Investigation of the Clinical Utility of the BRIEF2 in Youth With and Without Intellectual Disability.

OBJECTIVE: Executive function (EF) difficulties are commonly found in youth with intellectual disability (ID). Given mixed results from studies using performance-based EF measures, the EF profile has not been well characterized for this population. No published work has examined the clinical utility of the Behavior Rating Inventory of Executive Function, Second Edition (BRIEF2) in distinguishing EF in ID. We hypothesized that the BRIEF2 would show greater elevations in youth with ID compared to the Average IQ comparison group. METHODS: Participants included a large sample of 504 youth (157 in ID group; aged 8-18 years) referred for (neuro)psychological evaluation (2015-2019) and identified as meeting criteria for either ID or Average IQ comparison group. RESULTS: Significant elevations were found across BRIEF2 indices and scales. Only mild elevations were noted in selective cognitive regulation scales within the Average IQ group. Groups differed significantly across all EF dimensions, with greater differences observed in behavioral regulation (Self-Monitoring, Inhibition), Shift, and Working Memory. An elevated but less variable pattern of index scores was noted in ID, while the overall pattern of scaled scores appeared similar between groups. CONCLUSIONS: The less variable and consistently elevated profile may suggest fewer EF dimensions in individuals with ID than the model proposed in the test manual. Similar profiles between groups may reflect differences in severity, rather than differences in constructs measured by the EF factors, per se. Additional examination is needed to confirm potential structural differences in EF for youth with ID as measured by BRIEF2, with a clinical implication for greater efficiency of EF assessment in this population.

Respiratory mechanics and peripheral airway inflammation and dysfunction in asthma.

BACKGROUND: Clinical application of the forced oscillation technique (FOT) has progressed with the spread of commercially available FOT devices. The correlation between respiratory impedance and spirometry has been reported; however, the association with airway inflammation and pulmonary function, in the lung periphery in particular, is unclear. OBJECTIVE: To assess whether respiratory impedance is associated with peripheral airway inflammation and dysfunction in asthma. METHODS: Subjects included 78 patients with overall controlled asthma. We measured whole-breath or within-breath respiratory system resistance (Rrs) and reactance (Xrs) using a commercially available multi-frequency FOT device (MostGraph-01), and assessed the correlation with the fraction of exhaled nitric oxide (FeNO), alveolar nitric oxide concentration (CANO), maximal NO flux in the conductive airways (J'awNO), and the N2 phase III slope of single breath N2 washout (delta N2 ). RESULTS: The differences between inspiratory and expiratory phases of Xrs at 5 Hz (X5), resonant frequency (Fres), and a low-frequency reactance area (ALX) were significantly correlated with CANO; however, there was no correlation between respiratory impedance and FeNO or J'awNO. The delta N2 values were significantly correlated with whole-breath, inspiratory, and expiratory Rrs and Xrs, except for R20. CONCLUSIONS AND CLINICAL RELEVANCE: We conclude that respiratory impedance reflects peripheral airway inflammation and ventilation inhomogeneity.

D-Amino acid oxidase-induced oxidative stress, 3-bromopyruvate and citrate inhibit angiogenesis, exhibiting potent anticancer effects.

Angiogenesis is critical for cancer growth and metastasis. Steps of angiogenesis are energy consuming, while vascular endothelial cells are highly glycolytic. Glioblastoma multiforme (GBM) is a highly vascular tumor and this enhances its aggressiveness. D-amino acid oxidase (DAO) is a promising therapeutic protein that induces oxidative stress upon acting on its substrates. Oxidative stress-energy depletion (OSED) therapy was recently reported (El Sayed et al., Cancer Gene Ther, 19, 1-18, 2012). OSED combines DAO-induced oxidative stress with energy depletion caused by glycolytic inhibitors such as 3-bromopyruvate (3BP), a hexokinase II inhibitor that depleted ATP in cancer cells and induced production of hydrogen peroxide. 3BP disturbs the Warburg effect and antagonizes effects of lactate and pyruvate (El Sayed et al., J Bioenerg Biomembr, 44, 61-79, 2012). Citrate is a natural organic acid capable of inhibiting glycolysis by targeting phosphofructokinase. Here, we report that DAO, 3BP and citrate significantly inhibited angiogenesis, decreased the number of vascular branching points and shortened the length of vascular tubules. OSED delayed the growth of C6/DAO glioma cells. 3BP combined with citrate delayed the growth of C6 glioma cells and decreased significantly the number and size of C6 glioma colonies in soft agar. Human GBM cells (U373MG) were resistant to chemotherapy e.g. cisplatin and cytosine arabinoside, while 3BP was effective in decreasing the viability and disturbing the morphology of U373MG cells.

3-Bromopyruvate antagonizes effects of lactate and pyruvate, synergizes with citrate and exerts novel anti-glioma effects.

Oxidative stress-energy depletion therapy using oxidative stress induced by D-amino acid oxidase (DAO) and energy depletion induced by 3-bromopyruvate (3BP) was reported recently (El Sayed et al., Cancer Gene Ther., 19, 1-18, 2012). Even in the presence of oxygen, cancer cells oxidize glucose preferentially to produce lactate (Warburg effect) which seems vital for cancer microenvironment and progression. 3BP is a closely related structure to lactate and pyruvate and may antagonize their effects as a novel mechanism of its action. Pyruvate exerted a potent H(2)O(2) scavenging effect to exogenous H(2)O(2), while lactate had no scavenging effect. 3BP induced H(2)O(2) production. Pyruvate protected against H(2)O(2)-induced C6 glioma cell death, 3BP-induced C6 glioma cell death but not against DAO/D-serine-induced cell death, while lactate had no protecting effect. Lactate and pyruvate protected against 3BP-induced C6 glioma cell death and energy depletion which were overcome with higher doses of 3BP. Lactate and pyruvate enhanced migratory power of C6 glioma which was blocked by 3BP. Pyruvate and lactate did not protect against C6 glioma cell death induced by other glycolytic inhibitors e.g. citrate (inhibitor of phosphofructokinase) and sodium fluoride (inhibitor of enolase). Serial doses of 3BP were synergistic with citrate in decreasing viability of C6 glioma cells and spheroids. Glycolysis subjected to double inhibition using 3BP with citrate depleted ATP, clonogenic power and migratory power of C6 glioma cells. 3BP induced a caspase-dependent cell death in C6 glioma. 3BP was powerful in decreasing viability of human glioblastoma multiforme cells (U373MG) and C6 glioma in a dose- and time-dependent manner.

D-amino acid oxidase gene therapy sensitizes glioma cells to the antiglycolytic effect of 3-bromopyruvate.

Glioma tumors are refractory to conventional treatment. Glioblastoma multiforme is the most aggressive type of primary brain tumors in humans. In this study, we introduce oxidative stress-energy depletion (OSED) therapy as a new suggested treatment for glioblastoma. OSED utilizes D-amino acid oxidase (DAO), which is a promising therapeutic protein that induces oxidative stress and apoptosis through generating hydrogen peroxide (H2O2). OSED combines DAO with 3-bromopyruvate (3BP), a hexokinase II (HK II) inhibitor that interferes with Warburg effect, a metabolic alteration of most tumor cells that is characterized by enhanced aerobic glycolysis. Our data revealed that 3BP induced depletion of energetic capabilities of glioma cells. 3BP induced H2O2 production as a novel mechanism of its action. C6 glioma transfected with DAO and treated with D-serine together with 3BP-sensitized glioma cells to 3BP and decreased markedly proliferation, clonogenic power and viability in a three-dimensional tumor model with lesser effect on normal astrocytes. DAO gene therapy using atelocollagen as an in vivo transfection agent proved effective in a glioma tumor model in Sprague-Dawley (SD) rats, especially after combination with 3BP. OSED treatment was safe and tolerable in SD rats. OSED therapy may be a promising therapeutic modality for glioma.

Anti-tuberculosis drug susceptibility testing of Mycobacterium bovis BCG Tokyo strain.

SETTING: The Mycobacterium bovis bacille Calmette-Guérin (BCG) vaccine is the only vaccine against tuberculosis (TB), owing to its valuable protective effects and low virulence. However, it can occasionally cause systemic infection in immunocompromised hosts. Isoniazid (INH), rifampicin (RMP), streptomycin (SM) and ethambutol (EMB) are known to be effective anti-tuberculosis drugs and are used for the treatment of BCG infections. Unfortunately, there are few studies of the susceptibility of BCG vaccine strains to these drugs. OBJECTIVE: To measure the minimum inhibitory concentrations (MICs) of BCG Tokyo vaccine products for anti-tuberculosis drugs and assess vaccine safety in terms of drug susceptibility. DESIGN: We measured the MIC for one seed and five product lots of BCG Tokyo strain for INH, RMP, SM and EMB using Middlebrook 7H11 agar plates. RESULTS: The MIC results for INH were 0.06 and 0.125 mg/ml for the product and seed lots, respectively. The MIC results for RMP, SM and EMB were 0.25-0.5, 0.25 and 2-4 microg/ml, respectively. CONCLUSION: Our results indicate that the BCG Tokyo strain was susceptible to the major anti-tuberculosis drugs and treatable even in cases of severe adverse events, including systemic infection.

CD9 amino acids critical for upregulation of diphtheria toxin binding.

CD9 associates with a diphtheria toxin receptor (DTR) that is identical to the membrane-anchored form of heparin-binding EGF-like growth factor. We determined the region of CD9 important for upregulation activity. Human and monkey CD9 upregulates DT binding activity of DTR, while mouse CD9 has no upregulation activity. Transfection of chimeric constructs comprising monkey and mouse CD9s showed that the human sequence between Ala156 and Asp183 is essential for the upregulation activity. Studies of mutants, replacing a single amino acid within the region between Ala156 and Asp183 of monkey CD9 with the corresponding amino acid residue in mouse CD9, revealed that substitution of Gly158 is critical for the reduction of the upregulation activity and secondly for the substitution of Val159 and Thr175. These three amino acid residues were deduced to be located on the head domain of the second extracellular loop, suggesting that interactions of CD9 with DTR or DT at the domain containing these three amino acids were important for the upregulation of DT binding.

Effect of nordihydroguaiaretic acid on behavioral impairment and neuronal cell death after forebrain ischemia.

The purpose of this study was to evaluate the neuroprotective effect of nordihydroguaiaretic acid (NDGA), an antioxidant and/or 5-lipoxygenase inhibitor, on ischemia-reperfusion injury behavioral pharmacologically and histologically in vivo. First, the antioxidant activity of NDGA was evaluated in vitro by measuring the production of thiobarbituric acid reactive substances (TBARS) in rat brain homogenate. Second, the effect of NDGA on learning and memory impairment induced by rat four-vessel occlusion transient ischemia was investigated with the Morris water-maze task. Third, the effect of NDGA on pyramidal cell loss in the hippocampus after transient ischemia was examined. NDGA inhibited the production of TBARS with an IC(50) of 0.1 microM, and significantly attenuated postischemic learning and memory impairment at 10 mg/kg. Furthermore, consecutive 4-day administration of NDGA at 10 mg/kg significantly reduced the postischemic neuronal death. NDGA was found to be potent and effective as an anti-ischemia-reperfusion injury agent in terms of behavioral pharmacology and histology. The present results suggest that NDGA has beneficial effects on behavioral deficits and histological injury caused by ischemia-reperfusion.

Effects of arginine-vasopressin fragment 4-9 on rodent cholinergic systems.

Arginine-vasopressin fragment 4-9 (AVP4-9) has been demonstrated in animal studies to facilitate learning and memory. To clarify the mechanisms of this facilitation, we focused on the effects of AVP4-9 on rodent cholinergic systems. AVP4-9 (0.1 microM) enhanced the basal and the high-potassium-evoked acetylcholine (ACh) release from rat hippocampal slices (122.4 and 120.0% of control, respectively) in the presence of 1.3 mM calcium (physiological level) at 60 min after the incubation at 37 degrees C. The AVP4-9-stimulated basal ACh release was inhibited by a V1-selective antagonist ([(beta-mercapto-beta,beta-cyclopentamethylene propionic acid)1, O-methyl-Tyr2, Arg8] vasopressin), but not by a V2-selective antagonist ([adamantaneacetyl1, O-ethyl-D-Tyr2, Val4, aminobutyryl6, Arg8,9]-vasopressin). In addition, AVP4-9 did not affect the basal ACh release under the calcium-free condition at 37 degrees C or in the presence of 1.3 mM calcium at 4 degrees C. However, AVP4-9 facilitated the passive-avoidance response of scopolamine (a cholinergic blocker)-induced memory-deficient mice. These findings demonstrate that AVP4-9 stimulates ACh release via mediation by V1-like vasopressin receptors, and shows dependence on calcium ion and temperature. The results also suggest that the mechanism of the facilitative effects of AVP4-9 on learning and memory consist of the observed stimulation of cholinergic systems and other parallel pathways that would not be inhibited by cholinergic blocking.

Effects of prolyl endopeptidase inhibitors and neuropeptides on delayed neuronal death in rats.

We investigated the effects of the prolyl endopeptidase inhibitors 1-[1-(Benzyloxycarbonyl)-L-prolyl]prolinal (Z-Pro-Prolinal) and N-benzyloxycarbonyl-thioprolyl-thioprolinal-dimethylaceta l (ZTTA) on delayed neuronal death induced by four-vessel-occlusion transient ischemia in rats. We also examined the effects of [pGlu4, Cyt6, ArgS]vasopressin (vasopressin-(4-9)) and thyrotropin-releasing hormone (TRH) on the delayed neuronal death. Furthermore, we investigated the role of vasopressin receptors in the effects of vasopressin and prolyl endopeptidase inhibitors. Z-Pro-Prolinal, vasopressin-(4-9) and TRH protected pyramidal cells in the CA1 subfield of the rat hippocampus from delayed neuronal death after 10-min ischemia. The effect of vasopressin-(4-9) was abolished by vasopressin receptor antagonists. The effect of Z-Pro-Prolinal was also abrogated by the antagonists. These results suggest that the neuroprotective effect of prolyl endopeptidase inhibitors is mediated by neuropeptides such as [Arg8]vasopressin and TRH, and indicate the involvement of vasopressin receptors in the neuroprotective effect of vasopressin-(4-9) and prolyl endopeptidase inhibitors.

Peritoneal dialysis--associated peritonitis caused by gram-negative bacteria: characteristics similar to spontaneous bacterial peritonitis?

The aim of the study was to investigate the characteristics of PD-related peritonitis caused by gram-negative bacteria (GNP). We retrospectively studied the medical records of 164 patients (114 males, 50 females; mean age 46 +/- 15 years) who continued PD beyond 5 months between 1984 and 1998. The average observation time was 40 +/- 28 months (total of 6609 patient-months). A total of 166 episodes of peritonitis occurred during that time (mean incidence: 1 episode/40 patient-months). Of these, 35 were GNPs, and GNP incidence stayed almost constant over time. Most GNP patients (63%) recovered without complication with an average of 14 days' antibiotic treatment. In only 4 cases was PD abandoned. Clinical features of GNP were similar to those of spontaneous bacterial peritonitis (SBP). The unchanged incidence of GNP over time with advanced connection devices suggests that there are important mechanisms promoting micro-organisms of endogenous origin into the peritoneal cavity in PD patients.

Structure-function analysis of the diphtheria toxin receptor toxin binding site by site-directed mutagenesis.

Diphtheria toxin (DT) binds to the epidermal growth factor (EGF)-like domain of human membrane-anchored heparin-binding EGF-like growth factor (proHB-EGF), the human DT receptor (DTR). DT does not bind to mouse proHB-EGF because of amino acid substitutions within the EGF-like domain. We made 10 independent mutants, replacing a single amino acid within the EGF-like domain of human DTR/proHB-EGF with the corresponding amino acid residue in mouse proHB-EGF. The mutant proteins were transiently expressed in mouse L cells either expressing or not expressing DRAP27/CD9, and DT binding was measured. DT binding activity of GST fusion proteins containing the mutated EGF-like domain was also determined by a cell-free binding assay. The largest effect was seen with E141H, and second largest effects were seen with F115Y and L127F in all of the assay systems. We conclude that Phe115, Leu127, and Glu141 are critical amino acid residues for DT binding. A computer model of the tertiary structure of the EGF-like domain of human DTR/proHB-EGF was made. The model predicts that three amino acid residues critical for DT binding activity, Phe115, Leu127, and Glu141, are all located on the same face of the EGF-like domain, suggesting that this face of DTR/proHB-EGF interacts with the receptor-binding domain of DT.

Facilitation of passive avoidance response by newly synthesized cationized arginine vasopressin fragment 4-9 in rats.

The effects of a newly synthesized cationized arginine vasopressin fragment 4-9 analogue (C-AVP-(4-9)) on learning and memory in rats were studied by the passive avoidance test. C-AVP-(4-9) and its parent peptide, arginine vasopressin fragment 4-9 (AVP-(4-9)), a well known potent neuropeptide, were subcutaneously injected 1.5 hr prior to the retention test. The most effective doses of C-AVP-(4-9) and AVP-(4-9) were 8.6 x 10(-2) and 1.3 nmol/kg, respectively. To evaluate the distribution of C-AVP-(4-9) in the control nervous system (CNS), apparent tissue-plasma concentration rations (Kp.app) of intravenously administered radioiodinated C-AVP-(4-9) (125I-C-AVP-(4-9)) in the CNS in mice were determined. At the apparent steady state of plasma concentration of 125I-C-AVP-(4-9), the Kp.app values of the 125I-C-AVP-(4-9) in the cerebrum, cerebellum and spinal cord were over 12 times higher than that of the vascular space marker which slightly penetrates the BBB. Moreover, the rat cerebral homogenate converted C-AVP-(4-9) into its parent peptide AVP-(4-9). These results suggest that the potent effects of C-AVP-(4-9) on learning and memory may be due to AVP-(4-9) generated as a result of distribution and metabolism of peripherally administered C-AVP-(4-9) in the CNS.

Assembly of JC virus-like particles in COS7 cells.

JC virus lacks an appropriate cell line to support virus replication. The establishment of a JC pseudovirus assembly system could play an alternative role for a virus culture system. COS7 cells and a transfer vector, pcDL-SR alpha 296, were used to express JC viral structural genes. VP231-SR alpha, which encodes VP2/VP3 and VP1, but lacks 137 bp of the 5'-terminus of agnogene, showed both efficient nuclear migration and quantitative expression of the major capsid protein VP1. JC pseudovirus assembly was observed in the nucleus of VP231-SR alpha transfected cells. Evidence of JC pseudovirus assembly is presented. The further utilization of this system, which includes a study for the viral morphogenesis, serological diagnosis, as well as the potential application for gene transfer vector, is discussed.

LDH isoenzyme of the dialysate in noninfected PD patients.

We wished to investigate lactic dehydrogenase (LDH) and its isoenzymes in the dialysate of peritoneal dialysis (PD) patients under noninfected conditions. Twenty-seven continuous ambulatory peritoneal dialysis (CAPD) patients (20 males, 7 females; mean age 53 years; average duration of CAPD 29 months) were studied. Overnight dialysate and blood samples were collected. LDH activities, isoenzymes, glucose, creatinine of both specimens, and cancer antigen 125 (CA125) of the dialysate were determined. Serum and dialysate LDH was 470 +/- 163 and 10 +/- 4 U/L, respectively. Percentages of LDH1, LDH2, LDH3, LDH4, and LDH5 were 29 +/- 5, 37 +/- 4, 20 +/- 3, 9 +/- 4, and 6 +/- 2 (%) in the serum, 21 +/- 5, 30 +/- 5, 23 +/- 6, 16 +/- 4, and 8 +/- 3 (%) in the dialysate, respectively. There was no significant relationship between the serum and dialysate LDHs. There were no correlations between dialysate LDH, patient's age, duration of CAPD, peritoneal transport properties, and dialysate CA125. Although it was still difficult to determine the origin of dialysate LDH, a predominance of LDH5 was not observed in noninfected stable patients. Dialysate LDH activities were low, and the isoenzyme patterns were similar to those of the serum. Alteration of the isoenzyme patterns may suggest unfavorable events, such as infection, in the peritoneal cavity.

Effects of oral administration of soybean lecithin transphosphatidylated phosphatidylserine on impaired learning of passive avoidance in mice.

Soybean lecithin transphosphatidylated phosphatidylserine (SB-tPS) was investigated for its effect on the impaired learning of a passive avoidance task by mice induced by scopolamine or cycloheximide. SB-tPS (240, 360, 480 mg/kg) administered orally significantly prolonged the step-through latency shortened by scopolamine. SB-tPS (240 mg/kg) administered orally also prolonged the step-through latency shortened by cycloheximide. These results suggest that the effect of SB-tPS on the impaired learning behavior may be related not only to the cholinergic system but also the serotonergic system.

Immune system-related CD9 is expressed in mouse central nervous system myelin at a very late stage of myelination.

CD9 is a tetra-membrane-spanning glycoprotein involved in cell adhesion, migration, and proliferation in both the immune and the immature nervous system. In this study, CD9 expression was detected in myelin of mouse brain, starting at postnatal day 16. The amount of CD9 protein continuously increased with age and persisted in the adult brain. It appeared later than myelin proteolipid protein (PLP), a typical late myelin marker. Mature oligodendrocytes were abundant in CD9, although it was also detected in astrocytes and microglial cells in vitro. CD9 appeared at the end of the myelination process and was localized along the outermost membrane of compact myelin. CD9 is known to associate with integrins, which are candidate receptors for extracellular matrix and transmit extracellular signals into the cells. Taken together, CD9 at the surface of central nervous system (CNS) mature myelin may have a unique function to facilitate signal transduction and enhance myelin membrane adhesion to extracellular matrices at very late stages of development.

ZTTA, a postproline cleaving enzyme inhibitor, improves cerebral ischemia-induced deficits in a three-panel runway task in rats.

We investigated the effect of N-benzyloxycarbonyl-thioprolyl-thioprolinal-dimethylaceta l (ZTTA), a novel postproline cleaving enzyme (prolyl endopeptidase, PPCE) inhibitor, on the in vitro activity of rat brain PPCE and memory impairment induced by cerebral ischemia. ZTTA noncompetitively inhibited rat brain PPCE (ki = 2.9 microM). Cerebral ischemia for 5 min increased the number of errors in a working memory task with a three-panel runway paradigm. ZTTA at 6 mg/kg, administered immediately after blood flow reperfusion, significantly reduced the increase in working memory errors expected to occur 24 h after 5 min of ischemia. The antiamnesic action of ZTTA may be ascribable to a neuroprotective effect on the central nervous system due to some neuropeptides that are substrates of PPCE in the brain.