enterocytes. In a small number of patients appearing to have microvillous inclusion disease it has not proven possible to demonstrate the typical inclusions. The existence of another entity, termed intestinal microvillous dystrophy, has been proposed to account for such occurrences. This assertion was founded in large part upon the observation that the few subjects studied all displayed a slightly atypical clinical presentation. The case now being presented exhibited the morphologic features ascribed to intestinal microvillous dystrophy but had a clinical presentation that was entirely typical of microvillous inclusion disease. It serves thus to conceptually unite intestinal microvillous dystrophy with microvillous inclusion disease.

Cytomegalovirus Infections↗
PubMed1998

Elastostatics of a spherical inclusion in homogeneous biological media.

A three-dimensional spherical inclusion model that approximates a lesion bonded to a tissue matrix is proposed for biomedical elastography. Analytical formulae describing spatial strain and stress distributions generated in infinite media by uniform loading are given under a linear, homogeneous, isotropic elasticity assumption. Strain and stress distributions are also calculated using finite-element analysis (FEA) for a variety of cases to determine the effects of shear modulus distribution, external loading conditions (uniform stress versus uniform displacement), compressor size and matrix dimensions on the elastostatics of the tissue. Analytical strain and stress predictions are shown to agree with the FEA results to within 10% accuracy provided that the matrix dimensions are at least ten times that of the inclusion. Also for these cases, uniform-stress boundary conditions can be equivalently represented by uniform displacement of the boundary. Spherical inclusions exhibit a lower efficiency for transferring elastic shear modulus contrast into strain contrast than cylindrical or planar inclusions. Additional compression will increase the strain contrast. However, large compressions also lead to increases in ultrasonic signal decorrelation and strain and stress concentrations in the homogeneous matrix around the inclusion. Although strain concentrations may help describe the boundaries of the inclusion more clearly, they also increase the risk of damaging the tissue. Understanding the strain and stress distributions in a biological tissue containing a lesion is necessary for optimizing the experimental configurations and consequently improving the diagnostic values of elasticity imaging.

Cysts↗
PubMed1999

Formation of polyglutamine inclusions in non-CNS tissue.

Huntington's disease (HD) is one of a class of inherited progressive neurodegenerative disorders that are caused by a CAG/polyglutamine repeat expansion. We have previously generated mice that are transgenic for exon 1 of the HD gene carrying highly expanded CAG repeats which develop a progressive movement disorder and weight loss with similarities to HD. Neuronal inclusions composed of the exon 1 protein and ubiquitin are present in specific brain regions prior to onset of the phenotype, which in turn occurs long before specific neurodegeneration can be detected. In this report we have extended the search for polyglutamine inclusions to non-neuronal tissues. Outside the central nervous system (CNS), inclusions were identified in a variety of post-mitotic cells. This is consistent with a concentration-dependent nucleation and aggregation model of inclusion formation and indicates that brain-specific factors are not necessary for this process. To possibly gain insights into the wasting that is observed in the human disease, we have conducted a detailed analysis of the timing and progression of inclusion formation in skeletal muscle and an investigation into the cause of the severe muscle atrophy that occurs in the mouse model. The formation of inclusions in non-CNS tissues will be particularly useful with respect to in vivo monitoring of pharmaceutical agents selected for their ability to prevent polyglutamine aggregation in vitro, without the requirement that the agent can cross the blood-brain barrier in the first instance.

Animals↗
PubMed1993

Is ovarian cancer associated with an increased frequency of germinal inclusion cysts?

It has been proposed that epithelial ovarian cancers arise in germinal inclusion cysts of the ovary, which are thought to form as stigmata of ovulation. To evaluate whether the frequency of germinal inclusion cysts is associated with ovarian cancer, the authors counted the germinal inclusion cysts in single slides of sections from ovaries of 148 women who underwent incidental oophorectomy and from the contralateral ovaries of 37 women with unilateral ovarian cancer at Columbia-Presbyterian Medical Center, New York, New York, in 1985-1991. The mean number of germinal inclusion cysts was 2.7 for cases and 3.6 for controls. Conditional logistic regression analysis showed that germinal inclusion cysts were not associated with ovarian cancer (odds ratio = 0.98, 95% confidence interval 0.92-1.04). These findings do not support the hypothesis that increased formation of inclusion cysts is a risk factor for ovarian cancer.

Case-Control Studies↗
PubMed1994

Minimum number of water molecules required for inclusion of d-limonene in the cyclodextrin cavity.

The ratios of inclusion of d-limonene in alpha-, beta-, and gamma-cyclodextrin (CD) cavities were measured under low moisture conditions by a micro-aqueous method, and the minimum numbers of water molecules required for inclusion of d-limonene in alpha-, beta-, and gamma-CD cavities were determined by the auto-catalytic inclusion model. The minimum number of water molecules required for alpha-, beta-, and gamma-CD inclusion calculated from the proposed model were comparable with that in the CD cavity in the crystal state. The effect of ethanol upon the inclusion process was investigated, particularly with a low water content. This is the first study involving measurement of the minimum number of water molecules required for the inclusion of organic molecules (d-limonene) in the CD cavity.

Carbohydrate Conformation↗
PubMed1996

Proximal sciatic axotomy does not inhibit the induction of neurofilamentous inclusions following intracisternal aluminum chloride exposure.

We have previously demonstrated an acute, dose-dependent suppression of low molecular weight neurofilaments (NFL) and intermediate molecular weight neurofilaments (NFM) steady state mRNA levels while sparing those of high molecular weight (NFH) mRNA 48 hours (h) following the intracisternal inoculation of AlCl3 in young adult New Zealand white rabbits. To determine whether this alteration in NF steady state mRNA stoichiometry is a necessary prerequisite to the induction of neurofilamentous inclusions, we examined the response of spinal motor neurons to aluminum exposure in vivo following axotomy. Forty-eight h following a complete transection of the proximal sciatic nerve, rabbits were inoculated intracisternally with either 1000 microg AlCl3 in 100 microl 0.9% NaCl or 0.9% NaCl alone. Rabbits were killed at either 48 or 120 h post-inoculation, and the extent of neurofilamentous inclusion formation quantified in both the cervical and the lumbosacral cord. Following the axotomy, rabbits developed an ipsilateral hind-limb paralysis. In spinal motor neurons ipsilateral to the axotomy, chromatolytic changes were observed and both NFH and NFM mRNA levels were significantly reduced (p<0.001). At 48 h post-AlCl3 inoculation, 29% of motor neurons contralateral to the axotomy demonstrated inclusions, whereas 43% of ipsilateral motor neurons demonstrated inclusions (Fisher's test, two tailed, p = 0.0196). At 120 h post-axotomy 75% and 83%, respectively, of neurons were involved (p = 0.0212). Neurofilamentous inclusions did not form in NaCl-inoculated rabbits. These observations indicate that an altered stoichiometry of NF mRNA steady levels, with a relative overexpression of NFH mRNA, is not critical to the induction of neurofilamentous inclusions following AlCl3 exposure.

Aluminum Chloride↗
PubMedDate not supplied

Inclusion of women in nursing research: 1995-2001.

BACKGROUND: Although the inclusion of women in research has been examined in medical publications, little is known about how federal mandates have influenced the inclusion of women in research published in nursing journals. OBJECTIVE: This study aimed to examine the inclusion of women in published nursing research from 1995 to 2001, with a focus on the leading causes of mortality. METHOD: All the articles in each journal were reviewed, and all the research articles that focused on the top 10 causes of death were sampled to measure the inclusion of women, the characteristics of the women included, the funding source, and the topic. RESULTS: Of the 1,149 studies reviewed, 139 met the inclusion criteria, and 117 of these studies included women. Only 15 of the studies reported the age of the women in their samples, and 10 of these included women between the ages of 35 and 64 years. The most frequently reported race was White (n = 21), followed by African American (n = 17), Hispanic (n = 6), and Asian (n = 2). There were no associations among year of publication (p =.62), federal funding (p =.30), and inclusion of women. Among the studies including women, heart disease was the most frequent topic (n = 52), followed by Alzheimer's disease (n = 21), cancer (n = 14), respiratory illness (n = 14), and diabetes (n = 8). DISCUSSION: Although the majority of the research included women, continuing efforts must be made to include sufficient numbers of women.

Adult↗
PubMed1999

The clinical significance of cytomegaloviral inclusions in the allograft kidney.

BACKGROUND: Renal allografts are a frequent site of subclinical cytomegalovirus (CMV) infection diagnosed by culture, but histologic inclusions occur in less than 1% of biopsies. The natural history of this subgroup of patients has been reported only occasionally, mostly before the availability of ganciclovir therapy. METHODS: We analyzed the clinical parameters and pathologic findings in 10 patients with CMV inclusions at allograft biopsy. RESULTS: The patients were 29-72 years old, and 9 of 10 (90%) had previous episodes of acute rejection, 3 of whom needed OKT3 administration. Histopathologic examination of the allografts showed interstitial inflammation with tubulitis in 7 of 10 (70%) patients; in 3 of 10 (30%) patients, viral inclusions were present in the glomerular capillary endothelia without any associated inflammatory response. Morphologic criteria for acute transplant glomerulopathy or proliferative glomerulonephritis were not satisfied. Extrarenal viral inclusions were documented in the gastrointestinal tracts of 2 of 10 (20%) patients. The patients were treated with reduced immunosuppression and ganciclovir. Five patients lost their grafts 56.6+/-86.6 days (range, 4-210 days; median, 21 days) after initial diagnosis. The serum creatinine in the remaining five patients was 3.3+/-2.0 mg/dl (range, 1.2-6.5 mg/dl; median, 2.5 mg/dl) 77+/-16 days (range, 56-101 days; median, 77 days) after transplantation. Histopathologic examination showed no residual viral inclusions in 5 of 7 (71.4%) follow-up specimens available for examination. CONCLUSIONS: CMV inclusions in renal allograft biopsies typically occur after treatment for rejection. Ganciclovir eradicates replicative virus, but graft outcome is determined by coexisting acute rejection and chronic allograft nephropathy. Graft loss primarily attributable to CMV was not observed.

Adult↗
PubMed2002

Dynamic phase separation of fluid membranes with rigid inclusions.

Membrane shape fluctuations induce attractive interactions between rigid inclusions. Previous analytical studies showed that the fluctuation-induced pair interactions are rather small compared to thermal energies, but also that multibody interactions cannot be neglected. In this paper, it is shown numerically that shape fluctuations indeed lead to the dynamic separation of the membrane into phases with different inclusion concentrations. The tendency of lateral phase separation strongly increases with the inclusion size. Large inclusions aggregate at very small inclusion concentrations and for relatively small values of the inclusions' elastic modulus.

Journal Article↗
PubMed2006

Dynamic mechanical response of elastic spherical inclusions to impulsive acoustic radiation force excitation.

Acoustic radiation force impulse imaging has been used clinically to study the dynamic response of lesions relative to their background material to focused, impulsive acoustic radiation force excitations through the generation of dynamic displacement field images. Dynamic displacement data are typically displayed as a set of parametric images, including displacement immediately after excitation, maximum displacement, time to peak displacement, and recovery time from peak displacement. To date, however, no definitive trends have been established between these parametric images and the tissues' mechanical properties. This work demonstrates that displacement magnitude, time to peak displacement, and recovery time are all inversely related to the Young's modulus in homogeneous elastic media. Experimentally, pulse repetition frequency during displacement tracking limits stiffness resolution using the time to peak displacement parameter. The excitation pulse duration also impacts the time to peak parameter, with longer pulses reducing the inertial effects present during impulsive excitations. Material density affects tissue dynamics, but is not expected to play a significant role in biological tissues. The presence of an elastic spherical inclusion in the imaged medium significantly alters the tissue dynamics in response to impulsive, focused acoustic radiation force excitations. Times to peak displacement for excitations within and outside an elastic inclusion are still indicative of local material stiffness; however, recovery times are altered due to the reflection and transmission of shear waves at the inclusion boundaries. These shear wave interactions cause stiffer inclusions to appear to be displaced longer than the more compliant background material. The magnitude of shear waves reflected at elastic lesion boundaries is dependent on the stiffness contrast between the inclusion and the background material, and the stiffness and size of the inclusion dictate when shear wave reflections within the lesion will interfere with one another. Jitter and bias associated with the ultrasonic displacement tracking also impact the estimation of a tissue's dynamic response to acoustic radiation force excitation.

Acoustics↗
PubMed1986

Primary gastric lymphoma with intranuclear inclusions. An immunohistochemical and ultrastructural study.

Two patients with primary gastric lymphoma, diffuse lymphoma, a plasmacytoid variant of small cell type, with numerous intranuclear inclusions, are reported. In both cases, biopsy and surgical specimens showed diffuse proliferation of lymphoid cells having plasmacytoid features at different steps of maturation in lamina propria and submucosal layer. The cytoplasm of the lymphoma cells was stained with anti-IgM, anti-kappa chain, and anti-J chain antibodies by avidin-biotin peroxidase complex technique. A number of intranuclear inclusions in the lymphoma cells were diastase-resistant PAS positive, and also stained with anti-IgM, anti-kappa chain, and anti-J chain antibodies, suggesting a monoclonal situation. In an ultrastructural study of the first case, the inclusions were round and composed of amorphous materials surrounded by a single membrane. In many lymphoma cells, the perinuclear cisternal space was dilated and filled with materials similar to those within both inclusions and rough endoplasmic reticulum. A direct communication between the perinuclear space and the inclusion was demonstrated in some cells, suggesting the perinuclear cisternal origin of these inclusions.

Adult↗
PubMed1992

Cracks emanating from slipping inclusions near a bone-implant interface.

It has been hypothesized that mechanical fracture at the bone-cement-implant interface is the initial cause for loosening of orthopedic implants. Previous investigators have observed cracks to emanate from methacrylate beads, apparently acting as inclusions within the cement. It is believed that the bond between these inclusions and the surrounding matrix breaks prior to emanation of radial cracks from the inclusion. An analytical model is developed for radial cracks emanating from circular inclusions that allow slip along their interface. The solution to the interaction of a single dislocation and a slipping inclusion is used as a Green's Function to model the crack. The Mode I stress intensity factors are calculated for arbitrary orientations of the crack and for varying relative stiffness of the matrix and the inclusion to test feasibility of crack growth.

Bone Cements↗
PubMed2003

Interaction of a bounded ultrasonic beam with a thin inclusion inside a plate.

A theoretical study of the reflection of a two-dimensional Gaussian ultrasonic beam, incident at a Lamb angle of a plate containing a thin rectangular inclusion at an arbitrary position, is presented on the basis radiation mode theory. The inclusion is parallel to the plate surface and its thickness is assumed to be much smaller than the ultrasonic wavelength. It is shown that the amplitude and phase of the reflected beam profile can be used for accurate inclusion characterization. However, this only holds for certain internal positions of the inclusion and for material combinations that do not strongly perturb the excitation of Lamb waves in the plate. When these conditions are satisfied, it is possible to define the Lamb waves and the associated experimental conditions for which good estimates can be obtained of the position of the beginning point of the inclusion as well as of the length and the thickness of the inclusion.

Acoustics↗
PubMed2003

Evaluation of the antiulcerogenic activity of violacein and its modulation by the inclusion complexation with beta-cyclodextrin.

The effects of beta-cyclodextrin (betaCD) inclusion complexation on the ability of violacein to prevent gastric ulceration in mice were studied. Violacein-betaCD inclusion complexes were prepared in 1:1 and 1:2 molar ratios and analysed by differential scanning calorimetry and powder X-ray diffractometry. Violacein previously administered orally at 10 mg/kg significantly reduced indomethacin-induced gastric lesions, as well as 100 mg/kg of cimetidine (positive control). However, betaCD complexation in both molar ratios significantly potentiated the protective action of violacein. In the HCl--ethanol-induced gastric ulcer model, violacein and the 1:2 inclusion complex (10 mg/kg, p.o.) inhibited gastric damage by almost 85%, whereas a 63% reduction was observed for the positive control, lansoprazole, at 30 mg/kg. In contrast, treatment with the 1:1 inclusion complex resulted in almost total disappearance of the antiulcer activity in this model. No significant changes in stress-induced gastric injury were found. In addition, the 1:2 inclusion complex improved the antilipoperoxidant activity of violacein in rat liver cells exposed to t-butyl hydroperoxide, whereas the 1:1 complex was less active than violacein. In summary, the 1:2 betaCD inclusion complex has gastroprotective properties similar to or higher than that of violacein. An increase in mucosal defensive mechanisms and protection against peroxidative damage might be involved.

Animals↗
PubMed1997

Peritoneal inclusion cysts and their relationship to the ovaries: evaluation with sonography.

PURPOSE: To evaluate with sonography the relationship of the ovary to the peritoneal inclusion cyst. MATERIALS AND METHODS: The medical records, sonograms, and sonographic reports of 15 premenopausal women with surgically proved peritoneal inclusion cysts were examined. Twelve of these cases were examined prospectively. In another prospectively evaluated case, the sonogram was interpreted as revealing a peritoneal inclusion cyst; however, hydrosalpinx was confirmed with surgery. Thirteen women with peritoneal inclusion cysts had undergone pelvic surgery before sonography. Additional examination with computed tomography was performed in three women and with magnetic resonance imaging in another. RESULTS: The ovary was detected on sonograms in 12 of 15 women with peritoneal inclusion cysts. In nine of the 12 women, the ovary was demonstrated to be inside the cyst. Ten of 12 ovaries appeared normal. Cyst diameter ranged from 7.5 to 17.4 cm (mean, 11.6 cm). Sonograms of two cysts demonstrated internal echoes. Septa were seen in 11 of the 15 cysts. Cysts were ovoid in six women and irregular in nine; they were defined by adjacent pelvic structures. CONCLUSION: In premenopausal women with a history of pelvic surgery, the appearance on sonograms of the ovary inside a large, ovoid or irregular, anechoic cyst is characteristic of a peritoneal inclusion cyst.

Adult↗