An Rtn4/Nogo-A-interacting micropeptide modulates synaptic plasticity with age.

Micropeptides, encoded from small open reading frames of 300 nucleotides or less, are hidden throughout mammalian genomes, though few functional studies of micropeptides in the brain are published. Here, we describe a micropeptide known as the Plasticity-Associated Neural Transcript Short (Pants), located in the 22q11.2 region of the human genome, the microdeletion of which conveys a high risk for schizophrenia. Our data show that Pants is upregulated in early adulthood in the mossy fiber circuit of the hippocampus, where it exerts a powerful negative effect on long-term potentiation (LTP). Further, we find that Pants is secreted from neurons, where it associates with synapses but is rapidly degraded with stimulation. Pants dynamically interacts with Rtn4/Nogo-A, a well-studied regulator of adult plasticity. Pants interaction with Nogo-A augments its influence over postsynaptic AMPA receptor clustering, thus gating plasticity at adult synapses. This work shows that neural micropeptides can act as architectural modules that increase the functional diversity of the known proteome.

Changes in Object Relations over the Course of Psychodynamic Psychotherapy.

This study explores whether object relations (OR) functioning improves over the course of psychodynamic psychotherapy, and whether this improvement is related to symptom decrease as well as therapist technique. The sample consisted of 75 outpatients engaged in short-term psychodynamic psychotherapy at a university-based psychological service clinic. OR functioning was assessed pre- and post-treatment by independent raters using the Social Cognition and Object Relations Scale from in-session patient relational narratives. The Comparative Psychotherapy Process Scale was used to assess therapist activity and psychotherapy techniques early in treatment. Independent clinical ratings of global OR and psychotherapy techniques were conducted, and rater agreement was found to be in the excellent range. Regarding the results, global OR (overall quality and level of interpersonal functioning) significantly improved with large effect size after psychodynamic therapy. Change in global OR functioning was significantly and positively related to the incidence of psychodynamic techniques in early sessions, as were number of psychotherapy sessions attended. Patient self-reported reliable change in symptomatology and reliable change in global OR were significantly related as well. Multilevel model analyses confirmed pairwise correlations accounting for therapist effects on a variety of process-outcome measures, number of sessions attended, initial levels of psychiatric symptoms, employment of therapeutic techniques as well overall OR functioning at outcome. Limitations of the present study, future research directions and implications for clinical practice are also discussed. Copyright © 2016 John Wiley & Sons, Ltd. KEY PRACTITIONER MESSAGE: Psychodynamic psychotherapy seems to be effective in improving object relations functioning. Consider use of psychodynamic techniques early in treatment with patients expressing more pathological object representations. Improvements in object relations functioning during psychodynamic psychotherapy are also related to adaptive changes in patient self-reported symptomatology. Therapist effects were also present for the study. As such therapists should be mindful to assess patient change and their use of technique at several points in treatment and flexibly adjust their approach as necessary.

Neurodevelopmental disorders: mechanisms and boundary definitions from genomes, interactomes and proteomes.

Neurodevelopmental disorders such as intellectual disability, autism spectrum disorder and schizophrenia lack precise boundaries in their clinical definitions, epidemiology, genetics and protein-protein interactomes. This calls into question the appropriateness of current categorical disease concepts. Recently, there has been a rising tide to reformulate neurodevelopmental nosological entities from biology upward. To facilitate this developing trend, we propose that identification of unique proteomic signatures that can be strongly associated with patient's risk alleles and proteome-interactome-guided exploration of patient genomes could define biological mechanisms necessary to reformulate disorder definitions.

ERP correlates of a receptive language-switching task.

Previous research has shown large response time costs (in excess of 50 ms) when bilingual speakers switch predictably back and forth between naming items (a productive switching task) in their first (L1) and second languages (L2). A recent study using event-related potentials (ERPs) has shown that switching between languages is associated with activity over frontal (N2) and parietal (late positive complex) areas of cortex (Jackson, Swainson, Cunnington, & Jackson, 2001). Switching between naming in different languages requires a switch in both language representations and language-specific motor responses. The current study investigated a receptive (input) language-switching task with a common manual response. Number words were presented in L1 and L2, and participants were required to judge whether the words were odd or even (a parity judgement). Response costs were considerably reduced, and the frontal and parietal switch related activity reported in the productive switching task was absent. Receptive switching was associated with early switch-related activity over central sensors that were not language specific. These results are discussed in relation to the idea that there is no language-specific lexical selection mechanism. Instead the costs of receptive language switching may arise from outside the bilingual lexicon.

Organization, expression, and function of Caulobacter crescentus genes needed for assembly and function of the flagellar hook.

This paper reports on the organization, expression, and function of the divergently transcribed flbG and flaN operons in the hook gene cluster of Caulobacter crescentus. The transcription initiation site of flbG was determined previously, and in this work the transcription map was completed by locating the 3' end of the mRNA using nuclease S1 protection assays. A previous genetic study had suggested that the flbG operon is comprised of four genes; however, the nucleotide sequence revealed three tandemly arranged ORFs that correspond to 5'-flbG, flbH, and flgE. FlbG is similar to FliK proteins which are required for termination of hook synthesis, FlbH is similar to FlgD proteins which are essential scaffolding proteins that cap the hook during its assembly, and FlgE corresponds to the hook structural protein. The divergently transcribed flaN gene codes for a hook associated protein I homolog based on its inferred amino acid sequence similarity to FlgK proteins. Based on the amino acid sequence similarities and phenotypes of mutants, flbG, flbH, and flaN have been renamed fliK, flgD, and flgK, FlgD, FlgE, and FlgK proteins, with apparent molecular masses of 23, 68, and 41 kDa, respectively, were expressed from plasmids in a cell-free coupled transcription-translation system, and a protein corresponding to FliK was identified as part of a 190-kDa FliK-LacZ fusion protein. We present evidence showing that, in addition to its role in termination of hook synthesis, FliK is also required for initiation of hook assembly.

Inhaled benzene increases the frequency and length of lacI deletion mutations in lung tissues of mice.

This study investigated the frequency and pattern of mutations that arose in lacI transgenes in lung tissues of mice exposed to 300 p.p.m. of benzene for 6 h/day x 5 days/week for 12 weeks. The nucleotide sequence changes in 86 lacI- transgenes from lung tissues of eight benzene-exposed mice (BEM) and 78 spontaneous lacI- transgenes from lung tissues of eight unexposed control mice (UCM) were identified and compared. A total of 31% (27/86) of the lacI mutations in BEM are deletions compared with 9% (7/78) deletions in UCM. In BEM, 44% (12/27) of the deletions were longer than 10 bp, whereas only 14% (1/7) of the deletions in UCM exceeded 10 bp in length. Statistical tests supported the hypothesis that benzene exposure resulted in significant increases in both the frequency and length of deletions. Based on the lacI mutant frequency and fraction of unique mutations, lung tissues of BEM were estimated to have a 1.8-fold increase in lacI mutation frequency compared with lung tissues of UCM. The results presented in this paper demonstrate that inhaled benzene is a gene mutagen in lung tissues of mice.

Genetic organization and transposition properties of IS511.

IS511 is an endogenous insertion sequence (IS) of the bacterium Caulobacter crescentus strain CB15 and it is the first Caulobacter IS to be characterized at the molecular level. We determined the 1266-bp nucleotide sequence of IS511 and investigated its genetic organization, relationship to other ISs, and transposition properties. IS511 belongs to a distinct branch of the IS3 family that includes ISR1, IS476, and IS1222, based on nucleotide sequence similarity. The nucleotide sequence of IS511 encodes open reading frames (orfs) designated here as orfA and orfB, and their relative organization and amino acid sequences of the predicted protein products are very similar to those of orfAs and orfBs of other IS3 family members. Nuclease S1 protection assays identified an IS511 RNA, and its 5' end maps approximately 16 nucleotides upstream of orfA and about six nucleotides downstream of a sequence that is similar to the consensus sequence of C. crescentus housekeeping promoters. Evidence is presented that IS511 is capable of precise excision from the chromosome, and transposition from the chromosome to a plasmid. Transpositional insertions of IS511 occurred within sequences with a relatively high G + C content, and they were usually, but not always, flanked by a 4-bp direct repeat that matches a sequence at the site of insertion. We also determined the nucleotide sequence flanking the four endogenous IS511 elements that reside in the chromosome of C. crescentus. Our findings demonstrate that IS511 is a transposable IS that belongs to a branch of the IS3 family.

Inhalation of benzene leads to an increase in the mutant frequencies of a lacI transgene in lung and spleen tissues of mice.

The goal of this study was to determine if inhalation of benzene leads to an increase in the mutant frequencies in the tissues of male C57BL/6 mice. Mutant frequencies were measured using a previously described assay in which bacteriophage lambda lacI transgenes are rescued from mouse genomic DNA as infectious phage and scored for their LacI phenotype. Eight experimental mice were exposed to a target concentration of 300 ppm of benzene for 6 h/day x 5 days/week x 12 weeks, and eight control mice were treated similarly except that they were not exposed to benzene. Mutant frequencies were calculated as the ratio of LacI-/total phage recovered from organs of interest. The mean mutant frequency measured in lung tissues of mice exposed to benzene was (10.6 +/- 1.4) x 10(-5), which is about 1.7-fold higher than that of the unexposed controls. In spleen tissues from benzene-exposed mice the mean mutation frequency was (12.6 +/- 4.1) x 10(-5), which is about 1.5-fold higher than that of spleen tissues from unexposed controls. The differences in mean mutant frequencies between benzene-exposed and unexposed lung and spleen tissues are statistically significant. In liver tissues, however, the mean mutant frequencies of benzene-exposed mice and unexposed mice are not significantly different. These results demonstrate that inhaled benzene results in a statistically significant increase in the mutant frequencies in lung and spleen, but not in liver tissues of mice.

FlbD has a DNA-binding activity near its carboxy terminus that recognizes ftr sequences involved in positive and negative regulation of flagellar gene transcription in Caulobacter crescentus.

FlbD is a transcriptional regulatory protein that negatively autoregulates fliF, and it is required for expression of other Caulobacter crescentus flagellar genes, including flaN and flbG. In this report we have investigated the interaction between carboxy-terminal fragments of FlbD protein and enhancer-like ftr sequences in the promoter regions of fliF, flaN, and flbG. FlbDc87 is a glutathione S-transferase (GST)-FlbD fusion protein that carries the carboxy-terminal 87 amino acids of FlbD, and FlbDc87 binds to restriction fragments containing the promoter regions of fliF, flaN, and flbG, whereas a GST-FlbD fusion protein carrying the last 48 amino acids of FlbD failed to bind to these promoter regions. DNA footprint analysis demonstrated that FlbDc87 is a sequence-specific DNA-binding protein that makes close contact with 11 nucleotides in ftr4, and 6 of these nucleotides were shown previously to function in negative regulation of fliF transcription in vivo (S. M. Van Way, A. Newton, A. H. Mullin, and D. A. Mullin, J. Bacteriol. 175:367-376, 1993). Three DNA fragments, each carrying an ftr4 mutation that resulted in elevated fliF transcript levels in vivo, were defective in binding to FlbDc87 in vitro. We also found that a missense mutation in the recognition helix of the putative helix-turn-helix DNA-binding motif of FlbDc87 resulted in defective binding to ftr4 in vitro. These data suggest that the binding of FlbDc87 to ftr4 is relevant to negative transcriptional regulation of fliF and that FlbD functions directly as a repressor. Footprint analysis showed that FlbDc87 also makes close contacts with specific nucleotides in ftr1, ftr2, and ftr3 in the flaN-flbG promoter region, and some of these nucleotides were shown previously to be required for regulated transcription of flaN and flbG (D. A. Mullin and A. Newton, J. Bacteriol. 175:2067-2076, 1993). Footprint analysis also revealed a new ftr-like sequence, ftr5, at -136 from the transcription start site of flbG. Our results demonstrate that FlbD contains a sequence-specific DNA-binding activity within the 87 amino acids at its carboxy terminus, and the results suggest that FlbD exerts its effect as a positive and negative regulator of C. crescentus flagellar genes by binding to ftr sequences.

Identification of the promoter and a negative regulatory element, ftr4, that is needed for cell cycle timing of fliF operon expression in Caulobacter crescentus.

The fliF operon of Caulobacter crescentus, which was previously designated the flaO locus, is near the top of the flagellar-gene regulatory hierarchy, and it is one of the earliest transcription units to be expressed in the cell cycle. In this report, we have identified two cis-acting sequences that are required for cell cycle regulation of fliF transcription. The first sequence was defined by the effects of three 2-bp deletions and five point mutations, each of which greatly reduced the level of fliF operon transcript in vivo. These eight mutations lie between -37 and -22 within an 18-bp sequence that matches, at 11 nucleotides, sequences in the 5' regions of the flaQR (flaS locus) and fliLM operons, which are also expressed early and occupy a high level in the regulatory hierarchy (A. Dingwall, A. Zhuang, K. Quon, and L. Shapiro, J. Bacteriol. 174:1760-1768, 1992). We propose that this 18-bp sequence contains all or part of the fliF promoter. We have also identified a second sequence, 17 bp long and centered at -8, which we have provisionally designated ftr4 because of its similarity to the enhancer-like ftr sequences required for regulation of sigma 54 promoters flaN and flbG (D. A. Mullin and A. Newton, J. Bacteriol. 171:3218-3227, 1989). Six of the seven mutations in ftr4 examined resulted in a large increase in fliF operon transcript levels, suggesting a role for ftr4 in negative regulation. A 2-bp deletion at -12 and -13 in ftr4 altered the cell cycle pattern of fliF operon transcription; the transcript was still expressed periodically, but the period of its synthesis was extended significantly. We suggest that the ftr4 sequence may form part of a developmental switch which is required to turn off fliF operon transcription at the correct time in the cell cycle.

Cis- and trans-acting elements required for regulation of flagellar gene transcription in the bacterium Caulobacter crescentus.

The flagellar genes of the bacterium Caulobacter crescentus are organized into a regulatory hierarchy that consists of four classes or levels of genes, and expression of these genes is restricted to a discrete interval in the cell cycle that begins just prior to flagellum assembly. This paper summarizes data on the promoters and other cis-acting elements that are required for transcription of the level II gene fliF and the level III genes flaN and flbG. Regulated expression of flaN and flbG requires sigma 54 promoters, enhancer-like sequences called ftr, and sequences called ihf that conform to the consensus binding sequence for Escherichia coli integration host factor protein. The fliF regulatory region contains a new type of promoter sequence that is different from other known promoter motifs, and it has a sequence called ftr4 that is a site of negative regulation. ftr4 appears to function as part of a developmental switch that turns fliF transcription off at the correct time in the cell cycle. flbD, the last gene in the fliF operon is a negative regulator of fliF and an activator of both flaN and flbG expression. Evidence that FlbD protein plays a direct role as a transcriptional regulator comes from the finding that it has a DNA binding activity within its carboxy terminus that specifically recognizes ftr4 in fliF and four ftr elements in the flaN-flbG promoter region.

The importance of ABH antigens in platelet crossmatching.

In the present study, the frequency with which ABH incompatibility could be detected in platelet crossmatches was determined. The effect of chloroquine elution on ABH antigens was also evaluated, and a technique was developed to remove IgG anti-A from group O plasma using a chemically synthesized human blood group A trisaccharide antigen covalently linked to crystalline silica (Synsorb-A). Group O plasmas were found to be incompatible with 52 percent of group A platelets and 17 percent of group B platelets (p less than 0.05). In contrast, anti-A from group B plasmas rarely produced a positive crossmatch, and no anti-B that reacted with platelets could be demonstrated in group A plasmas. IgG anti-A reactions with group A platelets were eliminated in 100 percent of the group O plasmas tested after treatment with the synthetic solid-phase immunoadsorption technique. Synsorb-A may be a useful adjunct to platelet serologic testing when group O sera need to be tested against A platelets. Group A platelets bound less anti-A after exposure to chloroquine, but only 17 percent of platelets became negative when crossmatched with group O plasma. It was concluded that increased IgG binding occurs in a majority of platelet crossmatches using a k-ELISA technique when group O recipients are tested against group A donors. These results offer a potential explanation for conflicting results in studies of transfusion results with ABH-incompatible platelets. Transfusions of group B platelets to incompatible recipients may be more likely to yield satisfactory increments than incompatible transfusions of group A platelets, but this remains to be proven. There appear to be significant differences between red cells and platelets in regard to serologic reactivity in the ABH system.

Cardiovascular effects of desipramine in children.

The effect of desipramine hydrochloride was studied in children who were treated for eating disorders (5), attention deficit disorder (13), or affective disturbance (3). Serial heart rate, blood pressure, ECG, and 24-hour ambulatory monitoring were recorded before treatment and at 4 and 8 weeks during treatment. Maximum dose of desipramine was 5 mg/kg/day, average 4.25. A 21% increase in heart rate and 2.5% increase in QTc at 4 weeks were sustained at 8 weeks. No dysrhythmias or clinically significant changes in blood pressure occurred. Desipramine is safe in children who have normal cardiovascular examinations and ECGs when used within the limits of the study design. The cardiovascular effects of desipramine should be kept in mind and monitored when patients are starting tricyclic antidepressant therapy such as desipramine.

Effect of preoperative hemodynamic support on survival after cardiac transplantation.

The accessibility and success of cardiac transplantation promotes transplantation for a broad range of recipients, including those requiring intravenous inotropes or mechanical-assist devices. To determine if survival is dependent on preoperative requirements for hemodynamic support, we studied 230 patients who underwent transplant at the Loyola, Stanford, and UTAH programs from December 1, 1984 through November 30, 1986, and who were followed up for 34 months postoperatively. Group 1 (n = 132 of 230, 57%) patients required only oral medical therapy to maintain hemodynamic compensation; Group 2 (n = 69 of 230, 30%) patients were dependent on intravenous inotropic support; and Group 3 (n = 29 of 230, 13%) patients required mechanical assistance. Pretransplant characteristics showed that dilated cardiomyopathy was more common in Group 2 patients, and lower cardiac index and ejection fraction were more prevalent in Group 3 patients as expected. Although survival was lower in Group 3 only at 1 month (Group 1, 98.5%; Group 2, 92.8%; and Group 3, 86.2%; p less than 0.01), the survival advantage in Groups 1 and 2 was lost by 3 months, with 1-year survival rates of 88.6% in Group 1, 81.2% in Group 2, and 82.8% in Group 3. Allograft survival and cause of death were not different among the three groups. Acute rejection occurred at a lower monthly frequency in the first 4 months in Group 3 (Group 1, 0.47 +/- 0.03; Group 2, 0.47 +/- 0.05; and Group 3, 0.29 +/- 0.06; p less than 0.01), whereas infectious complications occurred at similar frequencies.(ABSTRACT TRUNCATED AT 250 WORDS)

Cardiac sarcoidosis: response to steroids and transplantation.

From 1976 to 1986, six cases of cardiac sarcoidosis have been documented by myocardial biopsy in three of five instances; on examination of the explanted heart after transplantation in two, and at autopsy in one patient. Right ventricular end-diastolic pressure was elevated in all four patients with right ventricular involvement with sarcoidosis. Of three patients treated with steroids, improvement in ventricular function and decrease in arrhythmia occurred in two, whereas failure to respond led to transplantation in the other patient. Two further patients have undergone heart transplantation, one for resistant ventricular arrhythmia and the other for congestive heart failure. No recurrence of sarcoidosis has occurred in the grafts. Because two of five patients had sarcoidosis diagnosed on gross examination, a negative endomyocardial biopsy does not exclude the diagnosis of myocardial sarcoidosis, which should therefore be pursued in the setting of unexplained heart failure, conduction abnormalities, and ventricular arrhythmia, particularly when right ventricular end-diastolic pressure is raised. Steroids may result in improvement in some patients even in the presence of severe morphological damage. Heart transplantation may be performed without increased risk of recurrence of sarcoidosis.