Isolation and characterization of a monoclonal anti-quadruplex DNA antibody from autoimmune "viable motheaten" mice

A cell line that produces an autoantibody specific for DNA quadruplex structures has been isolated and cloned from a hybridoma library derived from 3-month-old nonimmunized autoimmune, immunodeficient "viable motheaten" mice. This antibody has been tested extensively in vitro and found to bind specifically to DNA quadruplex structures formed by two biologically relevant sequence motifs. Scatchard and nonlinear regression analyses using both one- and two-site models were used to derive association constants for the antibody-DNA binding reactions. In both cases, quadruplexes had higher association constants than triplex and duplex molecules. The anti-quadruplex antibody binds to the quadruplex formed by the promoter-region-derived oligonucleotide d(CGCG4GCG) (Ka = 3.3 x 10(6) M-1), and has enhanced affinity for telomere-derived quadruplexes formed by the oligonucleotides d(TG4) and d(T2G4T2G4T2G4T2G4) (Ka = 5.38 x 10(6) and 1.66 x 10(7) M-1, respectively). The antibody binds both types of quadruplexes but has preferential affinity for the parallel four-stranded structure. In vitro radioimmunofilter binding experiments demonstrated that purified anti-DNA quadruplex antibodies from anti-quadruplex antibody-producing tissue culture supernatants have at least 10-fold higher affinity for quadruplexes than for triplex and duplex DNA structures of similar base composition and length. The antibody binds intramolecular DNA triplexes formed by d(G4T3G4T3C4) and d(C4T3G4T3G4), and the duplex d(CGCGCGCGCG)2 with an affinities of 6. 76 x 10(5), 5.59 x 10(5), and 8.26 x 10(5) M-1, respectively. Competition experiments showed that melted quadruplexes are not effective competitors for antibody binding when compared to native structures, confirming that the quadruplex is bound structure-specifically. To our knowledge, this is the first immunological reagent known to specifically recognize quadruplex structures. Subsequent sequence analysis demonstrates homologies between the antibody complementarity determining regions and sequences from Myb family telomere binding proteins, which are hypothesized to control cell aging via telomeric DNA interactions. The presence of this antibody in the autoimmune repertoire suggests a possible linkage between autoimmunity, telomeric DNA binding proteins, and aging.     

S-phase fractions of breast carcinomas. Relationships to morphology, estrogen and progesterone receptors, and early recurrence

Analysis of the S-phase fractions (SPF) measured by in vitro thymidine labeling, morphological appearances, and estrogen receptor (ER) assays of primary invasive breast carcinomas demonstrated several interrelationships. Lobular, mucinous, tubular, and adenocystic carcinomas consistently had low SPF and were usually positive for ER. The same was true for the carcinomas of no special histologic type [the not otherwise specified (NOS) group of E. R. Fisher including "infiltrating ductal" and undifferentiated carcinomas] with minimal anaplasia. Medullary, atypical medullary, and morphologically unclassifiable carcinomas with marked nuclear anaplasia nearly always had high SPF and were usually negative for ER. High SPF was associated with advanced stages of carcinoma initially or with early recurrence following mastectomy.     

Intravenous glucose tolerance test in middle-aged men with and without latent coronary heart disease

During a cardiovascular survey, aimed at detecting cases of latent coronary heart disease (CHD), glucose elimination was studied after i.v. loading in 1970 presumably healthy men aged 40-59 years. The aim was to throw light on the importance of deranged glucose tolerance for the development of CHD. Of the 1970 individuals, 1798 were defined as "normals", 33 had chronic, non-anginal chest pain, 34 had slight albeit typical angina pectoris. The remaining 105 had various symptoms/signs strongly suggestive of CHD, and underwent diagnostic coronary angiography (69 angiopositive, 36 angionegative). Plasma insulin was determined in relation to the test in 249 of the subjects. The following conclusions were reached: 1) Mean k-values were similar in all subgroups (p less than 0.10). 2) Low and borderline k-values were significantly more frequent in angiographed individuals compared with the group of normals (p less than 0.025). However, an almost identical frequency was seen in angiopositive and angionegative cases. 3) K-values did not change with age between 40 and 59 years. 4) K-values were unrelated to the severity of angiographic findings in individuals with proven CHD. 5) Significantly lower k-values were found in individuals with a positive diabetic heredity, and 6) in individuals with a high insulin response. 7) The i.v. glucose loading did not influence an exercise ECG recorded in relation to a near-maximal bicycle exercise test.     

Ultrastructure of ischemic contracture of the left ventricle ("stone heart")

Myocardial biopsies from two patients who had developed "stone heart" (myocardial rigor mortis; ischemic contracture of the left ventricle) were studied by electron microscopy. The ultrastructure of tissue in stone heart, though ischemic in nature, differed from that of classic myocardial infarction in some respects. Apart from depletion of glycogen and distension of the sarcoplasmic reticulum and T-tubules, myofibrillar degeneration was much more widespread. Mitochondrial degeneration with active lysosomal autodigestion, disruption of the microcirculation, and lymphedema were prominent changes also observed. In the light of known clinical and experimental observations, our findings suggest that stone heart is an accelerated form of ischemic injury occurring in vulnerable (hypertrophied) hearts and is probably related to ischemia-triggered release of endogenous catecholamines.     

Sympathetic response to stimulation of the pontine A5 region in conscious rabbits

OBJECT: This study is a retrospective analysis of clinical data obtained in 28 patients affected by obstructive hydrocephalus who presented with signs of midbrain dysfunction during episodes of shunt malfunction. METHODS: All patients presented with an upward gaze palsy, sometimes associated with other signs of oculomotor dysfunction. In seven cases the ocular signs remained isolated and resolved rapidly after shunt revision. In 21 cases the ocular signs were variably associated with other clinical manifestations such as pyramidal and extrapyramidal deficits, memory disturbances, mutism, or alterations in consciousness. Resolution of these symptoms after shunt revision was usually slow. In four cases a transient paradoxical aggravation was observed at the time of shunt revision. In 11 cases ventriculocistemostomy allowed resolution of the symptoms and withdrawal of the shunt. Simultaneous supratentorial and infratentorial intracranial pressure recordings performed in seven of the patients showed a pressure gradient between the supratentorial and infratentorial compartments, with a higher supratentorial pressure before shunt revision. Inversion of this pressure gradient was observed after shunt revision and resolution of the gradient was observed in one case after third ventriculostomy. In six recent cases, a focal midbrain hyperintensity was evidenced on T2-weighted magnetic resonance imaging sequences at the time of shunt malfunction. This rapidly resolved after the patient underwent third ventriculostomy. CONCLUSIONS: It is probable that in obstructive hydrocephalus, at the time of shunt malfunction, the development of a transtentorial pressure gradient could initially induce a functional impairment of the upper midbrain, inducing upward gaze palsy. The persistence of the gradient could lead to a global dysfunction of the upper midbrain. Third ventriculostomy contributes to equalization of cerebrospinal fluid pressure across the tentorium by restoring free communication between the infratentorial and supratentorial compartments, resulting in resolution of the patient's clinical symptoms.     

Issues and Techniques in Networked-Based Distributed Healthcare: Overview

For telemedicine to reach its potential in the Internet2 environment,significant progress must be made in a number of areas. The emergingtelecommunications will include the Internet2 between medicalinstitutions and emerging broadband loop and wireless technologiesfor interconnections to homes. Within the technical environment,problems that must be addressed include the development of robotic-controlled,remote-monitoring devices unique to the various medical specialties;the encoding, transmission, and decoding of raw data specificto the specialized needs of various medical application areas;and the reduction of data for storage and retrieval in patientrecords. Institutionally, human factor and information accessissues with respect to physicians, patients, and non-physicianhealthcare providers must be addressed; healthcare insuranceprovider policies, government regulation, and state licensinglaws must be adapted for a new paradigm of healthcare delivery;and the philosophy of capital investment for the delivery ofhealthcare must be revised.Many current practiceswill be supplanted by home healthcare alternatives, the use ofpatient-initiated preventive healthcare initiatives, and remote-clinicactivity. These applications will result in central record archivingsystems; the need for encryption schemes, virus protection, andvandalism protection; extensive, additional connectivity betweenhealthcare providers and professionals; and new commercial playersin the healthcare arena.The hot applicationareas will include women's health, children's health, healthcarefor senior citizens, home healthcare, well-patient monitoring,and early-detection of medical problems.     

Characterization of white matter damage in ischemic leukoaraiosis with diffusion tensor MRI

BACKGROUND AND PURPOSE: Information on the neuropathological changes underlying ischemic leukoaraiosis is only available postmortem, and there are limited data on histological appearances early in the disease. Diffusion tensor imaging allows determination of the directionality of diffusion, which is greater in the direction of white matter bundles. Therefore, the technique might be expected to show loss of anisotropy (directional diffusion) in leukoaraiosis. METHODS: Nine patients with ischemic leukoaraiosis (radiological leukoaraiosis and clinical lacunar stroke) and 10 age-matched controls were studied. Diffusion tensor imaging was performed, and maps of diffusion trace and fractional anisotropy were constructed. Mean values of trace and fractional anisotropy were determined in standard regions of the anterior and posterior white matter in both hemispheres. RESULTS: In all patients with ischemic leukoaraiosis, a characteristic abnormal pattern was found, with loss of anisotropy and increased trace in the white matter. For example, in the right anterior white matter mean (SD) trace/3 was 1.12 (0.33) x10(-3) mm2 s-1 in patients and 0.75 (0.11) in controls (P=0.001). In the same region, fractional anisotropy was 0.53 (0.11) in patients and 0.78 (0.09) in controls (P<0.001). Within the white matter regions, there was a strong negative correlation between mean diffusivity and anisotropy (r=-0.92, P<0.0001). CONCLUSIONS: The characteristic pattern found on diffusion tensor imaging in this patient group is consistent with axonal loss and gliosis leading to impairment to and loss of directional diffusion. The "in vivo histological" information obtained may be useful in monitoring disease progression and in investigating the pathogenesis of the cognitive impairment that may be present.     

Schizophrenia research: a biennium of progress. Proceedings from the Sixth International Congress on Schizophrenia Research. Colorado Springs, CO, April 12-16, 1997

The Sixth International Congress on Schizophrenia Research (ICOSR) took place in Colorado Springs, Colorado, April 12-16, 1997, where over 1,000 scientists presented and listened to the latest developments in the search for the cause and treatment of schizophrenia. The ICOSR is sponsored by Maryland Psychiatric Research Center, Case Western Reserve University, and the William K. Warren Foundation. The National Institute of Mental Health and several pharmaceutical companies contributed generously to the meeting. The ICOSR is co-organized by Dr. Carol A. Tamminga, Maryland Psychiatric Research Center, University of Maryland at Baltimore, and Dr. S. Charles Schulz, Case Western Reserve University, Cleveland, Ohio. The William K. Warren Research Award is given to a senior investigator, who has made outstanding contributions to our understanding of schizophrenia. The fifth William K. Warren Research Award was presented to Dr. Philip S. Holzman in recognition of his contributions to the identification of eye-tracking abnormalities as a potential phenotypic marker of the illness and also in recognition of his work as a lifelong mentor for schizophrenia researchers. The ICOSR Young Investigator Awards are presented to junior investigators who have demonstrated the potential to make significant contributions to research on schizophrenia. These awards promote scientific development by enabling these young researchers to attend the meeting. There were 30 Young Investigator Award winners. The ICOSR meeting is organized into four sessions: (1) a morning plenary session; (2) a plenary lecture; (3) a poster session; and (4) concurrent afternoon oral sessions. The morning plenary sessions are comprised of a set of 30-minute lectures, which provide an overview of a particular topic area relevant to schizophrenia research. The plenary lecture is an invited lecture on a basic topic related to current research efforts in schizophrenia. The poster sessions provide a forum for the presentation of prepublication reports of basic and clinical science projects. The afternoon sessions are a collection of approximately 10 focused presentations on current research projects related to a specific topic area. The purpose of this report is to provide an account of the proceedings from the plenary and afternoon oral sessions.     

Hinged Ilizarov external fixation for correction of antebrachial deformities

OBJECTIVE: To evaluate hinged circular external fixation for correction of antebrachial deformities in dogs. STUDY DESIGN: Uncontrolled clinical trial. ANIMAL POPULATION: Seven client-owned dogs. METHODS: Six dogs had one radius corrected and one dog had both radii corrected. Preoperative planning included measurement of the craniocaudal and mediolateral angular deformities, rotational deformity, length deficit, origin of deformity, graphical or mathematical determination of the amplitude and direction of the actual limb deformity, and frame assembly. RESULTS: Preoperatively, function and cosmesis were assessed to be fair to poor in all dogs. Deformity correction started 48 to 60 hours postoperatively and ranged from 0.46 mm to 1.36 mm twice daily. Hospitalization time ranged from 4 to 6 days. Corrections were mostly made by the owners, at home. Lengthening and angular correction ranged from 3 to 38 mm and 18 degrees to 48 degrees. Mean residual deformities were 2.7% of radial length and 2.7 degrees. The time duration with the circular external fixators in place ranged from 29 to 71 days. Two additional surgeries were necessary in one dog because of wire breakage. Mean follow-up was 40 months. Long-term function and cosmesis were good to excellent in all dogs. CONCLUSION: Although complications were present in six of seven dogs, the outcome of hinged Ilizarov external fixation was successful in all dogs treated for deformities of the antebrachium. CLINICAL RELEVANCE: Despite complex preoperative planning, the placement of hinged circular external fixators is straightforward, and allows precise correction of complex antebrachial deformities with minimal tissue trauma.