Role of G3BP1 in glucocorticoid receptor-mediated microRNA-15b and microRNA-23a biogenesis in endothelial cells.
MicroRNAs (miRNAs) are a family of non-coding RNAs that play crucial roles in regulating various normal cellular responses. Recent studies revealed that the canonical miRNA biogenesis pathway is subject to sophisticated regulation. Hormonal control of miRNA biogenesis by androgen and estrogen has been demonstrated, but the direct effects of the glucocorticoid receptor (GR) on miRNA biogenesis are unknown. This study revealed the role of GR in miRNA maturation. We showed that two GR agonists, dexamethasone and ginsenoside-Rg1 rapidly suppressed the expression of mature miR-15b, miR-23a, and miR-214 in human endothelial cells. RNA pulldown coupled with proteomic analysis identified GTPase-activating protein (SH3 domain) binding protein 1 (G3BP1) as one of the RNA-binding proteins mediating GR-regulated miRNA maturation. Activated GR induced phosphorylation of v-AKT Murine Thymoma Viral Oncogene Homologue (AKT) kinase, which in turn phosphorylated and promoted nuclear translocation of G3BP1. The nuclear G3BP1 bound to the G3BP1 consensus sequence located on primary miR-15b~16-2 and miR-23a~27a~24-2 to inhibit their maturation. The findings from this study have advanced our understanding of the non-genomic effects of GR in the vascular system.


USH2A is a Meissner's corpuscle protein necessary for normal vibration sensing in mice and humans.
Fingertip mechanoreceptors comprise sensory neuron endings together with specialized skin cells that form the end-organ. Exquisitely sensitive, vibration-sensing neurons are associated with Meissner's corpuscles in the skin. In the present study, we found that USH2A, a transmembrane protein with a very large extracellular domain, was found in terminal Schwann cells within Meissner's corpuscles. Pathogenic USH2A mutations cause Usher's syndrome, associated with hearing loss and visual impairment. We show that patients with biallelic pathogenic USH2A mutations also have clear and specific impairments in vibrotactile touch perception, as do mutant mice lacking USH2A. Forepaw rapidly adapting mechanoreceptors innervating Meissner's corpuscles, recorded from Ush2a-/- mice, showed large reductions in vibration sensitivity. However, the USH2A protein was not found in sensory neurons. Thus, loss of USH2A in corpuscular end-organs reduced mechanoreceptor sensitivity as well as vibration perception. Thus, a tether-like protein is required to facilitate detection of small-amplitude vibrations essential for the perception of fine-grained tactile surfaces.


Hyunsoonleella flava sp. nov., isolated from marine sediment.
A novel Gram-stain-negative, strictly aerobic, rod-shaped, non-motile and yellow-pigmented bacterial strain, designated T58T, was isolated from a marine sediment sample collected from the coastal area of Weihai, PR China. Strain T58T was most closely related to Hyunsoonleella pacifica SW033T with 97.1 % 16S rRNA gene sequence similarity, followed by Hyunsoonleella jejuensis KCTC 22242T (96.9 %). Based on 16S rRNA gene sequences, the phylogenetic analysis indicated that strain T58T represented a member of the genus Hyunsoonleella within the family Flavobacteriaceae of the phylum Bacteroidetes. Strain T58T was found to grow optimally at 30 °C, at pH 7.0-7.5 and in the presence of 2.0-3.0 % (w/v) NaCl. The major fatty acids were iso-C15 : 0, iso-C15 : 1 G, iso-C17 : 0 3-OH and iso-C15 : 0 3-OH. The major isoprenoid quinone was menaquinone 6 (MK-6). The major polar lipids were phosphatidylethanolamine, two unidentified aminolipids and two unidentified lipids. The DNA G+C content of the genomic DNA was 35.0 mol%. On the basis of its phylogenetic, phenotypic and chemotaxonomic characteristics, strain T58T is considered to represent a novel species of the genus Hyunsoonleella, for which the name Hyunsoonleella flava sp. nov. is proposed. The type strain is T58T (=KCTC 72081T=MCCC 1H00359T).


Antihyperglycemic and hypolipidemic activity of methanolic extract of Amaranthus viridis leaves in experimental diabetes.
To investigate the antihyperglycemic and hypolipidemic effects of methanolic extract of leaves of Amaranthus viridis (MEAV) in normal and Streptozotocin (STZ) induced diabetic rats. The anti-hyperglycemic and hypolipidemic activity of methanolic extract of leaves of Amaranthus viridis was evaluated by using normal and STZ induced diabetic rats at dose of 200 mg/kg and 400 mg/kg by mouth per day for 21 days. Blood glucose levels and body weight was monitored at specific intervals, and different biochemical parameters, serum cholesterol, serum triglyceride, high density lipoprotein, low density lipoprotein, very low density lipoprotein were also assessed in the experimental animals. Histology of pancreas was performed. The statistical data indicated a significant increase in the body weight, decrease in the blood glucose, total cholesterol and serum triglycerides after treatment with MEAV. High density lipoprotein (HDL) cholesterol level was significantly increased when treated with extract. Histologically, focal necrosis was observed in the diabetic rat pancreas; however, was less obvious in treated groups. The MEAV has beneficial effects in reducing the elevated blood glucose level and body weight changes, and improves the lipid profile of STZ induced rats.


Vector competence of Culex pipiens quinquefasciatus for Murray Valley encephalitis, Kunjin, and Ross River viruses from Australia.
Australian populations of Culex pipiens quinquefasciatus Say from Brisbane, Charleville, Cairns, Kowanyama (Queensland), Darwin (Northern Territory), Mildura (Victoria), and Port Hedland (Western Australia) proved to be either poorly susceptible or refractory to oral infection with low passage level Murray Valley encephalitis, Kunjin, and Ross River viruses. With respect to past or future epidemics of Murray Valley encephalitis or Kunjin (both known to cause clinical encephalitis in man), and of epidemic polyarthritis, this mosquito is viewed as an unlikely vector.


Recent findings on neurofeedback training for auditory hallucinations in schizophrenia.
To provide recent evidence on real-time neurofeedback (NFB) training for auditory verbal hallucinations (AVH) in schizophrenia patients. NFB is a promising technique that allows patients to gain control over their AVH by modulating their own speech-related/language-related networks including superior temporal gyrus (STG) and anterior cingulate cortex (ACC) using fMRI, fNIRS and EEG/MEG. A recent limited number of studies showed that while an EEG-based NFB study failed to regulate auditory-evoked potentials and reduce AVH, downregulation of STG hyperactivity and upregulation of ACC activity with fMRI-based NFB appear to alleviate treatment-resistant AVH in schizophrenia patients. A deeper understanding of AVH and development of more effective methodologies are still needed. Despite recent innovations in antipsychotics, many schizophrenia patients continue to suffer from treatment-resistant AVH and social dysfunctions. Recent studies suggested that real-time NFB shows promise in enabling patients to gain control over AVH by regulating their own speech-related/language-related networks. Although fMRI-NFB is suitable for regulating localized activity, EEG/MEG-NFB are ideal for regulating the ever-changing AVH. Although there are still many challenges including logistic complexity and burden on patients, we hope that such innovative real-time NFB trainings will help patients to alleviate severe symptoms and improve social functioning.


The chemistry of the collagen cross-links. The characterization of fraction C, a possible artifact produced during the reduction of collagen fibres with borohydride.
The present paper describes the isolation and identification of a major radioactive component of borotritide-reduced collagen, previously designated Fraction C. The derived structure for the compound confirms that it is identical with the ;post-histidine' component described by Tanzer et al. (1973) and given the trivial name histidino-hydroxymerodesmosine. Detailed studies of the effects of acid pH on the formation of Fraction C after borohydride reduction demonstrated the apparent lability of the non-reduced form, thus confirming our previous findings (Bailey & Lister, 1968). Inhibition of the formation of this component by the acid treatment appears to be due to protonation of the histidine imidazole group. Since the only new component formed on reduction of the acid-treated fibres was the reduced aldol condensation product, these results indicate that neither the histidine nor the hydroxylysine residues can be involved in covalent linkage with the aldol condensation product in the native fibre. It is suggested therefore that the proposed non-reduced aldimine form of Fraction C does not exist as an intermolecular cross-link in vivo. Thus the presence of histidino-hydroxymerodesmosine as a tetrafunctional cross-link in reduced collagen fibres is a result of a base-catalysed reaction promoted by the borohydride-reduction procedure and this component must therefore be considered as an artifact.


The First Global Physical Activity and Sedentary Behavior Guidelines for People Living With Disability.
The World Health Organization has released the first global public health guidelines on physical activity and sedentary behavior for people living with disability. This paper presents the guidelines, related processes, and evidence, and elaborates upon how the guidelines can support inclusive policy, practice, and research. Methods were consistent with the World Health Organization protocols for developing guidelines. Systematic reviews of the evidence on physical activity for health for people living with disability were appraised, along with a consideration of the evidence used to inform the general 2020 World Health Organization guidelines. Evidence supported the development of recommendations for people living with disability, stressing that there are no major risks to engaging in physical activity appropriate to an individual's current activity level, health status, and physical function, and that the health benefits accrued generally outweigh the risks. They also emphasize the benefits of limiting sedentary behavior. The guidelines mark a positive step forward for disability inclusion, but considerable effort is needed to advance the agenda. This paper highlights key considerations for the implementation of the new recommendations for people living with disability, in line with the human rights agenda underpinning the Global Action Plan on Physical Activity 2018-2030 and allied policies.


[Complaints and clinical findings six months after COVID-19: outpatient follow-up at the University Medical Center Freiburg].
Increasing evidence suggests that some patients suffer from persistent symptoms for months after recovery from acute COVID-19. However, the clinical phenotype and its pathogenesis remain unclear. We here present data on complaints and results of a diagnostic workup of patients presenting to the post-COVID clinic at the University Medical Center Freiburg. Retrospective data analysis of persistently symptomatic patients presenting to our clinic at least 6 months after onset of acute COVID-19. All patients were assessed by a doctor and routine laboratory analysis was carried out. Quality of life was assessed using SF-36 questionnaire. In case of specific persisting symptoms, further organ-specific diagnostic evaluation was performed, and patients were referred to respective departments/specialists. 132 Patients (58 male, 74 female; mean age 53.8 years) presented to our clinic at least 6 months after COVID-19. 79 (60 %) had been treated as outpatients and 53 (40 %) as inpatients. Most common complaints were persistent fatigue (82 %) and dyspnea on exertion (61 %). Further common complaints were impairments of concentration (54 %), insomnia (43 %), and impairments of smell or taste (35 %). Quality of life was reduced in all sections of the SF-36 questionnaire, yielding a reduced working capacity. Significant pathological findings in laboratory, echocardiographic and radiological work-up were rare. Impairments in lung function tests were more common in previously hospitalized patients. Patients presenting 6 months after onset of acute COVID-19 suffer from a diverse spectrum of symptoms with impaired quality of life, also referred to as Long COVID or Post-Acute Sequelae of SARS-CoV-2 infection (PASC). Further research is needed to determine the frequency of these post-COVID syndromes and their pathogenesis, natural course and treatment options. Evaluation and management should be multi-disciplinary.


[Bacteriophage induction in cultures of C1. botulinum type A].
The authors describe the mitomycin induction of bacteriophages in strains No. 4/2, 98, and 345 of Cl. botulinum, type A. All the strains under study produced phages of the same morphology with a head and a process capable of contraction. The phages detected by electron microscopy failed to express any lytic activity on the strains of type A and other Cl. botulinum types neither in the fluid nor in the hard nutrient media. The data obtained supplemented and widened the view on the incidence of phage carrier state in the cultures of botulism causative agent.


Adsorptive removal of phosphate from aqueous solutions using different types of red mud.
Red mud (RM) is an industrial waste generated during production of alumina from using the Bayer process or the sintering process. Four types of red mud from China were characterized for their diverse chemical and mineral compositions using inductively coupled plasma-atomic emission spectroscopy (ICP-AES), inductively coupled plasma-mass spectrometry (ICP-MS) and X-ray diffraction (XRD). Acid treatment was employed to obtain activated red mud (ARM), posing increased surface areas from 10-28 m2/g to 220-350 m2/g. RMs and ARMs were used to adsorb phosphate in solution to compare the adsorption capacity. Sample GZ3, a red mud from the sintering process, presented the highest adsorption capacity among the four raw RMs, posing an adsorption capacity of 0.37 mg P/g in the solution of 1 mg P/L with a solid/solution ratio of 0.5 g: 1 L. Whereas, activated GX (AGX), a high iron Bayer red mud from diaspore bauxite, showed the highest adsorption capacity of all the ARMs, with an adsorption capacity of 1.92 mg P/g in the same condition. The dynamic studies indicate that the adsorption mainly followed the pseudo second-order model. The models of Freundlich and Langmuir were used to simulate the sorption equilibrium on GZ3 and AGX. It suggests that the Freundlich model had a better correlation with GZ3 while the Langmuir model fitted well with AGX.


mbkmeans: Fast clustering for single cell data using mini-batch k-means.
Single-cell RNA-Sequencing (scRNA-seq) is the most widely used high-throughput technology to measure genome-wide gene expression at the single-cell level. One of the most common analyses of scRNA-seq data detects distinct subpopulations of cells through the use of unsupervised clustering algorithms. However, recent advances in scRNA-seq technologies result in current datasets ranging from thousands to millions of cells. Popular clustering algorithms, such as k-means, typically require the data to be loaded entirely into memory and therefore can be slow or impossible to run with large datasets. To address this problem, we developed the mbkmeans R/Bioconductor package, an open-source implementation of the mini-batch k-means algorithm. Our package allows for on-disk data representations, such as the common HDF5 file format widely used for single-cell data, that do not require all the data to be loaded into memory at one time. We demonstrate the performance of the mbkmeans package using large datasets, including one with 1.3 million cells. We also highlight and compare the computing performance of mbkmeans against the standard implementation of k-means and other popular single-cell clustering methods. Our software package is available in Bioconductor at https://bioconductor.org/packages/mbkmeans.


Breast-milk shedding of drug-resistant HIV-1 subtype C in women exposed to single-dose nevirapine.
Single-dose nevirapine reduces intrapartum human immunodeficiency virus 1 type (HIV-1) transmission but may also select for nonnucleoside reverse-transcriptase inhibitor (NNRTI) resistance in breast milk (BM) and plasma. Among 32 Zimbabwean women, median 8-week postpartum plasma and BM HIV-1 RNA levels were 4.57 and 2.13 log(10) copies/mL, respectively. BM samples from women with laboratory-diagnosed mastitis (defined as elevated BM Na(+) levels) were 5.4-fold more likely to have HIV-1 RNA levels above the median. BM RT sequences were not obtained for 12 women with BM HIV-1 RNA levels below the lower limit of detection of the assay used. In 20 paired BM and plasma samples, 65% of BM and 50% of plasma RT sequences had NNRTI-resistance mutations, with divergent mutation patterns.


Systemic toxicity of p,p'-DDE in aged male Wistar rats following oral administration.
Systemic toxicity of p,p'-DDE (DDE) in aged male rats was investigated in Wistar rats by oral administration of DDE. About 10 mg/kg DDE had been daily administered for 28 days from 48 weeks to 52 weeks of age and its effects were observed subsequently. The administration of DDE did not give rise to any overt signs of toxicity. Male reproductive organs were not affected and serum biochemistry was not impaired at the dose of this experiment. Organ weights of the liver and spleen slightly increased and the thymus weight reduced with DDE administration. Serum total cholesterol and free T4 levels slightly decreased with DDE administration, albeit statistically insignificant. Our results indicated that even aged male rats were resistant to DDE exposure at the daily dose of ca. 10 mg/kg. However, metabolic and immunological changes still remained uncertain. Further investigation should be performed to reveal the entire effects of DDE on aged male rats.


An evaluation of Mp1p antigen screening for talaromycosis in HIV-infected antiretroviral therapy-naïve population in Guangdong, China.
Talaromycosis is one of the most common opportunistic infections in human immunodeficiency virus (HIV) infected patients. However, few researches have explored the prevalence in Southern China and fully assessed the value of the Mp1p antigen screening for the diagnosis of talaromycosis. We performed a cross-sectional study of HIV-infected antiretroviral therapy (ART)-naïve adult patients who were seen in 2018 at Guangzhou Eighth People's Hospital, Guangzhou Medical University. Serum samples collected from all the 784 enrolled patients were tested for Mp1p antigen using double-antibody sandwich enzyme-linked immunosorbent assay. A culture of pathogen was conducted in 350 clinically suspected patients to confirm talaromycosis. The overall prevalence of talaromycosis based on the Mp1p antigen detection was 11.4% (89/784) and peaked at 32.2% (75/233) in patients with CD4+ ≤50 Nr/μl. Logistic regression analysis found Mp1p antigen positive rate decreased with the increase in CD4+ counts (OR 0.982, 95% CI 0.977-0.987, P<0.01). The optimal cut-off point of the CD4+ count was 50 Nr/μl or less. Among the 350 patients received both fungal culture and Mp1p antigen detection, 95/350 (27.1%) patients were culture-positive for a Talaromyces marneffei, 75/350 (21.4%) patients were Mp1p antigen positive. The Mp1p antigen assay showed a good agreement to the culture of pathogen, and the sensitivity, specificity, positive predictive value, negative predictive value and kappa value was 71.6% (68/95), 97.3% (248/255), 90.7% (68/75), 90.2% (248/275), and 0.737, respectively. The screening accuracy of the Mp1p antigen assay in patients with CD4+ counts of ≤50 Nr/μl was superior to that in those with higher CD4+ counts. Mp1p antigen screening can be an effective tool for more efficient diagnosis of Talaromycosis, especially in HIV/AIDS patients with low CD4+ counts. Future validation studies are needed.


Results of mechanical prosthetic valve replacement in active valvular endocarditis.
To evaluate the results of mechanical prosthetic valve replacement for active endocarditis 71 patients were reviewed. They were treated surgically between 1988 and 1993 in our institution. Mechanical valves were used in 54 patients (group 1) and bioprosthetic valves were used in 17 patients (group 2). In terms of demographic, clinical and surgical variables prior to operation groups were statistically the same. Follow up ranged from 2-63 months averaged 21 months. This study was carried out to: asses cardiac status postoperatively and assess the rate of mortality, recurrency and reoperations. Evaluation of cardiac status was assessed on the basis of symptoms and findings by examination, ECG, X-ray, echocardiography, and laboratory tests. Data analysis was done by means of statistical tests like: Student's "t"-test, Fisher exact test, one sided test of difference between two percentages, Kaplan Meyer survival analysis and Cox test. 4-year mortality was 20% in group 1 comparing to 28,6% in group 2, when early mortality were 13% in group 1 comparing to 17% in group 2. These differences were not significant. The recurrency rate was 8,5% in group 1 comparing to 28,6% in group 2 which was statistically significant (p=0.028). Especially early recurrency rates differed significantly between groups and were 4,2% in group 1 comparing to 21% in group 2 (p=0.022). The reoperations rate was 4,2% in group 1 comparing to 21% in group 2 which was statistically significant (p=0.022). Clinical status showed satisfactory values and significant improvement in both groups, slightly better after mechanical valve replacement. It is concluded that mechanical valve is recommended for valve replacement in active valvular endocarditis first of all due to low recurrency and reoperation rate.


Plastic occlusion stress test as a model to investigate the effects of skin delipidization on the stratum corneum water holding capacity in vivo.
The purpose of the study was to develop an in vivo model to study the effects of lipid removal on skin barrier. 16 subjects (age 41 +/- 8) were delipidized in vivo on the volar forearm using respectively ether/acetone (EA; 1:1) and chloroform/methanol (CM; 2:1). A third site served as control. Water holding capacity (WHC) was measured according to the plastic occlusion stress test (POST) procedure: the water desorption curve after removal of the occlusion was recorded in terms of skin surface water loss (SSWL) using an evaporimeter for 30 min. In the central part of the evaporation curve (bound water) the CM-treated site is significantly different from control and EA-treated sites (p < 0.01). The SSWL decay constants reflecting the desorption rate of water from SC are higher in the CM-treated site (p < 0.01). The data are consistent with the effect of CM delipidization (polar lipids) on bound water. No differences are recorded in the evaporation of free water. We conclude that polar lipids have a key role in modulating barrier function and WHC of the stratum corneum. The POST can represent a useful in vivo model to study the effects of lipid extraction on skin function.


[Effect of simvastatin on the oxygen transport function and prooxidant - antioxidant balance in blood].
The effects of simvastatin on the blood oxygen transport function and indices of prooxidant - antioxidant balance at incubation have been studied. Simvastatin at a concentration of 100 ng/ml increases p50 (the blood pO2 corresponding to its 50% oxygen saturation) at real values of pH and pCO2 from 39.53 + 2.41 (p <0.05) to 36.60 (36, 40, 37, 60) (p <0.05) mm Hg. An increase in the drug concentration led to a decrease in the level of this parameter, but in a dose-independent manner. In addition, the level of nitrates/nitrites in the blood plasma was also increased, which was evidence of increasing activity of the L-arginine-NO system. The indices of prooxidant - antioxidant balance exhibited no significant changes. The results demonstrate a new pleiotropic effect of simvastatin, which is realized via a change in the hemoglobin - oxygen affinity through modification of NO production. This effect must be taken into account in the treatment of pathology in the blood circulation.


Measuring Drug Response with Single-Cell Growth Rate Quantification.
Intratumoral heterogeneity is a substantial cause of drug resistance development during chemotherapy or other drug treatments for cancer. Therefore, monitoring and measuring cell exposure and response to drugs at the single-cell level are crucial. Previous research suggested that the single-cell growth rate can be used to investigate drug-cell interactions. However, currently established methods for quantifying single-cell growth are limited to isolated or monolayer cells. Here, we introduce a technique that accurately measures both 2D and 3D cell growth rates using label-free ratiometric stimulated Raman scattering (SRS) microscopy. We use deuterated amino acids, leucine, isoleucine, and valine, as tracers and measure the C-D SRS signal from deuterium-labeled proteins and the C-H SRS signal from unlabeled proteins simultaneously to determine the cell growth rate at the single-cell level. The technique offers single-cell level drug sensitivity measurement with a shorter turnaround time (within 12 h) than most traditional assays. The submicrometer resolution of the imaging technique allows us to examine the effects of chemotherapeutic drugs, including kinase inhibitors, mitotic inhibitors, and topoisomerase II inhibitors, on both the cell growth rate and morphology. The capability of quantifying 3D cell growth rates provides insight into a deeper understanding of the cell-drug interaction in the actual tumor environment.


Response to Kirton.
Further comment and analyses of data from 103 alcoholics indicate less internal consistency in alcoholics' responding than in normals, no tendency for innovative alcoholics to engage in more hostile/aggressive behaviors than adaptor alcoholics, and a likelihood that more than three factors for alcoholics would be required to account for variance comparable to that for nonalcoholic persons.


Altered intrinsic functional brain architecture in female patients with bulimia nervosa.
Bulimia nervosa is a severe psychiatric syndrome with uncertain pathogenesis. Neural systems involved in sensorimotor and visual processing, reward and impulsive control may contribute to the binge eating and purging behaviours characterizing bulimia nervosa. However, little is known about the alterations of functional organization of whole brain networks in individuals with this disorder. We used resting-state functional MRI and graph theory to characterize functional brain networks of unmedicated women with bulimia nervosa and healthy women. We included 44 unmedicated women with bulimia nervosa and 44 healthy women in our analyses. Women with bulimia nervosa showed increased clustering coefficient and path length compared with control women. The nodal strength in patients with the disorder was higher in the sensorimotor and visual regions as well as the precuneus, but lower in several subcortical regions, such as the hippocampus, parahippocampal gyrus and orbitofrontal cortex. Patients also showed hyperconnectivity primarily involving sensorimotor and unimodal visual association regions, but hypoconnectivity involving subcortical (striatum, thalamus), limbic (amygdala, hippocampus) and paralimbic (orbitofrontal cortex, parahippocampal gyrus) regions. The topological aberrations correlated significantly with scores of bulimia and drive for thinness and with body mass index. We reruited patients with only acute bulimia nervosa, so it is unclear whether the topological abnormalities comprise vulnerability markers for the disorder developing or the changes associated with illness state. Our findings show altered intrinsic functional brain architecture, specifically abnormal global and local efficiency, as well as nodal- and network-level connectivity across sensorimotor, visual, subcortical and limbic systems in women with bulimia nervosa, suggesting that it is a disorder of dysfunctional integration among large-scale distributed brain regions. These abnormalities contribute to more comprehensive understanding of the neural mechanism underlying pathological eating and body perception in women with bulimia nervosa.


Quantifying network heterogeneity.
Despite degree distributions give some insights about how heterogeneous a network is, they fail in giving a unique quantitative characterization of network heterogeneity. This is particularly the case when several different distributions fit for the same network, when the number of data points is very scarce due to network size, or when we have to compare two networks with completely different degree distributions. Here we propose a unique characterization of network heterogeneity based on the difference of functions of node degrees for all pairs of linked nodes. We show that this heterogeneity index can be expressed as a quadratic form of the Laplacian matrix of the network, which allows a spectral representation of network heterogeneity. We give bounds for this index, which is equal to zero for any regular network and equal to one only for star graphs. Using it we study random networks showing that those generated by the Erdös-Rényi algorithm have zero heterogeneity, and those generated by the preferential attachment method of Barabási and Albert display only 11% of the heterogeneity of a star graph. We finally study 52 real-world networks and we found that they display a large variety of heterogeneities. We also show that a classification system based on degree distributions does not reflect the heterogeneity properties of real-world networks.


Patients' Knowledge and Attitudes Towards Regular Alcohol Urine Screening: A Survey Study.
Despite its wide implementation, there is a paucity of data supporting the effectiveness of regular alcohol urine screening (RAUS) in maintaining abstinence. This study aims at investigating if RAUS serves other purposes, what attitudes patients display towards it, and patients' technical knowledge about basic screening notions. We conducted a cross-sectional survey among adults with alcohol dependence, attending outpatient alcohol-dependence treatment. It aimed at investigating patients' attitudes and beliefs towards RAUS, and technical notions of alcohol urine screening. For attitude assessment, we adapted the Drug Attitude Inventory (DAI-10) to the field of alcohol urine screening. Internal consistency, test-retest reliability, and concurrent validity were evaluated for the adapted questionnaire. In all, 128 patients completed the questionnaire. Patients rated RAUS as high. The DAI-10 mean score was 7.2 (SD = 3.6). Internal consistency analysis revealed a Cronbach alpha of 0.718. Test-retest reliability evaluation yielded an intraclass correlation coefficient of 0.932. The score of a single Likert-type question about overall perceived value was 8.5 (SD = 2). Their correlation with mean DAI-10 score was of r = 0.254, with P = 0.009. Apart from relapse prevention, patients frequently reported other functions such as showing professionals and family members that they do not drink, or having a closer contact with professionals. A majority of patients believed alcohol use goes undetected after 48 hours from last ingestion. Regular alcohol screening is highly valued by alcohol outpatients. It seems that apart from relapse prevention, other functions related to therapeutic alliance building, social desirability, and impression management also play a key role.


Left shift in the peripheral blood count at diagnosis in acute lymphocytic leukemia is significantly correlated with duration of complete remission.
The prognostic significance of a left shift in the peripheral blood at the time of diagnosis of acute lymphocytic leukemia was investigated by a retrospective analysis of 109 patients treated on the same protocol in a single institution. Left shift was defined as the presence of 1% or more of metamyelocytes, myelocytes, or promyelocytes. All peripheral blood films were checked at the time of diagnosis by one of the authors. It was found that the duration of complete remission at 92 mo was 74% in patients with left shift and 42% in those without left shift (p less than 0.05, log-rank test). By Cox regression analysis, only the total white cell count (p less than 0.001) and the presence or absence of left shift (p less than 0.01) were independently significant in determining the proportion of patients in complete remission. Patients with a left shift had a significantly higher granulocyte count at diagnosis (p less than 0.05). We postulate that left shift in the peripheral blood count at the time of diagnosis may be an indirect measure of the total leukemia cell load. It is a new prognostic factor of significance in determining the likely outcome of the disease.


Orthodontic Treatment of Bilateral Transposition of Maxillary Canines and Lateral Incisors.
Dental transposition is a severe disturbance of tooth position and its eruptive sequence. It may affect any tooth, and it may occur at any location within the dental arch, although some teeth may be more frequently affected than others. There are several types of dental transposition, and their classification depends on the type of teeth involved. The aetiology of transposition is not very clear and has always been the subject of controversies, and it is still not completely understood. The prevalence of dental transposition and the factors related to this dental anomaly have been well documented in the literature. When treating dental transposition, many factors which may affect the treatment results must be considered, such as aesthetics, occlusion, treatment duration, patient comfort, patient cooperation, and periodontal support. Various treatment options are currently available for the treatment of dental transposition. Successful treatment outcomes depend on the patient's age, concern about facial and dental aesthetics, motivation of the patient, functional requirements, type of malocclusion, and the position of the root apices. The present case report is aimed at outlining the orthodontic treatment of a young adult patient diagnosed with bilateral maxillary canine-lateral incisor transposition.


Cardiovascular function and treatment in β-thalassemia major: a consensus statement from the American Heart Association.
This aim of this statement is to report an expert consensus on the diagnosis and treatment of cardiac dysfunction in β-thalassemia major (TM). This consensus statement does not cover other hemoglobinopathies, including thalassemia intermedia and sickle cell anemia, in which a different spectrum of cardiovascular complications is typical. There are considerable uncertainties in this field, with a few randomized controlled trials relating to treatment of chronic myocardial siderosis but none relating to treatment of acute heart failure. The principles of diagnosis and treatment of cardiac iron loading in TM are directly relevant to other iron-overload conditions, including in particular Diamond-Blackfan anemia, sideroblastic anemia, and hereditary hemochromatosis. Heart failure is the most common cause of death in TM and primarily results from cardiac iron accumulation. The diagnosis of ventricular dysfunction in TM patients differs from that in nonanemic patients because of the cardiovascular adaptation to chronic anemia in non-cardiac-loaded TM patients, which includes resting tachycardia, low blood pressure, enlarged end-diastolic volume, high ejection fraction, and high cardiac output. Chronic anemia also leads to background symptomatology such as dyspnea, which can mask the clinical diagnosis of cardiac dysfunction. Central to early identification of cardiac iron overload in TM is the estimation of cardiac iron by cardiac T2* magnetic resonance. Cardiac T2* <10 ms is the most important predictor of development of heart failure. Serum ferritin and liver iron concentration are not adequate surrogates for cardiac iron measurement. Assessment of cardiac function by noninvasive techniques can also be valuable clinically, but serial measurements to establish trends are usually required because interpretation of single absolute values is complicated by the abnormal cardiovascular hemodynamics in TM and measurement imprecision. Acute decompensated heart failure is a medical emergency and requires urgent consultation with a center with expertise in its management. The first principle of management of acute heart failure is control of cardiac toxicity related to free iron by urgent commencement of a continuous, uninterrupted infusion of high-dose intravenous deferoxamine, augmented by oral deferiprone. Considerable care is required to not exacerbate cardiovascular problems from overuse of diuretics or inotropes because of the unusual loading conditions in TM. The current knowledge on the efficacy of removal of cardiac iron by the 3 commercially available iron chelators is summarized for cardiac iron overload without overt cardiac dysfunction. Evidence from well-conducted randomized controlled trials shows superior efficacy of deferiprone versus deferoxamine, the superiority of combined deferiprone with deferoxamine versus deferoxamine alone, and the equivalence of deferasirox versus deferoxamine.


Novel combination strategies to repair the injured mammalian spinal cord.
Due to the varied and numerous changes in spinal cord tissue following injury, successful treatment for repair may involve strategies combining neuroprotection (pharmacological prevention of some of the damaging intracellular cascades that lead to secondary tissue loss), axonal regeneration promotion (cell transplantation, genetic engineering to increase growth factors, neutralization of inhibitory factors, reduction in scar formation), and rehabilitation. Our goal has been to find effective combination strategies to improve outcome after injury to the adult rat thoracic spinal cord. Combination interventions tested have been implantation of Schwann cells (SCs) plus neuroprotective agents and growth factors administered in various ways, olfactory ensheathing cell (OEC) implantation, chondroitinase addition, or elevation of cyclic AMP. The most efficacious strategy in our hands for the acute complete transection/SC bridge model, including improvement in locomotion [Basso, Beattie, Bresnahan Scale (BBB)], is the combination of SCs, OECs, and chondroitinase administration (BBB 2.1 vs 6.6, 3 times more myelinated axons in the SC bridge, increased serotonergic axons in the bridge and beyond, and significant correlation between the number of bridge myelinated axons and functional improvement). We found the most successful combination strategy for a subacute spinal cord contusion injury (12.5-mm, 10-g weight, MASCIS impactor) to be SCs and elevation of cyclic AMP (BBB 10.4 vs 15, significant increases in white matter sparing, in myelinated axons in the implant, and in responding reticular formation and red and raphe nuclei, and a significant correlation between the number of serotonergic fibers and improvement in locomotion). Thus, in two injury paradigms, these combination strategies as well as others studied in our laboratory have been found to be more effective than SCs alone and suggest ways in which clinical application may be developed.


Concerns regarding the financial aspects of kidney transplantation: perspectives of pre-transplant patients and their family members.
African American and non-African American pre-transplant patients' and their families' concerns about the financial costs of kidney transplantation have not been well studied. We conducted structured group interviews among pre-transplant patients (seven African American, five non-African American) and their family members (six African American, five non-African American) to identify their concerns about transplant health insurance coverage, out-of-pocket expenses, and living donor expenses. We reviewed transcribed group audio recordings and identified common discussion themes. African American and non-African American patients and family members expressed uncertainty about which transplant-related costs were covered by health insurance and wanted information about how to choose insurance policies accordingly. Patients were particularly concerned about the impact of pre-existing illness on securing optimal health insurance, while family members wanted information about non-insurance-based financial resources. Both patients and family members expressed concern about paying for immunosuppressant medications and about gradual loss of insurance benefits after transplantation. Both patients and family members also expressed concern about potential financial hardships for living donors. African American and non-African American pre-transplant patients and families expressed a broad range of concerns about transplant health insurance policies, out-of-pocket expenses, non-insurance-based financial resources, and resources to address donors' financial burden. Efforts to improve education and develop more comprehensive transplant insurance policies are needed to facilitate informed decision-making for potential transplant recipients and donors.


Prenatal exposure to PFOS and PFOA in a pregnant women cohort of Catalonia, Spain.
This study was aimed at assessing the prenatal exposure to perfluorooctane sulfonic acid (PFOS) and perfluorooctanoic acid (PFOA) in a cohort of pregnant women living in Reus (Tarragona County, Catalonia, Spain). These chemicals were biomonitored in maternal plasma during the first trimester of pregnancy, at delivery, and in cord blood. The dietary exposure of PFOS and PFOA was estimated by using questionnaires of food frequency and water intake, as well as data on food levels previously reported in the same area. In addition, the exposure through air inhalation and indoor dust ingestion was also calculated. Finally, a physiologically-based pharmacokinetic (PBPK) model was applied in order to establish the prenatal exposure of the fetus/child and to adjust exposure assessment vs. biomonitoring results. Probabilistic calculations of fetal exposure were performed by forward internal dosimetry and Monte-Carlo simulation. Mean plasma levels of PFOA were 0.45, 0.13 and 0.12 ng/mL at the first trimester, at delivery and in cord plasma, while those of PFOS were 2.93, 2.21, and 1.17 ng/mL, respectively. Traces of PFOS were found in all samples in the trimester and at delivery, and almost in all cord blood samples. Transplacental transfers of PFOS and PFOA were estimated to be around 70% and 60%, respectively. A temporal decrease trend in plasma levels of PFOS and PFOA was noticed, when comparing current values with data obtained 10 years ago in the same area. In agreement with many other studies, dietary intake was the main route of exposure to PFOS and PFOA in our cohort of pregnant women. It is an important issue to establish the exposure in critical windows periods such as fetal development to perfluoroalkylated substances, but also to other endocrine disrupting chemicals.


Epsilon Class Glutathione S-Transferase Confers Phoxim Tolerance in the Black Cutworm Agrotis ipsilon (Lepidoptera: Noctuidae).
The black cutworm Agrotis ipsilon is a serious crop pest. Phoxim, an organophosphorus insecticide, has been widely used to control A. ipsilon. When phoxim is extensively applied, the susceptibility of A. ipsilon to insecticide is reduced. However, the mechanisms of tolerance of A. ipsilon to phoxim remain unclear. Herein, we report that an epsilon class glutathione S-transferase, AiGSTE1, confers phoxim tolerance in A. ipsilon. Exposure to a sublethal concentration (LC50) of phoxim caused oxidative stress and activated the transcription of AiGSTe1 genes in A. ipsilon larvae. Recombinant AiGSTE1 expressed in Escherichia coli could metabolize phoxim. Furthermore, E. coli cells overexpressing AiGSTE1 displayed significant tolerance to oxidative stress. Knockdown of AiGSTe1 by RNA interference significantly increased the mortality of A. ipsilon larvae to phoxim. These results demonstrate that AiGSTE1 confers phoxim tolerance in A. ipsilon by metabolizing the insecticide and preventing phoxim-induced oxidative stress.


Mesoscopic fluorescence molecular tomography of reporter genes in bioprinted thick tissue.
Three-dimensional imaging of thick tissue constructs is one of the main challenges in the field of tissue engineering and regenerative medicine. Optical methods are the most promising as they offer noninvasive, fast, and inexpensive solutions. Herein, we report the use of mesoscopic fluorescence molecular tomography (MFMT) to image function and structure of thick bioprinted tissue hosted in a 3-mm-thick bioreactor. Collagen-based tissue assembled in this study contains two vascular channels formed by green fluorescent protein- and mCherry-expressing cells. Transfected live cell imaging enables us to image function, whereas Flash Red fluorescent bead perfusion into the vascular channel allows us to image structure. The MFMT optical reconstructions are benchmarked with classical microscopy techniques. MFMT and wide-field fluorescence microscopy data match within 92% in area and 84% in location, validating the accuracy of MFMT reconstructions. Our results demonstrate that MFMT is a well-suited imaging modality for fast, longitudinal, functional imaging of thick, and turbid tissue engineering constructs.


Long-Term Incidence of Velopharyngeal Insufficiency and Other Sequelae following Uvulopalatopharyngoplasty.
Objective The aim of this study is to (1) assess incidence of long-term velopharyngeal insufficiency (VPI) and (2) determine other sequelae following classic and modified uvulopalatopharyngoplasty (UPPP and mUPPP) for treatment of obstructive sleep apnea (OSA). Data Sources Medline, PubMed, Cochrane Library database. Review Methods A systematic review was performed following standard Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) guidelines. Original research articles reporting on sequelae of UPPP and mUPPP for treatment of OSA, at a mean of 1 year follow-up, were included. Articles were retrieved using keywords UPPP complications and UPPP questionnaire. A random-effects model was used for pooling data. Results A total of 24 studies were included in this review. Complications included VPI (24 studies, n = 191), difficulty swallowing (7 studies, n = 83), taste disturbances (4 studies, n = 10), voice changes (7 studies, n = 46), foreign body (9 studies, n = 427), and dry pharynx (7 studies, n = 150). When pooling all studies together, VPI was the least common sequelae reported in 8.1% of the cases. Foreign body sensation was the most commonly reported sequelae at 31.2%, with difficulty swallowing (17.7%), dry pharynx (23.4%), voice changes (9.5%), and taste disturbances (8.2%) being the most to least likely. Conclusions The long-term effectiveness of UPPP and mUPPP is limited by the number of studies reporting short-term follow-up only. Despite this, long-term data suggest that complications such as VPI are more common than previously reported. Other sequelae, such as foreign body sensation, may be one of the most frequently expected complications after UPPP surgery.


Anxiolytic-antidepressant activity of Withania somnifera glycowithanolides: an experimental study.
The roots of Withania somnifera (WS) are used extensively in Ayurveda, the classical Indian system of medicine, and WS is categorized as a rasayana, which are used to promote physical and mental health, to provide defence against disease and adverse environmental factors and to arrest the aging process. WS has been used to stabilize mood in patients with behavioural disturbances. The present study investigated the anxiolytic and antidepressant actions of the bioactive glycowithanolides (WSG), isolated from WS roots, in rats. WSG (20 and 50 mg/kg) was administered orally once daily for 5 days and the results were compared by those elicited by the benzodiazepine lorazepam (0.5 mg/kg, i.p.) for anxiolytic studies, and by the tricyclic anti-depressant, imipramine (10 mg/kg, i.p.), for the antidepressant investigations. Both these standard drugs were administered once, 30 min prior to the tests. WSG induced an anxiolytic effect, comparable to that produced by lorazepam, in the elevated plus-maze, social interaction and feeding latency in an unfamiliar environment, tests. Further, both WSG and lorazepam, reduced rat brain levels of tribulin, an endocoid marker of clinical anxiety, when the levels were increased following administration of the anxiogenic agent, pentylenetetrazole. WSG also exhibited an antidepressant effect, comparable with that induced by imipramine, in the forced swim-induced 'behavioural despair' and 'learned helplessness' tests. The investigations support the use of WS as a mood stabilizer in clinical conditions of anxiety and depression in Ayurveda.


Desensitization to substance P-induced vasodilation in vitro is not shared by endogenous tachykinin neurokinin A.
Two mammalian tachykinins, substance P (SP) and neurokinin A (NKA), were measured by radioimmunoassay in canine cephalic blood vessels and tested for their vasoactivity in vitro. Levels of immunoreactive SP were approximately 2-3 times greater than those of immunoreactive NKA in common carotid, basilar, and middle cerebral arteries. Both endogenous tachykinins relaxed precontracted segments of common carotid and basilar arteries in a dose-dependent manner with an EC50 of 8.9 X 10(-11) M and 7 X 10(-10) M, respectively, when added cumulatively. Relaxation was endothelial dependent for both substances and not blocked or enhanced by pretreatment with indomethacin, propranolol, lithium chloride, or atropine. Neither SP nor NKA released 3H-inositol phosphates from phospholipid membranes of canine carotid segments after preincubation with 3H-inositol. SP but not NKA or the C-terminal fragments SP(4-11) caused desensitization to subsequent additions of itself but not to the relaxation induced by sodium nitroprusside, calcitonin gene-related peptide, or bradykinin. These studies demonstrate that at least 2 peptides derived from beta-preprotachykinin are contained within cephalic blood vessels and that these products share similar vasoactive properties but differ in their ability to desensitize vascular tachykinin receptors.


Potential for power deposition conformability using reflected-scanned planar ultrasound.
Specialized heating devices for the simultaneous delivery of hyperthermia and ionizing radiation are being developed by several investigators in an effort to increase thermal radiosensitization in clinical treatment. One particular device is the SURLAS (Scanning Ultrasound Refléctor-Linear Array System), which was designed specifically to operate concomitantly with medical linear accelerators. The technical feasibility of the SURLAS has been demonstrated, and a design optimization study has been performed. The main objective of this paper is to demonstrate the potential for power deposition conformability of the SURLAS. This has been done using a thermographic technique which provides qualitative, high spatial-resolution measurements of power deposition distributions. The technique consists of normally insonating one surface of a 1 cm layer of a Polyurethane phantom while the temperature field on the opposite air-exposed surface is recorded using an infrared camera during the first few minutes after power insult. The thermal fields measured in this way are good qualitative estimates of relative power deposition. To demonstrate conformability, a region of 10 cm (the length of the array) by 12 cm (the scanning distance) on the air-exposed phantom surface was divided into 24 sectors (24 subregions with independent power control). Each sector was 2.5 x 2 cm across and long the scanning direction respectively. Several sector insonation patterns were synthesized in an open-loop fashion by properly adjusting power levels to each of the elements of an array as a function of reflector position as the reflector was scanned continuously in a reciprocating fashion at a constant speed. The array was made of a single piezoelectric crystal with resonant frequency of 2.2 MHz and electrically segmented into four 2.5 x 2.5 cm elements. The reflector was made of a 0.5 mm thick Brass plate. Sufficient power was supplied to the array to induce peak temperature elevations of about 10 degrees C in 60s at a scanning speed of 2.4 cm/s. The results show that measured relative power deposition patterns agreed well with programmed insonation patterns demonstrating that the SURLAS possesses great potential for power conformability, and thus, for temperature feedback power deposition control.


Comparison between prospective and retrospective evaluation of Crohn's disease activity index.
The Crohn's disease activity index (CDAI) is the most widely used measure of clinical disease activity in patients entered into clinical trials. The prospective nature of the CDAI calculation precludes its use as a clinical assessment tool. We compared the retrospective evaluation of the CDAI with the prospective evaluation in a heterogeneous patient population of 100 patients with Crohn's disease. The correlation between the two assessment methods was good with an r-value of 0.84 (p < 0,0001). There was a tendency of patients with a high retrospective CDAI to have a lower prospective CDAI which is explained by intention to treat. This study shows that a retrospective assisted evaluation of the CDAI is as accurate as the traditional prospective evaluation.


Protein conformation of potato (Solanum tuberosum) lectin determined by circular dichroism.
The structure of potato (Solanum tuberosum) lectin, which is a hydroxyproline-rich glycoprotein, has been investigated by circular dichroism. The spectra of the native lectin, and of the oxidized, reduced and carboxymethylated and deglycosylated derivatives were examined, as was a hydroxyproline-rich glycopeptide and its deglycosylated derivative. It is concluded that the lectin contains about 35% polyproline II conformation, 34% type II beta-turn and 31% irregular conformation. No indications were found for the presence of alpha-helix or beta-sheet conformations. The polyproline II conformation is heat-stable, but is markedly destabilized by deglycosylation. The type II beta-turn is destabilized by cleavage of disulphide bonds.


Electrochemical advanced oxidation process using DiaChem electrodes.
The electrochemical advanced oxidation process (EAOP) using boron doped diamond (DiaChem, registered trademark of Condias GmbH) has been studied for wastewater treatment and drinking water disinfection. DiaChem electrodes consist of preferentially metallic base materials coated with a conductive polycrystalline diamond film by hot-filament chemical vapour deposition. They exhibit high overpotential for water electrolysis as well as high chemical inertness and extended lifetime. In particular the high overpotential for water decomposition opens the widest known electrochemical window, allowing the energy efficient production of hydroxyl radicals directly from aqueous solutions. The hydroxyl radicals on the other hand are effectively used for the oxidation of pollutants. The EAOP using DiaChem electrodes thus facilitates the direct and, if necessary, complete decomposition of even hazardous or persistent pollutants in different wastewaters. Current efficiencies of more than 90%, also without the use of additives for hydroxyl radical generation, have been demonstrated. Additionally, for drinking water preparation diamond electrodes facilitate disinfection with and without the support of chlorine.


Characterization of five different proteins produced by alternatively spliced mRNAs from the human cAMP-specific phosphodiesterase PDE4D gene.
We have isolated and characterized complete cDNAs for two isoforms (HSPDE4D4 and HSPDE4A5) encoded by the human PDE4D gene, one of four genes that encode cAMP-specific rolipram-inhibited 3',5'-cyclic nucleotide phosphodiesterases (type IVPDEs; PDE4 family). The HSPDE4D4 and HSPDE4D5 cDNAs encode proteins of 810 and 746 amino acids respectively. A comparison of the nucleotide sequences of these two cDNAs with those encoding the three other human PDE4D proteins (HSPDE4D1, HSPDE4D2 and HSPDE4D3) demonstrates that each corresponding mRNA transcript has a unique region of sequence at or near its 5'-end, consistent with alternative mRNA splicing. Transient expression of the five cDNAs in monkey COS-7 cells produced proteins of apparent molecular mass under denaturing conditions of 68, 68, 95, 119 and 105 kDa for isoforms HSPDE4D1-5 respectively. Immunoblotting of human cell lines and rat brain demonstrated the presence of species that co-migrated with the proteins produced in COS-7 cells. COS-cell-expressed and native HSPDE4D1 and HSPDE4D2 were found to exist only in the cytosol, whereas HSPDE4D3, HSPDE4D4 and HSPDE4D5 were found in both cytosolic and particulate fractions. The IC50 values for the selective PDE4 inhibitor rolipram for the cytosolic forms of the five enzymes were similar (0.05-0.14 microM), whereas they were 2-7-fold higher for the particulate forms of HSPDE4D3 and HSPDE4D5 (0.32 and 0.59 microM respectively), than for the corresponding cytosolic forms. Our data indicate that the N-terminal regions of the HSPDE4D3, HSPDE4D4 and HSPDE4D5 proteins, which are derived from alternatively spliced regions of their mRNAs, are important in determining their subcellular localization, activity and differential sensitivity to inhibitors.


Psychiatric involvement in adult patients with mitochondrial disease.
Mitochondrial disorders are caused by impairment of the respiratory chain. Psychiatric features often represent part of their clinical spectrum. However, the real incidence of psychiatric disorders in these diseases is unknown. The aim of this study was to evaluate psychiatric involvement in a group of patients with mitochondrial disorders and without already diagnosed mental illness. Twenty-four patients with mitochondrial disorder and without already diagnosed mental diseases have been studied by means of the mini-international neuropsychiatric interview (MINI) and the newcastle mitochondrial diseases adult scale (NMDAS). In patients with mitochondrial disease, psychiatric conditions were far more common than in general Italian population (about 60 vs. 20-25%), and included major depression, agoraphobia and/or panic disorder, generalized anxiety disorder, social anxiety disorder, psychotic syndromes. Psychiatric involvement did not seem to depend on disease progression. Large, multicenter studies are strongly needed to better characterize the natural history of mitochondrial disorders and of their psychiatric involvement. Moreover, the possibility of mitochondrial diseases should be considered in patients with psychiatric diseases. Finally, we encourage psychiatric evaluation as a routinary approach to mitochondrial patients.


Access to quality: evaluation of the allocation of deceased donor kidneys for transplantation.
Disparities in both access to the kidney transplant waiting list and waiting times for transplant candidates have been extensively documented with regard to ethnicity, gender, socioeconomic factors, and region. However, the issue of access to equivalent quality organs has garnered less attention. The principal aim of this study was to determine whether certain patient populations were more likely to receive lower quality organs. This was a retrospective cohort study of all deceased-donor adult renal transplant recipients in the United States from 1996 to 2002 (n = 45,832). Using previously reported categorization of donor quality (I to V), the propensity of transplant recipients to receive lower-quality kidneys in a cumulative logit model was evaluated. Older patients were progressively more likely to receive lower-quality organs (age > or = 65 yr, odds ratio [OR] = 2.1, P < 0.01) relative to recipients aged 18 to 24 yr. African American and Asian recipients had a greater likelihood of receiving lower-quality organs relative to non-Hispanic Caucasians. Regional allocation networks were highly variable with regard to donor quality. Neither recipient gender (OR = 1.00, P = 0.81) nor patient's primary diagnosis were associated with donor quality. Findings suggest that disparities in the quality of deceased donor kidneys to transplant recipients exist among certain patient groups that have previously documented access barriers. The extent to which these disparities are in line with broad policies of equity and potentially modifiable will have to be examined in the context of allocation policy.


Assessing relative humidity dependent photoacoustics to retrieve mass accommodation coefficients of single optically trapped aerosol particles.
Photoacoustic spectroscopy is widely used to measure the light absorption of aerosols. However, the impact of key factors such as the effect of relative humidity and mass exchange on photoacoustic measurements are still poorly understood. We assess such measurement biases and their physical origin by analysing the photoacoustic signal of single tetraethylene glycol (TEG) particles at varying relative humidities. Our results show a decrease in the photoacoustic signal at elevated relative humidities for small particles (0.8-1.5 μm), while for larger sizes (2.2-3.2 μm) the trend is reversed. We model the photoacoustic signal to interpret the observed behaviour in terms of mass and heat flux contribution. The single particle photoacoustic signal analysis presented in this paper additionally allows for the retrieval of the mass accommodation coefficient. Fitting our experimental data to the theoretical model reveals values of αM ≈ 0.02-0.005 for water on TEG in the temperature range 295-309 K.


Failure of neutrophil migration to infectious focus and cardiovascular changes on sepsis in rats: Effects of the inhibition of nitric oxide production, removal of infectious focus, and antimicrobial treatment.
Recently, we demonstrated that mice under lethal sepsis present failure of neutrophil migration (FNM) to infectious focus, which is mediated by nitric oxide. The aims of the present study were to investigate whether FNM is also observed in severe sepsis induced by cecal ligation and puncture in rats and the effects of the prevention of nitric oxide production and of the elimination of the infectious focus through peritoneal lavage or by antimicrobial treatment on FNM and disease outcome. Rats were submitted to several septic stimuli (low, moderate, and severe) by cecal ligation and puncture. Severe septic stimulus animals presented FNM to the peritoneal cavity that was accompanied by large numbers of bacteria in the peritoneal cavity, blood, and liver and lung tissues; high cytokines (tumor necrosis factor alpha, interleukin [IL] 1beta, IL-6, cytokine-induced neutrophil chemoattractant 1, and IL-10) concentrations in the infection site, sera, and lung tissues; marked hypotension; and high mortality rate. The exhaustive lavage of the peritoneal cavity to reduce the infectious focus did not ameliorate the disease outcome. The association of lavage procedure with aminoguanidine treatment re-established neutrophil migration, but only delayed the death of the animals. In contrast, the antimicrobial treatment of severe septic stimulus animals with sulfamethoxazole and trimethoprim significantly improved the survival rate of the severe septic stimulus but did not re-establish neutrophil migration. However, the association of aminoguanidine plus sulfamethoxazole and trimethoprim brought about a significant increase in the survival rate and re-established neutrophil migration to infectious focus; reduced the colony-forming units in the peritoneal cavity, blood, and lung tissues; and caused an improvement in the cardiovascular performance. The results showed, for the first time, that the pharmacological prevention of FNM to the infectious focus associated with the antimicrobial therapy could be a new beneficial strategy for the treatment of sepsis syndrome.


[Complication of arthroscopy of the knee].
Arthroscopy is an elementary surgical procedure in the management of the knee. Due to its minimal invasiveness the complications are not frequent. However, because of continuously increasing number of arthroscopies being performed worldwide (circa 1,000,000 per year), the total number of complications is significant. More frequent complications accompany particularly more complex and difficult procedures. The percentage of complications after arthroscopy oscilates about 2%. The most common complications are: intraarticular damage, neurological injury, vascular injury, compartment syndrome, instrument breakage, postoperative pain, haemarthrosis, synovitis, thromboembolism, infection, synovial fistula, complex regional pain syndrome 1, infrapatellar contracture syndrome. Current view on the complications of arthroscopy is presented in this paper basing on contemporary literature.


ROP18 is a key factor responsible for virulence difference between Toxoplasma gondii and Neospora caninum.
Toxoplasma gondii (T. gondii) and Neospora caninum (N. caninum) are both obligate intracellular protozoan parasites and share many common morphological and biological features. Despite these similarities the two parasites differ dramatically in virulence in mice, but the factors involved in virulence differences between the two parasites remain unknown. A secreted serine-threonine kinase called rhoptry protein 18 (ROP18) was identified to play a crucial role on virulence differences among different T. gondii clonal lineages. Intriguingly, we found that ROP18 in Nc1 strain of N. caninum (NcROP18) is a pseudogene due to several interrupting stop codons in the sequence in our previous studies. We assume that the difference of ROP18 leads to virulence difference between T. gondii and N. caninum. We constructed a transgenic N. caninum Nc1 stain by transfecting the TgROP18 from the T. gondii RH strain. Phenotype and virulence assays showed that the expression of TgROP18 in N. caninum did not affect the motility and cell invasion, but resulted in a significant increase in intracellular parasite proliferation and virulence in mice. Immunity-Related GTPase (IRG) phosphorylation assay showed that the transgenic parasite Nc1-TgROP18 was able to phosphorylate IRGs as T. gondii did. The present study indicated that the ROP18 plays a crucial role in virulence of the closely related parasites T. gondii and N. caninum and it is indeed a key factor responsible for the virulence difference between T. gondii and N. caninum.


Endothelin induced contractility of stellate cells from normal and cirrhotic rat liver: implications for regulation of portal pressure and resistance.
Hepatic stellate cells are similar to tissue pericytes and have been shown to be contractile. In this study, we examined the effects of known mediators of stellate cell contraction on portal pressure in rat livers after carbon tetrachloride induced injury (including cirrhosis) and investigated the contractility of stellate cells as a function of liver injury. Sarafotoxin S6C, an endothelin B (ETB) receptor agonist, had minor effects on portal pressure when perfused into normal livers at concentrations known to elicit stellate cell contraction (2 nmol/L). In contrast, both endothelin-1 (2 nmol/L) and angiotensin II (8.6 nmol/L) caused a rapid and pronounced rise in portal pressure. The effects of sarafotoxin S6C (a potent stellate cell contractile agonist) on portal pressure were greater in cirrhotic than normal liver, whereas those of angiotensin II were unchanged after liver injury. Endothelin-1 and sarafotoxin S6C induced contractility of stellate cells increased in proportion to the degree of liver injury. Contractility was greatest for stellate cells isolated from cirrhotic livers, a population of cells that displayed the most activated phenotype, as measured by immunoblot of smooth muscle alpha actin. Autoradiography of cirrhotic livers after perfusion with 125I-endothelin-1 revealed binding to cells in both sinusoidal spaces and collagenous fibrotic bands, consistent with known locations of stellate cells. Finally, the mixed endothelin-A (ETA) and ETB receptor antagonist, bosentan, reduced portal pressure in portal hypertensive animals, consistent with its inhibitory effect on stellate cell contraction. We conclude that stellate cell contractility increases with progressive liver injury and is proportional to the degree of stellate cell activation, becoming most prominent in the cirrhotic liver. Endothelin-stimulated contraction of stellate cells in cirrhotic liver may contribute to increased intrahepatic resistance and portal pressure.


The making of a mammalian peroxisome, version 2.0: mitochondria get into the mix.
A recent report from the Laboratory of Heidi McBride (McGill University) presents a role for mitochondria in the de novo biogenesis of peroxisomes in mammalian cells. Peroxisomes are essential organelles responsible for a wide variety of biochemical functions, from the generation of bile to plasmalogen synthesis, reduction of peroxides, and the oxidation of very-long-chain fatty acids. Like mitochondria, peroxisomes proliferate primarily through growth and division of pre-existing peroxisomes. However, unlike mitochondria, peroxisomes do not fuse; further, and perhaps most importantly, they can also be born de novo, a process thought to occur through the generation of pre-peroxisomal vesicles that originate from the endoplasmic reticulum. De novo peroxisome biogenesis has been extensively studied in yeast, with a major focus on the role of the ER in this process; however, in the mammalian system this field is much less explored. By exploiting patient cells lacking mature peroxisomes, the McBride laboratory now assigns a role to ER and mitochondria in de novo mammalian peroxisome biogenesis by showing that the formation of immature pre-peroxisomes occurs through the fusion of Pex3-/Pex14-containing mitochondria-derived vesicles with Pex16-containing ER-derived vesicles.


Intranasal insulin decreases circulating cortisol concentrations during early sleep in elderly humans.
Aging is associated with increases in hypothalamic-pituitary-adrenal (HPA) axis activity that can predispose to metabolic and cognitive impairments. We investigated in elderly and young subjects whether intranasal insulin administration to the human brain reduces early-sleep nadir concentrations of adrenocorticotropin and cortisol, that is, indicators of baseline HPA axis activity. In within-subject comparisons, intranasal insulin (160 IU) or placebo was administered to 14 elderly (mean age 70.0 years) and 30 young (23.6 years) healthy subjects before bedtime. Sleep was polysomnographically assessed and blood samples were repeatedly collected. Elderly compared with young participants displayed increased early-sleep cortisol concentrations (p < 0.04) and reductions in slow wave and REM sleep (p < 0.001). Insulin administration reduced cortisol levels between 2300 hours and 0020 hours in the elderly (p = 0.03) but not young participants (p = 0.56; p = 0.003 for interaction). Findings indicate that central nervous insulin acts as an inhibitory signal in basal HPA axis activity regulation and suggest that intranasal insulin may normalize sleep-associated stress axis activity in older age.


pckA-deficient Mycobacterium bovis BCG shows attenuated virulence in mice and in macrophages.
Phosphoenolpyruvate carboxykinase (PEPCK) catalyses the reversible decarboxylation and phosphorylation of oxaloacetate (OAA) to form phosphoenolpyruvate (PEP). In this study, the regulation of the PEPCK-encoding gene pckA was examined through the evaluation of green fluorescent protein expression driven by the pckA promoter. The results showed that pckA was upregulated by acetate or palmitate but downregulated by glucose. Deletion of the pckA gene of Mycobacterium bovis BCG led to a reduction in the capacity of the bacteria to infect and survive in macrophages. Moreover, mice infected with DeltapckA BCG were able to reduce the bacterial load much more effectively than mice infected with the parental wild-type bacteria. This attenuated virulence was reflected in the degree of pathology, where granuloma formation was diminished both in numbers and degree. The data indicate that PEPCK activity is important during establishment of infection. Whether its role is in the gluconeogenic pathway for carbohydrate formation or in the conversion of PEP to OAA to maintain the TCA cycle remains to be determined.


[Evaluation of the role of radical mastectomy, postoperative radiation and chemotherapy in stage IIb (TI-2NIMO) breast cancer].
The paper deals with the first results of a joint study carried out at a number of oncological institutions of the RSFSR. It was concerned with evaluation of the role of Holstead's radical operation, postoperative radiation and chemotherapy in patients with stage IIb (TI, 2NIMO) breast cancer. 822 history cases were analysed. Recurrences and metastases were detected in 12.6% of surgical patients (regional--3.2%, distant metastases--in 9.4%). A slightly lower frequency of recurrences and metastases (11.4%) was recorded in group II (Holstead's operation and postoperative radiation treatment). The lowest frequency was in group III: recurrences were 2.1 times less frequent than in study group 1 and 1.9 times less frequent than in group II. As for metastasis incidence, the figures were 2.2 and 2.1, respectively. The data obtained serve as a basement for further development of joint studies on the use of combined therapy of stage IIb breast cancer including chemotherapy.


Real-Time Emergency Department Electronic Notifications Regarding High-Risk Patients: A Systematic Review.
Background: To systematically review evidence on the feasibility and efficacy of real-time electronic notifications about patients at high risk of emergency department (ED) recidivism. Methods: Eight electronic databases were searched for empirical studies of real-time ED-based electronic tools, identifying adult patients at high risk of frequent utilization. Study selection and data extraction were performed independently by two reviewers. Qualitative data synthesis and assessment of strength of evidence were conducted through consensus discussion. Results: Of 2,256 records found through the search, 210 were duplicates, 2,004 were excluded based on abstract review, and 31 were excluded after full text review. The final sample consisted of 10 studies described in 11 articles describing the effect of real-time ED-based electronic notifications for high-risk patients. Three were randomized controlled trials (RCTs). All notifications were based on prespecified markers of risk. Seven studies integrated complex care plans into the electronic health record. Effect on ED use and length of stay (LOS) was mixed: nine studies reported decreased ED use, although results were statistically significant in only three studies; for LOS, one study reported a statistically significant reduction. Impact on cost and financial metrics was promising, with three (of three studies reporting this metric) showing improved organizational financial metrics. Three RCTs reported a reduction in opioid prescriptions. Conclusions: Real-time electronic notifications of ED providers regarding patients at high risk of ED recidivism are feasible. They may help reduce resource utilization and costs. Large knowledge gaps remain regarding patient- and provider-centered outcomes.


Attenuation of the oestrogen positive feedback mechanism with the age in postmenopausal women.
It has been reported that the positive feedback mechanism of oestrogens and progesterone is preserved, although attenuated, in late postmenopausal years. Whether this is also true for the positive feedback effect of oestrogens alone has not been investigated. Prospective intervention study. Thirty healthy postmenopausal women. The women were divided into three groups according to the years since menopause (group I: 2-8 years, group II: 9-17 years, group III: 18-25 years). They were studied during a period of 41 days. Two acute experiments (EP) of exogenous oestradiol, given via skin patches, were performed from days 1 to 7 (EP1) and from days 35 to 41 (EP2) to induce an LH surge. Between the two experiments (days 7-34), oestradiol was given at the dose of 100 μg every 3 days, while oral progesterone was added from day 21 to day 34 in order to simulate a luteal phase. Blood samples were taken every 6 h during EP1 and EP2 as well as on days 8, 13, 20, 21, 27 and 34. FSH, LH, oestradiol and progesterone were measured in all blood samples. An LH surge occurred as a result of the oestradiol positive feedback mechanism in group I and in group II, in both EP1 and EP2. Peak LH values during the surge were significantly lower in group II than in group I in both experiments. None of the patients in group III displayed an LH surge. These results demonstrate for the first time a gradual attenuation of the pituitary response to oestrogenic provocation over a certain period following the menopause, with complete abolition after 20 years. It is suggested that the reserves of pituitary gonadotrophs diminish with age.


More Relaxation by Deep Breath on Methacholine- Than on Exercise-Induced bronchoconstriction during the Routine Testing of Asthmatic Children.
Deep inspiration (DI) dilates normal airway precontracted with methacholine. The fact that this effect is diminished or absent in asthma could be explained by the presence of bronchial inflammation. The hypothesis was tested that DI induces more relaxation in methacholine induced bronchoconstriction-solely determined by the smooth muscle contraction-than in exercise induced bronchoconstriction, which is contributed to by both smooth muscle contraction and airway wall inflammation. The respiratory conductance (Grs) response to DI was monitored in asthmatic children presenting a moderately positive airway response to challenge by methacholine (n = 36) or exercise (n = 37), and expressed as the post- to pre-DI Grs ratio (GrsDI). Both groups showed similar change in FEV1 after challenge and performed a DI of similar amplitude. GrsDI however was significantly larger in methacholine than in exercise induced bronchoconstriction (p < 0.02). The bronchodilatory effect of DI is thus less during exercise- than methacholine-induced bronchoconstriction. The observation is consistent with airway wall inflammation-that characterizes exercise induced bronchoconstriction-rendering the airways less responsive to DI. More generally, it is surmised that less relief of bronchoconstriction by DI is to be expected during indirect than direct airway challenge. The current suggestion that airway smooth muscle constriction and airway wall inflammation may result in opposing effects on the bronchomotor action of DI opens important perspective to the routine testing of asthmatic children. New crossover research protocols comparing the mechanical consequences of the DI maneuver are warranted during direct and indirect bronchial challenges.


Pharmacokinetics of 18F-labeled fluconazole in rabbits with candidal infections studied with positron emission tomography.
[4-18F]Fluconazole was used to measure the pharmacokinetics of fluconazole in normal and infected animals. The biodistribution of fluconazole was determined after administration of the 18F-tracer with/without a pharmacological dose of unlabeled drug by radioactivity measurements on excised tissues. In normal rabbits and rabbits with candidal infection of the thigh, tissue concentrations of drug were determined by serial positron emission tomographic imaging. In rats, coinjection of tracer quantities of [4-18F]fluconazole with a pharmacological dose of unlabeled drug resulted in a relatively uniform distribution of radioactivity in most organs, whereas, when the 18F-tracer was injected alone, spleen, muscle and heart accumulation was decreased and liver accumulation was increased. In rabbits, this effect was less pronounced. Early accumulation of [4-18F]fluconazole was greater in infected muscle. The areas under the 2-hr uptake curves were 4.30 and 6.05 micrograms.hr.ml-1 for normal and infected tissue. A mathematical model was used to summarize the kinetics of fluconazole in normal and infected muscle. The model hypothesizes that fluconazole is compartmentalized in blood and tissue, with rate constants describing the transition between compartments. Direct measurement of the partition coefficient of fluconazole in muscle and predictions of the kinetic model were in close agreement, suggesting that fluconazole enters muscle via a passive transport mechanism. Transport rates of fluconazole, into (Kin) and out of tissue (kout), were increased in infected compared with normal muscle, possibly due to increased capillary permeability (Kin: 0.064 +/- 0.001 vs. 0.0270 +/- 0.0002, kout: 0.063 +/- 0.002 vs. 0.035 +/- 0.001).


[Individual approaches to monitoring and predicting rehabilitation recovery of stroke patients].
The article based on the results of our own research shows the importance of the development of objective options for the individual forecast to recovery of stroke patients, and control of the rehabilitation process. We examined 44 patients with stroke and 35 healthy volunteers as a control group. All patients and healthy volunteers underwent a neurological examination, neurophysiological testing, and extended neuropsychological survey. We evaluated the results of rehabilitation depending on the data obtained in the survey. The study had found that cognitive impairment reaching moderate severity can be regarded as a poor prognostic factor for recovery of hand function. The clear direct relationship was shown between the impaired hand (with decreased strength and agility) and severity of frontal-subcortical dysfunction, and visual-spatial disorders. We found that elongated latent periods of auditory evoked potentials, P300 latency prolongation (beyond the age norm) were inherent mainly for patients with moderate cognitive impairment with postural disorders and high risk of falls. These patients, according to our data, had almost no improvement of motor functions versus patients who had moderate cognitive impairment, in which the parameters of evoked potentials of the brain were within the age norm. Evoked potentials of the brain in conjunction with the results of the extended neuropsychological testing may be an objective indicator for prediction and possibly control of the dynamics of recovery of cognitive and motor functions in the rehabilitation of stroke patients.


The C-terminal region of Bfl-1 sensitizes non-small cell lung cancer to gemcitabine-induced apoptosis by suppressing NF-κB activity and down-regulating Bfl-1.
Gemcitabine is used to treat several cancers including lung cancer. However, tumor cells often escape gemcitabine-induced cell death via various mechanisms, which include modulating bcl-2 family members and NF-κB activation. We previously reported that the C-terminal region of Bfl-1 fused with GFP (BC) is sufficient to induce apoptosis in 293T cells. In the present study, we investigated the anti-tumor effect of combined BC gene therapy and gemcitabine chemotherapy in vitro and in vivo using non-small cell lung cancer cell lines and a xenograft model. Cell lines were resistant to low dose gemcitabine (4-40 ng/ml), which induced NF-κB activation and concomitant up-regulation of Bfl-1 (an NF-κB-regulated anti-apoptotic protein). BC induced the apoptosis of A549 and H157 cells with caspase-3 activation. Furthermore, co-treatment with BC and low dose gemcitabine synergistically and efficiently induced mitochondria-mediated apoptosis in these cells. When administered alone or with low dose gemcitabine, BC suppressed NF-κB activity, inhibited the nuclear translocation of p65/relA, and down-regulated Bfl-1 expression. Furthermore, direct suppression of Bfl-1 by RNA interference sensitized cells to gemcitabine-induced cell death, suggesting that Bfl-1 importantly regulates lung cancer cell sensitivity to gemcitabine. BC and gemcitabine co-treatment also showed a strong anti-tumor effect in a nude mouse/A549 xenograft model. These results suggest that lung cancer cells become resistant to gemcitabine via NF-κB activation and the subsequent overexpression of Bfl-1, and that BC, which has both pro-apoptotic and NF-κB inhibitory effects, could be harnessed as a gene therapy to complement gemcitabine chemotherapy in non-small cell lung cancer.


The relationship between the PD-L1 expression of surgically resected and fine-needle aspiration specimens for patients with pancreatic cancer.
Recently, therapeutic antibodies against programmed death-1/programmed death-ligand 1 (PD-1/PD-L1) have shown promising clinical results for several solid tumors, including pancreatic cancer. In this study, we evaluated the relationship between the PD-L1 expression of surgical resected and fine-needle aspiration (FNA) specimens for patients with pancreatic cancer. Of 121 patients who underwent endoscopic ultrasound-guided (EUS)-FNA before surgery for pancreatic cancer in an academic center, the 94 (78%) with adequate FNA specimens for a histological evaluation were retrospectively analyzed. All the patients had undergone upfront surgery without any chemotherapy or radiotherapy. We performed immunohistochemistry (IHC) staining to investigate the PD-L1 expression in both resected and FNA specimens. The positive-stained cells were counted, and their percentage was used for the investigation. Of the 94 patients, 16 (17%) and 11 (10%) were defined as positive on resected cancer specimens using cutoff points of 5% and 10% positively stained cancer cell counts, respectively. The concordance rates for the positive frequency of PD-L1 expression between resected and FNA specimens were 44% (7/16) and 55% (6/11) when the positivity was set to ≥ 5% and ≥ 10%, respectively. The concordance rates for the negative frequency of PD-L1 expression between two specimens were 97% (76/78) and 99% (82/83) when the positivity was set to ≥ 5% and ≥ 10%, respectively. Approximately, half of the patients with PD-L1 expression positive and almost all the patients with PD-L1 expression negative could be diagnosed on FNA specimens.


Quantitative susceptibility mapping as an indicator of subcortical and limbic iron abnormality in Parkinson's disease with dementia.
Late stage Parkinson's disease (PD) patients were commonly observed with other non-motor comorbidities such as dementia and psychosis. While abnormal iron level in the substantia nigra was clinically accepted as a biomarker of PD, it was also suggested that the increased iron deposition could impair other brain regions and induce non-motor symptoms. A new Magnetic Resonance Imaging (MRI) called Quantitative Susceptibility Mapping (QSM) has been found to measure iron concentration in the grey matter reliably. In this study, we investigated iron level of different subcortical and limbic structures of Parkinson's disease (PD) patients with and without dementia by QSM. QSM and volumetric analysis by MRI were performed in 10 PD dementia (PDD) patients (73 ± 6 years), 31 PD patients (63 ± 8 years) and 27 healthy controls (62 ± 7 years). No significant differences were observed in the L-Dopa equivalent dosage for the two PD groups (p = 0.125). Putative iron content was evaluated in different subcortical and limbic structures of the three groups, as well as its relationship with cognitive performance. One-way ANCOVA with FDR adjustment at level of 0.05, adjusted for age and gender, showed significant group differences for left and right hippocampus (p = 0.015 & 0.032, respectively, BH-corrected for multiple ROIs) and right thalamus (p = 0.032, BH-corrected). Post-hoc test with Bonferroni's correction suggested higher magnetic susceptibility in PDD patients than healthy controls in the left and right hippocampus (p = 0.001 & 0.047, respectively, Bonferroni's corrected), while PD patients had higher magnetic susceptibility than the healthy controls in right hippocampus and right thalamus (p = 0.006 & 0.005, respectively, Bonferroni's corrected). PDD patients also had higher susceptibility than the non-demented PD patients in left hippocampus (p = 0.046, Bonferroni's corrected). The magnetic susceptibilities of the left and right hippocampus were negatively correlated with the Mini-Mental State Examination score (r = -0.329 & -0.386, respectively; p < 0.05). This study provides support for iron accumulation in limbic structures of PDD and PD patients and its correlation with cognitive performance, however, its putative involvement in development of non-motor cognitive dysfunction in PD pathogenesis remains to be elucidated.


Effects of Pomegranate Juice Supplementation on Oxidative Stress Biomarkers Following Weightlifting Exercise.
The aim of this study was to test the hypothesis that pomegranate juice supplementation would blunt acute and delayed oxidative stress responses after a weightlifting training session. Nine elite weightlifters (21.0 ± 1 years) performed two Olympic-Weightlifting sessions after ingesting either the placebo or pomegranate juice supplements. Venous blood samples were collected at rest and 3 min and 48 h after each session. Compared to the placebo condition, pomegranate juice supplementation attenuated the increase in malondialdehyde (-12.5%; p < 0.01) and enhanced the enzymatic (+8.6% for catalase and +6.8% for glutathione peroxidase; p < 0.05) and non-enzymatic (+12.6% for uric acid and +5.7% for total bilirubin; p < 0.01) antioxidant responses shortly (3 min) after completion of the training session. Additionally, during the 48 h recovery period, pomegranate juice supplementation accelerated (p < 0.05) the recovery kinetics of the malondialdehyde (5.6%) and the enzymatic antioxidant defenses compared to the placebo condition (9 to 10%). In conclusion, supplementation with pomegranate juice has the potential to attenuate oxidative stress by enhancing antioxidant responses assessed acutely and up to 48 h following an intensive weightlifting training session. Therefore, elite weightlifters might benefit from blunted oxidative stress responses following intensive weightlifting sessions, which could have implications for recovery between training sessions.


Mental illness-related stigma among probation officers.
Probation officers tend to hold stigmatising attitudes towards individuals on their caseload with mental illnesses. This can result in poor outcomes for justice-involved people with mental illnesses. Though anti-stigma interventions in the general public and among other criminal-legal professionals hold promise, these interventions have not been tested among probation officers. To measure levels of stigma towards mental illnesses among probation officers before and after a brief online training related to mental illness. Two hundred and seventy-five probation and parole officers in one Midwest state completed a three-module online training programme designed to increase knowledge and skills for working with clients with mental illnesses. Officers completed a pre- and post-training assessment of level of stigmatising attitudes towards individuals with mental illnesses. Officer levels of stigma statistically significantly decreased from pre-test (8.22) to post-test (7.37; p < 0.001, t = 6.13). Nevertheless, pre-training stigma scores were the strongest predictor of stigma levels at post-test (p < 0.001, b = 0.90). While basic training can significantly decrease mental disorder stigmatising scores for probation officers, the practical effect of this brief online experience was very small. Nevertheless, the fact that such brief and generalised input can have impact at all is encouraging. Implementation could be a useful preliminary to improving the quality of probation supervision of clients with mental illnesses.


The short arm of laminin gamma2 chain of laminin-5 (laminin-332) binds syndecan-1 and regulates cellular adhesion and migration by suppressing phosphorylation of integrin beta4 chain.
The proteolytic processing of laminin-5 at the short arm of the gamma2 chain (gamma2sa) is known to convert this laminin from a cell adhesion type to a motility type. Here, we studied this mechanism by analyzing the functions of gamma2sa. In some immortalized or tumorigenic human cell lines, a recombinant gamma2sa, in either soluble or insoluble (coated) form, promoted the adhesion of these cells to the processed laminin-5 (Pr-LN5), and it suppressed their migration stimulated by serum or epidermal growth factor (EGF). Gamma2sa also suppressed EGF-induced tyrosine phosphorylation of integrin beta4 and resultant disruption of hemidesmosome-like structures in keratinocytes. Gamma2sa bound to syndecan-1, and this binding, as well as its cell adhesion activity, was blocked by heparin. By analyzing the activities of three different gamma2sa fragments, the active site of gamma2sa was localized to the NH(2)-terminal EGF-like sequence (domain V or LEa). Suppression of syndecan-1 expression by the RNA interference effectively blocked the activities of domain V capable of promoting cell adhesion and inhibiting the integrin beta4 phosphorylation. These results demonstrate that domain V of the gamma2 chain negatively regulates the integrin beta4 phosphorylation, probably through a syndecan-1-mediated signaling, leading to enhanced cell adhesion and suppressed cell motility.


High-throughput functional assays of IP3-evoked Ca2+ release.
This protocol describes procedures for high-throughput functional analyses of inositol 1,4,5-trisphosphate receptors (IP3Rs) in permeabilized cells. The methods are applicable to native IP3Rs in a variety of cells and to recombinant IP3Rs stably expressed in DT40 cells in which gene disruption has abolished expression of endogenous IP3Rs. A low-affinity Ca(2+)-indicator (Mag-Fluo-4) trapped within the endoplasmic reticulum (ER) of permeabilized cells is used to report changes in luminal free Ca(2+) concentration. A fluorescence plate-reader equipped to allow automated additions permits rapid measurements of the Ca(2+) release evoked by IP3R. The procedure can be completed in 2-3 h.


Effect of diet and intestinal AhR expression on fecal microbiome and metabolomic profiles.
Diet, loss of aryl hydrocarbon receptor (AhR) expression and their modification of the gut microbiota community composition and its metabolites affect the development of colorectal cancer (CRC). However, the concordance between fecal microbiota composition and the fecal metabolome is poorly understood. Mice with specific AhR deletion (AhRKO) in intestinal epithelial cell and their wild-type littermates were fed a low-fat diet or a high-fat diet. Shifts in the fecal microbiome and metabolome associated with diet and loss of AhR expression were assessed. Microbiome and metabolome data were integrated to identify specific microbial taxa that contributed to the observed metabolite shifts. Our analysis shows that diet has a more pronounced effect on mouse fecal microbiota composition than the impact of the loss of AhR. In contrast, metabolomic analysis showed that the loss of AhR in intestinal epithelial cells had a more pronounced effect on metabolite profile compared to diet. Integration analysis of microbiome and metabolome identified unclassified Clostridiales, unclassified Desulfovibrionaceae, and Akkermansia as key contributors to the synthesis and/or utilization of tryptophan metabolites. Akkermansia are likely to contribute to the synthesis and/or degradation of tryptophan metabolites. Our study highlights the use of multi-omic analysis to investigate the relationship between the microbiome and metabolome and identifies possible taxa that can be targeted to manipulate the microbiome for CRC treatment.


Health-related quality of life assessment in children followed in a cardiomyopathy clinic.
As our ability to diagnosis cardiomyopathy matures and genetic testing becomes more widespread, there has been an increase in the number of children followed for cardiomyopathy. The purpose of this study was to compare health-related quality of life (HRQoL) between children with cardiomyopathy and healthy controls and with children seen in clinic who are at risk for the development of cardiomyopathy. Patient and parent-proxy perspectives were obtained using the Pediatric Quality of Life Inventory (PedsQL(TM)) 4.0 Core Scales (ages 2-18 years) and the disease-specific Cardiac Module. Cardiomyopathy physicians' perceptions of the impact of cardiomyopathy on the functional status of the patients were collected. In addition, data regarding disease-specific medical and socioeconomic information were collected from chart review and parental report. The questionnaires were completed by 100 parent-proxies and 71 children. The PedsQL(TM) scores reported by children and their parent-proxies were compared to scores reported by pediatric norms. Compared to healthy controls, patients followed in a pediatric cardiomyopathy clinic scored lower in Total score when compared to pediatric norms (80.7 vs 86.4, p = 0.002). Interestingly, children with a family history of cardiomyopathy who are at risk for developing the disease scored similar to those children with a diagnosis of cardiomyopathy. Parental and patients perceptions were discrepant when compared, which may deter appropriate referral to behavioral health services. These results should encourage cardiomyopathy clinics to screen all patients for HRQoL impairments and to have behavioral services available to assist these children.