PLASMATIC LEVELS OF NEUROPEPTIDES, INCLUDING OXYTOCIN, IN CHILDREN WITH AUTISM SPECTRUM DISORDER, CORRELATE WITH THE DISORDER SEVERITY.

CONTEXT: Oxytocin has been investigated as a potential medication for psychiatric disorders. OBJECTIVE AND DESIGN: This study prospectively investigates correlations between oxytocin and other neuropeptides plasma levels in patients with autism spectrum disorders (ASD) according to severity and treatment, as compared to controls. SUBJECTS AND METHODS: Thirty-one children (6 neurotypical as control) participated in this study. The patients were classified into mildly and severely-affected, according to Autism Diagnostic Observation Schedule (ADOS) scores. Oxytocin, orexin A and B, α-MSH, ß-endorphins, neurotensin and substance P were investigated using a quantitative multiplex assay or a competitive-ELISA method. RESULTS: Plasma oxytocin levels differed between the groups (F (2, 24) =6.48, p=0.006, η2=0.35, observed power=86%): patients with the mild ASD had higher values of plasma oxytocin than those with the severe form (average difference=74.56±20.74pg/mL, p=0.004). CONCLUSIONS: These results show a negative correlation between plasma levels of oxytocin and the severity of ASD and support the involvement of oxytocinergic mechanisms in ASD.

Increased IRF4 expression in isolated B cells from common variable immunodeficiency (CVID) patients

Background: Common variable immunodeficiency (CVID) is a heterogeneous disorder characterized by low serum levels of immunoglobulins (Igs) and recurrent infection. In most CVID patients, a defect in the differentiation of B cells into plasma cells has been observed. Several factors play an important role in the proliferation and differentiation of B cells, including IRF4 and XBP1 transcription factors. Methods: In the present study we investigated the expression of IRF4 and XBP1 in the B-cells of CVID and healthy controls (HCs). For this purpose, we assessed the expression of IRF4 and XBP1 at both mRNA and protein levels by real time-PCR and flow cytometry, respectively. Results: We found that IRF4 expression was significantly increased in CVID patients compared with controls. Although the XBP1 protein level was lower in patients in comparison to controls, this difference was not significant. Conclusion: Taken together, increased IRF4 expression could be involved in defective functions of B cells in CVID patients

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Updated European Consensus Statement on diagnosis and treatment of adult ADHD.

Background Attention-deficit/hyperactivity disorder (ADHD) is among the most common psychiatric disorders of childhood that often persists into adulthood and old age. Yet ADHD is currently underdiagnosed and undertreated in many European countries, leading to chronicity of symptoms and impairment, due to lack of, or ineffective treatment, and higher costs of illness. Methods The European Network Adult ADHD and the Section for Neurodevelopmental Disorders Across the Lifespan (NDAL) of the European Psychiatric Association (EPA), aim to increase awareness and knowledge of adult ADHD in and outside Europe. This Updated European Consensus Statement aims to support clinicians with research evidence and clinical experience from 63 experts of European and other countries in which ADHD in adults is recognized and treated. Results Besides reviewing the latest research on prevalence, persistence, genetics and neurobiology of ADHD, three major questions are addressed: (1) What is the clinical picture of ADHD in adults? (2) How should ADHD be properly diagnosed in adults? (3) How should adult ADHDbe effectively treated? Conclusions ADHD often presents as a lifelong impairing condition. The stigma surrounding ADHD, mainly due to lack of knowledge, increases the suffering of patients. Education on the lifespan perspective, diagnostic assessment, and treatment of ADHD must increase for students of general and mental health, and for psychiatry professionals. Instruments for screening and diagnosis of ADHD in adults are available, as are effective evidence-based treatments for ADHD and its negative outcomes. More research is needed on gender differences, and in older adults with ADHD.

Increased IRF4 expression in isolated B cells from common variable immunodeficiency (CVID) patients.

BACKGROUND: Common variable immunodeficiency (CVID) is a heterogeneous disorder characterized by low serum levels of immunoglobulins (Igs) and recurrent infection. In most CVID patients, a defect in the differentiation of B cells into plasma cells has been observed. Several factors play an important role in the proliferation and differentiation of B cells, including IRF4 and XBP1 transcription factors. METHODS: In the present study we investigated the expression of IRF4 and XBP1 in the B-cells of CVID and healthy controls (HCs). For this purpose, we assessed the expression of IRF4 and XBP1 at both mRNA and protein levels by real time-PCR and flow cytometry, respectively. RESULTS: We found that IRF4 expression was significantly increased in CVID patients compared with controls. Although the XBP1 protein level was lower in patients in comparison to controls, this difference was not significant. CONCLUSION: Taken together, increased IRF4 expression could be involved in defective functions of B cells in CVID patients.

Validation of a HILIC/ESI-MS/MS method for the wood burning marker levoglucosan and its isomers in airborne particulate matter.

In the present study, a methodology involving hydrophilic interaction liquid chromatography (HILIC) and electrospray (ESI) tandem mass spectrometry (MS/MS) was developed for measurement of anhydrous monosaccharides as markers for wood burning in atmospheric aerosols, PM10. No extensive sample preparation, other than ultrasound-assisted solvent extraction and evaporation, was applied. A pentahydroxysilica column enabled separation of levoglucosan from mannosan and galactosan within 5 min and the quantitative performance was validated using the standard reference materials (SRM) 1649a and 1649b. The experimentally obtained results for SRMs were in agreement with values previously reported in other studies. Achieved instrumental limits of detection (LODs) were below 10 pg injected on column, corresponding to LODs in air lower than 0.10 ng/m3 for all measured isomers for 2-3 day sampling with 1.0 m-3 h-1 sampling rate. The validated method was used for the determination of levoglucosan and its isomers in atmospheric aerosols collected in three different Swedish urban areas during the winter and summer time in 2017. The total measured concentrations for levoglucosan and galactosan + mannosan were determined to be between 78 and 167 ng/m3 in January 2017, which is approximately 10-times higher compared to the levels detected in July, reflecting the higher frequency of wood burning for heating during the cold season. Calculated concentration ratios between levoglucosan and its isomers in the urban area samples indicated mostly mixed softwood/hardwood combustion in winter time; on the other hand, softwood burning was observed as the major emission in summer time.

Hydrogel nanoparticle encapsulated plasmid as a suitable gene delivery system.

To facilitate the delivery of genetic material, the use of appropriate carriers such as polymers is necessary. Nanoparticles comprising of chitosan-alginate polymers were formed through pregel preparation method. Chi/Alg nanoparticles had a mean Z-Average diameter of 161.8 nm and mean zeta 29.3 mV, respectively. The ability of plasmidcomplex in preventing DNA migration showed Chi/Alg nanoparticles have great capacity to maintain plasmid. The efficiency of nanoparticles for transfection of pEGFP-N1 plasmid in the cultured HEK 293 cells was measured by flow cytometry. Cell viability assays indicated that nanoparticles had no toxic effect on HEK 293 cells after 4 or 24 h. Our suitable candidate for gene delivery would be Alg/Chi nanoparticles.

Tamoxifen and tranilast show a synergistic effect against breast cancer in vitro.

OBJECTIVE: This study was aimed at examining a separate or combined effect of tamoxifen and tranilast drugs on growth and proliferation of breast cancer cells. BACKGROUND: Breast cancer is one of the most common cancers and the second leading cause of cancer death among women worldwide. Tamoxifen is the most widely used anti-estrogen for the treatment of breast cancer. Studies show that a combination therapy with other drugs enhances the activity of tamoxifen. Tranilast is an anti-inflammatory drug. We hypothesize that tranilast plus tamoxifen can work synergistically and help getting better result from this anticancer drug. METHODS: Two breast cancer cell lines, MCF-7 and MDA-MB-231, were treated with graduated concentrations of tamoxifen and tranilast alone or in combination at 24, 48 or 72 hours for MCF-7, and 48 hours for MDA-MB-231 cells. We used the MTT assay and lactate dehydrogenase leakage (LDH) assay to evaluate cell viability and cytotoxicity, respectively. RESULTS: In both ER-positive and ER-negative breast cancer cell lines, the combination of tranilast and tamoxifen was more effective in growth inhibition than single drug exposure. CONCLUSION: We have demonstrated that by means of a synergistic/additive inhibitory effect, tranilast was capable of enhancing the in vitro activity of tamoxifen on breast cancer cell lines. Based on the results obtained in this study, tranilast could be a candidate drug for combination therapy in resistant breast cancer patients (Fig. 9, Ref. 17). KEYWORDS: breast cancer, Tamoxifen, Tranilast, LDH release, MTT.

First isolation of Mycobacterium spp. in Mullus spp. in Turkey.

Ichthyozoonotic Mycobacterium spp. poses health risks both to fish and humans. In this study, the presence of ichthyozoonotic Mycobacterium spp. was investigated in red mullet (Mullus barbatus barbatus) and surmullet (Mullus surmuletus), widely caught species in the Mediterranean and the Aegean Sea. A total of 208 fish samples, provided from fishermen of Mersin province (Turkey) were studied. Using conventional methods, Mycobacterium spp. was isolated and identified at the genus level by PCR and at the species level by PCR-RFLP. Thirteen Mycobacterium spp. were detected in 13 (6.25%) fish samples. Four mycobacteria were identified as M. genavense, three as M. fortuitum, three as M. scrofulaceum, one as M. marinum, one as M. vaccae and one as M. aurum. No signs of mycobacteriosis were observed in fish samples. Findings of this study can contribute to future studies of onichthyozoonotic Mycobacterium spp. in seafood.

Distinct strains of Leishmania major induce different cytokine mRNA expression in draining lymph node of BALB/c mice.

Four genotypically distinct strains of L. major collected from persons residing in different endemic areas of cutaneous leishmaniasis in Iran were evaluated in BALB/c mice. Parasite virulence was evaluated by measuring the parasite burden in the lymph nodes. Immunogenicity of the strains was assessed by analysis of cytokines mRNA expression levels in popliteal lymph nodes of the mice in early (3, 16, 40 h) and late (week 1, W3, W5 and W8) time periods after infection. The expression of cytokines mRNA, namely Ifng, Il2,Il4,Il10 and Il12, was quantitated by real-time PCR. The lowest and the highest parasite loads were induced by Damghan (2·15 × 107) and Shiraz (9·59 × 109) strains, respectively. Moreover, Damghan strain elicited higher expression levels of Ifng and Il2 mRNA and the highest ratio of Ifng/Il4 mRNA expression compared with the other strains at 40 h and 8 weeks post-infection. The results indicate that the inoculation of BALB/c mice with different strains induced high diversity in parasite burden and cytokines gene expression. Amongst the four strains, Damghan strain showed the lowest parasite load and the highest tendency to induce expression of Th1 cytokines gene and might be considered as a safe and immunogenic strain.

Cloning, Expression and Characterization of Zebra Fish Ferroportin in Hek 293T Cell Line.

BACKGROUND: Ferroportin (Fpn), a regulator of iron homeostasis is a conserved membrane protein that exports iron across the enterocytes, macrophages and hepatocytes into the blood circulation. Fpn has also critical influence on survival of microorganisms whose growth is dependent upon iron, thus preparation of Fpn is needed to study the role of iron in immunity and pathogenesis of micoorganisms. METHODS: To prepare and characterize a recombinant ferroportin, total RNA was extracted from Indian zebrafish duodenum, and used to synthesize cDNA by RT-PCR. PCR product was first cloned in Topo TA vector and then subcloned into the GFP expression vector pEGFP-N1. The final resulted plasmid (pEGFP-ZFpn) was used for expression of Fpn-EGFP protein in Hek 293T cells. RESULTS: The expression was confirmed by appearance of fluorescence in Hek 293 T cells. Recombinant Fpn was further characterized by submission of its predicted amino acid sequences to the TMHMM V2.0 prediction server (hidden Markov model), NetOGlyc 3.1 and NetNGlyc 3.1 servers. The obtained Fpn from indian zebrafish also contained eight transmembrane domains with N- and C-termini inside the cytoplasm and harboured 78 O-glycosylated amino acids. CONCLUSION: The recombinant Fpn from Indian zebra fish was successfully expressed in Hek 293 cell line. Although the discrepancy in two amino acids was observed in our produced Fpn and resulted in an additional O-glycosylation site, but had no effect on the topology of the protein compared to other Fpn described by other researchers. Therefore this construct can be used in future iron studies.

Microstructural evolution of bainitic steel severely deformed by equal channel angular pressing.

High Si bainitic steel has been received much of interest because of combined ultra high strength, good ductility along with high wear resistance. In this study a high Si bainitic steel (Fe-0.22C-2.0Si-3.0Mn) was used with a proper microstructure which could endure severe plastic deformation. In order to study the effect of severe plastic deformation on the microstructure and properties of bainitic steel, Equal Channel Angular Pressing was performed in two passes at room temperature. Optical, SEM and TEM microscopies were used to examine the microstructure of specimens before and after Equal Channel Angular Pressing processing. X-ray diffraction was used to measure retained austenite after austempering and Equal Channel Angular Pressing processing. It can be seen that retained austenite picks had removed after Equal Channel Angular Pressing which could attributed to the transformation of austenite to martensite during severe plastic deformation. Enhancement of hardness values by number of Equal Channel Angular Pressing confirms this idea.

Soluble CD26/CD30 levels in visceral leishmaniasis: markers of disease activity.

Leishmania infantum is the causative agent of zoonotic visceral leishmaniasis (VL). If untreated the disease could be fatal; however, in some cases the infection can run a subclinical course. In subclinical infections a Th1-response predominates, while Th2-responses and/or probably Treg cells are related to unfavourable outcome of the disease in active VL. In the present study we determined the levels of soluble (s) CD26 and CD30 co-stimulatory molecules in sera from patients with active VL, asymptomatic individuals and healthy volunteers. Results showed a significant difference in both sCD26 and sCD30 between infected cases and normal individuals (P < or = 0.001). However, there was no significant difference in sCD26 levels between asymptomatic cases and patients, although the difference was not significant. sCD30 levels were significantly higher in VL patients than asymptomatic cases (P < or = 0.001). These findings suggest a possible association between sCD26 and sCD30 levels and the clinical manifestation of L. infantum infection.

Problem focused knowledge navigation: implementing the problem focused medical record and the O-HEAP note.

The current organization of most Computerized Medical Records (CMR) is based on the Problem Oriented Medical Record (POMR) and the SOAP (Subjective, Objective, Assessment and Plan) note. The organizational structure of the POMR and especially the SOAP note, does not allow for optimal use of computer capabilities in the follow up note. Since follow up visits are the most common office visit by far, this is a major flaw in the CMR. The authors propose a Problem Focused Medical Record and the OHEAP (Orientation, History, Exam, Assessment and Plan) note to resolve this problem. OHEAP starts with a powerful orientation structure that brings forward the timeline, last Assessment and Plan, and Plan Results for each problem along with the patient's historical tables as the starting point of every follow up visit. The Assessment and Plan portion brings problem specific differential diagnoses and their workups along with other relevant tables such as expert systems, treatments, instructions, medical literature or pathways. This leads to Problem Focused Knowledge Navigation that brings powerful efficiencies to the CMR. By recognizing the true workflow in the longitudinal diagnosis and management of any medical problem, the efficiency of the CMR is maximized. OHEAP allows for optimal use of both personal and external data elements in the medical record. Its powerful orientation attributes minimize the time spent in analyzing the current status of the problem while its connections to problem specific databases help resolve the problem.

Education review: applied medical informatics--informatics in medical education.

The importance of informatics training within a health sciences program is well recognized and is being implemented on an increasing scale. At Chicago Medical School (CMS), the Informatics program incorporates information technology at every stage of medical education. First-year students are offered an elective in computer topics that concentrate on basic computer literacy. Second-year students learn information management such as entry and information retrieval skills. For example, during the Introduction to Clinical Medicine course, the student is exposed to the Intelligent Medical Record-Entry (IMR-E), allowing the student to enter and organize information gathered from patient encounters. In the third year, students in the Internal Medicine rotation at Norwalk Hospital use Macintosh power books to enter and manage their patients. Patient data gathered by the student are stored in a local server in Norwalk Hospital. In the final year, we teach students the role of informatics in clinical decision making. The present senior class at CMS has been exposed to the power of medical informatics tools for several years. The use of these informatics tools at the point of care is stressed.

Intelligent Medical Record--entry (IMR-E).

This paper describes an automated medical record designed to allow providers to enter patient data at the point of care. The system runs on PCs and Macintoshes and uses a graphical user interface and object-oriented programming to take advantage of current mouse and pen technologies. The provider acquires all relevant patient data by pointing and clicking at selections on input screens, many of which contain anatomical drawings to help the provider quickly and accurately describe patient findings. The system also generates a grammatically correct progress note using the problem-oriented structure. Furthermore, items identified in the assessment and plans portion of the program can be ported to expert systems for medical decisions assistance or to billing systems. The system allows the provider to obtain the necessary information on a focused patient visit in less than 5 min or to enter a complete history and physical.

Activation patterns in experimental canine atrial flutter produced by right atrial crush injury.

OBJECTIVES: This study was designed to localize and characterize the atrial flutter reentrant circuit and the electrophysiologic effects of right atrial crush injury in a new canine model. BACKGROUND: In previous studies sustained atrial flutter was induced in the canine heart by rapid atrial pacing after a linear crush injury was placed in the right atrial free wall. METHODS: Eight dogs (group 1) with three electrode plaques on the right and left atria and Bachmann's bundle and seven dogs (group 2) with a single high density electrode plaque on the right atrium were studied with use of a 64-channel computerized mapping system. RESULTS: At baseline, during sinus rhythm and right and left atrial pacing, activation spread uniformly without areas of slow conduction. Crush injury produced marked conduction delay or complete block during sinus rhythm, increasing the mean difference in activation times across the injury compared with control values (group 1, 31 +/- 4 vs. 14 +/- 5 ms, p less than 0.01; group 2, 28 +/- 10 vs. 7 +/- 2 ms, p less than 0.01). Rapid atrial pacing (S1S1 200 ms) above and below the crush injury revealed a line of complete block across which adjacent electrodes recorded markedly different activation times (33 +/- 5 and 38 +/- 12 ms difference, respectively) and around which activation wave fronts proceeded, colliding opposite the stimulating electrodes. The mean atrial flutter cycle length of 11 episodes induced in group 1 and 14 episodes in group 2 was 157 +/- 16 and 140 +/- 16 ms, respectively (p = NS). Activation mapping revealed a reentrant circuit in the right atrium around the crush injury in all episodes. Although the reentrant circuit did not contain a discrete area of slow conduction, activation time below was longer than that above the crush injury (92 +/- 14 vs. 66 +/- 8 ms and 82 +/- 12 vs. 59 +/- 9 ms in groups 1 and 2, respectively, p less than 0.01 for both). Rapid atrial pacing or premature stimuli produced progressive conduction delay and unidirectional block between the crush injury and the tricuspid anulus, inducing atrial flutter directly in 9 of 25 episodes. In 16 episodes, atrial flutter developed after transient induction of atrial fibrillation. CONCLUSIONS: 1) Atrial flutter in this model is due to reentry in the right atrium; 2) the crush injury functions as an anatomic obstacle around which reentry may occur; and 3) the reentrant circuit does not contain a discrete area of slow conduction but, rather, generally slower conduction below the crush injury.

Mechanism of double potentials recorded during sustained atrial flutter in the canine right atrial crush-injury model.

BACKGROUND: During atrial flutter, double potentials may be recorded at specific sites in the atria. It has been suggested that double potentials represent sequential activations at the center of the reentrant circuit. An alternative hypothesis is that double potentials represent electrical activity in an area of slow conduction. Understanding their mechanism is important because double potentials have been considered a possible indicator of target sites for catheter ablation. METHODS AND RESULTS: We systematically studied double potentials in our canine model of atrial flutter produced by right atrial crush injury using a 64-channel computerized mapping system with 56 electrodes on the right atrium in seven mongrel dogs under general anesthesia. Activation maps were recorded during sinus rhythm before and after crush injury, during rapid pacing above and below the crush injury, and during sustained atrial flutter, entrainment of atrial flutter, and termination of atrial flutter induced with D-sotalol (2 mg/kg). During sinus rhythm before crush injury, activation was uniform, and double potentials were not recorded in any dog. After crush injury, activation proceeded up to and around the crush injury, and narrowly split double potentials were recorded in two of seven dogs. During rapid pacing above and below the crush injury, double potentials were recorded in five dogs. During 14 episodes of atrial flutter (mean cycle length, 140 +/- 16 msec), double potentials were recorded at electrodes along the crush injury. The activation time of the early x component of the double potentials (25 +/- 13 msec) was similar to that of adjacent electrodes above the crush injury (24 +/- 11 msec), and the activation time of the late y component (89 +/- 13 msec) was similar to that of adjacent electrodes below the crush injury (91 +/- 14 msec). The timing of the x and y components was dependent on the location of the recording electrode, with x and y widely spaced at the end of the crush injury near the area of earliest atrial activation during atrial flutter, more equally timed at the center of the crush injury, and more closely timed at the end of the crush injury opposite the area of earliest activation. During transient entrainment, double potentials were accelerated to the pacing rate, but their activation time relative to adjacent electrodes was maintained. During abrupt termination of atrial flutter, the early x component of the double potential was always recorded, but the late y component was not, because of conduction block below the posterior end of the crush injury. CONCLUSIONS: This study has shown in our canine model of atrial flutter that double potentials are recorded from the center of the reentrant circuit and that they represent sequential activations as the reentrant wave front passes on either side of the crush injury.

Informatics Workup.

We introduce the concept of a Medical Informatics Workup performed by fourth year medical students working in a busy inner-city Emergency Room. These students use portable computers (Macintosh PowerBook 170s connected to a removable cartridge hard drive and CD-ROM drive) to do the patient workups. The PowerBook 170 contains the automated medical record entry software (IMR-E), five expert system software packages, and a program that allows the PowerBook to emulate a PC-compatible computer. With this configuration the student has a portable system that allows for the creation of a computerized medical record at the patient's bedside, along with the ability to analyze the data and generate a list of differential diagnoses.