Studying biomolecule localization by engineering bacterial cell wall curvature.
In this article we describe two techniques for exploring the relationship between bacterial cell shape and the intracellular organization of proteins. First, we created microchannels in a layer of agarose to reshape live bacterial cells and predictably control their mean cell wall curvature, and quantified the influence of curvature on the localization and distribution of proteins in vivo. Second, we used agarose microchambers to reshape bacteria whose cell wall had been chemically and enzymatically removed. By combining microstructures with different geometries and fluorescence microscopy, we determined the relationship between bacterial shape and the localization for two different membrane-associated proteins: i) the cell-shape related protein MreB of Escherichia coli, which is positioned along the long axis of the rod-shaped cell; and ii) the negative curvature-sensing cell division protein DivIVA of Bacillus subtilis, which is positioned primarily at cell division sites. Our studies of intracellular organization in live cells of E. coli and B. subtilis demonstrate that MreB is largely excluded from areas of high negative curvature, whereas DivIVA localizes preferentially to regions of high negative curvature. These studies highlight a unique approach for studying the relationship between cell shape and intracellular organization in intact, live bacteria.


Hemodynamic effects of dexmedetomidine--fentanyl vs. nalbuphine--propofol in plastic surgery.
Dexmedetomidine has demonstrated to be useful in several clinical fields due to its respiratory safety and cardiovascular stability. We undertook this study to determine its usefulness in plastic surgery. Sixty patients were divided into two parallel groups. A group received dexmedetomidine--fentanyl and the comparison group received nalbuphine--propofol, both with same dose of midazolam. Blood pressure, heart rate and oxygen saturation were determined during the preoperative, intraoperative and recuperation periods. In both groups, hemodynamic constants decreased intraoperatively. Dexmedetomidine--fentanyl decreased more than in the nalbuphine--propofol (systolic blood pressure, p = 0.006; diastolic blood pressure, p = 0.01 and heart rate, p = 0.007). Comparatively, oxygen saturation was greater in the dexmedetomidine--fentanyl group vs. nalbuphine--propofol (p = 0.0001). Recovery time for the nalbuphine--propofol group was shorter than in the dexmedetomidine--fentanyl group (p = 0.0001). Dexmedetomidine shows the same cardiovascular stability but with absence of respiratory depression.


Transcranial direct current stimulation over left dorsolateral prefrontal cortex facilitates auditory-motor integration for vocal pitch regulation.
A growing body of literature has implicated the left dorsolateral prefrontal cortex (DLPFC) in the online monitoring of vocal production through auditory feedback. Specifically, disruption of or damage to the left DLPFC leads to exaggerated compensatory vocal responses to altered auditory feedback. It is conceivable that enhancing the cortical excitability of the left DLPFC may produce inhibitory influences on vocal feedback control by reducing vocal compensations. We used anodal transcranial direct current stimulation (a-tDCS) to modulate cortical excitability of the left DLPFC and examined its effects on auditory-motor integration for vocal pitch regulation. Seventeen healthy young adults vocalized vowel sounds while hearing their voice pseudo-randomly pitch-shifted by ±50 or ±200 cents, either during (online) or after (offline) receiving active or sham a-tDCS over the left DLPFC. Active a-tDCS over the left DLPFC led to significantly smaller peak magnitudes and shorter peak times of vocal compensations for pitch perturbations than sham stimulation. In addition, this effect was consistent regardless of the timing of a-tDCS (online or offline stimulation) and the size and direction of the pitch perturbation. These findings provide the first causal evidence that a-tDCS over the left DLPFC can facilitate auditory-motor integration for compensatory adjustment to errors in vocal output. Reduced and accelerated vocal compensations caused by a-tDCS over left DLPFC support the hypothesis of a top-down neural mechanism that exerts inhibitory control over vocal motor behavior through auditory feedback.


Whipple's disease: a review emphasizing immunology and microbiology.
Whipple's disease is an important and fascinating problem of local immunity in the gastrointestinal tract. Does the disease occur when an organism that is rare in nature infects an individual or do patients with Whipple's disease have a definable defect in their immune response that permits infection by a more common agent? Data in recent years indicate that there is only one type of microorganism that causes Whipple's disease. It is rod-shaped by electron microscopy and has a definable antigenic pattern by immunofluorescence. Paradoxically (considering its geometry), it reacts most strongly with antisera directed to streptococcal antigens. Patients with Whipple's disease do not have a disorder of immunoglobulin synthesis and do not have immune complexes present in their gut walls. Further, although earlier studies indicated that a defective response of T-lymphocytes to PHA is consistently present in these patients even years after therapy, more recent studies have found no consistent defect in mitogenic responses to PHA, CON A, or PWM. Also, recent studies indicate that mononuclear cells from patients with Whipple's disease usually mediate antibody-dependent cell-mediated cytotoxicity as well as controls, although spontaneous cell-mediated killing may be decreased. All these studies suggest that the defect is not primarily of lymphocytes, rather it is more likely that a defect exists in monocytes and macrophages. Future studies on Whipple's disease should add to existing knowledge of how the immune processes intracellular microorganisms.


[Rupture of voluminous atherosclerotic superficial femoral artery aneurysm].
Atherosclerotic aneurysms of the superficial femoral artery are very uncommon. We report a case of a ruptured superficial femoral artery aneurysm which was successfully treated surgically, and compare our data with those emerging from a review of the literature. We observed a 74-year-old man who presented with a suspected rupture of a superficial femoral artery aneurysm. After confirmation of the diagnosis with angio-CT, we treated the patient by performing an endoaneurysmectomy and replacing the arterial aneurysmatic segment with interposition of a polytetrafluoroethylene prosthetic graft. There has been no evidence of complications in the short to medium-term. In our review of literature, we collected 43 cases of superficial femoral artery aneurysms. These aneurysms were diagnosed after rupture (30-46%) or thrombosis and embolism (12-46%). Other aneurysms are frequently concomitant (27-69%). The best treatment is based on replacing the aneurysmatic segment with a prosthetic graft which may be autologous (saphenic vein) or heterologous (polytetra-fluoroethylene). In patients with obstructive arteriopathies exclusion of the aneurysm by ligation of the proximal and distal artery may be enough. The high incidence of complications suggests that resection and grafting should be performed electively, and the frequent association with aortoiliac or peripheral aneurysms means that thorough investigation and follow-up are mandatory.


Protective and Detoxifying Enzyme Activity and ABCG Subfamily Gene Expression in Sogatella furcifera Under Insecticide Stress.
Sogatella furcifera, an important migratory pest of rice, has substantial detrimental effects on rice production. To clarify the mechanism whereby S. furcifera responds to insecticide stress, we measured the activity of its protective [superoxide dismutase (SOD); peroxidase (POD); catalase (CAT)] and detoxifying [carboxylesterase (CarE); glutathione S-transferase (GST); mixed-function oxidase (MFO)] enzymes and the expression levels of its ATP-binding cassette subfamily G (ABCG) transporter genes in response to sublethal concentrations (LC10 and LC25) of the insecticides thiamethoxam, buprofezin, and abamectin. On the bases of the transcriptome data and the ABCG genes of Laodelphax striatellus, we obtained 14 full-length ABCG sequences for S. furcifera. RT-qPCR results showed that 13, 12, and 9 sfABCG genes were upregulated in the presence of thiamethoxam, buprofezin, and abamectin, respectively, at LC10. Moreover, 13 and 7 sfABCG genes were upregulated following treatment with thiamethoxam and abamectin, respectively, at LC25. Enzyme activity assays showed that although thiamethoxam, buprofezin, and abamectin induced GST, CarE, CAT, POD, and SOD activity, they did so at different concentrations and exposure times. The activity of MFO was generally inhibited with prolonged exposure to the three insecticides, with the inhibitory effect being most significant at 72 h. These results indicate that S. furcifera differs in its response to different types or concentrations of insecticides. Taken together, our results lay the foundations for gaining a deeper understanding of the mechanisms underlying the adaptation of S. furcifera to different types of insecticides, which would be of considerable significance for the development of effective pest management strategies.


Influence of short-term drought conditions and subsequent re-watering on the physiology and proteome of Lolium multiflorum/Festuca arundinacea introgression forms, with contrasting levels of tolerance to long-term drought.
Festuca arundinacea is a drought tolerant species. Lolium multiflorum has better forage quality but lower tolerance to abiotic stresses. Their hybrids offer an opportunity to perform research on the molecular basis of tolerance to drought. The aim of this work was to recognise the mechanisms of response to short-term drought (11 days) in a glasshouse in two L. multiflorum/F. arundinacea introgression forms with distinct levels of tolerance to long-term drought (14 weeks) in the field. Measurements of physiological parameters, analyses of protein accumulation profiles using two-dimensional gel electrophoresis, and mass spectrometry identification of proteins, which were accumulated differentially between the selected genotypes during short-term drought, were performed. Genotype 7/6, with lower yield potential during 14 weeks of drought, and lower ability to re-grow after watering, had a higher capacity for photosynthesis during 11 days of drought. Genotype 4/10, more tolerant to long-term drought, was able to repair damaged cell membranes after watering and was also characterised by lower transpiration during short-term drought. A total of 455 proteins were analysed, and the 17 that were differentially accumulated between the two genotypes were identified. The results of physiological and proteomic research led to a hypothesis that the higher photosynthetic capacity of genotype 7/6 could be due to a more efficient Calvin cycle, supported by higher accumulation of crucial proteins involving chloroplast aldolase.


Repetitive element DNA methylation levels in white blood cell DNA from sisters discordant for breast cancer from the New York site of the Breast Cancer Family Registry.
Global decreases in DNA methylation, particularly in repetitive elements, have been associated with genomic instability and human cancer. Emerging, though limited, data suggest that in white blood cell (WBC) DNA levels of methylation, overall or in repetitive elements, may be associated with cancer risk. We measured methylation levels of three repetitive elements [Satellite 2 (Sat2)], long interspersed nuclear element-1 (LINE-1) and Alu) by MethyLight, and LINE-1 by pyrosequencing in a total of 282 breast cancer cases and 347 unaffected sisters from the New York site of the Breast Cancer Family Registry (BCFR) using DNA from both granulocytes and total WBC. We found that methylation levels in all markers were correlated between sisters (Spearman correlation coefficients ranged from 0.17 to 0.55). Sat2 methylation was statistically significantly associated with increased breast cancer risk [odds ratio (OR) = 2.09, 95% confidence interval (CI) = 1.09-4.03; for each unit decrease in the natural log of the methylation level, OR = 2.12, 95% CI = 0.88-5.11 for the lowest quartile compared with the highest quartile]. These associations were only observed in total WBC but not granulocyte DNA. There was no association between breast cancer and LINE-1 and Alu methylation. If replicated in larger prospective studies, these findings support that selected markers of epigenetic changes measured in WBC, such as Sat2, may be potential biomarkers of breast cancer risk.


HIV risk behaviors and their correlates among HIV-positive adults with serious mental illness.
HIV risk behaviors and their correlates were examined in a sample of 154 HIV-seropositive and seriously mentally ill adults (83% male, 56% sexually active). Most sexually active participants engaged in HIV risk behavior during the past 6 months, although the rates of these behaviors were generally not higher, and in some cases were lower, than those reported in studies of otherwise comparable noninfected people. Variables significantly associated with one or more HIV risk behaviors in bivariate analyses included being female, any limitation in instrumental functioning, not having a bipolar disorder, more psychotic mental health symptoms, problem drinking, and not receiving HIV counseling. The latter three variables accounted for 22% of the variance in the total number of HIV risk behaviors. Interventions to reduce risky sexual practices may be most appropriately provided through public mental health systems, given that this is the primary setting in which seriously mentally ill adults receive formal treatment.


Passage rate of feed in very young chicks.
White Leghorn chicks were reared on diets in which soybean oil at 4% or corn oil at 5 or 10% was substituted isocalorically for carbohydrate calories. The transit time ( TRT ) of diets was determined in these chicks with ferric oxide as a marker. The average TRT for three experiments was 167 min. Overall, there was no significant effect (P greater than .05) for the addition of the vegetable oils to alter TRT . A significant interaction was detected for TRT to be shorter as chicks aged from 7 to 21 days of age when provided with 4 to 5% vegetable oil but not when provided with 10% corn oil. An extra metabolic effect was detected from the isocaloric substitution of lipid calories for carbohydrate calories but not necessarily only when TRT was longer. Thus, the chick was affected differently than the hen in which lipid produced longer TRT .


Erythropoietin ameliorates diabetes-associated cognitive dysfunction in vitro and in vivo.
Several studies indicate that erythropoietin (EPO) has remarkable neuroprotective effects in various central nervous system disorders, while little is known about the effects of EPO in diabetes-associated cognitive dysfunction. Therefore, the present study aimed to investigate whether EPO ameliorates diabetes-associated cognitive dysfunction in vivo and in vitro. We investigated the protective effects of EPO on high-glucose (HG)-induced PC12 cell death and oxidative stress. The effects of EPO (300 U/kg administered three times a week for 4 weeks) on diabetes-associated cognitive decline were investigated in diabetic rats. EPO significantly increased cell viability, increased the activity of superoxide dismutase, decreased the production of malondialdehyde and reactive oxygen species, and decreased the apoptosis rate. Additionally, LY294002, a phosphatidylinositol 3-kinase (PI3K) inhibitor, abolished the protective effects of EPO in HG-treated PC12 cells. In diabetic rats, EPO prevented deficits in spatial learning and memory in the Morris water maze test. The results of real-time PCR and Western blotting showed that EPO upregulated EPO receptor, PI3K, and phosphorylated Akt2 relative to unphosphorylated Akt2 (p-Akt2/Akt2) and downregulated glycogen synthase kinase-3β (GSK-3β). These studies demonstrate that EPO is an effective neuroprotective agent in the context of diabetes-associated cognitive dysfunction and show that this effect involves the PI3K/Akt/GSK-3β pathway.


Gastrointestinal stromal tumor in a patient with undiagnosed neurofibromatosis type I: an uncommon cause of extrahepatic cholestasis.
In this case report we present a 61-year-old patient with obstructive jaundice. Bile duct obstruction was caused by a tumor at the duodenal papilla and bile flow was restored by a plastic stent. Using endoscopic ultrasound and computed tomography imaging two additional tumors of the same morphology were found in the stomach wall and the pelvic region suggesting a multilocular gastrointestinal stroma tumor (GIST). Diagnosis of GIST was confirmed cytologically from the gastric lesion. Based on typical cutaneous manifestations (café-au-lait spots, several tiny dermal neurofibromata and Lisch nodules in the iris), a thus far unidentified neurofibromatosis type I was diagnosed which is known to promote multilocular GIST formation. Tumor resection failed because of cardiac decompensation due to a Takotsubo cardiomyopathy during induction of anesthesia. The patient has been started on imatinib instead and shows so far a stable disease over 6 months.


Transforming growth factor-beta1 induces apoptosis of rat microglia without relation to bcl-2 oncoprotein expression.
Transforming growth factor-beta (TGF-beta) plays a role as an immunosuppressive cytokine within the central nervous system (CNS). The CNS cells targeted by action of TGF-beta1 have not been defined. In this study, we tested the effect of TGF-beta1 on microglia, astrocytes and oligodendrocytes from newborn rats. TGF-beta1 selectively induced apoptosis of microglia, and not of astrocytes or oligodendrocytes. To study the apoptotic mechanism, bcl-2 oncoprotein expression in microglia, astrocytes and oligodendrocytes was measured. Bcl-2 was mainly expressed in microglia, indicating that microglial bcl-2 does not prevent TGF-beta1-mediated microglial apoptosis. The relative protein expression of bcl-2 in microglia was not related to frequency of microglial apoptosis. Thus, TGF-beta1-induced microglial apoptosis was regulated by a bcl-2-independent mechanism. Expression of cytokine (IL-1beta, IL-6, IL-12, IFN-gamma and TNF-alpha) mRNA on microglia was not influenced by treatment with TGF-beta1.


Right Phrenic Nerve Palsy following Long-gap Esophageal Atresia and Tracheoesophageal Fistula Repair.
Esophageal atresia (EA) and tracheoesophageal fistula (TEF) are surgically correctable congenital anomalies with reported surgical common complications such as anastomotic leaks, recurrent TEF, and esophageal strictures; however, phrenic nerve injury (PNI) is a very rare but possible complication which we have highlighted in our case report. Here, we report a baby girl operated for long-gap EA and TEF having respiratory distress and failed attempts to wean off oxygen support. Serial chest X-rays showed elevated right hemidiaphragm, whereas ultrasound thorax confirmed our diagnosis of right PNI causing diaphragmatic palsy. Conservative management with the hope of spontaneous recovery failed, so diaphragmatic plication was done at 5 weeks from index surgery. Postplication, the baby was weaned off oxygen and pressure support the very 1st day and had improved respiratory physiology.


MiR-520b inhibits the development of glioma by directly targeting MBD2.
MicroRNAs play important roles in the process of cancer, which microRNA-520b (miR-520b) has been reported to play critical roles in tumor progression in many types of cancers. However, its role in glioma remains unknown. In this study, we found that miR-520b could inhibit growth and progression in glioma by targeting methyl-CpG-binding domain 2 (MBD2). First, we analyzed the expression of miR-520b in different glioma grades and different cell lines (U87, U251 and astrocyte). Then we assessed the effect of miR-520b on glucose metabolism, invasion, angiogenesis and chemosensitivity in U87 and U251 cells. By using an online database, miR-520b was found to directly bind to the 3'-untranslated regions (3'-UTR) of MBD2 and reduce its expression at the protein level, which further inhibits the development of glioma. MBD2 was also found to be over-expressed in human glioma tissues and in U87 and U251 cells and its level was inversely correlated with that of miR-520b. Furthermore, restoration of MBD2 partially rescued the miR-520b-induced inhibitory effect on glucose metabolism, invasion, angiogenesis and chemosensitivity in glioma cells. In summary, to date, this is the first study to demonstrate that miR-520b functions as a tumor suppressor in glioma by directly targeting MBD2, suggesting that MBD2 may be a potential therapeutic target for glioma.


Dissemination of results from medical student public health research training and factors associated with publication.
To examine factors associated with successful publications resulting from mandatory public health research training attachment, the Trainee Intern Health Care Evaluation (TIHE) projects, at the University of Otago, Dunedin School of Medicine, New Zealand. A total of 227 TIHE projects completed during the period from January 1985 to December 2013 were included in the study. In February 2016, Medline and Google Scholar databases were searched independently by both authors for publications using predefined search criteria. Overall, 25 (11.1%) out of 227 projects resulted in 19 articles, 3 conference presentations/abstracts and 4 cited report abstracts. Nineteen (8.4%) projects resulted in 22 peer-reviewed journal publications, the majority of which were original articles (86.4%). The number of projects commissioned by a client was independently associated with the likelihood of publication, conference abstract or citation of the project report (OR 1.40; P<0.01, 95% CI 1.14 to 1.71). The number of authors and the number of non-student authors were positively associated with publication in higher impact journals, while student first-authored articles were more likely to be published in lower impact journals. Projects completed in more recent years were more likely to be published. Mandatory medical student research experiences promote tangible research output. These findings may help to influence policy around the introduction of required medical school research and facilitate encouraging academic careers among medical students. Future research could focus on examining how different student-related, supervisor-related and programme-related factors influence publication rates from mandatory medical student research attachments.


Therapeutic biomaterials based on extracellular vesicles: classification of bio-engineering and mimetic preparation routes.
Extracellular vesicles (EVs) are emerging as novel theranostic tools. Limitations related to clinical uses are leading to a new research area on design and manufacture of artificial EVs. Several strategies have been reported in order to produce artificial EVs, but there has not yet been a clear criterion by which to differentiate these novel biomaterials. In this paper, we suggest for the first time a systematic classification of the terms used to build up the artificial EV landscape, based on the preparation method. This could be useful to guide the derivation to clinical trial routes and to clarify the literature. According to our classification, we have reviewed the main strategies reported to date for their preparation, including key points such as: cargo loading, surface targeting strategies, purification steps, generation of membrane fragments for the construction of biomimetic materials, preparation of synthetic membranes inspired in EV composition and subsequent surface decoration.


Engineering a Native Inducible Expression System in Shewanella oneidensis to Control Extracellular Electron Transfer.
Shewanella oneidensis MR-1 is a model organism for understanding extracellular electron transport, in which cells transfer intracellular electrons to an extracellular terminal electron acceptor such as insoluble minerals or poised electrodes. Biotechnological applications exploiting the respiratory capabilities of Shewanella species have led to their proposed use in wastewater treatment, bioremediation, and remote sensors. Transcriptional regulation tools can be used to rationally engineer S. oneidensis, optimizing performance in biotechnological applications, introducing new capabilities, or investigating physiology. Engineered gene expression in S. oneidensis has primarily involved the use of foreign regulatory systems from Escherichia coli. Here we characterize a native S. oneidensis pathway that can be used to induce gene expression with trimethylamine N-oxide, then engineer strains in which extracellular electron transfer is controlled by this compound. The ability to induce this pathway was assessed by measuring iron reduction over time and by analyzing anodic current produced by cells grown in bioreactors.


Glucose transporter 3 is a rab11-dependent trafficking cargo and its transport to the cell surface is reduced in neurons of CAG140 Huntington's disease mice.
Huntington's disease (HD) disturbs glucose metabolism in the brain by poorly understood mechanisms. HD neurons have defective glucose uptake, which is attenuated upon enhancing rab11 activity. Rab11 regulates numerous receptors and transporters trafficking onto cell surfaces; its diminished activity in HD cells affects the recycling of transferrin receptor and neuronal glutamate/cysteine transporter EAAC1. Glucose transporter 3 (Glut3) handles most glucose uptake in neurons. Here we investigated rab11 involvement in Glut3 trafficking. Glut3 was localized to rab11 positive puncta in primary neurons and immortalized striatal cells by immunofluorescence labeling and detected in rab11-enriched endosomes immuno-isolated from mouse brain by Western blot. Expression of dominant active and negative rab11 mutants in clonal striatal cells altered the levels of cell surface Glut3 suggesting a regulation by rab11. About 4% of total Glut3 occurred at the cell surface of primary WT neurons. HD(140Q/140Q) neurons had significantly less cell surface Glut3 than did WT neurons. Western blot analysis revealed comparable levels of Glut3 in the striatum and cortex of WT and HD(140Q/140Q) mice. However, brain slices immunolabeled with an antibody recognizing an extracellular epitope to Glut3 showed reduced surface expression of Glut3 in the striatum and cortex of HD(140Q/140Q) mice compared to that of WT mice. Surface labeling of GABAα1 receptor, which is not dependent on rab11, was not different between WT and HD(140Q/140Q) mouse brain slices. These data define Glut3 to be a rab11-dependent trafficking cargo and suggest that impaired Glut3 trafficking arising from rab11 dysfunction underlies the glucose hypometabolism observed in HD.


Safety and efficacy of vardenafil in patients with erectile dysfunction: result of a bridging study in Japan.
Vardenafil is a selective and highly potent phosphodiesterase type 5 (PDE5) inhibitor for the treatment of erectile dysfunction (ED), with improved selectivity for PDE5 and demonstrated efficacy for improving sexual function in men with ED. The current study investigated the safety and efficacy of this new PDE5 inhibitor in Japanese men with ED. This was a prospective, double blind, randomized clinical trial designed to evaluate the efficacy and safety of vardenafil. Following a 4-week treatment-free observation period, 283 eligible patients were randomized to 12 weeks treatment with vardenafil 5 mg, 10 mg, 20 mg, or placebo. Primary efficacy responses were assessed using the scores of Q3 and Q4 of the international index of erectile function (IIEF). All three vardenafil doses showed significantly better improvement than the placebo group in Q3 and Q4 scores of the IIEF questionnaire, either at 12 weeks or at the 'last observation carried forward' (LOCF, P < 0.0001). Q3 scores were improved to 4.06 with vardenafil 5 mg, 4.53 with vardenafil 10 mg, and 4.64 with vardenafil 20 mg, versus 3.17 with placebo. Comparable scores for Q4 were 3.47, 4.15 and 4.31 versus 2.31 for placebo. Up to 86% of patients achieved improved erections as assessed by the global assessment question (GAQ). Reported adverse event rates were 35.3%, 45.3% and 54.5% with vardenafil 5 mg, 10 mg and 20 mg, respectively, versus 21.1% in the placebo group. No serious adverse drug reactions were reported. The most common treatment-emergent adverse events were transient headache, flushing and rhinitis, which were mostly mild. Vardenafil is an effective and well-tolerated treatment for ED and provides improvement in key indices of erectile function among Japanese men with ED. The results of our trial show that up to nearly 90% of patients achieve improved erections with the administration of vardenafil.


Simultaneous determination of toxins in algae and water samples by high-performance liquid chromatography with triple quadrupole mass spectrometry.
A facile method based on liquid chromatography coupled with electrospray ionization tandem triple quadrupole mass spectrometry working in selected reaction monitoring mode has been established to analyze toxins in the algae and water samples. Twelve types of toxins (anatoxin, cylindrospermopsin, dinophysistoxin-1, nodularin, okadaic acid, microcystins) were efficiently separated under optimized liquid chromatography coupled with mass spectrometry conditions in the selected reaction monitoring mode. Correlation coefficients of the calibration curves, all felt in the range of 0.9958-0.9998, indicated good linearity. The detection limits of toxins in this method were all lower than 0.20 ng/mL and the quantification limits were in the range from 0.04 to 0.60 ng/mL. Except for anatoxin, cylindrospermopsin, and nodularin, the other toxins' recoveries varied from 55.45 to 140.85%. And the relative standard deviations of interday and intraday precision were at 8.61% (n = 5). The high-performance liquid chromatography (HPLC)/electrospray ionization (ESI)-mass spectrometery (MS) method was also successfully applied to analyze the algae and water samples. Owing to its exclusive selectivity and excellent sensitivity, the developed method is a tool for comprehensive analyses of the 12 types of toxins at nanogram levels.


Role of mitogen-activated protein kinase activation in the production of inflammatory mediators: differences between primary rat alveolar macrophages and macrophage cell lines.
Stimulation of macrophages has been shown to activate all three families of mitogen activated protein kinases (MAPKs). However, variable results are reported in the literature with respect to the particular kinases activated with any given stimulus. In this study, the role of activation of MAPKs was examined in the production of inflammatory mediators by measuring the phosphorylation of the kinases and their ability to phosphorylate specific substrates in rat primary alveolar macrophages, a rat alveolar macrophage cell line (NR8383), and two mouse monocytic cell lines (RAW 264.7 and J774A.1). In the three cell lines examined, all three families of MAPKs were activated upon stimulation with either lipopolysaccharide (LPS) or LPS plus interferon-gamma; in contrast, only ERK1/2 was activated in primary rat alveolar macrophages upon stimulation with LPS. Inhibition of ERK1/2 activation by the MEK inhibitor PD98059 abrogated nitric oxide and tumor necrosis factor-alpha (TNF-alpha) production in primary rat alveolar macrophages, but the p38 inhibitor SB203580 had no effect on the production of these two inflammatory mediators. These observations indicate that MAPK activation is cell specific and explain some of the conflicting results reported in the literature. These studies emphasize the need to exercise caution in extrapolating data from cell lines to primary cells.


[Use of filter paper to identify antibodies against Myxovirus influenzae].
A total of 165 sera and 142 eluates from filtration papers impreggnated with whole blood were examined in reaction of complement fixation against A/Hong Kong 1/68 (H3N2) and B/Johannesburg 3/56 virus. To perform HIT 152 sera and 155 eluates were used. The results obtained were elaborated statistically. The method of filtration papers does not seem to be suitable for HIT, whereas the CFT was in several cases practicable.


Transjugular intrahepatic portosystemic shunt in cavernomatous portal vein occlusion.
A 72-year-old patient with liver cirrhosis and cavernomatous portal vein occlusion presented with refractory ascites. We treated the patient with transjugular intrahepatic portosystemic shunt: transjugular transhepatic puncture of an intrahepatic hilar collateral vein was performed; transjugular intrahepatic portosystemic shunt was created between the right hepatic vein and the patent superior mesenteric vein using this hilar collateral vein as the connecting pathway. The ascites was resolved and the patient remains asymptomatic, while shunt patency is maintained 16 months after the intervention.


Catecholamines, angiotensin II and sodium concentrations in cerebrospinal fluid in young men with borderline hypertension.
To evaluate the role of central nervous mechanisms and their relationships to the peripheral sympathetic nervous system in borderline hypertension, we measured catecholamines, angiotensin II (AII) and sodium (Na) concentrations in cerebrospinal fluid (CSF) with plasma catecholamines concomitantly in 12 young men with borderline hypertension and 7 age-matched healthy normotensive men on ordinary salt intake. Plasma norepinephrine (NE) and epinephrine (E) were higher in the borderline hypertensives than in the normotensives (NE: 239 +/- 15 vs 190 +/- 11 pg/ml, p less than 0.05, E: 83 +/- 9 vs 43 +/- 6 pg/ml, p less than 0.01). NE levels in CSF were also higher in the borderline hypertensives than in the normotensives (200 +/- 15 vs 150 +/- 18 pg/ml, p less than 0.05). In most of the subjects, CSF E and plasma and CSF dopamine levels were below the sensitivity of the assay. CSF NE correlated positively with both plasma NE (p less than 0.01) and mean blood pressure (p less than 0.05) in all subjects. Immunoreactive AII and Na concentrations in CSF did not differ between the borderline hypertensives and normotensives. These results suggest that peripheral sympathoadrenal overactivity in young subjects with borderline hypertension may be related to an altered function of central noradrenergic neurons. AII and Na in the central nervous system do not appear to have an important role in borderline hypertension.


Effect of tea catechins on the change of glutathione levels caused by Pb(++) in PC12 cells.
Our previous research showed that tea catechins could significantly increase the viability of lead-exposed PC12 cells. Whereas the cellular thiol status is known to be responsible for protecting against lead-induced toxicity, whether the role of tea catechins on lead-induced PC12 cell toxicity is related to the metabolism of intracellular thiol compounds remained vague. In the present study, it was found that Pb(2+) significantly decreased reduced glutathione (GSH)/oxidative glutathione (GSSG) and protein sulfhydryl groups (PSH)/glutathione-protein mixed disulfide (GSSP) ratios as well as glutathione reductase activities in a concentration-dependent manner. Both (-)-epicatechin and (-)-epicatechin gallate (ECG) supplementation resulted in an increased GSH/GSSG ratio and glutathione reductase activities. The galloylated catechins (ECG or (-)-epigallocatechin gallate) treatment significantly decreased the GSSP levels and increased the intracellular PSH/GSSP ratio in lead-exposed PC12 cells. To our surprise, as compared with the group treated by lead acetate, 100 microM EGC treatment following lead exposure significantly decreased GSH/GSSG and PSH/GSSP ratios, as well as glutathione reductase activities. The results suggested that the effect of tea catechins on the intracellular thiols status in PC12 cells was different, which may be related to their chemical structures and/or regulation of special gene expression properties.


Autoamputation of a pedunculated, subserosal uterine leiomyoma presenting as a giant peritoneal loose body.
Peritoneal loose bodies (PLBs) are defined as fibrotic or calcified-free bodies within the peritoneal cavity; they commonly autoamputate from appendices epiploicae that have undergone torsion. Pedunculated, subserosal uterine leiomyomas (PSULs) are subserosal uterine leiomyomas connected to the uterus via a pedicle. In the present report, we describe the case of a PLB that originated from the autoamputation of a PSUL, confirmed based on histological evidence consistent with a uterine leiomyoma and the laparoscopic findings of a broken pedicle. This case clearly demonstrates the potential for a uterine leiomyoma to be the source of a PLB. Our findings contribute to the understanding of the etiological relationship between PLBs and uterine leiomyomas.


Combining domain filling with a self-organizing map to analyze multi-species hydrocarbon signatures on a regional scale.
For the period of the Barnett Coordinated Campaign, October 16-31, 2013, hourly concentrations for 46 volatile organic compounds (VOCs) were recorded at 14 air monitoring stations within the Barnett Shale of North Texas. These measurements are used to identify and analyze multi-species hydrocarbon signatures on a regional scale through the novel combination of two techniques: domain filling with Lagrangian trajectories and the machine learning unsupervised classification algorithm called a self-organizing map (SOM). This combination of techniques is shown to accurately identify concentration enhancements in the lightest measured alkane species at and downwind of the locations of active-permit oil and gas facilities, despite the model having no a priori knowledge of these source locations. Site comparisons further identify the SOM's ability to distinguish between signatures with differing influences from oil- and gas-related processes and from urban processes. A random forest (a machine learning supervised classification) analysis is conducted to further probe the sensitivities of the SOM classification in response to changes in any hydrocarbon species' concentration values. The random forest analysis of four representative classes finds that the SOM classification is appropriately more sensitive to changes in certain urban-related species for urban-related classes, and to changes in oil- and gas-related species for oil- and gas-related classes.


Further characterization of a non-essential mutator gene in Escherichia coli K-12.
The properties of mutR, a mutator closely linked to thyA, have been further characterized. We have found that the mutator gene is carried on a specialized transducing phage (lambdapcI857 thyA) generated by the excision of lambdacI857 integrated at a secondary attachment site between lysA and thyA. We present three lines of evidence indicating that mutR is a nonessential gene. (i) Deletions of the mutator can be found amoung survivors of heat induction of lambdacI857 when the phage is integrated between lysA and thyA. (ii) Mutations in mutR can be induced with the frameshift mutagen ICR-191. (iii) An amber mutant in mutR has been found. Viable strains could be made by combining the mutator with polB, polA polR, ligts7, and uvrA mutations. The mutator was still able to increase the spontaneous mutation frequency in these genetic backgrounds. When the reversion patterns of a series of well-characterized trpA mutations were analyzed, the results suggested that mutR is more efficient at causing transitions than transversion mutations.


Reverse engineering human neurodegenerative disease using pluripotent stem cell technology.
With the technology of reprogramming somatic cells by introducing defined transcription factors that enables the generation of "induced pluripotent stem cells (iPSCs)" with pluripotency comparable to that of embryonic stem cells (ESCs), it has become possible to use this technology to produce various cells and tissues that have been difficult to obtain from living bodies. This advancement is bringing forth rapid progress in iPSC-based disease modeling, drug screening, and regenerative medicine. More and more studies have demonstrated that phenotypes of adult-onset neurodegenerative disorders could be rather faithfully recapitulated in iPSC-derived neural cell cultures. Moreover, despite the adult-onset nature of the diseases, pathogenic phenotypes and cellular abnormalities often exist in early developmental stages, providing new "windows of opportunity" for understanding mechanisms underlying neurodegenerative disorders and for discovering new medicines. The cell reprogramming technology enables a reverse engineering approach for modeling the cellular degenerative phenotypes of a wide range of human disorders. An excellent example is the study of the human neurodegenerative disease amyotrophic lateral sclerosis (ALS) using iPSCs. ALS is a progressive neurodegenerative disease characterized by the loss of upper and lower motor neurons (MNs), culminating in muscle wasting and death from respiratory failure. The iPSC approach provides innovative cell culture platforms to serve as ALS patient-derived model systems. Researchers have converted iPSCs derived from ALS patients into MNs and various types of glial cells, all of which are involved in ALS, to study the disease. The iPSC technology could be used to determine the role of specific genetic factors to track down what's wrong in the neurodegenerative disease process in the "disease-in-a-dish" model. Meanwhile, parallel experiments of targeting the same specific genes in human ESCs could also be performed to control and to complement the iPSC-based approach for ALS disease modeling studies. Much knowledge has been generated from the study of both ALS iPSCs and ESCs. As these methods have advantages and disadvantages that should be balanced on experimental design in order for them to complement one another, combining the diverse methods would help build an expanded knowledge of ALS pathophysiology. The goals are to reverse engineer the human disease using ESCs and iPSCs, generate lineage reporter lines and in vitro disease models, target disease related genes, in order to better understand the molecular and cellular mechanisms of differentiation regulation along neural (neuronal versus glial) lineages, to unravel the pathogenesis of the neurodegenerative disease, and to provide appropriate cell sources for replacement therapy. This article is part of a Special Issue entitled SI: PSC and the brain.


The effect of reproductive compensation on recessive disorders within consanguineous human populations.
We investigate the effects of consanguinity and population substructure on genetic health using the UK Asian population as an example. We review and expand upon previous treatments dealing with the deleterious effects of consanguinity on recessive disorders and consider how other factors, such as population substructure, may be of equal importance. For illustration, we quantify the relative risks of recessive lethal disorders by presenting some simple calculations that demonstrate the effect 'reproductive compensation' has on the maintenance of recessive alleles. The results show how reproductive compensation can effectively counteract the purging of deleterious alleles within consanguineous populations. Whereas inbreeding does not elevate the equilibrium frequency of affected individuals, reproductive compensation does. We suggest this effect must be built into interpretations of the incidence of genetic disease within populations such as the UK Asians. Information of this nature will benefit health care workers who inform such communities.


Perception of English Stress of Synthesized Words by Three Chinese Dialect Groups.
This study investigated the possible prosodic transfer influences native regional dialects may have in the perception of English lexical stress by speakers of three Chinese dialects [Beijing (BJ), Changsha (CS), and Guangzhou (GZ)] compared to 20 American English (AE) speakers. F0, duration, intensity, and vowel reduction were manipulated in nonce disyllabic words. Participants performed four-word sequence recall tasks to identify lexical stress location. They performed better with natural sounds than with manipulated words. This study focused on the performance differences in manipulating words. The results showed that all four-group members performed similarly processing F0 condition nonce words. BJ and CS participants were more accurate than GZ participants in duration and vowel reduction cues. Reaction time (RT) suggested that the processing time of acoustic cues differed significantly across language groups. The findings indicate that first language (L1) dialect effect is robust in second language (L2) stress perception tasks.


Bisphenol A induces cell cycle arrest in primary and prostate cancer cells through EGFR/ERK/p53 signaling pathway activation.
Bisphenol A (BPA) belongs to the class of chemicals known as endocrine disruptors and has been also involved in the pathogenesis and progression of endocrine related cancer such as breast and prostate cancers. Here, we have investigated the effect of BPA in human prostate cancer LNCaP cells and in human non-transformed epithelial prostate EPN cells. Our data showed that BPA induces the down regulation of cyclin D1 expression and the upregulation of the cell cycle inhibitors p21 and p27, leading to cell cycle arrest. Interestingly, we found that the BPA anti-proliferative response depends on a strong and rapid activation of epidermal growth factor receptor (EGFR), which stimulates ERK-dependent pathway. This, in turn, induces expression of p53 and its phosphorylation on residue Ser15, which is responsible for cell cycle arrest. EGFR activation occurs upon a cross talk with androgen (AR) and estradiol receptor-β (ERβ) which are known to bind BPA. Altogether, these findings show a novel signaling pathway in which EGFR activation plays a key role on BPA-induced cell cycle inhibition through a pathway involving AR and ERβ/EGFR complexes, ERK and p53. Our results provide new insights for understanding the molecular mechanisms in human prostate cancer. On the other, they could allow the development of new compounds that may be used to overcome human prostate cancer resistance to endocrine therapy in promising target therapeutic approaches.


Job mobility, its determinants, and its effects: longitudinal data from the Maastricht Cohort Study.
This prospective study of 12,140 employees examined the effects of work characteristics and situational, psychological, and health aspects on job mobility. Before job change, the mobility group reported significantly more conflicts with the supervisor, higher physical and emotional strain, higher degree of job insecurity, lower job satisfaction, and lower degree of commitment compared with employees who did not change jobs. After job change, the mobility group reported improved autonomy, task diversity, decreased occurrence of conflicts with the supervisor, decreased physical and emotional strain, and improved training possibilities and job security than before the change. Changing jobs had a positive effect on employees with respect to job perception and job satisfaction and led to reduced fatigue and need for recovery.


Induction of unresponsiveness to skin allografts in adult mice disparate at defined regions of the H-2 complex. I. Effect of donor-specific bone marrow in ALS-treated mice.
The induction of unresponsiveness to skin allografts was studied in adult congenic B10 mice disparate either at isolated K, I, or D regions of the H-2 complex or at combined K/D and I regions. Recipient mice were treated with antilymphocyte serum (ALS) and injected with donor bone marrow after skin grafting. Unresponsiveness to antigens of the isolated K or D regions (class I) could not be induced, as shown by the inability of bone marrow to prolong graft survival in these mice compared with ALS-treated controls. Significant unresponsiveness to antigens of the isolated I region could be induced by ALS immunosuppression alone, and it was not possible to determine the additive effects of bone marrow. Unresponsiveness to antigens of the K or D regions, or both, could be induced when they were presented with antigens of the I region. This indicates that the I region exerts a tolerogenic effect on the K and D regions. The requirement for the concomitant presentation of antigens of the K/D and I regions to achieve unresponsiveness is consistent with our previous observations that active suppression is one of the mechanisms of the induction of unresponsiveness in ALS-treated, marrow injected mice.


Oral manifestations of human T-cell lymphotropic virus infection in adult patients from Brazil.
Human T-cell lymphotropic virus type 1 (HTLV-1) was the first human retrovirus discovered and its pathogenesis is related to T cells infection. This study aimed to verify the presence of oral manifestations in a Brazilian population of patients who was seropositive for HTLV, and identify risk factors for oral manifestations. An assessment was made of 139 patients at the Emilio Ribas Institute of Infectious Diseases. A total of 112 (80.5%) patients were HTLV-1, 26 (18.7%) were HTLV-2+. About 35.2% of patients had myelopathy/tropical spastic paraparesis (HAM/TSP), with 48 of them being HTLV-1+ and one patient was seropositive for HTLV-1 and -2. The most common oral manifestations were: xerostomia (26.8%), candidiasis (20.8%), fissured tongue (17.9%), and loss of tongue papillae (10.0%). A multivariate logistic regression analysis showed that HAM/TSP is an independent risk factor for xerostomia (P = 0.02). The patients who were HAM/TSP+ were three times more likely to develop xerostomia when compared with patients without HAM/TSP (odds ratio = 2.69, 95% confidence interval = 1.17-6.17). Despite the fact that the findings of this study suggest a relationship between xerostomia and HAM/TSP, more studies should be developed to show what the association would be between xerostomia presented by HTLV patients and pathogenesis of the virus.


Flt-3 ligand and sequential FL/interleukin-2 in patients with metastatic renal carcinoma: clinical and biologic activity.
Flt3 (fms-like tyrosine kinase 3) ligand, a cytokine that stimulates increases in the number of dendritic cells (DC) in vivo, has been shown to have antitumor activity in murine models via an immune-mediated mechanism. Therefore, we examined the clinical activity of this cytokine in patients with an immunologic-responsive cancer, metastatic renal cell carcinoma. Flt3 ligand (25 microg/kg subcutaneous) was administered daily for the first 14 days of a 28-day cycle. Although the treatment was well tolerated and was confirmed to induce expansion of lineage (Lin)-/HLA-DR+/CD11c+ myeloid DC and Lin-/HLA-DR+/CD123+ plasmacytoid DC, no clinical activity was observed. Reasoning that DC expanded by Flt3 ligand might potentiate the clinical activity of low-dose interleukin-2, a second study was conducted of sequential administration of 25 microg/kg of Flt3 ligand daily for 7 days was followed by 11 x 10(6) IU of subcutaneous interleukin-2 for 4 consecutive days x 4 weeks. In this study, increased numbers of circulating DC were again observed, which was followed by increased numbers of activated T cells, confirming a biologic effect of each cytokine. However, toxicity and clinical efficacy were similar to what has been seen with low-dose interleukin-2 alone, with two minor responses observed. These results demonstrate that Flt3 ligand, although capable of inducing expansion of circulating myeloid and plasmacytoid DC in patients with metastatic renal cell carcinoma, lacks significant clinical activity at the doses and schedules examined.


Cocaine Self-Administration Elevates GluN2B within dmPFC Mediating Heightened Cue-Elicited Operant Responding.
Cue-elicited drug-craving correlates with hyperactivity within prefrontal cortex (PFC), which is theorized to result from dysregulated excitatory neurotransmission. The NMDA glutamate receptor is highly implicated in addiction-related neuroplasticity. As NMDA receptor function is regulated critically by its GluN2 subunits, herein, we assayed the relation between incubated cue-elicited cocaine-seeking following extended access to intravenous cocaine (6 h/d; 0.25 mg/infusion for 10 d) and the expression of GluN2A/B receptor subunits within PFC sub regions during early versus late withdrawal (respectively, 3 vs. 30 days). Cocaine-seeking rats exhibited elevated GluN2B expression within the dorsomedial aspect of the PFC (dmPFC); this effect was apparent at both 3 and 30 days withdrawal and occurred in cocaine-experienced rats, regardless of experiencing an extinction test or not. Thus, elevated dmPFC GluN2B expression appears to reflect a pharmacodynamic response to excessive cocaine intake that is independent of the duration of drug withdrawal or re-exposure to drug-taking context. The functional relevance of elevated dmPFC GluN2B expression for drug-seeking was assessed by the local infusion of the prototypical GluN2B-selective antagonist ifenprodil (1.0 µg/side). Ifenprodil did not alter cue-elicited responding in animals with a history of saline self-administration. In contrast, ifenprodil lowered cue-elicited cocaine-seeking, while potentiating cue-elicited sucrose-seeking. Thus, the effects of an intra-dmPFC ifenprodil infusion upon cue reactivity are reinforcer-specific, arguing in favor of targeting GluN2B-containing NMDA receptors as a pharmacological strategy for reducing behavioral reactivity to drug-associated cues with the potential benefit of heightening the reinforcing properties of cues associated with non-drug primary rewards.


Accuracy Assessment of Three-dimensional Surface Reconstructions of In vivo Teeth from Cone-beam Computed Tomography.
The accuracy of three-dimensional (3D) reconstructions from cone-beam computed tomography (CBCT) has been particularly important in dentistry, which will affect the effectiveness of diagnosis, treatment plan, and outcome in clinical practice. The aims of this study were to assess the linear, volumetric, and geometric accuracy of 3D reconstructions from CBCT and to investigate the influence of voxel size and CBCT system on the reconstructions results. Fifty teeth from 18 orthodontic patients were assigned to three groups as NewTom VG 0.15 mm group (NewTom VG; voxel size: 0.15 mm; n = 17), NewTom VG 0.30 mm group (NewTom VG; voxel size: 0.30 mm; n = 16), and VATECH DCTPRO 0.30 mm group (VATECH DCTPRO; voxel size: 0.30 mm; n = 17). The 3D reconstruction models of the teeth were segmented from CBCT data manually using Mimics 18.0 (Materialise Dental, Leuven, Belgium), and the extracted teeth were scanned by 3Shape optical scanner (3Shape A/S, Denmark). Linear and volumetric deviations were separately assessed by comparing the length and volume of the 3D reconstruction model with physical measurement by paired t- test. Geometric deviations were assessed by the root mean square value of the imposed 3D reconstruction and optical models by one-sample t-test. To assess the influence of voxel size and CBCT system on 3D reconstruction, analysis of variance (ANOVA) was used (μ = 0.05). The linear, volumetric, and geometric deviations were -0.03 ± 0.48 mm, -5.4 ± 2.8%, and 0.117 ± 0.018 mm for NewTom VG 0.15 mm group; -0.45 ± 0.42 mm, -4.5 ± 3.4%, and 0.116 ± 0.014 mm for NewTom VG 0.30 mm group; and -0.93 ± 0.40 mm, -4.8 ± 5.1%, and 0.194 ± 0.117 mm for VATECH DCTPRO 0.30 mm group, respectively. There were statistically significant differences between groups in terms of linear measurement (P < 0.001), but no significant difference in terms of volumetric measurement (P = 0.774). No statistically significant difference were found on geometric measurement between NewTom VG 0.15 mm and NewTom VG 0.30 mm groups (P = 0.999) while a significant difference was found between VATECH DCTPRO 0.30 mm and NewTom VG 0.30 mm groups (P = 0.006). The 3D reconstruction from CBCT data can achieve a high linear, volumetric, and geometric accuracy. Increasing voxel resolution from 0.30 to 0.15 mm does not result in increased accuracy of 3D tooth reconstruction while different systems can affect the accuracy.


Selection Criteria for Small-Molecule Inhibitors in Area-Selective Atomic Layer Deposition: Fundamental Surface Chemistry Considerations.
ConspectusAtomically precise and highly selective surface reactions are required for advancing microelectronics fabrication. Advanced atomic processing approaches make use of small molecule inhibitors (SMI) to enable selectivity between growth and nongrowth surfaces. The selectivity between growth and nongrowth substrates is eventually lost for any known combinations, because of defects, new defect formation, and simply because of a Boltzmann distribution of molecular reactivities on surfaces. The selectivity can then be restored by introducing etch-back correction steps. Most recent developments combine the design of highly selective combinations of growth and nongrowth substrates with atomically precise cycles of deposition and etching methods. At that point, a single additional step is often used to passivate the unwanted defects or selected surface chemical sites with SMI. This step is designed to chemically passivate the reactive groups and defects of the nongrowth substrates both before and/or during the deposition of material onto the growth substrate. This approach requires applications of the fundamental knowledge of surface chemistry and reactivity of small molecules to effectively block deposition on nongrowth substrates and to not substantially affect deposition on the growth surface. Thus, many of the concepts of classical surface chemistry that had been developed over several decades can be applied to design such small molecule inhibitors. This article will outline the approaches for such design.This is especially important now, since the ever-increasing number of applications of this concept still rely on trial-and-error approaches in selecting SMI. At the same time, there is a very substantial breadth of surface chemical reactivity analysis that can be put to use in this process that will relate the effectiveness of a potential SMI on any combination of surfaces with the following: selectivity; chemical stability of a molecule on a specific surface; volatility; steric hindrance, geometry, packing, and precursor of choice for material deposition; strength of adsorption as detailed by interdisplacement to determine the most stable SMI; fast attachment reaction kinetics; and minimal number of various binding modes.The down-selection of the SMI from the list of chemicals that satisfy the preliminary criteria will be decided based on optimal combinations of these requirements. Although the specifics of SMI selection are always affected by the complexity of the overall process and will depend drastically on the materials and devices that are or will be needed, this roadmap will assist in choosing the potential effective SMIs based on quite an exhaustive set of "SMI families" in connection with general types of target surfaces.


Endogenous leptin receptor signaling in the medial nucleus tractus solitarius affects meal size and potentiates intestinal satiation signals.
Leptin receptor (LepRb) signaling in the hindbrain is required for energy balance control. Yet the specific hindbrain neurons and the behavioral processes mediating energy balance control by hindbrain leptin signaling are unknown. Studies here employ genetic [adeno-associated virally mediated RNA interference (AAV-RNAi)] and pharmacological methodologies to specify the neurons and the mechanisms through which hindbrain LepRb signaling contributes to the control of food intake. Results show that AAV-RNAi-mediated LepRb knockdown targeting a region encompassing the mNTS and area postrema (AP) (mNTS/AP LepRbKD) increases overall cumulative food intake by increasing the size of spontaneous meals. Other results show that pharmacological hindbrain leptin delivery and RNAi-mediated mNTS/AP LepRb knockdown increased and decreased the intake-suppressive effects of intraduodenal nutrient infusion, respectively. These meal size and intestinally derived signal amplification effects are likely mediated by LepRb signaling in the mNTS and not the AP, since 4th icv and mNTS parenchymal leptin (0.5 μg) administration reduced food intake, whereas this dose did not influence food intake when injected into the AP. Overall, these findings deepen the understanding of the distributed neuronal systems and behavioral mechanisms that mediate the effects of leptin receptor signaling on the control of food intake.


Quantitative Analysis of Lipid-Rich Necrotic Core in Carotid Atherosclerotic Plaques by In Vivo Magnetic Resonance Imaging and Clinical Outcomes.
BACKGROUND The aim of this study was to explore the accuracy of in vivo magnetic resonance imaging (MRI) in the quantitative evaluation of lipid-rich necrotic core (LRNC) in carotid atherosclerotic plaques compared with histopathology, and to assess the association of LRNC size with cerebral ischemia symptoms. MATERIAL AND METHODS Thirty patients were enrolled and 19 patients (16 men and 3 women) were analyzed. All the patients were submitted to MRI on a Siemens Avanto (1.5-Tesla) device before carotid endarterectomy (CEA). The scanning protocol included three-dimensional time of flight (3D TOF), T1-weighted image (T1WI), T2-weighted image (T2WI), turbo spin-echo T2-weighted (T2-TSE), and contrast-enhanced T1-weighted image. MRI images were reviewed for quantitative measurements of LRNC areas. LRNC specimens were collected for histology. Percentages of LRNC area to total vessel area were assessed to determine the association of MRI with histological findings. RESULTS There were 151 pairs of matched MRI and pathological sections. LRNC area percentages (LRNC area/vessel area) measured by MRI and histology were 20.6±9.0% and 18.7±9.5%, respectively (r=0.69, p<0.001). Twelve out of 19 patients had symptoms (S-group; 3 had recent stroke, 3 had a recent stroke and a history of transient ischemic attack (TIA), and 6 had TIA); the remaining 7 subjects showed no symptoms (NS-group). LRNC area percentages in the S- and NS-groups were 22.2±5.8% and 12.6±10.7%, respectively (p<0.05). CONCLUSIONS MRI can quantitatively measure LRNC in carotid atherosclerotic plaques, and may be useful in predicting the rupture risk of plaques. These findings provide a basis for imaging use in individualized treatment plan.


Plasma oxidative stress parameters in men and women with early stage Alzheimer type dementia.
It is well known that oxidative stress is one of the earliest events in Alzheimer's disease pathogenesis, indicating that may play a key role in this disease. In our study, we measured the levels of oxidative stress indicators (TBARS and protein carbonyls content) and the non-enzymatic (glutathione (GSH) and oxidized glutathione (GSSG)) and enzymatic (glutathione peroxidase (GPx), catalase (CAT) and superoxide dismutase (SOD)) defense systems in the plasma of 46 patients diagnosed of ATD and 46 age-matched controls. We found decreased levels in total GSH in ATD patients, although healthy control women showed lower levels of total GSH than healthy control men. On the contrary, we found increased levels of TBARS and carbonyl groups content in ATD patients in both genders. The activity of the plasma antioxidant enzymes showed no changes for SOD activity in ATD patients, independently of the gender, although western blot analysis showed an increase in SOD-1 protein. CAT activity was also decreased in ATD patients, although this decrease is mainly due to the decrease found in men but not in women. However, western blot analysis did not show differences in CAT protein between controls and ATD patients. Finally, a decrease of GPx activity was found in ATD patients in both genders. However, as with CAT protein, western blot analysis did not show differences in GPx protein between controls and ATD patients. Our results suggest that there is a defect in the antioxidant defense system that is incapable of responding to increased free radical production, which may lead to oxidative damage and the development of the pathological alterations that characterize the neurodegenerative disorder of patients with ATD. Thus, oxidative damage could be one important aspect for the onset of ATD and oxidative stress markers could be useful to diagnose the illness in their earliest stages through both non-invasive, reliable and cost-affordable methods.


Diagnosing neuroendocrine dysfunction in patients after aneurysmal subarachnoid hemorrhage in clinical practice - does basal hormone screening make sense?
Recent studies indicate that neuroendocrine dysfunction is a more frequent sequel of aneurysmal subarachnoid hemorrhage (SAH), than has so far been recognized. However, from the available data it remains unclear whether certain subgroups of SAH patients carry a higher risk to sustain endocrine sequelae due to the hemorrhage than others and should be specifically followed up in terms of hormone assessment. To investigate whether a basal hormone screening is a practical method in clinical routine to single out patients in whom endocrine function testing is warranted, we established a screening protocol, based on the findings from a cohort of 40 SAH patients (study group) who had all been investigated by basal hormone para meters as well as standardized endocrinological function testing, within the framework of a previously published clinical study. We then applied this protocol to 45 newly investigated SAH-patients (screening group). According to the thus established protocol, 20 of the 45 screened patients (44.4 %) were recommended further investigations, 12 of whom agreed to undergo dynamic endocrine function testing. Altogether, the percentage of test-confirmed neuroendocrine dysfunction was only 13.3 % (6/ 45) in the screening group as compared to 55 % in the study group. Low IGF-I (2 SD below normal) did not serve to predict growth hormone deficiency, whereas low 9 am serum cortisol was of limited value to single out ACTH-deficiency in SAH-patients. In summary we conclude that basal hormone screening is not sufficient to identify SAH patients with impaired hypothalamo-pituitary function, at least not in the context of clinical routine practice.


The role of deoxycorticosterone in the biosynthesis of cardenolides in Digitalis lanata.
The role of deoxycorticosterone in the biosynthesis of digitoxigenin was investigated by the simultaneous administration of deoxy[1,2-(3)H(2)]corticosterone and [4-(14)C]progesterone to a Digitalis lanata plant. The biosynthetically formed [(3)H,(14)C]digitoxigenin and deoxy[(3)H,(14)C]corticosterone were isolated and the distribution of the two isotopes in these products was determined. The transformation of progesterone into deoxycorticosterone in vivo was established. The biosynthetic route from progesterone via deoxycorticosterone to cardenolides was found to be of little significance.


Adiabatic ekpyrosis: scale-invariant curvature perturbations from a single scalar field in a contracting universe.
The Universe can be made flat and smooth by undergoing a phase of ultraslow (ekpyrotic) contraction, a condition achievable with a single, canonical scalar field and conventional general relativity. It has been argued, though, that generating scale-invariant density perturbations requires at least two scalar fields and a two-step process that first produces entropy fluctuations and then converts them to curvature perturbations. In this Letter we identify a loophole in the argument and introduce an ekpyrotic model based on a single, canonical scalar field that generates nearly scale-invariant curvature fluctuations through a purely "adiabatic mechanism" in which the background evolution is a dynamical attractor. The resulting spectrum can be slightly red with distinctive non-Gaussian fluctuations.


Systems-level questions in Drosophila oogenesis.
This paper describes computational and experimental work on pattern formation in Drosophila egg development (oogenesis), an established experimental model for studying cell fate diversification in developing tissues. Epidermal growth factor receptor (EGFR) is a key regulator of pattern formation and morphogenesis in Drosophila oogenesis. EGFR signalling in oogenesis can be genetically manipulated and monitored at many levels, leading to large sets of heterogeneous data that enable the formulation of increasingly quantitative models of pattern formation in these systems.


Posterior reversible encephalopathy syndrome in a patient with systemic lupus erythematosus/systemic sclerosis overlap syndrome.
Posterior reversible encephalopathy syndrome is a clinicoradiologic entity associated with diverse medical conditions. It is very important to properly recognize this condition because early diagnosis and treatment usually result in its complete resolution, whereas a delay in giving an adequate therapy may lead to permanent neurologic sequelae. A case of posterior reversible encephalopathy syndrome in a female patient with an overlap syndrome of systemic lupus erythematosus and systemic sclerosis is presented here.


The Role of SHI/STY/SRS Genes in Organ Growth and Carpel Development Is Conserved in the Distant Eudicot Species Arabidopsis thaliana and Nicotiana benthamiana.
Carpels are a distinctive feature of angiosperms, the ovule-bearing female reproductive organs that endow them with multiple selective advantages likely linked to the evolutionary success of flowering plants. Gene regulatory networks directing the development of carpel specialized tissues and patterning have been proposed based on genetic and molecular studies carried out in Arabidopsis thaliana. However, studies on the conservation/diversification of the elements and the topology of this network are still scarce. In this work, we have studied the functional conservation of transcription factors belonging to the SHI/STY/SRS family in two distant species within the eudicots, Eschscholzia californica and Nicotiana benthamiana. We have found that the expression patterns of EcSRS-L and NbSRS-L genes during flower development are similar to each other and to those reported for Arabidopsis SHI/STY/SRS genes. We have also characterized the phenotypic effects of NbSRS-L gene inactivation and overexpression in Nicotiana. Our results support the widely conserved role of SHI/STY/SRS genes at the top of the regulatory network directing style and stigma development, specialized tissues specific to the angiosperm carpels, at least within core eudicots, providing new insights on the possible evolutionary origin of the carpels.


Electroconvulsive therapy, brain-derived neurotrophic factor, and possible neurorestorative benefit of the clinical application of electroconvulsive therapy.
Treatment-resistant depression (TRD) is a growing problem in psychiatry. A recent meta-analysis has estimated TRD to be as high as 40%. Just over a decade ago, TRD was estimated to be as low as 10% to 15%. The causes of TRD are not fully understood. Finding ways to bring patients to remission faster may be part of the solution, but increasing our understanding of how depression works and how the brain responds to treatment may shed some light on this growing problem. Patients with TRD have been shown to have decreased volumes in gray matter structures, particularly in the hippocampus. Hippocampal volumes are correlated with decreased expression of neurotrophic factors (most notably, brain-derived neurotrophic factor [BDNF]), and decreased expression of BDNF correlates with the presence of depression. Increased expression of BDNF has a strong association with increased volumes in the hippocampus. Electroconvulsive therapy (ECT), a safe and effective treatment of severe depression, has been shown to be effective in TRD. Patients who undergo ECT have also had measurable increases in BDNF, indicating that ECT may be modulating intracellular processes in the patients with depression. Taken together, ECT may have a positive effect on restoring gray matter volume in patients with depression and especially TRD.


Internal and external carotid contributions to near-infrared spectroscopy during carotid endarterectomy.
The internal carotid (ICA) and external carotid (ECA) contributions to changing concentrations of oxyhemoglobin (Hbo2) and deoxyhemoglobin (Hb) during carotid endarterectomy were assessed with the use of near-infrared spectroscopy (NIRS). NIRS optodes were placed on the forehead with an interoptode distance of 6 cm, and laser-Doppler flowmetry (LDF) was used to monitor the change in skin blood flow between the optodes. Hb, Hbo2, LDF, arterial blood pressure, and middle cerebral artery flow velocity were recorded continuously. The ECA was clamped 2 minutes before the ICA was clamped. Suitable multimodal recordings were achieved in 44 patients. When the ECA was clamped, 76% of patients showed a fall in Hbo2 and 65% an increase in Hb. When corrected for changes in arterial blood pressure, an accompanying fall in cutaneous LDF predicted the fall in Hbo2 with high sensitivity (100%) and specificity (100%). Among those with no NIRS changes during ECA clamping, 56% had severe ECA stenosis or occlusion; none of these showed an accompanying fall in LDF. In contrast, when the ICA was clamped, substantial additional changes in NIRS occurred in 55% of cases, all of which were associated with a fall in flow velocity, but none with a change in LDF. Patients with a constant flow velocity after ICA clamping also showed no change in NIRS. Both the ECA and ICA vascular territories contribute to NIRS changes during carotid endarterectomy. The external carotid contribution to NIRS can be monitored with cutaneous LDF.


Recognition of species groups of Naupactus Dejean (Coleoptera: Curculionidae) from Argentina and neighboring countries.
Naupactus Dejean is the most diverse genus of the tribe Naupactini (Curculionidae: Entiminae), with more than 200 species occurring in South America, of which about 40 range in Argentina and neighboring countries. The Argentinean species treated herein were classified into nine groups having different biogeographic patterns: (1) the groups of Naupactus xanthographus, N. delicatulus and N. auricinctus mainly occur in northeastern Argentina (Misiones province) and reach the highest species diversity in the Atlantic and Parana forests of Brazil; (2) the groups of N. hirtellus, N. cinereidorsum, N. rivulosus and N. tarsalis show the highest species diversity in the Chacoan biogeographic province and also occur in the Yungas, Espinal, Monte, Parana forest (Argentina) and Cerrado (Brazil); (3) the groups of N. leucoloma and N. purpureoviolaceus have the highest species diversity in the Pampean biogeographic province, being also present in adjoining areas, mainly Chaco, Espinal, Monte and Parana forest. We provide descriptions, a dichotomous key, habitus photographs and line drawings of genitalia for the identification of the nine species groups, and a list of the Argentinean species from each group, together with their abbreviated synonymies, updated geographic distributions (including six new country records and several state/province records) and host plant associations. We discuss the characters that allow the separation of the species groups in a geographic distribution context, and provide information on species reassigned to genera other than Naupactus; among these, we transferred N. cephalotes (Hustache) to the tribe Tanymecini, genus Eurymetopellus, establishing the new combination Eurymetopellus cephalotes.


Vasopressin and blood pressure regulation.
The vasoconstrictor actions of arginine vasopressin (AVP) have been shown to occur in concentrations much lower than previously thought. Pressor responses to AVP are a poor index of vasoconstrictor activity since, in contrast to other vasoconstrictor agents, the expected rise of pressure is offset by dose-dependent decreases of cardiac output. The mechanisms for this appear to be, in large part, modulation of the autonomic nervous system whereby AVP enhances vagal nerve activity and reduces peripheral sympathetic nerve activity. AVP enhancement of baroreceptor reflex gain is in part responsible for these changes in some species (dog and rabbit), but not in others (rat). The release of AVP appears to contribute significantly to the normalization of arterial pressure in volume-depleted and hypotensive states. The link between plasma AVP and hypertension remains unclear, but it appears likely that it has an important permissive action in the development of sodium-dependent forms of hypertension.


Sexual risk-taking behaviors among young migrant population in Sweden.
Migration is a complex process of high uncertainty with adjustments to new contexts and experiences influencing individuals' health. This study aims to assess the prevalence of self-reported sexual risk-taking behaviors among migrant youth population in Sweden, fulfilling the research gap in that field. A pre-tested, web-based self-administered cross-sectional survey was used to collect data among 1563 migrant youth (15-25 years old) in Sweden. The survey was conducted in high schools and Swedish language schools for foreigners between December 2018 and November 2019. Pearson chi-square and t-tests were used to compare whether sociodemographic characteristics and migration status varied between those engaging in sexual risk behaviors or not. Multivariate logistic regression was used to determine the adjusted odds ratio of the key outcome variable and independent variables. There is a profound heterogeneity in migrant youth characteristics related to engagement in different sexual risk-taking behaviors. Those engaging in condomless sex were older, coming from the Americas and Europe, living longer in Sweden and came to live with their family. Belonging to the Islamic religion was a protective factor. Sex under the influence of drugs was related to those from Europe, and Middle East and North Africa (MENA) and coming to Sweden to work/study, where age was a protective factor. Living longer in Sweden, coming for work/study or to live with family had higher odds to engage in sex in exchange for gifts/money. The results highlight the needed reconsideration of the broader system response that can influence migrant youth health outcomes and public health implications. The approach should consider and relate to sexual risk-taking behavior's long-term consequences. Migrant youth background needs and knowledge should guide this response.


Heparin surface immobilization through hydrophilic spacers: thrombin and antithrombin III binding kinetics.
The immobilization of heparin onto polymeric surfaces using hydrophilic spacer groups has been effective in curtailing surface induced thrombus formation. In this study, the effect of hydrophilic spacers (PEO) on the binding kinetics of immobilized heparin with antithrombin III (ATIII) and thrombin was investigated. Monodispersed, low molecular weight heparin was fractionated on an ATIII affinity column to isolate high-ATIII affinity heparin. This high-ATIII affinity fraction was immobilized onto a styrene/p-amino styrene random copolymer surface using hydrophilic poly(ethylene oxide) (PEO) spacer groups. Styrene/p-amino styrene random copolymer was chosen as the model surface to provide quantitative and reproducible surface concentrations of available amine groups, grafted PEO spacers, and immobilized heparin. The polymer substrate was coated onto glass beads, tolylene diisocyanate modified PEO was covalently coupled to the surface, followed by heparin immobilization. The bioactivity of immobilized heparin was 16.2%, relative to free heparin, and a 1:1 binding ratio between heparin and PEO was achieved. The binding of ATIII and thrombin to control surfaces (no heparin), soluble heparin, heparin immobilized directly onto the surface, and heparin immobilized via spacer groups, were compared. Soluble heparin bound both thrombin and ATIII, while heparin immobilized directly onto the surface bound only thrombin. Spacer-immobilized heparin bound both ATIII and thrombin, although to a lesser extent than soluble heparin. Thus, the enhanced bioactivity of spacer-immobilized heparin, compared to direct-immobilization, may be attributed to the retention of ATIII binding.


Coordination copolymerization of severely encumbered isoalkenes with ethylene: enhanced enchainment mediated by binuclear catalysts and cocatalysts.
This contribution describes the implementation of the binuclear organotitanium "constrained geometry catalysts" (CGCs), (mu-CH(2)CH(2)-3,3'){(eta(5)-indenyl)[1-Me(2)Si((t)()BuN)](TiMe(2))}(2)[EBICGC(TiMe(2))(2); Ti(2)] and (mu-CH(2)-3,3'){(eta(5)-indenyl)[1-Me(2)Si((t)BuN)](TiMe(2))}(2)[MBICGC(TiMe(2))(2); C1-Ti(2)], in combination with the bifunctional bisborane activator 1,4-(C(6)F(5))(2)BC(6)F(4)B(C(6)F(5))(2) (BN(2)) in ethylene + olefin copolymerization processes. Specifically examined are the classically poorly responsive 1,1-disubstituted comonomers, methylenecyclopentane (C), methylenecyclohexane (D), 1,1,2-trisubstituted 2-methyl-2-butene (E), and isobutene (F). For the first three comonomers, this represents the first report of their incorporation into a polyethylene backbone via a coordination polymerization process. C and D are incorporated via a ring-unopened pathway, and E is incorporated via a novel pathway involving 2-methyl-1-butene enchainment in the copolymer backbone. In ethylene copolymerization, Ti(2) + BN(2) enchains approximately 2.5 times more C, approximately 2.5 times more D, and approximately 2.3 times more E than the mononuclear catalyst analogue [1-Me(2)Si(3-ethylindenyl)((t)BuN)]TiMe(2) (Ti(1)) + B(C(6)F(5))(3) (BN) under identical polymerization conditions. Polar solvents are found to weaken the catalyst-cocatalyst ion pairing, thus influencing the comonomer enchainment selectivity.


Non-catalytic heteroepitaxial growth of aligned, large-sized hexagonal boron nitride single-crystals on graphite.
Although van der Waals heterostructures composed of graphene and hexagonal boron nitride (h-BN) have attracted wide interest, it is still challenging to prepare them with high quality and controllability. Since contamination induced by transfer cannot be avoided in the case of growth on a metal catalyst, the non-catalytic growth of graphene and h-BN is highly desired. However, unlike graphene, few studies have reported the non-catalytic growth of h-BN, and the lack of controllability in terms of crystal orientation and nucleation density, and size of h-BN has hindered the practical applications of the heterostructures. In this work, we demonstrate the heteroepitaxial growth of aligned monolayer h-BN single-crystals on exfoliated graphite by chemical vapour deposition (CVD) without a metal catalyst. Triangular shaped domains were aligned with each other, which suggests the epitaxy between h-BN and the underlying graphite. Characterisation by Raman spectroscopy, Auger electron spectroscopy, and X-ray photoelectron spectroscopy also confirmed that the h-BN/graphite samples were of high quality. A growth kinetics study over different temperatures indicated an increase in the growth rate at high temperature. Control of nucleation density was realised by changing the hydrogen pressure during CVD or by the heating temperature in air before CVD. Under the optimised growth conditions, the edge length of h-BN single-crystals grew to ∼1 μm, which is the largest size to date for non-catalytic growth. These results will help to obtain structure-controlled, large-area, and impurity-free heterostructures based on h-BN and graphene.


Prospects for therapeutic mitochondrial transplantation.
Mitochondrial dysfunction has been implicated in a multitude of diseases and pathological conditions- the organelles that are essential for life can also be major players in contributing to cell death and disease. Because mitochondria are so well established in our existence, being present in all cell types except for red blood cells and having the responsibility of providing most of our energy needs for survival, then dysfunctional mitochondria can elicit devastating cellular pathologies that can be widespread across the entire organism. As such, the field of "mitochondrial medicine" is emerging in which disease states are being targeted therapeutically at the level of the mitochondrion, including specific antioxidants, bioenergetic substrate additions, and membrane uncoupling agents. New and compelling research investigating novel techniques for mitochondrial transplantation to replace damaged or dysfunctional mitochondria with exogenous healthy mitochondria has shown promising results, including tissue sparing accompanied by increased energy production and decreased oxidative damage. Various experimental techniques have been attempted and each has been challenged to accomplish successful transplantation. The purpose of this review is to present the history of mitochondrial transplantation, the different techniques used for both in vitro and in vivo delivery, along with caveats and pitfalls that have been discovered along the way. Results from such pioneering studies are promising and could be the next big wave of "mitochondrial medicine" once technical hurdles are overcome.


Deferoxamine mesylate is toxic for retinal pigment epithelium cells in vitro, and its toxicity is mediated by p38.
Deferoxamine mesylate is clinically used as a chelating agent but might induce retinopathy. To evaluate its effect on the retinal pigment epithelium (RPE), porcine RPE cells were stimulated with deferoxamine. Cell death was assessed with trypan blue exclusion assay. To investigate the pathway of cell death, the mitogen-activated protein kinases (MAPKs) Erk, JNK, and p38 were inhibited with U0126, SP600125, and SB203580, respectively. Their activity was determined by Western blot. Deferoxamine induces significant cell death in RPE cells, accompanied by phosphorylation of p38 and Erk. Inhibition of p38 attenuates cell death. In conclusion, deferoxamine is directly toxic on RPE cells, its toxicity depending on p38.


Personal values and mortality: Power, benevolence and self-direction predict mortality risk.
Although personal values guide one's life in several important aspects, no study has examined how personal values relate to mortality. The current study strives to fill this gap. I ask: Which personal values prospectively predict mortality? A large population-based sample of middle-aged and older adults in Germany was used (N = 6089). Personal values were measured in 2008 according to Schwartz using the Portraits Value Questionnaire. Mortality was monitored up to 2017, resulting in a maximum follow-up period of about 10 years. Using Cox survival regression analyses, I found that Self-direction and, in part, Benevolence predicted a decreased mortality risk. Conversely, Power predicted an increased mortality risk. Thus, valuing independent thought and action (Self-Direction) and valuing the well-being of close others (Benevolence) seem to be associated with improved health, whereas valuing dominance and materialism (Power) is associated with diminished health. Future research should examine the mechanisms by which these personal values relate to mortality.


KMT2C methyltransferase domain regulated INK4A expression suppresses prostate cancer metastasis.
Frequent truncation mutations of the histone lysine N-methyltransferase KMT2C have been detected by whole exome sequencing studies in various cancers, including malignancies of the prostate. However, the biological consequences of these alterations in prostate cancer have not yet been elucidated. To investigate the functional effects of these mutations, we deleted the C-terminal catalytic core motif of Kmt2c specifically in mouse prostate epithelium. We analysed the effect of Kmt2c SET domain deletion in a Pten-deficient PCa mouse model in vivo and of truncation mutations of KMT2C in a large number of prostate cancer patients. We show here for the first time that impaired KMT2C methyltransferase activity drives proliferation and PIN formation and, when combined with loss of the tumour suppressor PTEN, triggers loss of senescence, metastatic dissemination and dramatically reduces life expectancy. In Kmt2c-mutated tumours we show enrichment of proliferative MYC gene signatures and loss of expression of the cell cycle repressor p16INK4A. In addition, we observe a striking reduction in disease-free survival of patients with KMT2C-mutated prostate cancer. We identified truncating events of KMT2C as drivers of proliferation and PIN formation. Loss of PTEN and KMT2C in prostate cancer results in loss of senescence, metastatic dissemination and reduced life expectancy. Our data demonstrate the prognostic significance of KMT2C mutation status in prostate cancer patients. Inhibition of the MYC signalling axis may be a viable treatment option for patients with KMT2C truncations and therefore poor prognosis.


Fingerprinting Acinetobacter strains from clinical sources by numerical analysis of electrophoretic protein patterns.
A total of 57 strains of Acinetobacter calcoaceticus was fingerprinted by SDS-PAGE of cellular protein. All strains were also examined by conventional, API and N/F-Tek methods, and antibiotic sensitivity patterns were determined. In general, using the API 20E and N/F-Tek methods, it was possible to assign isolates of A. calcoaceticus to the two accepted "biotypes" or "varieties", A.c. anitratus and A.c. lwoffi, but these methods did not offer useful subdivision of the biotypes. Gel electrophoresis permitted subdivision of the strains into clusters in a manner which suggests that the technique may be valuable in typing strains isolated during outbreaks of infection in hospital.


[Changes of glutamate and γ-aminobutyric acid of extracellular fluid in hippocampus during electrical stimulation of anterior nucleus thalamus in rats].
To explore the changes of extracellular glutamate (Glu) and γ-aminobutyric acid (GABA) in hippocampus during the high-frequency stimulation of anterior nucleus thalamus (ANT) in epileptic rats. A total of 30 rats were divided equally into 2 groups:epilepsy model and normal control (n = 15 each). Concentric bipolar electrodes were stereotaxically implanted in bilateral ANT. A high-frequency stimulation of 130 Hz was applied in the epilepsy group. Microdialysis probes were unilaterally lowered into hippocampus. The concentrations of Glu and GABA in dialysate samples were determined by high-performance liquid chromatography (HPLC). Electrical stimulation of ANT induced an increase of GABA and a decrease of Glu in hippocampus. An increase of GABA and a decrease of Glu during the electrical stimulation of ANT play an important role in the therapy of epilepsy.


Renal stone composed of ritonavir.
Nephrolithiasis is a well-known side effect of many HIV protease inhibitors. However, there have not been reports of stones associated with ritonavir use. Here, we report the case of a 33-year-old woman with HIV on antiretroviral therapy who presented with sharp left flank pain and passed a stone that was later found to contain only ritonavir. Of note, the patient's treatment regimen had not included ritonavir for 2 years prior to this incidence. This case is notable both for the novel finding of a renal calculus composed entirely of ritonavir and the development of nephrolithiasis years after cessation of the aggravating drug. This finding suggests that patients on ritonavir should be more closely monitored and for longer periods of time for potential lithiasis formation.


Magnetic resonance imaging findings in chronic inflammatory demyelinating polyneuropathy with intracranial findings and enhancing, thickened cranial and spinal nerves.
Chronic inflammatory demyelinating polyneuropathy is a rare autoimmune disorder characterized by chronically progressive or relapsing symmetric sensorimotor involvement. We describe the imaging findings in our patient. Magnetic resonance imaging showed presence of an intracranial white matter lesion and enhancing, thickened cranial and spinal nerves. This disorder has been described very infrequently in the radiology literature.


Bacteriophage chi sensitivity and motility of Escherichia coli K-12 and Salmonella typhimurium Fla- mutants possessing the hook structure.
The production of hook protein and flagellin in 29 Fla- mutants of Escherichia coli K-12 was determined by the complement fixation assay. Six mutants produced hook protein, and four of them also produced flagellin. A flaE mutation was introduced into these fla mutants carrying the hook structure. All of these mutants made polyhooks and were used as hosts for a newly isolated host-range mutant of chi phage that has a high affinity for the hook structure. All except one mutant produced significant amounts of progeny phages. A flaD flaE double mutant was that exception which did not yield significant amounts of progeny by the phage propagation method. All of the flaE double mutants produced comparable amounts of polyhooks, and no qualitative difference was detected between chi-sensitive and chi-insensitive mutants by the complement fixation assay. Accordingly, it was thought that the polyhook of the flaD flaE mutant had a mechanical defect for chi phage infection. This assumption was confirmed by tethered-cell experiments; the flaD flaE mutant did not rotate. These results are well explained by a proposed regulation pathway of flagellar genes. flaE mutants can express other genes which govern the final step of the flagellar morphogenesis, whereas flaD mutants cannot rotate, possibly because the mocha operon is not expressed. The results obtained in E. coli were also found to be applicable to Salmonella typhimurium.


Methemoglobinemia and transesophageal echo.
Methemoglobin (MHb) is the oxidized form of Hemoglobin (Hb) containing iron in its ferric (Fe3+) rather than ferrous (Fe2+) state. Under physiologic conditions, diaphorase II accounts for only a small percentage of the red blood cell reducing capacity but can be pharmacologically activated by exogenous cofactors. Methemoglobinemia is an abnormal elevation of MHb levels resulting in impaired oxygen delivery to tissues as well as a left shift of the oxygen-Hb dissociation curve. We present the case of a 56-year-old female patient who underwent transesophageal echocardiography (TEE) prior to elective cardioversion. Prep for TEE included use of Hurricane spray. Twenty min after receipt, the patient's O2 saturation by pulse oximetry dropped from 100% to 86%; heart rate and blood pressure were unchanged. Physical exam revealed pallor, perioralcyanosis and acrocyanosis without tachypnea, respiratory distress, or jugular venous distension. A 100% non-rebreather mask provided no improvement. MHb was suspected and arterial blood gasses were drawn which was dark chocolate in appearance. Methylene Blue at 1 mg/kg over 5 minutes was administered empirically. ABG results were: pH 7.44/ CO2 40/ O2 315/ HCO2 26/ O2 sat 69%; MHb levels were 30.1% confirming the diagnosis of methemoglobinemia. This condition resolved within minutes. Though uncommon, MHb is the most Sported adverse event associated with topical benzocaine use. Untreated, it can lead to significant cardiopulmonary compromise, neurologic sequelae, and even death. Prompt recognition of this potentially life-threatening side-effect is essential in order to provide opportune treatment.


Secondary metabolism of olive secoiridoids. New microcomponents detected in drupes by electrospray ionization and high-resolution tandem mass spectrometry.
Olive drupe tissues have been selected as a possible source of information about the secondary metabolism of oleaceae secoiridoids. The structures of a number of new microcomponents, detected and isolated by a combined high-performance liquid chromatography (HPLC)-fraction collector/ultraviolet electrospray ionization mass spectrometry (UV/ESI-MS) apparatus, can be inferred from the data obtained by high-resolution tandem mass spectrometry in a QqTOF instrument. Some of them (1, 2, 3 and 4) are closely correlated to oleuropein. Others, such as neo-nuzhenide (5) and 2''-hydroxyoleuropein (6), typical of other oleaceae families, have never been found in olive tissues.


Two cases of acute respiratory distress syndrome resulting from adult-onset Still's disease.
Abstract Adult-onset Still's disease (AOSD) is a systemic inflammatory disorder of unknown origin. Acute respiratory distress syndrome (ARDS) is a rare complication of AOSD, with only nine cases having been reported in the literature. Here, we describe two cases of AOSD complicated with ARDS that were successfully treated with immunosuppressive therapy, including corticosteroids. Although ARDS is a life-threatening complication in AOSD, early commencement of high-dose corticosteroids and mechanical ventilation improve the prognosis.