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Biomedical subjects

N Roos

Publications and source records attributed to N Roos.

At least 19 recordsLinked to original sources
PubMed2002

Coexistence of non-Hodgkin's lymphoma and non-small cell lung carcinoma: diagnosis and treatment.

Abstract. In this communication, we will present a very rare case of the coexistence of non-Hodgkin's lymphoma (NHL; low malignant lymphocytic lymphoma of the B-cell type) and a non-small-cell lung carcinoma (NSCLC). A patient with a 15-year history of NHL developed a generalized relapse of the lymphoma with an additional tumor mass in the left lower lobe of the lung. Bronchoscopy showed the evidence of the NHL. Due to non-responding chemotherapy on the lung tumor, the coexistence of a second malignancy was histologically proved in a second bronchoscopy. Resection of the lung tumor with complex lobectomy and lymphadenectomy was performed. After that, chemotherapy with four cycles of carboplatin supplemented with taxol was induced. The patient was discharged from the hospital with a stable remission of both tumor diseases. Restaging after six months showed no evidence of a tumor relapse. This is a very rare case of the coexistence of NHL and NSCLC; we will discuss the difficulty of diagnostic and treatment of both tumor diseases.

Aged↗
PubMed2001

Hypertensive patients and their general practitioners.

This study examined the relative contribution of hypertensive patients and their physicians in selecting total annual physician visit frequencies and made regional comparisons between two Canadian cities. We found that the frequency of physician visits was primarily influenced by physician referrals and physician practice patterns, which accounted for about 80 percent of the total explainable variance in physician visits. The relative contribution of other available patient and physician characteristics in determining visit frequency was rather small.

Adult↗
PubMed2001

Performance of the ACG case-mix system in two Canadian provinces.

BACKGROUND: While the adjusted clinical group (ACG) system has been extensively validated in the United States, its use in other developed nations has been limited. This article examines the performance of the system in 2 Canadian provinces and assesses the extent to which ACGs can account for same-year and next-year health care expenditures. METHODS: The study population included all residents of Manitoba and British Columbia who were continuously enrolled in the provincial health plans from April 1, 1995, to March 31, 1997. ACGs were assigned through diagnoses from fee-for-service physician claims and hospital separation records. "Physician" costs were calculated from the fee-for-service tariffs, and for Manitobans, "total" costs were also computed by combining physician and hospital costs. Linear regression was used to examine the ability of the ACG system to explain variation in individual costs (truncated at the 99th percentile). RESULTS: The British Columbia and Manitoba data were generally acceptable, with fewer than 2% rejected diagnoses. Higher costs were associated with both the accumulation of morbidities and their relative severity. For physician costs, the ACG system explained approximately 50% and approximately 25% of the variation in same-year and next-year truncated costs, respectively. For total costs, the system explained approximately 40% and approximately 14% of these respective costs. CONCLUSIONS: The application of ACGs in Canada is feasible using existing data. The ability of the ACG system to explain variation in costs is similar to that found in US health systems. While application of ACGs in Canada shows promise, further research is required to examine how closely they reflect population morbidity burdens and health care needs.

Adolescent↗
PubMed2001

Structure and assembly of intracellular mature vaccinia virus: isolated-particle analysis.

In a series of papers, we have provided evidence that during its assembly vaccinia virus is enveloped by a membrane cisterna that originates from a specialized, virally modified, smooth-membraned domain of the endoplasmic reticulum (ER). Recently, however, Hollinshead et al. (M. Hollinshead, A. Vanderplasschen, G. I. Smith, and D. J. Vaux, J. Virol. 73:1503-1517, 1999) argued against this hypothesis, based on their interpretations of thin-sectioned material. The present article is the first in a series of papers that describe a comprehensive electron microscopy (EM) analysis of the vaccinia Intracellular Mature Virus (IMV) and the process of its assembly in HeLa cells. In this first study, we analyzed the IMV by on-grid staining, cryo-scanning EM (SEM), and cryo-transmission EM. We focused on the structure of the IMV particle, both after isolation and in the context of viral entry. For the latter, we used high-resolution cryo-SEM combined with cryofixation, as well as a novel approach we developed for investigating vaccinia IMV bound to plasma membrane fragments adsorbed onto EM grids. Our analysis revealed that the IMV is made up of interconnected cisternal and tubular domains that fold upon themselves via a complex topology that includes an S-shaped fold. The viral tubules appear to be eviscerated from the particle during viral infection. Since the structure of the IMV is the result of a complex assembly process, we also provide a working model to explain how a specialized smooth-ER domain can be modulated to form the IMV. We also present theoretical arguments for why it is highly unlikely that the IMV is surrounded by only a single membrane.

Dithiothreitol↗
PubMed2001

Structure and assembly of intracellular mature vaccinia virus: thin-section analyses.

In the preceding study (see accompanying paper), we showed by a variety of different techniques that intracellular mature vaccinia virus (vaccinia IMV) is unexpectedly complex in its structural organization and that this complexity also extends to the underlying viral core, which is highly folded. With that analysis as a foundation, we now present different thin-section electron microscopy approaches for analyzing the IMV and the processes by which it is assembled in infected HeLa cells. We focus on conventional epoxy resin thin sections as well as cryosections to describe key intermediates in the assembly process. We took advantage of streptolysin O's ability to selectively permeabilize the plasma membrane of infected cells to improve membrane contrast, and we used antibodies against bone fide integral membrane proteins of the virus to unequivocally identify membrane profiles in thin sections. All of the images presented here can be rationalized with respect to the model put forward for the assembly of the IMV in the accompanying paper.

Bacterial Proteins↗
PubMed2001

Nuclear deviation in hepatic parenchymal cells on sinusoidal surfaces in Arctic animals.

In normal rat and human, most of the nuclei of hepatic parenchymal cells are centrally located in the cytoplasm. However, it is reported that the nuclei of hepatic parenchymal cells are situated at a deviated position on sinusoidal surfaces under pathological situations such as chronic hepatitis, hepatocellular carcinoma, adenomatous hyperplasia, or regeneration. During a study on the mechanism of extreme vitamin A-accumulation in hepatic stellate cells of arctic animals including polar bears, arctic foxes, bearded seals, and glaucous gulls, we noticed that these arctic animals displayed the nuclear deviation in hepatic parenchymal cells on sinusoidal surfaces. In this study, we assessed the frequency of hepatic parenchymal cells showing the nuclear deviation on the sinusoidal surfaces in arctic animals. A significantly higher frequency of the nuclear deviation in hepatic parenchymal cells was seen in polar bears (89.8+/-3.4%), arctic foxes (68.6+/-10.5%), bearded seals (63.6+/-8.4%), and glaucous gulls (24.2+/-5.8%), as compared to that of control rat liver (9.8+/-3.5%). However, no pathological abnormality such as fibrosis or necrosis was observed in hepatic parenchymal and nonparenchymal cells of arctic animals, and there were no differences in the intralobular distribution of parenchymal cells displaying the nuclear deviation in the livers from either arctic animals and control rats. The hepatic sinusoidal littoral cells such as stellate cells or extracellular matrix components in the perisinusoidal spaces may influence the nuclear positioning and hence the polarity and intrinsic physiological function of parenchymal cells.

Animals↗
PubMed2001

What is the potential for formal patient registration in Canadian primary care? The scale of 'informal registration' in Manitoba.

OBJECTIVES: Registration of patients with a physician is increasingly being considered across Canada as part of a reform of the primary health care system. This study aims to determine how much 'informal registration' currently exists; that is, to identify the number/proportion of patients who, given existing utilization patterns, already receive the majority of their care from one practice, in order to assess the potential for formal registration. METHODS: Administrative data were used to classify patients (n = 528,905) as being informally registered with a clinic if they received the majority of their care (75% or more of their total ambulatory visits) from the same practice over a three-year period (1994-1996). The proportion and number of informally registered patients were examined. RESULTS: Substantial variability emerged across practices in the proportion of informally registered patients (15-68%) and the number of informally registered patients per physician (544-1378 patients per full-time equivalent physician). Informal registration was higher among rural practices (60%) than among urban practices (38%). CONCLUSIONS: While formal registration of patients with physicians is increasingly being considered in Canada as a means to improve the primary care system, the potential disruption to both patients and physicians in moving towards registration should not be underestimated. The relatively low levels of existing informal registration suggest a need to enhance access by, for example, providing after-hours services.

Cohort Studies↗
PubMed2000

Minimal exclusion of plasma membrane proteins during retroviral envelope formation.

The retrovirus forms its envelope by budding at the plasma membrane (PM). This process is primarily driven by its cytoplasmic core-precursor protein, Gag, as shown by the efficient formation of virus-like Gag particles in the absence of its envelope protein, Env. Most interestingly, several studies have demonstrated incorporation of various PM proteins into retrovirus, but the underlying mechanism of this phenomenon has remained elusive. We have purified Moloney murine leukemia virus Gag particles by sedimentation in an iodixanol gradient and donor PMs by flotation in a sucrose gradient and compared their protein compositions at equal lipid basis. We found that most PM proteins are present at similar density in both membranes. The inclusion of PM proteins was unaffected by incorporation of Env protein into the envelope of the Gag particles and whether these were produced at high or low level in the cells. These findings indicate that most PM proteins become incorporated into the retrovirus envelope without significant sorting. This feature of retrovirus assembly should be considered when studying retrovirus functions and developing retrovirus vectors.

Animals↗
PubMed2000

Protein-bound D-amino acids, and to a lesser extent lysinoalanine, decrease true ileal protein digestibility in minipigs as determined with (15)N-labeling.

Heat and alkali treatment of food may increase the concentrations of protein-bound D-amino acids and cross-links such as lysinoalanine (LAL). To examine how protein treatment affects digestibility, purified test meals [total protein 150 g/kg dry matter (DM), 0.44 MJ/(kg BW(0.75). d)] were prepared, containing (g/kg DM) casein, 75; beta-lactoglobulin, 50; or wheat protein, 40. Each was (15)N-labeled. Test proteins were used either in their native form or after treatment for 6 or 24 h at 65 degrees C, pH 10.5-11.5. Each meal was fed to nine adult miniature pigs (twofold complete cross-classification). Chyme was collected continuously over 33 h postprandially via T-fistulas in the terminal ileum, and digestibilities of test proteins and individual L- and D-amino acids were calculated on the basis of recovery of (15)N and the respective amino acids in the chyme. Treatment of casein, beta-lactoglobulin or wheat protein for 24 h increased levels of D-amino acid residues. L-Asparagine and aspartate (L-Asx) were particularly susceptible; 14. 7 +/- 0.4, 11.7 +/- 0.2 and 11.0 +/- 0.9%, respectively, underwent racemization. LAL levels increased in parallel; 11.7 +/- 0.3, 13.6 +/- 0 and 14.8 +/- 0.0%, respectively, of total lysine was converted to LAL. At the same time, prececal protein digestibility was decreased by 13.4 +/- 2.3, 15.3 +/- 1.4 and 17.8 +/- 1.2% units, respectively (P < 0.05; mean +/- SEM, n = 9). Digestibility of individual L-amino acids decreased by 10-15%, but L-amino acids prone to peptic cleavage, such as L-phenylalanine and L-tyrosine, were not affected. Digestibilities of D-amino acids and LAL were approximately 35%. It seems that mainly D-amino acids, and to a lesser extent LAL, were responsible for lower digestibility by interfering with peptic cleavage.

Amino Acids↗
PubMed2000

Characterization of vaccinia virus intracellular cores: implications for viral uncoating and core structure.

The entry of vaccinia virus (VV) into the host cell results in the delivery of the double-stranded DNA genome-containing core into the cytoplasm. The core is disassembled, releasing the viral DNA in order to initiate VV cytoplasmic transcription and DNA replication. Core disassembly can be prevented using the VV early transcription inhibitor actinomycin D (actD), since early VV protein synthesis is required for core uncoating. In this study, VV intracellular cores were accumulated in the presence of actD and isolated from infected cells. The content of these cores was analyzed by negative staining EM and by Western blotting using a collection of antibodies to VV core and membrane proteins. By Western blot analyses, intracellular actD cores, as well as cores prepared by NP-40-dithiothreitol treatment of purified virions (NP-40/DTT cores), contained the core proteins p25 (encoded by L4R), 4a (A10L), 4b (A3L), and p39 (A4L) as well as small amounts of the VV membrane proteins p32 (D8L) and p35 (H3L). While NP-40/DTT cores contained the major putative DNA-binding protein p11 (F17R), actD cores entirely lacked this protein. Labeled cryosections of cells infected for different periods of time in the presence or absence of actD were subsequently used to follow the fate of VV core proteins by EM. These EM images confirmed that p11 was lost at the plasma membrane upon core penetration. The cores that accumulated in the presence of actD were labeled with antibodies to 4a, p39, p25, and DNA at all times examined. In the absence of the drug the cores gradually lost their electron-dense inner part, concomitant with the loss of p25 and DNA labeling. The remaining core shell still labeled with antibodies to p39 and 4a/4b, implying that these proteins are part of this structure. These combined data are discussed with respect to the structure of VV as well as core disassembly.

Blotting, Western↗
PubMed2000

Immunocytochemical localization of Shc and activated EGF receptor in early endosomes after EGF stimulation of HeLa cells.

After binding of epidermal growth factor (EGF), the EGF receptor (EGFR) becomes autophosphorylated via tyrosine. The ligand-activated receptor is internalized by endocytosis and subsequently degraded in the lysosomal pathway. To follow EGFR activation after EGF stimulation, we generated antisera to the EGFR phosphotyrosine sites pY992 and pY1173. The SH2 region of Shc binds to both these sites. Both antisera identified EGFR after EGF binding and did not crossreact with the unactivated receptor. The intracellular distribution of phosphorylated EGFR after ligand binding was traced by two-color immunofluorescence confocal microscopy and immunoelectron microscopy. Before EGF stimulation EGFR was primarily located along the cell surface. When internalization of activated EGFR was inhibited by incubation with EGF on ice, Y992- and Y1173-phosphorylated EGFR were located along the plasma membrane. Ten minutes after internalization at 37C, Y992- and Y1173-phosphorylated EGFR were almost exclusively located in early endosomes, as shown by co-localization with EEA1. Immunoelectron microscopy confirmed that phosphorylated EGFR was located in intracellular vesicles resembling early endosomes. After EGF stimulation, the adaptor protein Shc redistributed to EGFR-containing early endosomes. Our results indicate that EGFR activation of Shc via tyrosine-phosphorylated Y992 and Y1173 occurred in early endocytic compartments, and support a role for membrane trafficking in intracellular signaling.

Adaptor Proteins, Signal Transducing↗
PubMed1999

[Intra- and peri-renal gas formation. Bilateral emphysematous pyelonephritis].

Bilateral emphysematous pyelonephritis is an extremely rare, rapidly progressive and life-threatening intrarenal and perirenal infection. It is associated with diabetes mellitus, obstruction of the upper urinary tract and the presence of gas-forming coliform bacteria. As it presents as a severe acute pyelonephritis, a high index of suspicion and computer-tomographic imaging are essential to make the diagnosis. Only immediate combined medical and (bilateral) surgical therapy can reduce the high patient mortality. Postoperatively, computer-tomographic monitoring is recommended to follow the response to therapy.

Aged↗
PubMed1999

Detection of pulmonary nodules with overlapping vs non-overlapping image reconstruction at spiral CT.

The aim of this study was to analyze whether overlapping image reconstruction increases numbers of pulmonary nodules detected at helical CT. Forty-eight helical CT scans (21 with a slice thickness of 10 mm; 27 with a slice thickness of 5 mm) of patients with known pulmonary nodules were reconstructed both with overlapping and non-overlapping image reconstruction. Two readers recorded number and size of pulmonary nodules as well as diagnostic confidence. With overlapping image reconstruction each reader diagnosed more pulmonary nodules (slice thickness 10 mm: +24.0 and +26.7%, both p < 0.01; slice thickness 5 mm: +9.5 and +11.9%, both not significant) and more "definite" nodules (slice thickness 10 mm: +20.3%, p < 0.05, and +30.8%, p < 0.005; slice thickness 5 mm: +18.0 and +17.0%, both p < 0.05). Nodules diagnosed with overlapping image reconstruction only were almost exclusively smaller than the slice thickness. The increase in number of nodules detected was not associated with a decrease in diagnostic confidence. Overlapping image reconstruction improves detection of pulmonary nodules smaller than the slice thickness at spiral CT.

Adolescent↗
PubMed1999

Recovery of 15N-lactoferrin is higher than that of 15N-casein in the small intestine of suckling, but not adult miniature pigs.

Performance of biological functions of lactoferrin in the small intestine requires at least some resistance to degradation. Therefore, we studied prececal digestibility of lactoferrin in comparison to casein both in suckling and adult miniature pigs, applying 15N-labeled proteins. In study 1, 43 piglets (10-d-old), deprived of food for 12 h received 10 mL of sow's milk supplemented with 120 mg of 15N-labeled protein (porcine or bovine lactoferrin or bovine casein). Piglets were anesthetized 150 min later, after which the small intestine was excised, cut into three sections, and chyme was collected. In study 2, nine food-deprived boars fitted with T-canulae at the terminal ileum were given two semisynthetic experimental meals (204 g) in a cross-over design, 2 wk apart. One contained 7.5% (g/100 g) 15N-labeled bovine casein, the other 1.25% 15N-labeled bovine lactoferrin. Both were adjusted to 15% total protein with nonlabeled casein. Ileal chyme was collected from the canula over 33 h postprandially. All diets contained the indigestible marker chromic oxide. 15N-digestibility of lactoferrin, both porcine (84.4 +/- 3.2%) and bovine (82.3 +/- 4.8%), was significantly lower than casein digestibility (97.6 +/- 0.5%) in the distal small intestine of suckling piglets (P < 0.05). Based on immunoblotting after acrylamide electrophoresis, 4.5% of non- and partially digested lactoferrin was found in the last third of the small intestine of piglets. In adult miniature pigs there was no difference in 15N-digestibility of bovine lactoferrin compared to bovine casein (90.7 +/- 1.9% vs. 93.9 +/- 1.0%, P > 0.05). In suckling miniature pigs, the reduced digestibility of lactoferrin may provide the prerequisite for biological actions along the whole intestinal tract. The source of lactoferrin, porcine or bovine, made no difference in this respect.

Aging↗
PubMed1999

Open reading frame 1a-encoded subunits of the arterivirus replicase induce endoplasmic reticulum-derived double-membrane vesicles which carry the viral replication complex.

The replicase of equine arteritis virus (EAV; family Arteriviridae, order Nidovirales) is expressed in the form of two polyproteins (the open reading frame 1a [ORF1a] and ORF1ab proteins). Three viral proteases cleave these precursors into 12 nonstructural proteins, which direct both genome replication and subgenomic mRNA transcription. Immunofluorescence assays showed that most EAV replicase subunits localize to membranes in the perinuclear region of the infected cell. Using replicase-specific antibodies and cryoimmunoelectron microscopy, unusual double-membrane vesicles (DMVs) were identified as the probable site of EAV RNA synthesis. These DMVs were previously observed in cells infected with different arteriviruses but were never implicated in viral RNA synthesis. Extensive electron microscopic analysis showed that they appear to be derived from paired endoplasmic reticulum membranes and that they are most likely formed by protrusion and detachment of vesicular structures with a double membrane. Interestingly, very similar membrane rearrangements were observed upon expression of ORF1a-encoded replicase subunits nsp2 to nsp7 from an alphavirus-based expression vector. Apparently, the formation of a membrane-bound scaffold for the replication complex is a distinct step in the arterivirus life cycle, which is directed by the ORF1a protein and does not depend on other viral proteins and/or EAV-specific RNA synthesis.

Animals↗
PubMed1999

Pulmonary nodules: experimental and clinical studies at low-dose CT.

PURPOSE: To compare the number of pulmonary nodules detected at helical low- and standard-dose computed tomography (CT) and to investigate the diagnostic value of low-dose CT with a radiation exposure equivalent to that used at chest radiography. MATERIALS AND METHODS: Two radiologists recorded pulmonary nodules at standard-dose (250 or 100 mA, pitch of 1; 200 mA, pitch of 2) or low-dose CT (50 or 25 mA, pitch of 1 or 2) in five postmortem specimens and 75 patients. Nodules were assessed by size (5 mm or smaller, 6-10 mm, or larger than 10 mm) and by diagnostic confidence ("definite nodule," "definite lesion, not classic nodule," or "questionable lesion, possibly representing a vessel") with the Wilcoxon signed rank test. Artifacts depicted at low-dose CT were recorded. RESULTS: There were no statistically significant differences in the number of nodules detected at standard- or low-dose CT except in nodules 5 mm or smaller that were assessed as definite nodules at standard- or low-dose CT (25 mA, pitch of 2) (472 vs 397, P < .05). Artifacts that possibly interfered with nodule detection were observed exclusively at CT with 25 mA and a pitch of 2. CONCLUSION: Pulmonary nodules were detected reliably at CT with 50 mA and pitch of 2 or with 25 mA and a pitch of 1. However, further reduction of the dose to that used at chest radiography was associated with a significant decrease in the number of nodules 5 mm or smaller that were detected, possibly due to artifacts.

Adult↗
PubMed1999

Helical CT of pulmonary nodules in patients with extrathoracic malignancy: CT-surgical correlation.

OBJECTIVE: Our aim was to assess the sensitivity of helical CT for revealing pulmonary nodules. Thoracotomy with palpation of the deflated lung, resection, and histologic examination of palpable nodules was used as the gold standard. SUBJECTS AND METHODS: Thirteen patients underwent helical CT (slice thickness, 5 mm; reconstruction intervals, 3 mm and 5 mm; interpreted by two independent observers). Subsequently, patients underwent unilateral (n = 6) or bilateral (n = 7) surgical exploration, and CT-surgical correlation of 20 lungs was performed. RESULTS: Ninety nodules were resected (61 were smaller than 6 mm; 13 were 6-10 mm; 11 were larger than 10 mm; in five nodules, the size was not recorded at surgery). Sixty-nine nodules were located in the pulmonary parenchyma and 21 in the visceral pleura. Of the 90 lesions, 43 (48%) were found on histology to represent metastases. For lesions detected by at least one observer, the sensitivity of helical CT was 69% for intrapulmonary nodules smaller than 6 mm, 95% for intrapulmonary nodules larger than or equal to 6 mm, and 100% for histologically proven intrapulmonary metastases larger than or equal to 6 mm. For lesions smaller than or equal to 10 mm, sensitivity was better using a reconstruction interval of 3 mm rather than of 5 mm. CONCLUSION: In this study, the sensitivity of helical CT exceeded the sensitivity of conventional CT in previous reports. However, because of limitations in the detection of intrapulmonary nodules smaller than 6 mm and of pleural lesions, complete surgical exploration should remain the procedure of choice in patients undergoing pulmonary metastasectomy. Preoperative helical CT should be used to guide the surgeon to lesions that are difficult to palpate.

Adult↗