Molecular dynamics study of water channels in natural and synthetic amyloid-ß fibrils.

We compared all-atom explicit solvent molecular dynamics simulations of three types of Aß(1-40) fibrils: brain-seeded fibrils (2M4J, with a threefold axial symmetry) and the other two, all-synthetic fibril polymorphs (2LMN and 2LMP, made under different fibrillization conditions). Fibril models were constructed using either a finite or an infinite number of layers made using periodic images. These studies yielded four conclusions. First, finite fibrils tend to unravel in a manner reminiscent of fibril dissolution, while infinite fibrils were more stable during simulations. Second, salt bridges in these fibrils remained stable in those fibrils that contained them initially, and those without salt bridges did not develop them over the time course of the simulations. Third, all fibrils tended to develop a "stagger" or register shift of ß-strands along the fibril axis. Fourth and most importantly, the brain-seeded, 2M4J, infinite fibrils allowed bidirectional transport of water in and out of the central longitudinal core of the fibril by rapidly developing gaps at the fibril vertices. 2LMP fibrils also showed this behavior, although to a lesser extent. The diffusion of water molecules in the fibril core region involved two dynamical states: a localized state and directed diffusion in the presence of obstacles. These observations provided support for the hypothesis that Aß fibrils could act as nanotubes. At least some Aß oligomers resembled fibrils structurally in having parallel, in-register ß-sheets and a sheet-turn-sheet motif. Thus, our findings could have implications for Aß cytotoxicity, which may occur through the ability of oligomers to form abnormal water and ion channels in cell membranes.

Ionic strength independence of charge distributions in solvation of biomolecules.

Electrostatic forces enormously impact the structure, interactions, and function of biomolecules. We perform all-atom molecular dynamics simulations for 5 proteins and 5 RNAs to determine the dependence on ionic strength of the ion and water charge distributions surrounding the biomolecules, as well as the contributions of ions to the electrostatic free energy of interaction between the biomolecule and the surrounding salt solution (for a total of 40 different biomolecule/solvent combinations). Although water provides the dominant contribution to the charge density distribution and to the electrostatic potential even in 1M NaCl solutions, the contributions of water molecules and of ions to the total electrostatic interaction free energy with the solvated biomolecule are comparable. The electrostatic biomolecule/solvent interaction energies and the total charge distribution exhibit a remarkable insensitivity to salt concentrations over a huge range of salt concentrations (20 mM to 1M NaCl). The electrostatic potentials near the biomolecule's surface obtained from the MD simulations differ markedly, as expected, from the potentials predicted by continuum dielectric models, even though the total electrostatic interaction free energies are within 11% of each other.

Increased endoplasmic reticulum stress in decidual tissue from pregnancies complicated by fetal growth restriction with and without pre-eclampsia.

OBJECTIVES: Endoplasmic reticulum (ER) stress has been implicated in both pre-eclampsia (PE) and fetal growth restriction (FGR), and is characterised by activation of three signalling branches: 1) PERK-pEIF2α, 2) ATF6 and 3) splicing of XBP1(U) into XBP1(S). To evaluate the contribution of ER stress in the pathogenesis of PE relative to FGR, we compared levels of ER stress markers in decidual tissue from pregnancies complicated by PE and/or FGR. STUDY DESIGN: Whole-genome transcriptional profiling was performed on decidual tissue from women with PE (n = 13), FGR (n = 9), PE+FGR (n = 24) and controls (n = 58), and used for pathway and targeted transcriptional analyses of ER stress markers. The expression and cellular localisation of ER stress markers was assesses by Western blot and immunofluorescence analyses. RESULTS: Increased ER stress was observed in FGR and PE+FGR, including both the PERK-pEIF2α and ATF6 signalling branches, whereas ER stress was less evident in isolated PE. However, these cases demonstrated elevated levels of XBP1(U) protein. ATF6 and XBP1 immunoreactivity was detected in most (>80%) extravillous trophoblasts, decidual cells and macrophages. No difference in the proportion of immunopositive cells or staining pattern was observed between study groups. CONCLUSIONS: Increased PERK-pEIF2α and ATF6 signalling have been associated with decreased cellular proliferation and may contribute to the impaired placental growth characterising pregnancies with FGR and PE+FGR. XBP1(U) has been proposed as a negative regulator of ER stress, and increased levels in PE may reflect a protective mechanism against the detrimental effects of ER stress.

Objective prioritization of positional candidate genes at a quantitative trait locus for pre-eclampsia on 2q22.

Pre-eclampsia/eclampsia (PE/E) is a common, serious medical disorder of human pregnancy. Familial association of PE/E has been recognized for decades, but the genetics are complex and poorly understood. In an attempt to identify PE/E susceptibility genes, we embarked on a positional cloning strategy using 34 Australian and New Zealand PE/E pedigrees. An initial 10-cM resolution genome scan revealed a putative susceptibility locus spanning a broad region on chromosome 2 that overlaps an independently determined linkage signal seen in Icelandic PE pedigrees. Subsequent fine mapping using 25 additional short tandem repeat (STR) markers in this region and non-parametric multipoint linkage analysis did not change the overall position. Under a strict diagnosis of PE, we obtained significant evidence of linkage on 2q with a peak log-of-odds ratio score (LOD) of 3.43 near marker D2S151 at 155 cM. To prioritize positional candidate genes at the 2q locus for detailed analysis, we applied an objective prioritization strategy that integrates quantitative bioinformatics, assessment of differential gene expression and association analysis of single-nucleotide polymorphisms (SNPs). Highest priority was assigned to the activin receptor gene ACVR2. This gene also showed >10-fold differential gene expression in human decidual tissue from normotensive and PE individuals. We genotyped five known SNPs in this gene in our pedigrees and performed tests for association and linkage disequilibrium. One SNP (rs1424954) showed strong preliminary evidence of association with PE (P = 0.007), whereas two others (rs1364658 and rs1895694) exhibited nominal evidence (P < 0.05). Haplotype analysis revealed no additional association information. There was evidence of weak linkage disequilibrium among these SNPs. The highest observed LD occurred between the two strongest associated SNPs, suggesting that the observed signals may be the signature of an observed functional variant.

Gene expression of the constant region of the heavy chain of immunoglobulin G (IgG CRHC) is down-regulated in human decidua in association with preeclampsia.

An aberrant interaction at the maternal/fetal interface between the genetically distinct fetal trophoblast cells and cells of the maternal decidua has been proposed as an initiating factor in one of the major complications of human pregnancy, preeclampsia. Biochemical and epidemiological studies suggest that the immune system plays an important role in preeclampsia. Thus, the aim of this study was to determine the decidual gene expression status in preeclampsia of one of the key components of the adaptive immune system. Total RNA was extracted from decidua collected from women with normal pregnancies and those complicated by preeclampsia. Reverse Northern analysis was performed on 72 cDNAs from human decidua and differentially expressed genes identified were analysed further using semi-quantitative RT-PCR and Northern blot analysis. Expression of the gene encoding the constant region of the heavy chain of immunoglobulin G (IgG CRHC) was shown to be down-regulated in association with preeclampsia. These data support the hypothesis that immune maladaptation may play an important role in the pathogenesis of preeclampsia.

Detection of CAG repeats in pre-eclampsia/eclampsia using the repeat expansion detection method.

Pre-eclampsia/eclampsia is a serious disorder of human pregnancy that has a worldwide incidence of 2-10% and carries a severe morbidity and mortality risk for both mother and child. Its precise cause remains unknown. However, there is increasing evidence of an underlying complex maternal genetic susceptibility. Its high population incidence in the face of strong negative selection pressure suggests that the gene(s) involved have a selective advantage and/or a high mutation rate. One class of genetic diseases that involve a high mutation rate are the trinucleotide repeat expansion diseases. Thus, the aim of this study was to determine whether there is an association between a trinucleotide (CAG) repeat expansion and pre-eclampsia/eclampsia. We have used the repeat expansion detection (RED) method, which was developed to directly identify clinically significant repeat expansions, to analyse genomic DNA from an Australian and New Zealand population. The maximal CAG repeat length for each individual was recorded and the Mann-Whitney U and Wilcoxon rank sum test for independent samples were used to compare distributions for CAG/CTG repeats between two populations. There were no statistically significant differences between the distribution of CAG repeats in normotensive (n = 59) and severe pre-eclampsia (n = 69) (Mann-Whitney U = 1732; P = 0.14), and normotensive (n = 59) and eclamptic (n = 15) populations (Mann-Whitney U = 417, P = 0.726). Therefore, these RED results do not support a role for a large CAG expansion in pre-eclampsia/eclampsia. However, these data do not preclude the possibility that a small CAG expansion is associated with the disorder nor do they negate the hypothesis that a highly mutable gene contributes to the genetic component of pre-eclampsia/eclampsia.

Breakthrough adverse reactions to low-osmolar contrast media after steroid premedication.

OBJECTIVE: The purpose of this study was to review the nature of adverse reactions, or "breakthrough reactions," experienced by patients who received steroid premedication and low-osmolar contrast media. We compared the demographics of patients having these breakthrough reactions with those of patients who did not develop these reactions. MATERIALS AND METHODS: We retrospectively reviewed our radiology quality improvement database to identify patients with breakthrough reactions that occurred from January 1, 1994, through October 1, 1999, and we reviewed their medical records. We compared these patients with a control cohort of patients who had a history of prior adverse reaction to contrast media but no breakthrough reaction after administration of low-osmolar contrast media and premedication with corticosteroids. RESULTS: Over the 6-year period, 52 patients experienced 61 breakthrough reactions. The breakthrough reaction was mild in 76% of the patients. The breakthrough reaction was similar to the patient's initial adverse reaction in 85% of the patients. A history of seafood allergy or hay fever was statistically more likely to be identified in the breakthrough group than the control group. CONCLUSION: Breakthrough reactions occur in a substantial number of patients despite premedication with steroids and use of low-osmolar contrast agents. Typically the breakthrough reaction is of similar severity to the patient's initial reaction. Severe or life-threatening reactions are seen in 24% of patients.

Isolated pelvic deep venous thrombosis: relative frequency as detected with MR imaging.

PURPOSE: To determine the relative frequency of deep venous thrombosis (DVT) isolated to the pelvic veins, as demonstrated with magnetic resonance (MR) imaging. MATERIALS AND METHODS: The reports of 769 MR examinations performed from June 1993 through December 1999 in patients with suspected DVT were reviewed retrospectively. MR venography was performed by using a two-dimensional gradient-recalled-echo sequence (typically repetition time, 34 msec; echo time, 13 msec; flip angle, 60 degrees ). The presence of DVT was categorized by location in the pelvis, thigh, or calf. RESULTS: DVT was identified in 167 (21.7%) of the 769 MR examinations. Thirty-four (20.4%) of the 167 studies demonstrated DVT isolated to the pelvic veins. CONCLUSION: The relative frequency of isolated pelvic DVT detected with MR venography was higher than that reported in prior studies with ultrasonography (US) or ascending venography. MR venography should be performed in patients with suspected pelvic DVT or when clinical suspicion persists despite a negative US study.

The anterior iliac separation: alternative index for pelvic morphometry in fetuses with Down syndrome.

OBJECTIVE: The goal of this study was to assess the diagnostic use of an anterior iliac separation measurement as an alternative index for the iliac angle in the assessment of fetal pelvic morphometry. SUBJECTS AND METHODS: In 358 fetuses, the anterior iliac separation, iliac length, and iliac angle were prospectively measured on antenatal sonography. All measurements were obtained at two axial levels (superior and inferior). The gestational age of the fetus was recorded. The anterior iliac separation was normalized by iliac length, and coefficients of variation were calculated for all measurements. The effects of axial level and gestational age were assessed in a linear regression model. The diagnostic use of the anterior iliac separation relative to that of the iliac angle was assessed in a comparison of 24 fetuses with Down syndrome and 247 non-Down syndrome fetuses. RESULTS: The anterior iliac separation was less variable than the iliac angle at both superior and inferior levels. There were statistically significant effects for gestational age and axial level on both the anterior iliac separation and the iliac angle, but there was no significant effect for either factor when the anterior iliac separation was normalized by the iliac length. Comparing Down and non-Down syndrome fetuses, we found that the normalized anterior iliac separation had discriminating power similar to the iliac angle. CONCLUSION: The linear measurement of the anterior iliac separation has diagnostic properties similar to the iliac angle and is subject to less measurement variability. This simpler measurement may be particularly useful when normalized by the iliac length.

MRI of conjoined twins illustrating advances in fetal imaging.

Complex congenital fetal abnormalities are optimally evaluated with both ultrasound and magnetic resonance imaging. With the advent of ultrafast imaging sequences, fetal imaging has significantly improved. We present two cases of conjoined thoracoomphalopagus twins to discuss this unusual anomaly and to illustrate the recent advances in magnetic resonance imaging of the fetus.

Staging of pancreatic cancer before and after neoadjuvant chemoradiation.

Neoadjuvant chemoradiation therapy is used at many institutions for treatment of localized adenocarcinoma of the pancreas. Accurate staging before neoadjuvant therapy identifies patients with distant metastatic disease, and restaging after neoadjuvant therapy selects patients for laparotomy and attempted resection. The aims of this study were to (1) determine the utility of staging laparoscopy in candidates for neoadjuvant therapy and (2) evaluate the accuracy of restaging CT following chemoradiation. Staging laparoscopy was performed in 98 patients with radiographically potentially resectable (no evidence of arterial abutment or venous occlusion) or locally advanced (arterial abutment or venous occlusion) adenocarcinoma of the pancreas. Unsuspected distant metastasis was identified in 8 (18%) of 45 patients with potentially resectable tumors and 13 (24%) of 55 patients with locally advanced tumors by CT. Neoadjuvant chemoradiation therapy and restaging CT were completed in a total of 103 patients. Thirty-three patients with potentially resectable tumors by restaging CT underwent surgical exploration and resections were performed in 27 (82%). Eleven (22%) of 49 patients with locally advanced tumors by restaging CT were resected, with negative margins in 55%; the tumors in these 11 patients had been considered locally advanced because of arterial involvement on restaging CT. Staging laparoscopy is useful for the exclusion of patients with unsuspected metastatic disease from aggressive neoadjuvant chemoradiation protocols. Following neoadjuvant chemoradiation, restaging CT guides the selection of patients for laparotomy but may overestimate unresectability to a greater extent than does prechemoradiation CT.

Nonenhanced helical CT and US in the emergency evaluation of patients with renal colic: prospective comparison.

PURPOSE: To compare nonenhanced helical computed tomography (CT) with ultrasonography (US) for the depiction of urolithiasis. MATERIALS AND METHODS: During 9 months, 45 patients (mean age, 44 years; mean weight, 92.5 kg) prospectively underwent both nonenhanced helical CT (5-mm collimation; pitch of 1.5) and US of the kidneys, ureters, and bladder. US evaluation included a careful search for ureteral calculi. Presence of calculi and obstruction and incidental diagnoses were recorded. Clinical, surgical, and/or imaging follow-up data were obtained in all patients. The McNemar test was used to compare groups. RESULTS: Diagnoses included 23 ureteral calculi and one each of renal cell carcinoma, appendicitis, ureteropelvic junction obstruction, renal subcapsular hematoma, cholelithiasis, medullary calcinosis, and myelolipoma. CT depicted 22 of 23 ureteral calculi (sensitivity, 96%). US depicted 14 of 23 ureteral calculi (sensitivity, 61%). Differences in sensitivity were statistically significant (P: =.02). Specificity for each technique was 100%. When modalities were compared for the detection of any clinically relevant abnormality (eg, unilateral hydronephrosis and/or urolithiasis in patients with an obstructing calculus), sensitivities of US and CT increased to 92% and 100%, respectively. One case of appendicitis was missed at US, whereas medullary calcinosis and myelolipoma were missed at CT. CONCLUSION: Nonenhanced CT has a higher sensitivity for the detection of ureteral calculi compared with US.

A genome scan in families from Australia and New Zealand confirms the presence of a maternal susceptibility locus for pre-eclampsia, on chromosome 2.

Epidemiological studies have shown that genetic factors contribute to the etiology of the common and serious pregnancy-specific disorder pre-eclampsia (PE)/eclampsia (E). Candidate-gene studies have provided evidence (albeit controversial) of linkage to several genes, including angiotensinogen on 1q42-43 and eNOS on 7q36. A recent medium-density genome scan in Icelandic families identified significant linkage to D2S286 (at 94.05 cM) on chromosome 2p12 and suggestive linkage to D2S321 (at 157.5 cM) on chromosome 2q23. In the present article, the authors report the results of a medium-density genome scan in 34 families, representing 121 affected women, from Australia and New Zealand. Multipoint nonparametric linkage analysis, using the GENEHUNTER-PLUS program, showed suggestive evidence of linkage to chromosome 2 (LOD=2.58), at 144.7 cM, between D2S112 and D2S151, and to chromosome 11q23-24, between D11S925 and D11S4151 (LOD=2.02 at 121.3 cM). Given the limited precision of estimates of the map location of disease-predisposing loci for complex traits, the present finding on chromosome 2 is consistent with the finding from the Icelandic study, and it may represent evidence of the same locus segregating in the population from Australia and New Zealand. The authors propose that the PE/E-linked locus on chromosome 2p should be designated the "PREG1" (pre-eclampsia, eclampsia gene 1) locus.

Radiologic manifestations of sclerosing cholangitis with emphasis on MR cholangiopancreatography.

Magnetic resonance cholangiopancreatography (MRCP) is a relatively new, noninvasive cholangiographic technique that is comparable with invasive endoscopic retrograde cholangiopancreatography (ERCP) in the detection and characterization of extrahepatic bile duct abnormalities. The role of MRCP in evaluation of the intrahepatic bile ducts, especially in patients with primary or secondary sclerosing cholangitis, is under investigation. The key cholangiographic features of primary sclerosing cholangitis are randomly distributed annular strictures out of proportion to upstream dilatation. As the fibrosing process worsens, strictures increase and the ducts become obliterated, and the peripheral ducts cannot be visualized to the periphery of the liver at ERCP. In addition, the acute angles formed with the central ducts become more obtuse. With further progression, strictures of the central ducts prevent peripheral ductal opacification at ERCP. Cholangiocarcinoma occurs in 10%-15% of patients with primary sclerosing cholangitis; cholangiographic features that suggest cholangiocarcinoma include irregular high-grade ductal narrowing with shouldered margins, rapid progression of strictures, marked ductal dilatation proximal to strictures, and polypoid lesions. Secondary sclerosing and nonsclerosing processes can mimic primary sclerosing cholangitis at cholangiography. These processes include ascending cholangitis, oriental cholangiohepatitis, acquired immunodeficiency syndrome-related cholangitis, chemotherapy-induced cholangitis, ischemic cholangitis after liver transplantation, eosinophilic cholangitis, and metastases.

Normal fetal pelvis: important factors for morphometric characterization with US.

PURPOSE: To prospectively evaluate iliac angle and iliac length in a large number of normal fetuses and to identify factors that may influence these measurements. MATERIALS AND METHODS: At antenatal ultrasonography (US) in 356 fetuses, the iliac angle and iliac length were measured at two axial levels (superior and inferior). In mixed linear models, the statistical significance and magnitude of effect on the measurement of iliac angle and iliac length were estimated for gestational age, fetal sex, maternal diabetes status, axial level, and spine position relative to the transducer. RESULTS: Statistically significant effects were found for gestational age, axial level, and spine orientation but not for fetal sex or maternal diabetes status. The iliac angle was found to decrease by 15.7 degrees from the superior to inferior portion of the pelvis, decrease by approximately 0.37 degrees /wk, and decrease by as much as 15.6 degrees when the spine is directed to the side. Iliac length was found to increase by 0.8 mm/wk from 13 weeks to term, decrease by 1.2 mm from the superior to the inferior portion of the pelvis, and increase by as much as 1.29 mm when the spine is not directly subjacent to the transducer. CONCLUSION: The axial level of measurement, gestational age, and spine orientation must be accounted for if these morphometric indexes are used to discriminate fetuses with and those without Down syndrome.

Pelvic CT morphometry in Down syndrome: implications for prenatal US evaluation--preliminary results.

PURPOSE: To characterize pelvic morphometric differences in patients with and those without Down syndrome by using computed tomography (CT) and to determine useful indexes for ultrasonographic (US) evaluation. MATERIALS AND METHODS: Pelvic CT scans in seven patients with Down syndrome and in 27 patients without Down syndrome were reviewed. Iliac angle, iliac length, sacroiliac joint angle, and anterior iliac wing separation were measured at superior, middle, and inferior transverse sacral levels. The effects of chromosomal status and transverse level were evaluated statistically. RESULTS: Significant differences were found for mean iliac angle (P < .007) and length (P < .005) between patients without Down syndrome (angle, 75 degrees; length, 8.4 cm) and those with Down syndrome (angle, 82 degrees; length, 7.5 cm). Depending on the level of measurement, variations in iliac angle between patients without and those with Down syndrome were as much as 13 degrees and 15 degrees, respectively, and variations in length were as much as 1.6 cm and 0.9 cm, respectively. The greatest differences were at the middle sacral level. Sacroiliac joint angle and the anterior iliac wing separation were not different between groups. CONCLUSION: Patients with Down syndrome had a larger mean iliac angle and a shorter mean iliac length. The most pronounced differences were at the middle sacral level, which suggests that this may be the optimal level for measuring these parameters at prenatal US.

Imaging-guided percutaneous biopsy of focal splenic lesions: update on safety and effectiveness.

OBJECTIVE: The purpose of this study is to determine the safety and effectiveness of percutaneous imaging-guided biopsy in the diagnosis of focal splenic lesions. MATERIALS AND METHODS: From May 1995 to November 1997, 20 imaging-guided biopsies of focal splenic lesions were performed in 18 patients, including seven patients with a prior diagnosis of extrasplenic malignancy (breast cancer, n = 3; lymphoma, n = 2; ovarian cancer, n = 1; and osteogenic sarcoma, n = 1), three immunosuppressed patients (cause of immunosuppression: AIDS, n = 1; liver transplantation, n = 1; and bone marrow transplantation, n = 1), two patients with anemia, one patient with a recent history of IV drug abuse, and five patients with incidentally discovered splenic lesions. Biopsies were performed with an 18-gauge (n = 1), a 20-gauge (n = 8), or a 22-gauge (n = 14) self-aspirating needle or an 18-gauge cutting needle (n = 1). Biopsies were considered successful if a specific diagnosis of benign or malignant disease was made. RESULTS: A specific diagnosis was made in 16 (88.9%) of 18 patients, and no complications occurred. Malignancy was diagnosed in six patients including three patients with lymphoma. Benign conditions were diagnosed in 10 patients: a cyst in two patients; hamartoma in one; lipogranuloma in one; infarct in one; and infection in four, including one case each of Candida albicans, Pneumocystis carinii, Mycobacterium tuberculosis, and mixed flora. The tenth benign diagnosis was a pseudotumor of the spleen related to a bulbous tail of the pancreas that was inseparable from the splenic hilum. Biopsy did not establish a diagnosis in one patient with lymphoma and in one patient with presumed splenic candidiasis. A mean of 1.5 needle passes was made per biopsy. CONCLUSION: Imaging-guided splenic biopsy is a safe technique that provides a specific diagnosis in most patients with focal splenic lesions.