Fatty acid synthase cooperates with glyoxalase 1 to protect against sugar toxicity.
Fatty acid (FA) metabolism is deregulated in several human diseases including metabolic syndrome, type 2 diabetes and cancers. Therefore, FA-metabolic enzymes are potential targets for drug therapy, although the consequence of these treatments must be precisely evaluated at the organismal and cellular levels. In healthy organism, synthesis of triacylglycerols (TAGs)-composed of three FA units esterified to a glycerol backbone-is increased in response to dietary sugar. Saturation in the storage and synthesis capacity of TAGs is associated with type 2 diabetes progression. Sugar toxicity likely depends on advanced-glycation-end-products (AGEs) that form through covalent bounding between amine groups and carbonyl groups of sugar or their derivatives α-oxoaldehydes. Methylglyoxal (MG) is a highly reactive α-oxoaldehyde that is derived from glycolysis through a non-enzymatic reaction. Glyoxalase 1 (Glo1) works to neutralize MG, reducing its deleterious effects. Here, we have used the power of Drosophila genetics to generate Fatty acid synthase (FASN) mutants, allowing us to investigate the consequence of this deficiency upon sugar-supplemented diets. We found that FASN mutants are lethal but can be rescued by an appropriate lipid diet. Rescued animals do not exhibit insulin resistance, are dramatically sensitive to dietary sugar and accumulate AGEs. We show that FASN and Glo1 cooperate at systemic and cell-autonomous levels to protect against sugar toxicity. We observed that the size of FASN mutant cells decreases as dietary sucrose increases. Genetic interactions at the cell-autonomous level, where glycolytic enzymes or Glo1 were manipulated in FASN mutant cells, revealed that this sugar-dependent size reduction is a direct consequence of MG-derived-AGE accumulation. In summary, our findings indicate that FASN is dispensable for cell growth if extracellular lipids are available. In contrast, FA-synthesis appears to be required to limit a cell-autonomous accumulation of MG-derived-AGEs, supporting the notion that MG is the most deleterious α-oxoaldehyde at the intracellular level.


Detailed analysis of estrous-related changes in wheel running and self-stimulation.
Previous studies have demonstrated that a number of behaviors change their probability of occurrence during the estrous cycle. Wheel running is known to show estrous-related changes while reports upon intracranial reinforcement (self-stimulation, ICS) have been equivocal. The present studies examined both behaviors to further resolve the underlying behavioral determinants of these estrous effects. Data for wheel running and intracranial self-stimulation behaviors of adult female Holtzman rats were collected nightly on a minute by minute basis across the estrous cycle. It was found that: (a) both behaviors showed significant estrous-related changes in frequency; (b) the two behaviors occurred in discontinuous episodes (bursts) during the dark phase of the diurnal cycle; and (c) of three burst parameters (number/session, average length, rate of response), number proved the best predictor of estrous-related behavioral change. Burst number may reflect an important motivational parameter underlying estrous-related changes in behavior.


[Evaluation of the basic characteristics of rat behavior in the "open field" test].
Heart rate, motor activity and defecation were studied in rats of the Wistar strain in 10 successive one-hour "open field" tests. Defecation and heart rate parameters decreased monotonously both from test to test and throughout one session. In motor activity a second rise was recorded at 6--12th minutes against the background of a gradual decline. A positive correlation between the total value of defecation and the heart rate in 2nd-3rd minutes of the first test was observed. At the 13th minute the motor activity positively correlated with the heart rate. Significant negative correlation between motor activity and total value of defecation appeared only at 10--11th minutes of the first test. A conclusion is drawn on twofactorial motivational basis of rats' behaviour in the "open field" test.


Visual disturbance following sclerotherapy for varicose veins, reticular veins and telangiectasias: a systematic literature review.
The objective of the study was to review the literature reporting visual disturbance (VD)following sclerotherapy for varicose veins. Underlying mechanisms will be discussed. A literature search of the databases Medline and Google Scholar was performed. Original articles including randomized trials, case series and case reports reporting VD in humans following sclerotherapy for varicose veins were included. Additional references were also obtained if they had been referenced in related publications. The search yielded 4948 results of which 25 reports were found to meet the inclusion criteria. In larger series with at least 500 included patients the prevalence of VD following sclerotherapy ranges from 0.09% to 2%. In most reports foam sclerotherapy was associated with VD (19); exclusive use of liquid sclerosant was reported in two cases, some reports included foam and liquid sclerosant (4). There were no persistent visual disorders reported. VD occurred with polidocanol and sodium tetradecyl sulphate in different concentrations (0.25–3%). Various forms of foam preparation including various ways of foam production and the liquid –air ratio (1 or 2 parts of liquid mixed with 3, 4 or 5 parts of air) were reported in association with the occurrence of VD. VDs following sclerotherapy for varicose veins are rare and all reported events were transient. Bubble embolism or any kind of embolism seems unlikely to be the only underlying mechanism. A systemic inflammatory response following sclerotherapy has been suggested. Further research to clarify the mechanism of action of sclerosants is required.


UV-ozone ashing of cells and tissues for spatially resolved trace element analysis.
UV/ozone ashing of thin tissue sections and cell cultures is a simple technique to enhance relative elemental concentrations, while maintaining their spatial location at the sub-micron level. This approach may enhance the capability of spatially resolved analysis techniques to detect the distribution of trace elements in biological matrices. We present results from light microscopy and x-ray spectromicroscopy studies of tissues and cells demonstrating that the micro-structure is very well conserved. We show the signal enhancement resulting from the removal of carbon, which allows otherwise undetectable gadolinium to be mapped in cancer tissue for a novel neutron capture therapy.


Functional Assessment of Cardiac Responses of Adult Zebrafish (Danio rerio) to Acute and Chronic Temperature Change Using High-Resolution Echocardiography.
The zebrafish (Danio rerio) is an important organism as a model for understanding vertebrate cardiovascular development. However, little is known about adult ZF cardiac function and how contractile function changes to cope with fluctuations in ambient temperature. The goals of this study were to: 1) determine if high resolution echocardiography (HRE) in the presence of reduced cardiodepressant anesthetics could be used to accurately investigate the structural and functional properties of the ZF heart and 2) if the effect of ambient temperature changes both acutely and chronically could be determined non-invasively using HRE in vivo. Heart rate (HR) appears to be the critical factor in modifying cardiac output (CO) with ambient temperature fluctuation as it increases from 78 ± 5.9 bpm at 18°C to 162 ± 9.7 bpm at 28°C regardless of acclimation state (cold acclimated CA- 18°C; warm acclimated WA- 28°C). Stroke volume (SV) is highest when the ambient temperature matches the acclimation temperature, though this difference did not constitute a significant effect (CA 1.17 ± 0.15 μL at 18°C vs 1.06 ± 0.14 μl at 28°C; WA 1.10 ± 0.13 μL at 18°C vs 1.12 ± 0.12 μl at 28°C). The isovolumetric contraction time (IVCT) was significantly shorter in CA fish at 18°C. The CA group showed improved systolic function at 18°C in comparison to the WA group with significant increases in both ejection fraction and fractional shortening and decreases in IVCT. The decreased early peak (E) velocity and early peak velocity / atrial peak velocity (E/A) ratio in the CA group are likely associated with increased reliance on atrial contraction for ventricular filling.


A promoter sequence involved in cell-specific expression of the pea glutamine synthetase GS3A gene in organs of transgenic tobacco and alfalfa.
The DNA sequence of the pea cytosolic glutamine synthetase GS3A gene promoter has been determined and the start of transcription mapped using S1 nuclease. The full-length promoter and a series of 5' deletions were fused to beta-glucuronidase (GUS) and introduced into transgenic tobacco and alfalfa. In transgenic tobacco the GS3A promoter directed GUS expression in the phloem cells of the vasculature in leaves, stems and roots. GUS expression was also detected in the vasculature of cotyledons and the root tips of germinating T1 seedlings. The promoter conferred a similar expression pattern in transgenic alfalfa, and expression was also observed in root nodules. Nodule expression was located in nodule primordia, as well as the meristem, symbiotic zone, and vasculature of mature nodules. The promoter was found to be active even when deleted to -132 relative to the start of transcription. DNA mobility-shift analysis identified a protein present in nuclear and whole-cell plant extracts which bound to a 17 bp DNA element contained within the minimal -132 promoter required for expression.


[Enhancement of the antitumor effect of cyclophosphane in an experiment].
A significant antitumor effect of acetylcellulose microsphere-encapsulated cyclophosphamide was observed in the course of the studies carried out in 105 male Wistar rats bearing Walker's carcinosarcoma and PC-1 carcinoma. The effect is due to a high tissue level of the drug maintained for a long time as the drug uniformly passes through the porous microsphere wall. Local (intratumoral) treatment with cyclophosphamide is more effective than standard methods. The inhibitory effect of microsphere-enclosed cyclophosphamide on neoplastic growth is potentiated by tumor ischemia.


Immediate Early Genes Anchor a Biological Pathway of Proteins Required for Memory Formation, Long-Term Depression and Risk for Schizophrenia.
While the causes of myriad medical and infectious illnesses have been identified, the etiologies of neuropsychiatric illnesses remain elusive. This is due to two major obstacles. First, the risk for neuropsychiatric disorders, such as schizophrenia, is determined by both genetic and environmental factors. Second, numerous genes influence susceptibility for these illnesses. Genome-wide association studies have identified at least 108 genomic loci for schizophrenia, and more are expected to be published shortly. In addition, numerous biological processes contribute to the neuropathology underlying schizophrenia. These include immune dysfunction, synaptic and myelination deficits, vascular abnormalities, growth factor disruption, and N-methyl-D-aspartate receptor (NMDAR) hypofunction. However, the field of psychiatric genetics lacks a unifying model to explain how environment may interact with numerous genes to influence these various biological processes and cause schizophrenia. Here we describe a biological cascade of proteins that are activated in response to environmental stimuli such as stress, a schizophrenia risk factor. The central proteins in this pathway are critical mediators of memory formation and a particular form of hippocampal synaptic plasticity, long-term depression (LTD). Each of these proteins is also implicated in schizophrenia risk. In fact, the pathway includes four genes that map to the 108 loci associated with schizophrenia: GRIN2A, nuclear factor of activated T-cells (NFATc3), early growth response 1 (EGR1) and NGFI-A Binding Protein 2 (NAB2); each of which contains the "Index single nucleotide polymorphism (SNP)" (most SNP) at its respective locus. Environmental stimuli activate this biological pathway in neurons, resulting in induction of EGR immediate early genes: EGR1, EGR3 and NAB2. We hypothesize that dysfunction in any of the genes in this pathway disrupts the normal activation of Egrs in response to stress. This may result in insufficient electrophysiologic, immunologic, and neuroprotective, processes that these genes normally mediate. Continued adverse environmental experiences, over time, may thereby result in neuropathology that gives rise to the symptoms of schizophrenia. By combining multiple genes associated with schizophrenia susceptibility, in a functional cascade triggered by neuronal activity, the proposed biological pathway provides an explanation for both the polygenic and environmental influences that determine the complex etiology of this mental illness.


Across-subjects classification of stimulus modality from human MEG high frequency activity.
Single-trial analyses have the potential to uncover meaningful brain dynamics that are obscured when averaging across trials. However, low signal-to-noise ratio (SNR) can impede the use of single-trial analyses and decoding methods. In this study, we investigate the applicability of a single-trial approach to decode stimulus modality from magnetoencephalographic (MEG) high frequency activity. In order to classify the auditory versus visual presentation of words, we combine beamformer source reconstruction with the random forest classification method. To enable group level inference, the classification is embedded in an across-subjects framework. We show that single-trial gamma SNR allows for good classification performance (accuracy across subjects: 66.44%). This implies that the characteristics of high frequency activity have a high consistency across trials and subjects. The random forest classifier assigned informational value to activity in both auditory and visual cortex with high spatial specificity. Across time, gamma power was most informative during stimulus presentation. Among all frequency bands, the 75 Hz to 95 Hz band was the most informative frequency band in visual as well as in auditory areas. Especially in visual areas, a broad range of gamma frequencies (55 Hz to 125 Hz) contributed to the successful classification. Thus, we demonstrate the feasibility of single-trial approaches for decoding the stimulus modality across subjects from high frequency activity and describe the discriminative gamma activity in time, frequency, and space.


Increased threshold for TCR-mediated signaling controls self reactivity of intraepithelial lymphocytes.
To examine the effect of self Ag on activation requirements of TCR-alphabeta intestinal intraepithelial lymphocytes (IELs), we utilized the 2C transgenic (Tg) mouse model specific for a peptide self Ag presented by class I MHC, H-2Ld. CD8alpha alpha and CD4-CD8- IELs from syngeneic (H-2b, self Ag-) and self Ag-bearing (H-2b/d, self Ag+) strains were examined for their ability to respond in vitro to P815 (H-2d) cell lines expressing the endogenous antigenic peptide, p2Ca. Proliferation, cytokine production, and CTL activity were elicited in IEL T cells isolated from self Ag- H-2b mice when stimulated with P815 cells expressing basal levels of self Ag. These responses were enhanced following the addition of exogenous p2Ca peptide and ectopic expression of the costimulatory molecule, B7-1. By comparison, IEL from self Ag-bearing mice failed to respond to basal levels of self Ag presented by P815 cells even in the presence of B7-1-mediated costimulation. However, the addition of increasing amounts of exogenous p2Ca peptide induced a response from the in vivo "tolerized" T cells. These results suggest that exposure to self Ag in vivo increased the threshold of TCR activation of Ag-exposed self-reactive IELs. The dependence of increased signal 1 to activate self-reactive IELs suggests a defect in TCR signaling that may maintain self tolerance in vivo. These data suggest that conditions that overcome signal 1 IEL defects may initiate autoreactive responses in the intestine.


A comparison of two empirical models for the enzymatic hydrolysis of pretreated poplar wood.
Two empirical models for the enzymatic hydrolysis of cellulose are used to analyze the same set of experimental data to determine if one model is superior to the other. Both models adequately describe the data. The parameters in both models may be correlated to the hydrolysis conditions of enzyme, substrate, and product inhibitor concentrations. Both empirical models have features which are consistent with theoretical models of cellulose hydrolysis.


Coupling fractionation and batch desorption to understand arsenic and fluoride co-contamination in the aquifer system.
The present work is an attempt to study As and F+ coevality using laboratory based assays which couples fractionation and batch dissolution experiments. Sequential extraction procedure (SEP) resulting into five "operationally defined phases", was performed on sediment and soil samples collected from the Brahmaputra flood plains, Assam, India. High correlation between the Fe (hydr)oxide fraction and total As content of the soil/sediment sample indicates the involvement of Fe (hydr)oxides as the principal source of As. F- being an anion has high potential to be sorbed onto positively charged surfaces. Findings of the SEP were used to design the batch desorption experiments by controlling the Fe (hydr)oxide content of the soil/sediment. Desorption of As and F- was observed under acidic, neutral and alkaline pH from untreated and Fe (hydr)oxide removed samples. Highest amount of As and F- were found to be released from untreated samples under alkaline pH, while the amount leached from samples with no Fe (hydr)oxide was low. The study showed that the Fe (hydr)oxide fraction commonly found in the soils and sediments, had high affinity for negatively charged species like F- oxyanions of As, AsO43- (arsenate) and AsO33- (arsenite). Fe (hydr)oxide fraction was found to play the major role in co-evolution of As and F-. Two sorption coefficients were proposed based on easily leachable fraction and As present in the groundwater of sampling location for understanding of contamination vulnerability from the leaching.


Tensions in Maternal Care for Children, Youth, and Adults With Autism Spectrum Disorder.
This study explored the experiences of mothers caring for an individual with autism spectrum disorder (ASD) ranging from 5 to 25 years of age, and examined pervasive tensions in caregiving. Guided by ethnographic methods, a retrospective cross-sectional study was conducted. Interviews with 85 mothers were analyzed inductively. Prevalent tensions in maternal caregiving were identified: (a) difficulties obtaining, yet resistance to, an ASD diagnosis; (b) identified giftedness of the child versus notions of deficit imposed by others; (c) disability-related behaviors erroneously interpreted as 'poor parenting'; (d) contradictory considerations in diagnosis disclosure; (e) the invisibility yet pervasiveness of ASD; (f) extensive need for, yet the lack of, accessible services; (g) ASD-related care demands versus other pressing responsibilities; (h) arguments for inclusive versus exclusive services; and (i) aims of nurturing independence versus managing safety risk. Tensions were heightened by insufficient supports relative to need. Implications and recommendations for practice and policy are offered.


Streptococcus pyogenes type 12 M protein gene regulation by upstream sequences.
A partial nucleotide sequence that included 1,693 base pairs of the M12 (emm12) gene of group A streptococci (strain CS24) and adjacent upstream DNA was determined. Type 12 M protein-specific mRNA of strain CS24 is transcribed from two promoters (P1 and P3) separated by 30 bases. The transcription start sites of the emm12 gene were located more than 400 bases downstream of a deletion that causes decreased M-protein gene transcription in strain CS64. Deletion analysis of M protein-expressing plasmids indicated that an upstream region greater than 1 kilobase is required for M-protein gene expression. The M-protein gene transcriptional unit appears to be monocistronic. Analysis of the emm12 DNA sequence revealed three major repeat regions. Two copies of each repeat, A and B, existed within the variable 5' end of the gene; repeat C demarcated the 5' end of the constant region shared by emm12 and emm6.


Estrogen regulation of microRNAs, target genes, and microRNA expression associated with vitellogenesis in the zebrafish.
Estrogen is a steroid hormone that has been implicated in a variety of cellular and physiological processes and in the development of diseases such as cancer. Here we show a remarkable widespread microRNA (miRNA) downregulation in the zebrafish (Danio rerio) liver following 17β-estradiol (E2) treatment. This unique miRNA expression signature in the fish liver was further supported by a combination of computational predictions with gene expression microarray data, showing a significant bias toward upregulation of miRNA target genes after E2 treatment. Using pathway analysis of target genes, their involvement in the processes of cell cycle, DNA replication, and proteasome was observed, suggesting that miRNAs are incorporated into robust regulatory networks controlled by estrogen. In oviparous vertebrates, including fish, the formation of yolky eggs during a process known as vitellogenesis is regulated by estrogen. Microarrays were used to compare miRNA expression profiles between the livers of vitellogenic and nonvitellogenic zebrafish females. Among the upregulated miRNAs in vitellogenic females, were five members of the miR-17-92, a polycistronic miRNA cluster with a role in cell proliferation and cancer. Furthermore, a number of miRNA target genes related to fish vitellogenesis were revealed, including vtg3, a putative target of miR-122; the most abundant miRNA in the liver. Moreover, several of the differentially expressed miRNAs were only conserved in oviparous animals, which suggest an additional novel level of regulation during vitellogenesis by miRNAs and consequently, improves our knowledge of the process of oocyte growth in egg-laying animals.


Familial occurrence of dysembryoplastic neuroepithelial tumor-like neoplasm of the septum pellucidum: case report.
Dysembryoplastic neuroepithelial tumor (DNT)-like neoplasms of the septum pellucidum are extremely rare. In this article, we report the familial occurrence of these neoplasms. We report two cases of such neoplasms: Patient 1, a 42-year-old woman, and Patient 2, the 20-year-old nephew of Patient 1. Patient 1 experienced headache and worsening dizziness; Patient 2 experienced headache and worsening dizziness and also had partial seizures. In both cases, magnetic resonance imaging (MRI) revealed an intraventricular tumor adjacent to the septum pellucidum. Both tumors appeared as a hypointense region on T1-weighted MRI, and both appeared as a hyperintense region on T2-weighted MRI without gadolinium enhancement. Interestingly, both tumors had a high apparent diffusion coefficient. Both tumors were subtotally removed and had common histological findings, such as alveolar structures with oligodendroglia-like cells and "specific glioneuronal element." These findings are consistent with a dysembryoplastic neuroepithelial tumor-like neoplasm. After tumor removal, the symptoms disappeared. The postoperative course was uneventful, and the patients did not require adjuvant therapy. MRI showed no regrowth of residual tumors at 4 years (Patient 1) and 2 years (Patient 2) postoperatively. The familial occurrence of this rare tumor suggests that both of these cases arose from a common germline mutation. Identification of this rare tumor in this rare location is important to avoid unnecessary adjuvant therapy. A markedly high apparent diffusion coefficient and histological findings of specific glioneuronal element can facilitate the differential diagnosis of dysembryoplastic neuroepithelial tumor-like neoplasms. Genetic study of affected patients in this family may provide clues to its molecular pathogenesis.


A new tandem mass spectrometer for photofragment spectroscopy of cold, gas-phase molecular ions.
We present here the design of a new tandem mass spectrometer that combines an electrospray ion source with a cryogenically cooled ion trap for spectroscopic studies of cold, gas-phase ions. The ability to generate large ions in the gas phase without fragmentation, cool them to approximately 10 K in an ion trap, and perform photofragment spectroscopy opens up new possibilities for spectroscopic characterization of large biomolecular ions. The incorporation of an ion funnel, together with a number of small enhancements, significantly improves the sensitivity, signal stability, and ease of use compared with the previous instrument built in our laboratory.


Identifying IBD Providers' Knowledge Gaps Using a Prospective Web-based Survey.
As treatments, management strategies, and the role of advanced practice providers (APPs) have evolved in recent years, the Crohn's & Colitis Foundation sought to understand the educational and resource needs of clinicians caring for patients with inflammatory bowel diseases (IBDs). The aim of this study was to describe the self-identified IBD knowledge and resource gaps of clinicians to inform the development of future programming. A survey containing 19 questions created by the foundation's Professional Education Committee, a subset of its National Scientific Advisory Committee, was conducted from September 7, 2018 to October 15, 2018. Responses were included from providers if they were currently seeing any IBD patients in a clinical setting. The foundation distributed the survey by email and various social media channels to encourage a diverse response. The survey included questions on comfort levels around diagnosis, treatment, and management of patients with IBD, in addition to preferences and utilization of educational resources. The × 2 test was used to evaluate significant differences among respondents in the various domains surveyed. There were 197 eligible responses, of which 75% were from MD/Dos, followed by 25% APN/PA/RN/MSN/PhD/other; and 70% of respondents provide care for adult patients. The amount of time in practice was divided evenly among respondents. Fifty-seven percent of respondents practice in an academic/university setting, and approximately 75% indicated that ≥21% of their practice consisted of patients with IBD. Forty-four percent and 46% of respondents reported access to IBD based mental health providers and social workers in their practice, respectively. Seventy-two percent reported access to radiologists, 69% had access to dietitians, and 62% had access to advance practice providers. The areas of greatest educational need were prescribing medical cannabis (if approved locally) for pain management (62%); caring for patients with prior malignancy (35%); caring for pregnant patients and family planning (33%); caring for elderly patients (30%); and therapy decisions, including use of JAK inhibitors (29%), drug holidays (25%), and use of biosimilars (24%). More than 50% of respondents stated they do not participate in shared decision-making, citing time as the most common limiting factor. The majority of providers cited live education as their preferred learning format, and they wish to earn continuing medical education (CME) hours. This survey helped identify current IBD educational needs in our professional community. With a rapidly changing treatment landscape and an increase in the diversity of providers delivering care, additional opportunities to keep abreast of practice changes are critical to providing comprehensive, quality care in IBD. Our survey demonstrated that shared decision-making is underutilized in practice due to a need for resources that aid in its efficient integration into practice. Based on our results, a focus on creating live learning opportunities that offer CME are needed in the areas of therapeutic decision-making and treating IBD in special subsets (eg, prior malignancy, pregnancy, elderly).


Ultrasonicated sour Jujube seed flavonoids extract exerts ameliorative antioxidant capacity and reduces Aβ-induced toxicity in Caenorhabditis elegans.
Sour Jujube seed from Ziziphus jujuba Mill. var. Spinosa (Bunge) Hu ex H. F. Chow is a traditional Chinese herb. It was demonstrated with significant activities in anti-depression and antioxidant by numerous pharmacological studies. Flavonoids is one of the main constituents in sour Jujube seed. The aim of this study was to propose a green ultrasound-assisted extraction (UAE) process of flavonoids from sour Jujube seed. The extraction parameters were investigated and optimized using single factor experiments, Plackett-Burman design (PBD) and response surface methodology (RSM). Moreover, a comparative analysis between ultrasound-assisted extraction and heat reflux extraction was performed to verify the ameliorating effects of ultrasound-assisted extraction on the flavonoids yield, the composition, antioxidant capacities in vitro and ROS scavenging capacity in PC12 cells. Meanwhile, the effects of flavonoids extract (FE) on Aβ transgenic Caenorhabditis elegans (GMC101) behavior were investigated. The optimal extracting conditions of total flavonoids were as follows: ethanol concentration 70.60 (v/v%), liquid-solid ratio 15.02:1 mL/g, ultrasonic power 404 W, extraction time 60.03 min. The highest extraction yield was 1.59%. When compared to Heat reflux extraction (HRE) that only has gained a yield of 1.356%. Approximately, the UAE method was able to increase the yield by 17.11%. Moreover, FE extracted by UAE displayed larger capacity of scavenging ABTS, DPPH, superoxide, and hydroxyl radicals and reducing the level of ROS accumulation in PC12 cells, suggesting the biological functions of these compounds could be also better protected under UAE conditions. Furthermore, FE could also increase the chemotaxis and heat stress resistance ability, delay the paralysis and extend the lifespan of Caenorhabditis elegans. UAE is a green and efficient technique for the preparation of flavonoids from sour Jujube seed. The flavonoids extract can reduce Aβ-induced toxicity in Caenorhabditis elegans.


Automatic beam angle selection in IMRT planning using genetic algorithm.
The selection of suitable beam angles in external beam radiotherapy is at present generally based upon the experience of the human planner. The requirement to automatically select beam angles is particularly highlighted in intensity-modulated radiation therapy (IMRT), in which a smaller number of modulated beams is hoped to be used, in comparison with conformal radiotherapy. It has been proved by many researchers that the selection of suitable beam angles is most valuable for a plan with a small number of beams (< or = 5). In this paper an efficient method is presented to investigate how to improve the dose distributions by selecting suitable coplanar beam angles. In our automatic beam angle selection (ABAS) algorithm, the optimal coplanar beam angles correspond to the lowest objective function value of the dose distributions calculated using the intensity-modulated maps of this group of candidate beams. Due to the complexity of the problem and the large search space involved, the selection of beam angles and the optimization of intensity maps are treated as two separate processes and implemented iteratively. A genetic algorithm (GA) incorporated with an immunity operation is used to select suitable beam angles, and a conjugate gradient (CG) method is used to quickly optimize intensity maps for each selected beam combination based on a dose-based objective function. A pencil-beam-based three-dimensional (3D) full scatter convolution (FSC) algorithm is employed for the dose calculation. Two simulated cases with obvious optimal beam angles are used to verify the validity of the presented technique, and a more complicated case simulating a prostate tumour and two clinical cases are employed to test the efficiency of ABAS. The results show that ABAS is valid and efficient and can improve the dose distributions within a clinically acceptable computation time.


Redescription of Proteocephalus glanduligerus (Cestoda: Proteocephalidea), a parasite of clariid catfishes in Africa with a unique glandular apical organ.
The tapeworm Proteocephalus glanduligerus (Janicki, 1928) Fuhrmann, 1933 (Cestoda: Proteocephalidea), a parasite of clariid catfishes (Clarias spp.) in Africa, is redescribed on the basis of an evaluation of voucher material and newly collected specimens from Clarias gariepinus and C. cf. anguillaris from the Sudan, Ethiopia, and South Africa. The most typical characteristic of P. glanduligerus (previously misspelled as P. glanduligera or P. glanduliger) is the presence of an extremely large glandular apical organ, the size of which is 1.6-3.5 times larger than that of the suckers. In addition, other morphological characteristics unreported in previous accounts are provided, such as an unusual position of osmoregulatory canals (situated close to each other, with the dorsal canal latero-ventral to testes), the presence of a vaginal sphincter, several (usually 3-5) uterine pores, and eggs with paired lateral auricular swellings (extensions) of the outer envelope.


Effect of highly active antiretroviral therapy use on oral manifestations in pediatric patients infected with HIV.
The aim was to assess the prevalence of oral lesions in HIV-infected children undergoing highly active anti-retroviral therapy (HAART), and the association between the duration of HAART usage and oral lesions. Totally, 111 medical and dental records of HIV-infected children, aged from 2 to 16 years old were reviewed for medical data, presence of oral lesions, and caries prevalence. According to the type of medication, the children were grouped as follows: 51 were under HAART (G1), 46 were using anti-retroviral medication (G2), and 14 were using no medication (G3). The majority of the HIV children had AIDS (65.8%), of which 86.3% were in G1, 63% in G2, and 0% in G3. The mean length of therapy was 34.4 months, with no difference between groups (Kruskal-Wallis; P = 0.917). The prevalence of the oral lesions was 23.4%, namely, G1 was 27.5%, G2 was 21.7%, and G3 was 14.3% (P > 0.05). Gingivitis was the most common oral manifestation (15.3%) seen in the three groups, followed by gingival linear erythema and pseudomembranous candidiasis in G1 and G2. The mean values regarding deft and DMFT indexes were, respectively, 3.2 and 1.9 (G1), 2.8 and 1.6 (G2), and 3.8 and 3.0 (G3). For the patients without AIDS (n = 38), oral manifestations were seen in 29.4% of G2 compared to G1, with 0% (Chi-square; P > 0.05). In terms of therapy duration, 47.65% of the patients who had been under HAART for 18 months or less had oral manifestations, compared to 13.3% of those who had been treated for a longer time (Chi-square; P = 0.007). Although the prevalence of oral lesions was similar between the groups, it was less in patients without AIDS and those under HAART. The duration of HAART usage had a significant influence on the prevalence of these lesions.


Fas ligand expression in the germinal centre.
Whereas the importance of the Fas/FasL system in the regulation of T-cell homeostasis is well established, it is not yet clear if FasL is involved in B-cell regulation, especially in the clonal selection of B lymphocytes in the germinal centre (GC). This study therefore investigated the expression of FasL protein in tonsils and lymph nodes with lymphofollicular hyperplasia by western blotting and immunohistochemistry. In all the samples examined, western blot analysis showed FasL proteins of 33 and 52 kD, which presumably correspond to membrane-bound and soluble forms of the FasL protein. Immunohistochemically, FasL was found in a limited number of cells confined to a cluster in the light zone of the GC. The signal showed a delicate meshwork-like pattern of branching processes enmeshing the centrocytes and the few centroblasts of the light zone. In serial sections, the immunostaining pattern for FasL was found largely to coincide with the CD23 staining of follicular dendritic cells (FDCs), which are typically located in the light zone. In contrast, the FasL signal did not correspond to the distribution of the CD4-positive GC T-cells. In conclusion, expression of FasL in lymphofollicular hyperplasia seems to be largely confined to the light zone of the GCs, where selection of FDC-associated centrocytes is known to occur. These observations thus suggest that FasL is involved in selection processes of the B-cell system.


Cardiopulmonary resuscitation in the very low birth weight infant: the Vermont Oxford Network experience.
The limited literature available to date suggests that the use of delivery room cardiopulmonary resuscitation (DR-CPR) is associated with very poor outcomes, especially for extremely low birth weight infants. We reviewed the cumulative experience of the Vermont Oxford Network to determine the actual utilization of DR-CPR and the neonatal outcomes of such infants. A retrospective review of information available in the Vermont Oxford Network Database for the years 1994 to 1996. The data set was collected from 196 neonatal units who participate in the Network (data for infants 401 to 500 g were from 1996 only). Infants were eligible for study if they received DR-CPR defined as the administration of chest compressions and/or epinephrine in the delivery room as noted on the Vermont Oxford Network Database record. Information regarding survival was available for 27 707 newborns with birth weights from 501 to 1500 g, and 497 infants with birth weights from 401 to 500 g. There were 24 001 (86.6%) survivors. Overall DR-CPR was given to 9.3% of infants from 401 to 500 g and 6% of infants from 501 to 1500 g, 82.1% receiving chest compressions, and 66.7% receiving epinephrine. Survival of infants receiving DR-CPR was 23. 9% for infants of 401 to 500 g, and 63.3% for infants of 501 to 1500 g, compared with 16.7% and 87.9% for infants in these weight groups not receiving DR-CPR. Survival was greater for infants of 501 g or greater without DR-CPR compared with those who received this intervention within each 250-g birth weight subgroup. For infants of <1000 g, survival was 53.8% with DR-CPR compared with 74.9% without. Head ultrasounds were available for 95.5% of all surviving infants and 96.7% of infants who received DR-CPR. Overall, any grade of intraventricular hemorrhage (IVH) occurred more frequently in infants who received DR-CPR (38%) than in those who did not (21%). Grade 3 or 4 (severe) IVH was seen in 15.3% of infants who received DR-CPR compared with 4.9% of the infants who did not. Overall, survival without severe IVH occurred in 52.2% of DR-CPR infants compared with 81.3% of infants who did not require this intervention. The majority of very low birth weight and extremely low birth weight infants who receive DR-CPR survive, and at least half of such infants who survive do not have evidence of severe IVH. Further follow-up studies are required to determine the long-term neurodevelopmental outcome of such infants. The current study does not support the previously noted poor outcome in extremely low birth weight infants who receive DR-CPR.


A triad of lys12, lys41, arg78 spatial domain, a novel identified heparin binding site on tat protein, facilitates tat-driven cell adhesion.
Tat protein, released by HIV-infected cells, has a battery of important biological effects leading to distinct AIDS-associated pathologies. Cell surface heparan sulfate protoglycans (HSPGs) have been accepted as endogenous Tat receptors, and the Tat basic domain has been identified as the heparin binding site. However, findings that deletion or substitution of the basic domain inhibits but does not completely eliminate Tat-heparin interactions suggest that the basic domain is not the sole Tat heparin binding site. In the current study, an approach integrating computational modeling, mutagenesis, biophysical and cell-based assays was used to elucidate a novel, high affinity heparin-binding site: a Lys12, Lys41, Arg78 (KKR) spatial domain. This domain was also found to facilitate Tat-driven β1 integrin activation, producing subsequent SLK cell adhesion in an HSPG-dependent manner, but was not involved in Tat internalization. The identification of this new heparin binding site may foster further insight into the nature of Tat-heparin interactions and subsequent biological functions, facilitating the rational design of new therapeutics against Tat-mediated pathological events.


A 3-D reconstruction system for the human jaw using a sequence of optical images.
This paper presents a model-based vision system for dentistry that will assist in diagnosis, treatment planning, and surgical simulation. Dentistry requires an accurate three-dimensional (3-D) representation of the teeth and jaws for diagnostic and treatment purposes. The proposed integrated computer vision system constructs a 3-D model of the patient's dental occlusion using an intraoral video camera. A modified shape from shading (SFS) technique, using perspective projection and camera calibration, extracts the 3-D information from a sequence of two-dimensional (2-D) images of the jaw. Data fusion of range data and 3-D registration techniques develop the complete jaw model. Triangulation is then performed, and a solid 3-D model is reconstructed. The system performance is investigated using ground truth data, and the results show acceptable reconstruction accuracy.


Mechanisms and management of paroxysmal supraventricular tachycardia.
Paroxysmal supraventricular tachycardia (PSVT) is defined as a regular rapid heart beat, which initiates and terminates suddenly. PSVT may have a variety of electrophysiologic mechanisms, including atrial tachycardia, atrioventricular (AV) nodal reentry, and tachycardia involving an accessory AV connection. Atrial tachycardia may be reentrant or may be caused by abnormal automaticity or triggered activity. AV nodal reentry, in which the reentrant circuit is confined to the AV node or the region around the AV node, is the most common type of PSVT in adults. Orthodromic supraventricular tachycardia is the most frequently found tachycardia in patients with accessory AV connections. During orthodromic supraventricular tachycardia, antegrade conduction occurs via the AV node, and retrograde conduction occurs via an accessory AV connection. Other types of PSVT, including junctional tachycardia, are less common. A definitive diagnosis of the PSVT mechanism usually requires electrophysiologic testing, but clinical and electrocardiographic clues may be present. Understanding the mechanism of PSVT can alter therapy because the response to antiarrhythmic drugs may be somewhat different depending on the PSVT type. In addition, the risks and efficacy of catheter ablation for curing PSVT may differ depending on the PSVT type. A better understanding of PSVT mechanisms, which has developed over the past 20 years, has led to dramatic improvements in therapy.


Reversible left ventricular dysfunction associated with raised troponin I after subarachnoid haemorrhage does not preclude successful heart transplantation.
Transient abnormalities in ECGs, echocardiograms, and cardiac enzymes have been described in the acute setting of subarachnoid haemorrhage. In addition, left ventricular dysfunction has been reported at the time of brain death. A patient with an acute subarachnoid haemorrhage who presented with raised troponin I (TnI) concentrations and diffuse left ventricular dysfunction is described. After declaration of brain death 32 hours later, the heart was felt initially not suitable for transplantation. A normal cardiac catheterisation, however, lead to successful transplantation of the donor heart. Raised catecholamine concentrations and metabolic perturbations have been proposed as the mechanisms leading to the cardiac dysfunction seen with brain death. This may be a biphasic process, allowing time for myocardial recovery and reversal of the left ventricular dysfunction. Awareness of this phenomenon in the acutely ill neurologic population needs to be raised in order to prevent the unnecessary rejection of donor hearts.


Measurements of variable capacitance using single port radio frequency reflectometry.
A radio frequency (RF) reflectometry technique is presented to measure device capacitances using a probe station. This technique is used to characterize micro-electromechanical system (MEMS) variable capacitor devices that can be connected to create pull-up and pull-down networks used in digital gates for reversible computing. Adiabatic reversible computing is a promising approach to energy-efficient computing that can dramatically reduce heat dissipation by switching circuits at speeds below their RC time constants, introducing a trade-off between energy and speed. The variable capacitors in this study will be measured using single port RF reflectometry achieved with a custom-made RF probe. The RF probe consists of a micromanipulator with an on-board matching network and is calibrated by measuring a capacitive bank that shows a clearly visible frequency shift with the increase in capacitance. The RF probe worked well when measuring static capacitors with no parasitic resistance; however, the frequency shift is masked when measuring the MEMS variable capacitors due to their high in-series parasitic resistance (around 80 kΩ). Therefore, RF reflectometry has the potential to measure MEMS variable capacitors in the range of 0-30 fF when not masked by a high in-series parasitic resistance, creating a fast and versatile method for characterizing variable capacitors that can be used in energy-efficient computing.


Resolving the Three-Dimensional Rotational and Translational Dynamics of Single Molecules Using Radially and Azimuthally Polarized Fluorescence.
We report a radially and azimuthally polarized (raPol) microscope for high detection and estimation performance in single-molecule orientation-localization microscopy (SMOLM). With 5000 photons detected from Nile red (NR) transiently bound within supported lipid bilayers (SLBs), raPol SMOLM achieves 2.9 nm localization precision, 1.5° orientation precision, and 0.17 sr precision in estimating rotational wobble. Within DPPC SLBs, SMOLM imaging reveals the existence of randomly oriented binding pockets that prevent NR from freely exploring all orientations. Treating the SLBs with cholesterol-loaded methyl-β-cyclodextrin (MβCD-chol) causes NR's orientational diffusion to be dramatically reduced, but curiously NR's median lateral displacements drastically increase from 20.8 to 75.5 nm (200 ms time lag). These jump diffusion events overwhelmingly originate from cholesterol-rich nanodomains within the SLB. These detailed measurements of single-molecule rotational and translational dynamics are made possible by raPol's high measurement precision and are not detectable in standard SMLM.


Chronic myelomonocytic leukemia revealed by uncontrollable hematuria.
Nephrectomy was performed for uncontrollable unilateral hematuria in an apparently healthy 72-year-old man. The suburothelial connective tissue of the kidney was infiltrated by primitive myeloid cells with associated acute vasculitis and foci of extramedullary hematopoiesis. Subsequently, the patient was shown to have chronic myelomonocytic leukemia. Although renal involvement and vasculitis have been recorded previously in chronic myelomonocytic leukemia, this is the first occasion, to our knowledge, where their concurrence resulted in such a spectacular presentation.


Validation of AGA clinical care pathway and AASLD practice guidance for nonalcoholic fatty liver disease in a prospective cohort of patients with type 2 diabetes.
Recently, the American Gastroenterological Association (AGA) and American Association for the Study of Liver Diseases (AASLD) developed clinical pathways to evaluate populations at high-risk for nonalcoholic fatty liver disease (NAFLD). We assessed the diagnostic performance of the new guidance in a well-phenotyped cohort of patients with Type 2 diabetes mellitus (T2DM). This prospective study enrolled adults age ≥50 years with T2DM. Participants underwent a standardized clinical research visit with magnetic resonance imaging (MRI) and ultrasound-based assessment of liver fat and stiffness and ELF testing. Of 417 participants (36% men) with T2DM with FIB-4 and MRE data, the prevalence of NAFLD was 64% and 12% had advanced fibrosis (MRE≥3.63 kPa). Applying the AGA pathway of FIB-4 and vibration controlled transient elastography (VCTE) the false negative rate was 3.3% and 18% would qualify for specialty referral. Applying FIB-4 + ELF AASLD pathway the false negative rate was 4.5% but 50% would qualify specialty referral. Applying higher ELF cut points improved the pathway yielding a similar false negative rate of 4.9% but decreased specialty referral to 27%. Validation of the AGA clinical pathway in a prospectively recruited cohort with T2DM revealed a low false negative rate and avoided specialty referral in a large percentage of patients. The AASLD pathway with FIB-4 + ELF resulted in a high rate of specialty referral, which improved with utilization of higher ELF cut-points and may serve as an alternative for primary care and endocrinology clinics without access to VCTE.


Diagnosis & management of imported malaria in pregnant women in non-endemic countries.
Malaria in pregnancy is an important cause of maternal and foetal morbidity and is a potentially life-threatening infection. With ever-growing global exchanges, imported malaria in pregnancy is becoming an issue of concern in non-endemic countries where women, because of low immunity, have higher risk of severe diseases and death. Malaria in pregnancy is a dangerous condition which can be associated with important consequences for both mother and child such as stillbirth, low birth weight, maternal anaemia. In non-endemic-countries it is more frequent in its severe form which can lead to maternal death if not treated adequately. Specific anti-malarial interventions such as the use of repellents and insecticide treated bed nets in addition to chemoprophylaxis should be used by pregnant women if they are travelling to endemic areas. In cases of confirmed infection, specific treatment regimens vary according to gestational age and the presence of complications. Malaria should be considered a global health problem, increasingly involving western countries. Clinicians all over the world need to be prepared for this emerging disease both in terms of prevention and therapy.


The surface antigens of orthopoxviruses detected by cross-neutralization tests on cross-absorbed antisera.
Cross-neutralization tests were done on accepted species and recently isolated members of the genus Orthopoxvirus using antisera which had been separately absorbed with the various viruses. The results provided evidence for the involvement of four neutralizing antigens, and their distribution among 13 virus strains was determined. Monkeypox (Congo-8-Lombe), camelpox (Gorgan), ectromelia (Mill Hill), 'Lenny' and elephant poxviruses had distinctive antigenic formulae. Lister and Wyeth vaccines were indistinguishable but different from Copenhagen and EM63 vaccines which were themselves distinct. Cowpox (Brighton), buffalopox (BP4), MK 10, and Moscow poxviruses were indistinguishable. Examples were found where viruses shared surface antigens but were not all neutralized by antibody to them. this reduced the practical value of the technique for virus identification. Evidence was also obtained for the existence in some viruses of a fifth antigen, antibody to which could block neutralization by antibody to one particular antigen.


The uncrowded window of object recognition.
It is now emerging that vision is usually limited by object spacing rather than size. The visual system recognizes an object by detecting and then combining its features. 'Crowding' occurs when objects are too close together and features from several objects are combined into a jumbled percept. Here, we review the explosion of studies on crowding--in grating discrimination, letter and face recognition, visual search, selective attention, and reading--and find a universal principle, the Bouma law. The critical spacing required to prevent crowding is equal for all objects, although the effect is weaker between dissimilar objects. Furthermore, critical spacing at the cortex is independent of object position, and critical spacing at the visual field is proportional to object distance from fixation. The region where object spacing exceeds critical spacing is the 'uncrowded window'. Observers cannot recognize objects outside of this window and its size limits the speed of reading and search.


Imaging orbitals with attosecond and Ångström resolutions: toward attochemistry?
The recently developed attosecond light sources make the investigation of ultrafast processes in matter possible with unprecedented time resolution. It has been proposed that the very mechanism underlying the attosecond emission allows the imaging of valence orbitals with Ångström space resolution. This controversial idea together with the possibility of combining attosecond and Ångström resolutions in the same measurements has become a hot topic in strong-field science. Indeed, this could provide a new way to image the evolution of the molecular electron cloud during, e.g. a chemical reaction in 'real time'. Here we review both experimental and theoretical challenges raised by the implementation of these prospects. In particular, we show how the valence orbital structure is encoded in the spectral phase of the recombination dipole moment calculated for Coulomb scattering states, which allows a tomographic reconstruction of the orbital using first-order corrections to the plane-wave approach. The possibility of disentangling multi-channel contributions to the attosecond emission is discussed as well as the necessary compromise between the temporal and spatial resolutions.


DSB proteins and bacterial pathogenicity.
If DNA is the information of life, then proteins are the machines of life--but they must be assembled and correctly folded to function. A key step in the protein-folding pathway is the introduction of disulphide bonds between cysteine residues in a process called oxidative protein folding. Many bacteria use an oxidative protein-folding machinery to assemble proteins that are essential for cell integrity and to produce virulence factors. Although our current knowledge of this machinery stems largely from Escherichia coli K-12, this view must now be adjusted to encompass the wider range of disulphide catalytic systems present in bacteria.


Autosomal Dominant Alzheimer Disease: A Unique Resource to Study CSF Biomarker Changes in Preclinical AD.
Our understanding of the pathogenesis of Alzheimer disease (AD) has been greatly influenced by investigation of rare families with autosomal dominant mutations that cause early onset AD. Mutations in the genes coding for amyloid precursor protein (APP), presenilin 1 (PSEN-1), and presenilin 2 (PSEN-2) cause over-production of the amyloid-β peptide (Aβ) leading to early deposition of Aβ in the brain, which in turn is hypothesized to initiate a cascade of processes, resulting in neuronal death, cognitive decline, and eventual dementia. Studies of cerebrospinal fluid (CSF) from individuals with the common form of AD, late-onset AD (LOAD), have revealed that low CSF Aβ42 and high CSF tau are associated with AD brain pathology. Herein, we review the literature on CSF biomarkers in autosomal dominant AD (ADAD), which has contributed to a detailed road map of AD pathogenesis, especially during the preclinical period, prior to the appearance of any cognitive symptoms. Current drug trials are also taking advantage of the unique characteristics of ADAD and utilizing CSF biomarkers to accelerate development of effective therapies for AD.


A 17-Year-Old Boy With High-Functioning Autism, Gastrointestinal Illness, and Seizures.
A healthy 17-year-old boy with a high-functioning pervasive developmental disorder presented to the emergency department after having a 4-minute episode of seizure-like activity in the setting of presumed viral gastroenteritis. Within an hour of emergency department arrival, he developed a forehead-sparing facial droop, right-sided ptosis, and expressive aphasia, prompting stroke team assessment and urgent neuroimaging. Laboratory results later revealed a serum sodium of 119 mmol/L. Neurologic deficits self-resolved, and a full physical examination revealed diffuse abdominal tenderness in the lower abdomen with rebound tenderness in the right-lower quadrant. The patient was admitted to the PICU for electrolyte management and monitoring. A computed tomography (CT) scan of the abdomen obtained the following morning revealed the patient's final diagnosis.


Molecular diagnosis of Helicobacter pylori.
H. pylori infection can be diagnosed with many different tests. If the patient is undergoing endoscopy with gastric biopsies, culture and histology remain the diagnostic methods of choice. Indirect tests include rapid urease tests, urea breath tests, and serology. Molecular methods such as PCR offer marginal improvements when done on biopsy material, but has the advantage of being able to accurately identify H. pylori in areas outside the stomach where cultures usually fail. PCR can detect low numbers of organisms in gastric juice, bile, stool and oral secretions. Because of its high sensitivity it can also be used for epidemiologic investigations of environmental sources. However, the largest role for PCR may be in molecular fingerprinting. Arbitrary Primer PCR (RAPD) on the whole bacterial genome can reliably and accurately distinguish between isolates. PCR-based RFLP analysis can separate isolates based on restriction fragment sizes in a smaller amplified genome segment. REP-PCR can group isolates into clusters that appear to have different clinical expressions. These methods promise to shed new light on the transmission and pathogenicity of H. pylori.


Rapid HIV testing of clients of a mobile STD/HIV clinic.
HIV rapid testing may enhance the effectiveness of a mobile HIV/sexually transmitted disease (STD) screening clinic in at-risk populations who normally do not seek care. Our goal was to determine the usability and post-test counseling rates of rapid HIV testing services for clients tested on a mobile clinic. HIV Oraquick rapid HIV-1 testing (OraSure Technologies, Inc., Bethlehem, PA) (blood) was offered to clients seeking HIV/STI counseling and testing services from the street at predetermined locations in areas of high STD morbidity, drug use, and commercial sex work. Rapid test results were available on the same day at the van within 10 minutes. Disease intervention specialists (DIS) attempted to locate and counsel positive clients who did not stay for results. By comparison, when offered at the same time, 64.5% of clients preferred Oraquick to traditional serologic testing. The post-test counseling rate for clients tested for Oraquick was 89% for infected and 93% for uninfected. By comparison, 11% of infected clients and 40% of uninfected clients tested for traditional test were post-test counseled. Clients who tested for the traditional enzyme immunoassay (EIA) test were told to return to the van in 14 days for results and post-test counseling. In the adjusted model, we also found statistically significant differences comparing clients who choose Oraquick to traditional serologic tests. These data suggest that rapid HIV testing services may enhance the effectiveness of mobile STD/HIV clinics.


Effect of leishmanial excreted factor on the activities of adenylate cyclase from hamster liver and Leishmania tropica.
The carbohydrate segment of the excreted factor of culture cells from L. tropica and L. donovani was shown to interact with the adenylate cyclase from hamster liver. At a concentration of 1 mg/ml the excreted factor inhibited the stimulated activity of the mammalian enzyme by about 50%, independent of the activation by glucagon, 5'-guanylylimidodiphosphate and forskolin, respectively. In contrast to the adenylate cyclase system from hamster liver the enzyme from L. tropica-culture cells was not stimulated by addition of glucagon, 5'-guanylylimidodiphosphate and forskolin. In addition, the activity of the adenylate cyclase from L. tropica was not affected by addition of its endogenous excreted factor.


Diagnostic values of edema-sensitive T2-weighted imaging, TSE T1-weighted early contrast-enhanced imaging, late gadolinium enhancement, and the Lake Louise criteria in assessing acute myocarditis: A single-center cardiac magnetic resonance study.
The aim of this study was to evaluate the diagnostic accuracy of the Lake Louise consensus criteria using cardiac magnetic resonance (CMR) imaging assessment of edema, hyperemia, and late gadolinium enhancement (LGE) in the diagnostic determination of acute myocarditis. A total of 44 patients with acute myocarditis and 24 healthy controls were included in this retrospective study. The presence of edema was defined as a myocardial mean signal intensity >1.9 times that of the skeletal muscle in the same slice on T2-weighted short tau inversion-recovery sequences. Hyperemia was defined as an early gadolinium enhancement ratio (EGEr) ≥4 calculated using the contrast-enhancement of the myocardium and skeletal muscle on TSE T1-weighted sequences, and LGE was assessed by visual examination. The reference methods used to determine the presence of myocarditis were based on the European Society of Cardiology Working Group on Myocardial and Pericardial Diseases guidelines for clinical and biochemical findings. The diagnostic accuracy of edema, hyperemia, LGE, and the Lake Louise criteria (at least 2 of 3 components) was 75.7%, 64.2%, 88.5%, and 84.2%, respectively. Among the 3 components of the Lake Louise criteria, edema had the highest specificity (100%), and LGE had the highest sensitivity (86%). The use of LGE and/or edema as a criterion for acute myocarditis revealed the highest diagnostic accuracy (92.8%) among the CMR sequences and combinations of components examined. LGE and/or edema as a sole criterion for the diagnosis of acute myocarditis demonstrated better diagnostic accuracy than the Lake Louise criteria. The use of EGEr did not improve the performance of CMR-based diagnosis. Alternatives to the use of EGEr are recommended for the diagnosis of acute myocarditis.


Corneal parameters, ocular biometers, and retinal and choroidal thickness in acromegaly patients.
To compare ocular findings of acromegaly patients with healthy individuals and investigate the relation of serum levels of insulin-like growth factor (IGF-1) along with growth hormone (GH) and pituitary tumor (adenoma) dimensions (TD) with specific ocular parameters. The ocular parameters of acromegaly patients (n = 38) were compared with those of healthy subjects (n = 36). These parameters were intraocular pressure, keratometric (K1-K2) values, central corneal thickness (CCT), total axial length along with anterior chamber-lens-vitreous length, retinal nerve fiber layer (RNFL) thickness, central foveal thickness (CFT), choroidal thickness (CT), ganglion cell layer thickness (GCLT), and inner plexiform layer thickness (IPLT). Also investigated was whether there was a correlation between disease duration, TD, GH, IGF-I, CCT, RNFL, CFT, GCLT, IPLT, and CT. The lens length of the acromegaly group was increased (p = 0.014). GH and IGF-1 levels were positively correlated with CT and CCT, respectively (p = 0.041, r = 0.343) (p = 0.03, r = 0.347). Analysis of TD also found a highly negative correlation with the mean RNFL thickness of the acromegaly patients (p < 0.01, r = -0.603). The mean value of the inner parts of GCLT and IPLT was negatively correlated with TD (p = 0.041, r = -0.343 and p = 0.025, r = -0.379, respectively). Serum IGF-1 and GH levels might be determinant factors in CCT and CT, respectively. The pituitary adenoma size increasing may be prone to lead RNFL, ganglion cell layer, inner plexiform layer thinning. Increased lens thickness was found in the acromegaly group.


[Heavy metals in water of drinking fountains].
The purpose of this study was to analyze the levels of lead (Pb), cadmium (Cd), aluminum (Al), zinc (Zn), cooper (Cu) and chromium (Cr) in the water of drinking fountains distributed in the Campus of the University of São Paulo. Ribeirão Preto, Brazil. Thirty random samples were collected in different parts of the Campus that were analyzed by Atomic Absorption Spectrophotometry. According to WHO's Drinking Water Guidelines; lead, cadmium and zinc were found in concentrations higher than those recommended in 40%, 20% and 13% of the samples, respectively. The results were analyzed considering nutritional and toxicological aspects related to the presence of essential and toxic elements for the human being. Reviewing the regulatory limits established in ten countries of America, authors focus in the necessity to find a consensus in the establishment of the higher levels of heavy metals in potable water. The tolerable daily intake, have to be the basis to assure the prevention of diseases associated with a long-term ingestion of these elements through foods.


Airway delivery of soluble mycobacterial antigens restores protective mucosal immunity by single intramuscular plasmid DNA tuberculosis vaccination: role of proinflammatory signals in the lung.
Protection by parenteral immunization with plasmid DNA vaccines against pulmonary tuberculosis (TB) is very modest. In this study, we have investigated the underlying mechanisms for the poor mucosal protective efficacy and the avenues and mechanisms to improve the efficacy of a single i.m. immunization with a monogenic plasmid DNA TB vaccine in a murine model. We show that i.m. DNA immunization fails to elicit accumulation of Ag-specific T cells in the airway lumen despite robust T cell responses in the spleen. Such systemically activated T cells cannot be rapidly mobilized into the airway lumen upon Mycobacterium tuberculosis exposure. However, airway deposition of low doses of soluble mycobacterial Ags in previously immunized mice effectively mobilizes the systemically activated T cells into the airway lumen. A fraction of such airway luminal T cells can persist in the airway lumen, undergo quick, robust expansion and activation and provide marked immune protection upon airway M. tuberculosis exposure. Airway mucosal deposition of soluble mycobacterial Ags was found to create a tissue microenvironment rich in proinflammatory molecules including chemokines and hence conducive to T cell recruitment. Thus, in vivo neutralization of MIP-1alpha or IFN-inducible protein-10 markedly inhibited the accumulation of Ag-specific T cells in the airway lumen. Our data suggest that immunoprotective efficacy on the mucosal surface by i.m. plasmid DNA immunization could be substantially improved by simple mucosal soluble Ag inoculation and restoration of mucosal luminal T cells. Our study holds implication for the future design of DNA vaccination strategies against intracellular infections.


Truly intensive clinical ethics immersion at the Washington Hospital Center.
Opportunities for practical, hospital-based training in those skills demanded by clinical ethics consultation (CEC) have been limited. Given the number of individuals who provide part-time CEC, greater access to condensed, practical training such as the clinical ethics immersion course offered by the Washington Hospital Center, is necessary. Two participants in the initial cohort evaluate their CE training at a busy, urban referral center, exploring prior expectations, perceptions of its utility and suggestions for improvement. Such training will prove valuable not only for bioethicists who lack practical CEC experience "at the bedside" but also for ethics consultants whose ethics services have a low consult volume who wish to sharpen their skills.


Ambulatory monitoring and electronic measurement of urinary leakage in the diagnosis of detrusor instability and incontinence.
The ability of conventional artificial filling urodynamic studies (CMG) and ambulatory monitoring during natural bladder filling (AM) to detect phasic detrusor activity (detrusor instability) and incontinence was studied in 52 patients suspected on clinical grounds of having bladder dysfunction, but in whom a CMG had not provided an adequate explanation of their symptoms. Detrusor instability (DI) was found on AM in 31 patients who were not unstable on conventional CMG using the criteria of the International Continence Society: DI was diagnosed on filling in 20 patients and on provocation in a further 11. Incontinence was demonstrated by electronic nappy testing in 23 patients: 13 had urge incontinence due to DI, 7 had genuine stress incontinence alone and 3 had both genuine stress incontinence and detrusor instability. Thus DI was diagnosed significantly more frequently by AM than by CMG. Ambulatory monitoring was more sensitive in the diagnosis of instability and incontinence and it may prove to be a valuable aid in the diagnosis of bladder dysfunction not detected during conventional cystometry.


The UK's dysfunctional relationship with medical migrants: the Daniel Ubani case and reform of out-of-hours services.
In 2008, a patient died in the UK after being given an excessive dose of diamorphine by an overseas-trained doctor working in out-of-hours (OOH) primary care. This incident led to a debate on the recourse to international medical graduates and on the shortcomings of the OOH system. It is argued here that a historical reflection on the ways in which the NHS uses migrant labour can serve to reframe these discussions. The British Medical Association, the General Medical Council, and the House of Commons Health Committee have emphasised the need for more regulation of overseas graduates. Such arguments fit into a well-established pattern of dependency on and denigration of overseas graduates. They give insufficient weight to the multiple systemic failings identified in reports on OOH provision by the Department of Health and the Care Quality Commission. Medical migrants are often found in under-resourced and unpopular parts of healthcare systems, in the UK and elsewhere. Their presence provides an additional dimension to Julian Tudor Hart's inverse care law: the resources are fewer where the need is greatest, and the practitioner dealing with the consequences is more likely to be a migrant. The failings of the UK OOH system need to be understood in this context. Efforts to improve OOH care should be focused on controlling quality rather than the movement of doctors. A wider reflection on the nature of the roles that international medical graduates are asked to play in healthcare systems is also required.


Ovarian metastases in colorectal cancer.
The aims of this study were to determine the incidence, clinicopathologic features, and prognostic factors of ovarian involvement in primary colorectal cancer (CRC) and also to clear the role of prophylactic oophorectomy. Data from women with primary CRC treated between 1990 and 2004 were retrieved, and clinical and pathologic features of those who had undergone oophorectomy during CRC surgery were reviewed. One hundred eighty cases (mean age, 48 years) were included. In 120 cases, ovaries were preserved, and 60 cases underwent bilateral oophorectomy in addition to primary CRC resection. Reasons for oophorectomy were prophylactic in 22 (36.6%), abnormal morphology in 35 (58.3%), and undetermined in 3 cases (5%). There were 5 metastatic carcinomas, 8 primary ovarian tumors, and 47 normal ovaries in the pathologic evaluation. No complication directly related to oophorectomy was noted. Patients with ovarian metastases had higher stages of tumor. Ovarian metastases were not related to menstrual status, CRC location, size, differentiation, and mucin production, as well as abnormal morphology of the ovary. The global prevalence of ovarian metastases in CRCs was 2.7%, and isolated ovarian metastases occurred in fewer than half of them. Of 120 women who underwent colectomy alone, 8 (6.6%) developed ovarian metastasis during 2 years of follow-up. Only 3 cases had isolated ovarian metastases. No patient with synchronous or metachronous ovarian metastases from CRC survived 5 years. Isolated ovarian metastases from primary CRC occur with a low frequency, and this may partially explain the debate regarding prophylactic oophorectomy at the time of curative resection for primary CRC.


The effects of alpha - tocopherol and verapamil on mucosal functions after gut ischemia / reperfusion.
We have previously shown that in the canine small bowel autotransplantation model, adding alpha - tocopherol to Euro Collins solution for perfusion enhanced the integrity of the small bowel mucosa for up to 12 hours in the posttransplantation period. The purpose of this study was to investigate the effects of verapamil and a tocopherol on reperfusion injury in the canine small bowel autotransplantation model. The study consisted of four groups of six animals each. In Group 1 (control group) grafts were perfused with Euro Collins solution while those of Group 2 were perfused with Euro Collins and alpha- tocopherol. Group 3 grafts were perfused with Euro Collins + verapamil and Group 4 with Euro Collins, alpha- tocopherol and verapamil. Autotransplantation model was used to avoid immunological injury. Graft function and mucosal integrity were assessed by the analysis of enzymatic activities (mucosal glutaminase, maltase and lactase) and histopathological examination. Malonyldialdehyde (per gram tissue) was used as an indicator of lipid peroxidation. Glutaminase activity of Group 4 was found to be higher than that of the other groups at all time points (p<0.05). In the verapamil group (Group 3), the amount of lipid peroxidation showed the greatest decrease in comparison to other groups at the 12 hour time point (p<0.05). Maltase activity in Group 3 was significantly different at all time points (p<0.05). Lactase activity showed no significant difference between groups at each time point. The microscopic appearance of tissue injury shown by histopathological examination of tissue samples obtained at different time points was related to decrease in mucosal enzymatic activities. alpha- tocopherol is a promising agent for the prevention of tissue injury caused by free oxygen radicals during organ transplantation. Verapamil, as an antioxidant, also has preventive effects in organ preservation. Using verapamil together with alpha- tocopherol in the same preservation solution did not increase the preventive effect of alpha- tocopherol.


Binding of the human cytomegalovirus 80-kDa immediate-early protein (IE2) to minor groove A/T-rich sequences bounded by CG dinucleotides is regulated by protein oligomerization and phosphorylation.
The 80-kDa immediate-early regulatory protein IE2 of human cytomegalovirus (HCMV) functions as an essential positive transactivator of downstream viral promoters, but it also specifically down-regulates transcription from the major immediate-early promoter through a 14-bp DNA target motif known as the cis-repression signal (CRS) located at the transcription start site. The IE2 protein purified from bacteria as a fusion product of either staphylococcal Protein A/IE2(290-579) or glutathione-S-transferase (GST)/IE2(346-579) bound specifically to a [32P]-labeled CRS oligonucleotide probe in an in vitro electrophoretic mobility shift assay (EMSA). In contrast, no direct interaction with the CRS probes could be detected with IE2 wild-type protein in extracts from infected or transfected mammalian cells or when synthesized by in vitro translation. However, in vitro phosphorylation of GST/IE2(346-579) by incubation with either the catalytic subunit of protein kinase A (PKA) or a HeLa cell nuclear extract strongly inhibited its DNA-binding activity. This process required ATP hydrolysis and could be reversed by subsequent incubation with bacterial alkaline phosphatase. Importantly, dephosphorylation of the constitutively expressed native IE2 protein present in a nuclear extract from the U373(A45) cell line unmasked a specific CRS DNA-binding activity that could be supershifted with anti-IE2 monoclonal antibody (mAb). A series of high-molecular-weight hetero-oligomeric DNA-bound structures of intermediate mobility were formed in EMSA assays when a mixture of staphylococcal Protein A/IE2 and GST/IE2 was coincubated with the CRS probe. Coincubation with a DNA-binding negative but dimerization-competent GST/IE2 deletion mutant competitively inhibited DNA-binding by staphylococcal Protein A/IE2, whereas coincubation with a GST/IE2 deletion mutant that lacked the ability to both dimerize and bind to DNA failed to influence the mobility of the DNA-bound staphylococcal Protein A/IE2 protein. Therefore, IE2 appears to bind to DNA as a higher-order oligomer in which the presence of subunits with mutant DNA-binding domains interferes with the overall DNA-binding function. A series of point mutations introduced into each of nine conserved motifs throughout the DNA-binding and dimerization domain, all of which abolish the ability of the transfected intact IE2 protein to autoregulate the MIE promoter, also all lacked the ability to bind to CRS sequences as GST/IE2(346-379) fusion proteins. Detailed analysis of point mutations in the 14-bp CRS target DNA binding motif revealed that IE2 binds in a relatively sequence-independent manner to 10-bp-long A/T-rich DNA elements bounded on each side by CG dinucleotides. Moreover, the A/T-rich minor groove binding agent distamycin, but not the G/C-rich minor groove binding agent chromomycin-A3, actively competed with IE2 for binding to the CRS motif in a dose-dependent fashion. In conclusion, IE2 binds preferentially as multimerized dimers to A/T-rich sequences in the minor groove that are flanked on both sides by appropriately spaced CG dinucleotides, and inhibition of the DNA-binding or oligomerization activity by PKA phosphorylation probably accounts for the inactivity of the mammalian and in vitro translated forms of the protein.


Effects of Shuanghuangbu on the total protein content and ultrastructure in cultured human periodontal ligament cells.
Successful periodontal regeneration depends on the migration, proliferation and differentiation of periodontal ligament cells in periodontal defects. The total protein content and the ultrastructure demonstrate the metabolizability and activity of periodontal ligament cells. This study was conducted to observe the effects of Shuanghuangbu, a mixture of medicinal herbs, on the total protein content and the ultrastructure of human periodontal ligament cells. Periodontal ligament cells were grown to confluence and then cultured in Dulbecco's modified eagle medium (DMEM) supplemented with Shuanghuangbu over the concentration range of 0 to 1000 microg/ml. The total protein content in cultured cells was determined by using Coommasie brilliant blue technique. Periodontal ligament cells were incubated in 0 and 100 microg/ml Shuanghuangbu decoction for 5 days, then observed through transmission electron microscope. The total protein content of human periodontal ligament cells increased in each experiment group added 10 - 1000 microg/ml Shuanghuangbu respectively, and the effect of 100 microg/ml was excellent. Under the transmission electron microscope, there were more rough endoplasmic reticulums and mitochodrias in the experiment group than those in the control group. Shuanghuangbu stimulates the protein synthesis of human periodontal ligament cells and improves human periodontal ligament cells' metabolizability and activity.


DFT study on the standard electrode potentials of imidazole, tetrathiafulvalene, and tetrathiafulvalene-imidazole.
Extensive DFT calculations on the standard electrode potentials of imidazole (Im), tetrathiafulvalene (TTF), and 2-, 4-, and 5-TTF-Im were carried out. Geometries and Gibbs free energies of H-bonded dimer, anion, protonated cation, and neutral structures of Im, mono- and dication, and neutral structures of TTF in gas and acetonitrile solvent were computed by using 10 hybrid density functionals (B3LYP, TPSSH, PBEH1PBE, M06, M062X, X3LYP, BMK, B1B95, M05, M052X) combined with the TZVP basis set. CPCM and SMD solvation models were applied to predict the Gibbs free energies of molecules in acetonitrile solvent. Frequency calculations were carried out for all structures, and none of them has been found to exhibit any imaginary frequency. Finally, the BMK hybrid functional was selected for computation of the standard electrode potential of TTF-Im, because it gives the most accurate values in both Im and TTF, differing by 0.05 V from the experimental ones. Moreover, frequencies from the BMK functional are reasonably close to the experimental ones. The standard electrode potentials of 2-, 4-, and 5-TTF-Im predicted for two-electron oxidation are 0.946, 0.870, and 0.839 V in CPCM and 0.927, 0.866, and 0.824 V in SMD. For one-electron oxidation these are 0.491, 0.421, and 0.400 V in CPCM and 0.476, 0.377, and 0.360 V in SMD, respectively.


Advance Care Planning in palliative care: a qualitative investigation into the perspective of Paediatric Intensive Care Unit staff.
The majority of children and young people who die in the United Kingdom have pre-existing life-limiting illness. Currently, most such deaths occur in hospital, most frequently within the intensive care environment. To explore the experiences of senior medical and nursing staff regarding the challenges associated with Advance Care Planning in relation to children and young people with life-limiting illnesses in the Paediatric Intensive Care Unit environment and opportunities for improvement. Qualitative one-to-one, semi-structured interviews were conducted with Paediatric Intensive Care Unit consultants and senior nurses, to gain rich, contextual data. Thematic content analysis was carried out. UK tertiary referral centre Paediatric Intensive Care Unit. Eight Paediatric Intensive Care Unit consultants and six senior nurses participated. Four main themes emerged: recognition of an illness as 'life-limiting'; Advance Care Planning as a multi-disciplinary, structured process; the value of Advance Care Planning and adverse consequences of inadequate Advance Care Planning. Potential benefits of Advance Care Planning include providing the opportunity to make decisions regarding end-of-life care in a timely fashion and in partnership with patients, where possible, and their families. Barriers to the process include the recognition of the life-limiting nature of an illness and gaining consensus of medical opinion. Organisational improvements towards earlier recognition of life-limiting illness and subsequent Advance Care Planning were recommended, including education and training, as well as the need for wider societal debate. Advance Care Planning for children and young people with life-limiting conditions has the potential to improve care for patients and their families, providing the opportunity to make decisions based on clear information at an appropriate time, and avoid potentially harmful intensive clinical interventions at the end of life.


Eye movement impairment recovery in a Gaucher patient treated with miglustat.
In Gaucher Disease (GD) the enzyme (imiglucerase) replacement therapy (ERT) is not able to stop the progression of the neurological involvement, while the substrate reduction therapy (SRT), performed by N-Butyldeoxynojirimycin (miglustat), is an alternative that should be evaluated. Two sisters, presenting the same genotype (R353G/R353G), were diagnosed as suffering from GD; one of them later developed neurological alterations identified by quantitative saccadic eye movements analysis. The aim of the study was to quantitatively measure the miglustat effects in this GD neurological patient. Eye movement analysis during subsequent controls was performed by estimating the characteristic parameters of saccadic main sequence. The study demonstrates that the SRT alone can be effective in GD3. Moreover, it confirms that quantitative eye movement analysis is able to precociously identify also slight neurological alterations, permitting more accurate GD classification.


UroLift and Rezum: minimally invasive surgical therapies for the management of benign prostatic hyperplasia.
INTRODUCTION Minimally invasive surgical therapies for benign prostatic hyperplasia (BPH) are popular alternatives to the gold standard transurethral resection of the prostate (TURP). These procedures have fewer discernable side effects on urinary and sexual function, when compared to TURP, making it a desirable option for many patients. We provide an updated literature review on the current landscape of minimally invasive modalities, specifically the prostatic urethral lift (UroLift) and water vapor thermal therapy (Rezum), for the surgical treatment of BPH. Both UroLift and Rezum have demonstrated excellent efficacy and durability in relieving lower urinary tract symptoms (LUTS) in the BPH patient. When compared to TURP, these minimally invasive therapies can be performed in an outpatient setting, with decreased hospitalization, operative and catheterization times, which minimizes overall healthcare costs. Moreover, these therapies have no discernable adverse effects on sexual function (both ejaculatory and erectile) or sexual satisfaction, making it a desirable option for many patients. Both the UroLift and Rezum are office-based, minimally invasive techniques capable of providing durable, and significant relief of LUTS secondary to BPH. In select patients, they demonstrate comparable efficacy to TURP with the added advantage of preserving sexual function and minimizing patient morbidity and healthcare cost. An individualized, shared decision-making approach is essential in selecting the optimal treatment option for each patient.


Effects of chronic diazepam exposure on the behaviors and oxidative stress homeostasis in the eyes and brains of female Japanese medaka.
Diazepam (DZP) residue has been frequently detected in wastewater, surface water, and groundwater due to its extensive use over the decades. In this study, we exposed female Japanese medaka (Oryzias latipes) to environmentally relevant doses of DZP (800 and 8000 ng/L) for 4 weeks, aimed to investigate their behavioral responses and possible links with ocular and brain oxidative stress homeostasis. As a result, DZP exposure could significantly reduce swimming activity (800 ng/L) and anxiety (800 and 8000 ng/L), indicating a sedative effect on medaka. The DZP exposure also significantly increased the social interaction in medaka at 8000 ng/L. Furthermore, exposure to DZP could alter the ocular and brain oxidative stress homeostasis in medaka. The ocular CAT activities significantly increased in the 800 ng/L-DZP groups, and the brain SOD, CAT, GST and MDA levels also significantly increased in both DZP exposure groups. Correlation analysis revealed that the ocular and brain oxidative stress induced by DZP exposure might play an important role in their behavioral toxicity to medaka. Our findings highlight the necessity to clarify the exact link between DZP exposure-induced oxidative stress in the neural and sensor systems and its behavioral toxicity to better assess the risks on nontarget aquatic species.


Super-Resolution Imaging of the Actin Cytoskeleton in Living Cells Using TIRF-SIM.
Super-resolution (SR) imaging techniques have advanced rapidly in recent years, but only a subset of these techniques is gentle enough to be used by cell biologists to study living cells with minimal photodamage. Our research is focused on studies of the dynamic remodeling of the actin cytoskeleton in living pancreatic beta cells during insulin secretion. These studies require super-resolution light microscopic techniques that are gentle enough to record rapid changes of the actin cytoskeleton in real time. In this chapter, we describe an SR technique that breaks the diffraction limit of the conventional light microscope called TIRF-SIM. Using this SR techniques, we have been able to show that (1) microvilli on pancreatic beta cells translocate in the plane of the plasma membrane and (2) the cortical actin network reorganizes when cells are stimulated to secrete insulin. We describe the FIJI plugins that were used to process and analyze the TIRF-SIM images to obtain quantitative data.


A male with unilateral microphthalmia reveals a role for TMX3 in eye development.
Anophthalmia and microphthalmia are important birth defects, but their pathogenesis remains incompletely understood. We studied a patient with severe unilateral microphthalmia who had a 2.7 Mb deletion at chromosome 18q22.1 that was inherited from his mother. In-situ hybridization showed that one of the deleted genes, TMX3, was expressed in the retinal neuroepithelium and lens epithelium in the developing murine eye. We re-sequenced TMX3 in 162 patients with anophthalmia or microphthalmia, and found two missense substitutions in unrelated patients: c.116G>A, predicting p.Arg39Gln, in a male with unilateral microphthalmia and retinal coloboma, and c.322G>A, predicting p.Asp108Asn, in a female with unilateral microphthalmia and severe micrognathia. We used two antisense morpholinos targeted against the zebrafish TMX3 orthologue, zgc:110025, to examine the effects of reduced gene expression in eye development. We noted that the morphant larvae resulting from both morpholinos had significantly smaller eye sizes and reduced labeling with islet-1 antibody directed against retinal ganglion cells at 2 days post fertilization. Co-injection of human wild type TMX3 mRNA rescued the small eye phenotype obtained with both morpholinos, whereas co-injection of human TMX3(p.Arg39Gln) mutant mRNA, analogous to the mutation in the patient with microphthalmia and coloboma, did not rescue the small eye phenotype. Our results show that haploinsufficiency for TMX3 results in a small eye phenotype and represents a novel genetic cause of microphthalmia and coloboma. Future experiments to determine if other thioredoxins are important in eye morphogenesis and to clarify the mechanism of function of TMX3 in eye development are warranted.


[Can we talk about addiction for excessive indoor tanning?].
Tanning is socially highly valued and, like other forms of behaviour, can take on an addictive aspect. This finding is based on the observations of many dermatologists who report an addictive relationship with tanning cabins in their patients, despite the announcement of the diagnosis of malignant melanoma. This article attempts to synthesize the existing literature on excessive indoor tanning and addiction to investigate possible associations. A literature review was conducted from 1974 to 2012, using PubMed, Google Scholar, EMBASE, PsycInfo, and the following keywords alone or combined : Tanning, Addiction, Dependence, and Sunbeds, to explore the possible relevance of the addictive model. Excessive tanning is not present in the international classifications of Psychiatry, but may be related to addiction. There is so far no consensus on the definition of the concept, or recommendations on the management of this disorder. No study on psychopathology was found in the literature, although some hypotheses can be advanced. Further studies must be performed, especially on clinical psychopathology, neurobiology and management to improve our understanding of excessive indoor tanning.


Improving the Lifestyle of Adolescents Through Peer Education and Support in Vietnam: Protocol for a Pilot Cluster Randomized Controlled Trial.
In Ho Chi Minh City, Vietnam, recent studies found a rapid increase in overweight and obesity in adolescents. There is a need for effective health promotion interventions to support healthy diets and encourage a physically active lifestyle. This study will help fill an evidence gap on effective interventions to prevent excess weight gain in adolescents and generate new insights about peer-led education to promote healthy lifestyles. We aim to assess the feasibility and acceptability of a combined peer-led and peer support intervention among junior high school students in Ho Chi Minh City. Additionally, the efficacy of the intervention on adolescents' dietary practices and time spent on physical activity will also be measured in this pilot study. The Peer Education and Peer Support (PEPS) project is a pilot cluster randomized controlled trial with 2 intervention and 2 control schools. The intervention consists of 4 weekly education sessions of why and how to choose healthy food and drinks and how to be more physically active. Additionally, the intervention includes a school-based and online support system to help maintain student engagement during the intervention. We will use in-depth interviews with students, peer leaders, teachers, and parents; focus group discussions with peer educators; and direct observation of the school environment and peer leaders' interactions with the students. Acceptability and feasibility of the intervention will be assessed. We will also quantitatively assess limited efficacy by measuring changes in student' physical activity levels and dietary behaviors. We delivered the peer education intervention at the start of each school year over 3 months for all new grade 6 adolescents in the selected schools, followed by peer support and home engagement activities over 6 months until the end of the school year. There was a baseline assessment and 2 post-intervention assessments: the first immediately after the intervention to assess the short-term impact and the second at the end of the school year to assess the sustained impact on changes in adiposity, diet, and physical activity. The findings of this study will be used to develop a larger-scale cluster randomized controlled trial to examine the impact of a multicomponent, school- and home-based health promotion intervention. The trial will use innovative peer education methods to reduce overweight and obesity and improve dietary choices and physical activity levels in Vietnamese adolescents. Australian New Zealand Clinical Trials Registry ACTRN12619000421134; https://www.anzctr.org.au/Trial/Registration/TrialReview.aspx?id=376690&isReview=true. DERR1-10.2196/15930.


Serum cytokine profiles in acute primary HIV-1 infection and in infectious mononucleosis.
Serum cytokine profiles, T-cell subsets, and general parameters of immune activation were evaluated in 15 patients with acute primary HIV-1 infection, and compared with those obtained from 18 patients with acute primary Epstein-Barr virus (EBV) infection and from 18 control subjects in order to elucidate possible defects of immune response to HIV in early phases of virus-host interaction. Mean CD4+ cell count, serum concentrations of interleukin (IL)-2, IL-4, soluble IL-2 receptor (sIL-2R), tumor necrosis factor (TNF)-alpha, 5'-neopterin, and beta 2-microglobulin were significantly lower in acute HIV-1 infection than in EBV infection. Both acute HIV-1 and EBV infections were characterized by significantly higher mean CD8+ cell count and soluble CD8 antigen (sCD8) levels compared to control subjects, while acute HIV-1 infection was accompanied by the highest interferon (IFN)-gamma serum concentrations. In primary HIV-1 infection, significant impairment of CD4+- mediated T-helper function may lead to viral escape and persistence of infection despite an early and vigorous CD8+ T-lymphocyte activation.


Congenital malformations induced by laetrile.
Laetrile administered orally ot pregnant hamsters caused skeletal malformations in the offspring, but intravenous laetrile filed to result in embryopathic effects. Oral laetrile significantly increased in situ cyanide concentrations, while intravenous laetrile did not. Thiosulfate administration protected embryos from the teratogenic effects of oral laetrile. The embryopathic effects of oral laetrile appear to be due to cyanide released by bacterial beta-glucosidase activity.


Evaluation of diffusion of triamcinolone acetonide from the distal interphalangeal joint into the navicular bursa in horses.
To determine whether triamcinolone acetonide diffuses from the distal interphalangeal joint (DIPJ) to the navicular bursa, diffusion is direct or systemic, and addition of sodium hyaluronan has an effect on diffusion in horses. 11 adult horses without forelimb lameness. 1 randomly chosen forelimb DIPJ of each horse received an injection of 10 mg of triamcinolone acetonide plus 20 mg of sodium hyaluronan (group 1), and the contralateral forelimb DIPJ received an injection of 10 mg of triamcinolone acetonide plus 2 mL of lactated Ringer's solution (group 2). Synovial fluid samples were taken from both forelimb navicular bursae and 1 hind limb navicular bursa (systemic control group) at 6 hours. Triamcinolone acetonide concentrations in synovial fluid were quantified by use of high-performance liquid chromatography plus tandem mass spectrometry. Data were logarithmically transformed, and contrast analysis was performed on the 3 groups. Triamcinolone acetonide was detected in navicular bursal samples in all groups. Groups 1 and 2 had significantly greater concentrations of triamcinolone acetonide than the systemic control group. There was no significant difference between groups 1 and 2. Triamcinolone acetonide diffused directly from the DIPJ into the navicular bursa in clinically normal horses, and diffusion was not affected by addition of hyaluronan. Injection into the DIPJ with triamcinolone acetonide or a triamcinolone acetonide-hyaluronan combination can potentially be used for treatment of navicular syndrome, but further studies are needed to determine whether triamcinolone acetonide diffuses similarly in horses with navicular syndrome.