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The metabolism of gluconate in Escherichia coli. The subsidiary system and the nature of the gntS gene.

The transport and phosphorylation of gluconate in E. coli occurs through two systems (GntI and GntII) which duplicate activities. bioH-asd deletion mutants do not grow on media with gluconate as sole carbon source because they lack the GntI system and do not express GntII. Although E. coli c177 is a delta (bioH-asd) mutant, it carries the pyrB linked mutation gnt177 that enables it to metabolize this substrate through inducible expression of the GntII system. Several gntS derivatives which are unable to grow on gluconate were isolated from E. coli C177 by spontaneous curing of the transposon Tn10 previously inserted at the gntS locus (zjf::Tn10, min 95.3). A representative gntS mutant, E. coli TI141A retained the ability to take up gluconate but had lost the thermosensitive gluconokinase activity (gene gntV, min 96.9). Furthermore, it could be demonstrated that gntV is repressed in E. coli TI141A. The results indicate that gntS might specify a trans-acting positive regulator involved in the control of at least the expression of the thermosensitive gluconokinase (GntII), instead of a gluconate uptake system as it was previously postulated. Likewise, these results can be used to reconsider whether the locus altered by the gnt177 lesion is allelic with that of the GntII permease instead of a regulator, as it was originally postulated.

Alleles↗

Relative bioavailability estimation of carbamazepine crystal forms using an artificial stomach-duodenum model.

The in vitro dissolution of carbamazepine (CBZ) was investigated using an automated artificial stomach-duodenum (ASD) model. Successful simulation of the dog physiology in the fasted state showed that the rank order of the ASD estimated bioavailabilities is as follows: Form III > Form I > dihydrate. This result is in excellent agreement with those found in literature. Additional simulations comparing different gastric transit times during fasted and fed states are also discussed.

Animals↗

Traumatic brain injury, dissociation, and posttraumatic stress disorder in road traffic accident survivors.

This study investigated the symptom profiles of acute stress disorder (ASD) and posttraumatic stress disorder (PTSD) in participants who did and did not sustain traumatic brain injury (TBI), following a road traffic accident. The participants were assessed at three time points: as soon as possible posttrauma as well as at 6 weeks and 3 months posttrauma. At the first assessment, fewer participants from the TBI group recalled feeling fear and helplessness at the time of the trauma, fewer TBI participants reported recurrent intrusive thoughts and images, and more TBI participants reported dissociation since the trauma, relative to the non-TBI group. At the second assessment, fewer participants from the TBI group recalled feeling intense helplessness at the time of the trauma. Fewer TBI participants also reported reliving and physiological reactions on trauma reminders relative to the non-TBI group. At 3 months posttrauma, there was no difference in PTSD symptom profile between non-TBI and TBI groups. Our findings indicate that the presence of TBI is likely to influence the distribution of certain symptoms, but need not be a significant barrier to diagnosing ASD and PTSD.

Accidents, Traffic↗

The Acute Stress Checklist for Children (ASC-Kids): development of a child self-report measure.

This project aimed to develop and validate a brief, practical self-report measure of acute stress disorder (ASD) reactions (including ASD diagnostic criteria) in children and adolescents. A draft measure was reviewed for content validity by an expert panel, piloted with youth, and revised for clarity and coverage of key constructs. The new 29-item Acute Stress Checklist for Children (ASC-Kids) was administered to 176 youth age 8 to 17 who had a recent injury or intensive care unit admission. The ASC-Kids demonstrated strong test-retest reliability and internal consistency, as well as concurrent and predictive validity with other traumatic stress measures. Results of exploratory factor analyses were consistent with current conceptualizations of acute traumatic stress reactions. The ASC-Kids is a promising measure of child acute stress reactions.

Adolescent↗

Simple model of double adiabatic inversion (DAI) efficiency.

Multislice continuous arterial spin labeling (CASL) using alternating single adiabatic inversion (SAI) and double adiabatic inversion (DAI) (ASD) suffers from control inefficiencies that reduce the perfusion signal-to-noise ratio (SNR). A simple analytic model for DAI efficiency (chi) is presented, and compared with simulation and experimental data. The DAI efficiency (defined as the ratio of uninverted to equilibrium spins) depends primarily on the effective adiabaticity (beta') and the inversion time (both determined from the labeling RF field and gradient amplitudes, and flow velocities). In this study, chi behaved like a damped oscillator, starting from 1 at beta'=0 (no spin inversion), dropping to 0.5 at beta'=0.5 (spin saturation), rising again to a peak at beta' approximately 3, and subsequently oscillating at higher beta'. The simple model matched the flow phantom data for beta' < 2, with an accuracy of better than 10%, but it diverged from the flow phantom data as the data oscillated at higher beta'. The relative inversion efficiencies of ASD and simultaneous proximal and distal irradiation (SPDI) were measured in 28 elderly volunteers. The ratio of CASL difference (control-label) signals measured in gray matter (= 1.16 +/- 0.05), and the simple efficiency models (= 1.13 +/- 0.02) had a correlation coefficient of 0.974.

Aged↗

Prevalence of acute and post-traumatic stress disorder and comorbid mental disorders in breast cancer patients during primary cancer care: a prospective study.

This study aimed at the identification of acute and post-traumatic stress responses, and comorbid mental disorders in breast cancer patients. Structured clinical interviews for DSM-IV (SCID) were conducted post-surgery with 127 patients (t1). Screening measures were used to assess post-traumatic stress responses, anxiety, and depression at t1 and at 6 months follow-up (t2). Based on the SCID, prevalence rates were 2.4% for both, cancer-related ASD and PTSD. Experiences most frequently described as traumatic were the cancer diagnosis itself and subsequent feelings of uncertainty. Patients with lifetime PTSD (8.7%) were more likely to meet the criteria for cancer-related ASD or PTSD (OR=14.1). Prevalence estimates were 7.1% for Adjustment Disorder, 4.7% for Major Depression, 3.1% for Dysthymic Disorder and 6.3% for Generalized Anxiety Disorder. Using the screening instruments, IES-R, PCL-C and HADS, we found PTSD in 18.5% at t1 and 11.2-16.3% at t2. The estimates of anxiety and depression reveal rates of 39.6% (t1) and 32.7% (t2) for anxiety, as well as 16.0% (t1) and 13.3% (t2) for depression (t1) (cut-off> or =8). The diagnosis of a life-threatening illness has been included as a potential trauma in the DSM-IV. However, it has to be critically evaluated whether subjective feelings of uncertainty like fears of treatment count among traumatic stressors, and thus, whether the diagnosis of PTSD is appropriate in this group of cancer patients. However, a large number of women with emotional distress illustrate the need for psychosocial counseling and support in this early treatment phase.

Acute Disease↗

Paternal age and the risk of congenital heart defects.

The effect of paternal age on the risk of birth defects among offspring is less well studied than the effect of maternal age, with few comprehensive epidemiologic studies having been conducted. Advanced paternal age has been shown to be associated with an increase in new dominant mutations that result in particular congenital anomalies. The relationship between paternal age and more common birth defects, for example, cardiac defects, has not been as extensively evaluated. Therefore, a total of 4,110 cases of congenital heart defects was identified from the British Columbia Health Surveillance Registry. Matched controls were obtained from the birth files of British Columbia for the years 1952-1973. Prevalence odds ratios for paternal age, adjusted for maternal age and other factors, were estimated for 8 cardiac defect groups. A suggestive general pattern of increasing risk with increasing age among cases (excluding chromosomal anomalies) relative to controls was found for ventricular septal defects (VSD), atrial septal defects (ASD), and patent ductus arteriosus (PDA). In addition, an increased risk among men younger than 20 yr was found for VSD and ASD. These findings are consistent with the results of some previous epidemiologic studies. Based on the results of this study it is estimated that for cardiac defects such as VSD, approximately 5% of cases may be due to advanced paternal age (> 35 yr), possibly through dominant mutations.

Adult↗

Analysis of blood lymphocyte subsets in children living on territory that received high amounts of fallout from Chernobyl accident.

The major lymphocyte subsets in the peripheral blood were assessed in 120 children 6-13 years old living on areas that received high levels of radioactivity as fallout after the Chernobyl nuclear power plant accident. Seventy-one of the children were suffering from recurrent respiratory disease (RRDC) and 49 were not (non-RRDC). As controls, a total of 87 RRDC and non-RRDC living on noncontaminated areas were evaluated. We did not find significant differences in major lymphocyte subsets between the values in non-RRDC living on radionuclide-contaminated areas and noncontaminated areas. However, RRDC living on radionuclide-contaminated areas had a significantly lower percentage of CD3+ T and CD3+CD4+ T-helper/ inducer cells compared to control RRDC. Furthermore, the decrease in percentage of CD3+CD4+ cells was more profound in RRDC living in radiation-contaminated settlements with an average summary dose (ASD) Cs-137(134) and Sr-90 for the population > 1.0 mSv than in RRDC living in contaminated settlements with an ASD Cs-137(134) and Sr-90 < 1.0 mSv. These data indicated that long-time exposure to small doses of radiation could affect the immune system in children living around Chernobyl.

CD3 Complex↗

Catalytic antibody model and mutagenesis implicate arginine in transition-state stabilization.

To probe the mechanism of the catalytic antibody NPN43C9, we have constructed a three-dimensional model of the NPN43C9 variable region using our antibody structural database (ASD), which takes maximal advantage of immunoglobulin sequence and structural information. The ASD contains separately superimposed variable light and variable heavy chains, which reveal not only conserved backbone structure, but also structurally conserved side-chain conformations. The NPN43C9 model revealed that the guanidinium group of light chain Arg L96 was positioned at the bottom of the antigen-binding site and formed a salt bridge with the antigen's phosphonamidate group, which mimics the negatively charged, tetrahedral transition states in the hydrolysis reaction. Thus, the model predicts both binding and catalytic functions for Arg L96, which previously had not been implicated in either. First, Arg L96 should enhance antigen binding by electrostatically complementing the negative charge of the antigen, which is buried upon complex formation. Second, Arg L96 should promote catalysis by electrostatically stabilizing the negatively charged transition states formed during catalysis. These hypotheses were tested experimentally by design and characterization of the R-L96-Q mutant, in which Arg L96 was replaced with Gln by site-directed mutagenesis. As predicted, antigen binding in the R-L96-Q mutant was decreased relative to that in the parent NPN43C9 antibody, but binding of antigen fragments lacking the phosphonamidate group was retained. In addition, the R-L96-Q mutant had no detectable esterase activity. Thus, the computational model and experimental results together suggest a mechanism by which the catalytic antibody NPN43C9 stabilizes high-energy transition states during catalysis.

Amino Acid Sequence↗

Expression and purification of aspartate beta-semialdehyde dehydrogenase from infectious microorganisms.

l-Aspartate-beta-semialdehyde dehydrogenase (ASA DH) lies at the first branch point in the aspartate metabolic pathway that leads to the formation of the amino acids lysine, isoleucine, methionine, and threonine in most plants, bacteria, and fungi. Since the aspartate pathway is not found in humans, but is necessary for bacterial cell wall biosynthesis, the enzymes in this pathway are potential targets for the development of new antibiotics. The asd gene that encodes for ASA DH has been obtained from several infectious organisms and ligated into a pET expression vector. ASA DHs from Haemophilus influenza, Pseudomonas aeruginosa, and Vibrio cholerae were expressed as soluble proteins in Escherichia coli, while ASA DH from Helicobacter pylori was obtained primarily as inclusion bodies. The V. cholerae genome contains two asd genes. Both enzymes have been expressed and purified, and each displays significant ASA DH activity. The purification of highly active ASA DH from each of these organisms has been achieved for the first time, in greater than 95% purity and high overall yield. Kinetic parameters have been determined for each purified enzyme, and the values have been compared to those of E. coli ASA DH.

Animals↗

Papillary fibroelastoma of the tricuspid valve in association with an atrial septal defect: report of a case.

Although papillary fibroelastoma is rare, it is the most common primary tumor of the heart valves. We describe herein the case of a 64-year-old woman scheduled to undergo atrial septal defect (ASD) repair, in whom a papillary fibroelastoma of the tricuspid valve was diagnosed by transesophageal echocardiography (TEE). Surgical resection of the papillary fibroelastoma at the time of ASD repair prevented the fatal embolization sometimes associated with this lesion. Thus, intraoperative TEE played an important role in identifying the location of the tumor and its anatomic attachment, and in assessing the adequacy of surgical treatment.

Echocardiography, Transesophageal↗

Children with congenital heart disease: probability of natural survival.

The age distribution of death in all children with congenital heart disease (CHD), who died in a 27-year period in Central Bohemia (population of 1.2 million), and the data on the incidence of CHD in children born in Bohemia (population of 6.3 million) in 1980 were used to calculate the probability of survival of a child born with CHD. Eighty-six percent of these children survived to the first month of life--mostly those with pulmonary stenosis (PS, 99%), aortic stenosis (AS, 95%), ventricular septal defect (VSD, 92%), and atrioventricular septal defect (AVSD, 91%). Seventy-one percent of patients survived the first year of life--mostly those with PS (97%), AS (91%), atrial septal defect (ASD, 89%), VSD (80%), and persistent ductus arteriosus (PDA, 78%). In total, 67% of CHD patients can be expected to survive childhood. The highest survival rates were found in PS (94%), AS and ASD (84%), VSD and PDA (70-80%), and coarctation of the aorta (COA, 68%). The survival rate for the remaining forms of CHD was less than 50%. The highest mortality rate (10% of all children born with CHD) can be expected in the first postnatal week. The lowest survival in the first week was found among those with hypoplastic left heart (HLHS, 39%), double-outlet right ventricle (DORV, 50%), truncus arteriosus (TrA, 57%), pulmonary atresia (PA, 70%), and transposition of the great arteries (TGA, 83%). In addition, total anomalous pulmonary venous connection (TAPVC) and single ventricle had the highest risk of death in the first year of life.

Adolescent↗

Scintigraphic determination of severity in pulmonary parenchymal damage in patients with atrial septal defect.

According to the pattern observed on a lung perfusion scan, patients with atrial septal defect (ASD) were classified into three stages: stage I, normal perfusion (n = 15); stage II, apical prone perfusion (n = 22); and stage III, mottled perfusion (n = 23). These stages were compared with hemodynamic data obtained from cardiac catheterization and with the right ventricular ejection fraction (RVEF). Staging from I to III correlated well with pulmonary arterial pressure, and patients in stage III showed a significant increase in pulmonary arteriolar resistance and a significant decrease in Qp/Qs. There was no significant difference in RVEF between the stages. We concluded that the scintigraphic severity determined by lung perfusion scan could be valid for evaluating the extent of pulmonary parenchymal damage in patients with ASD.

Cardiac Catheterization↗

Immunohistochemical differential diagnosis of granulocytic sarcomas and malignant lymphomas on formalin-fixed material.

A panel of monoclonal antibodies (anti-CD45 [common leukocyte antigen], Ki-B3, L26, MT1, UCHL1, anti-CD15 [X-hapten], anti-neutrophil granule protein elastase [NP57]), anti-lysozyme, and the naphthol-ASD-chloroacetate reaction were applied to two cases of granulocytic sarcoma (GS) for evaluation of their utility in differentiating GS from malignant lymphoma. Lysozyme and naphthol-ASD-chloroacetate esterase were found to be the most reliable markers for detection of the myeloid nature of the tumour cells. GS infiltrated solely the mucosa of the nasal cavity in one case, while in the other it involved both the nasal cavity and maxillary sinus with simultaneous eruptions on the skin of the trunk. In both cases, peripheral blood and bone marrow findings were inconspicuous at the time of diagnosis of GS.

Child↗

Slit-ventricle syndrome in shunt operated children.

Chronic intracranial hypotension is considered as a frequent complication in shunted hydrocephalus, besides obstruction and shunt-infections. In the last twenty years 32 cases of slit-ventricle were diagnosed among the more than one thousand operations on hydrocephalic children at the Paediatric Department of the National Institute of Neurosurgery, Budapest, Hungary. Most of them have been operated on in infancy. Time from the first operation to the development of slit-ventricle ranged from one to twelve years, the mean was 6.5 years. Seven patients were symptomless (22%), while 25 patients (78%) had more or less severe slit-ventricle syndrome with headache (25 cases), nausea/vomiting (23 cases), altered consciousness (21 cases), brainstem signs (12 cases), and epileptic fits (2 cases). Ten patients with moderate clinical signs improved under conservative treatment. In 15 cases an anti-siphon device (ASD) was implanted. In five of them the clinical result was good, but in the remaining 10 cases typical hypertensive signs were seen. In these cases low flow rate valves were implanted instead of the middle flow rate valve and ASD. In one case the intracranial hypertension persisted, so a middle flow rate shunt system was "reimplanted" and finally the patient improved. In this study the experiences with these 32 cases will be analysed and discussed. The authors stress the primary use of combined valves to avoid the slit-ventricle syndrome.

Cerebrospinal Fluid Shunts↗

Interventional pediatric cardiology: device closures.

Surgical treatment of various septal defects has been long established. With the advances in transcatheter therapy dilatation techniques for valvular stenosis and vascular obstruction have become established procedures. Closure of septal defects in the catheterization laboratory has also been introduced; some of these have come into regular use in current practice. In 1967, Porstmann et al reported the use of Ivalon plug to close patent ductus arteriosus (PDA). Since then, several devices have been used including Rashkind PDA ocluder (not being used now), Gianturco coils, detachable coils (for small PDA), CardioSEAL and other umbrella devices and Amplatzer PDA occluder. Closure rates vary from 95-98% in most series, however, some of these devices are very expensive, more so, when compared to the cost of surgical ligation of PDA. Catheter closure of secundum atrial septal defect (ASD) has also been done by various devices like clamshell device, Sideris Buttoned device, ASDOS device, Amplatzer device and cardioSEAL. So far no device has been accepted as ideal for every case, however, Amplatzer device has been used most extensively. Issues such as completeness of endothelialisation, incidence of late arrhythmias, endocarditis remain uncertain. However, in select population of ASD cases with a central secundum defect, device closure is being used increasingly. Device closure of ventricular septal defect remains challenging and controversial and is probably available to a small group of children with defects that are difficult to close surgically and involve higher risk.

Cardiac Catheterization↗

[Surgical treatment for mitral regurgitation associated with secundum atrial septal defect].

We reviewed 25 patients who underwent a mitral valvuloplasty, from 1984 to 1996, for mitral regurgitation (MR) associated with atrial septal defect (ASD). Mean grade of MR was 2.3 +/- 0.7. The locations of mitral valve lesion were as follows; Postero-medial side of the anterior leaflet (AML) (11 patients: 44%), posteromedial side to center of the AML (7 patients: 28%), whole of the AML (5 patients: 20%), center of the AML (1 patient: 4%), posteromedial side of the posterior leaflet (PML) (1 patient: 4%). In summary, the mitral valve lesion was located in the AML in 96% patients and were seen in the postero-medial side of 96% patients. Mitral valve repair was performed as follows; chordae shortening only (3 patients: 12%), chordae shortening + Kay's annuloplasty (9 patients: 36%), Kay's anuloplasty (10 patients: 40%), using artificial chordae only (1 patient: 4%), using artificial chordae + Kay's annuloplasty (1 patient: 4%), using artificial chordae + ring annuloplasty (1 patient: 4%). In 24 patients, the grade of MR was less than 2/4 in the early postoperative period. In one patient, the grade of 3/4 MR was still remained. Reoperation were required in 2 patients, because of gradual increase of MR, 9 years and 10 years after the initial operation, respectively. In another patient, the grade 3/4 MR recurrently occurred at 6 months after the operation. He has been well maintained medically. In all 4 patients who had more than the grade 3/4 MR postoperatively, the annuloplasty was performed with Kay's method and the cause of MR was poor coaptation around the center of the AML. The mitral valve lesion associated with ASD seemed to be the dislocation of the AML which cause the discrepancy of the coaptation zone between both leaflets, without any prominent prolapse and chordae elongation. We put a particular emphasis on that the mitral valve repair should be performed with the recognition of the etiology of the mitral valve lesion. Especially, if the lesion extends around the center of the AML, sufficient coaptation area of both leaflets at the center of the AML should be obtained by anuloplasty.

Adult↗