Frizzled-6 Regulates Hematopoietic Stem/Progenitor Cell Survival and Self-Renewal.
Adult hematopoietic stem/progenitor cell (HSPC) numbers remain stable in the absence of external stressors. After bone marrow (BM) transplant, HSPCs need to expand substantially to repopulate the BM and replenish the peripheral blood cell pool. In this study, we show that a noncanonical Wnt receptor, Frizzled-6 (Fzd6), regulates HSPC expansion and survival in a hematopoietic cell-intrinsic manner. Fzd6 deficiency increased the ratio of Flt3(hi) multipotent progenitors to CD150(+) stem cells in the mouse BM, suggesting defective stem cell maintenance. Competitive transplantation experiments demonstrated that Fzd6(-) (/) (-) HSPCs were able to home to the BM but were severely impaired in their capacity to reconstitute a lethally irradiated host. Lack of Fzd6 resulted in a strong activation of caspase-3 and a gradual loss of donor HSPCs and peripheral blood granulocytes. Fzd6 was also necessary for the efficient HSPC expansion during emergency hematopoiesis. Mechanistically, Fzd6 is a negative regulator of Cdc42 clustering in polarized cells. Furthermore, β-catenin-dependent signaling may be disinhibited in Fzd6(-) (/) (-) HSPCs. Collectively, our data reveal that Fzd6 has an essential role in HSPC maintenance and survival. Noncanonical Wnt-Fzd6 signaling pathway could thus present an interesting target for promoting HSPC expansion and multilineage hematopoietic recovery after transplant.


Predicting the Effect of Variants of Unknown Significance in Molecular Tumor Boards with the VUS-Predict Pipeline.
Precision oncology utilizing molecular biomarkers for targeted therapies is one of the hopes to treat cancer. The availability of patient specific molecular profiling through next-generation sequencing, though, increases the amount of available data per patient to an extent that computational support is required to identify potential driver alterations for targeted therapies and rational decision-making in molecular tumor boards (MTBs). For some genetic variants evidence-based drug recommendations are available in public databases, but for the majority, the variants of unknown significance (VUS), this clinical information is missing. Additionally, for most of these variants no information about the functional impact on the protein is accessible. To acquire maximal functional evidence for VUS, the VUS-Predict pipeline collects estimations about the effect of a VUS by integrating multiple pre-existing tools. Pre-existing tools implement different approaches for their predictions, which are summarized by our newly developed tool with a common score and classification in neutral or deleterious variants. The primary tools are chosen based on their sensitivity and specificity on well-known variants of the transcription factor TP53. Resulting negative and positive predictive values are used to calibrate the VUS-Predict pipeline. Further, the pipeline is evaluated using data from public cancer databases and cases of the MTB in Göttingen, both also in comparison with the ensemble method REVEL. The results show that VUS-Predict has clear advantages in a clinical setting due to clear and traceable predictions. In particular, VUS outperforms REVEL in the real-life setting of a MTB. Likewise, an evaluation on variants of public cancer databases confirms the good results of VUS-Predict and shows the need for a reliable gold standard and unambiguous results of the tools under test.


pH regulation of endogenous acid production in subjects with chronic ketoacidosis.
In a previous study of starvation-induced acute ketoacidosis, net ketoacid production was inhibited by acid and facilitated by base ingestion. To determine whether hydrogen ion modifies net ketoacid production during chronic ketoacidosis, six over-weight volunteers ingested NaHCO3, NaCl, or NH4Cl (2 mmol X kg-1 X day-1), each for 7 days, during weeks 6-8 of ketogenic dieting. During days 4-7 of each phase, blood bicarbonate was stable but lower in the NH4Cl (19.6 +/- 0.7 mM) than the NaHCO3 (23.7 +/- 0.7 mM) phases. Throughout these periods, acid intake differed by 216 mmol/day, whereas acid output differed by 129 mmol/day between the NaHCO3 and the NH4Cl phases. The major contribution to this difference in acid balance was a difference in net organic acid (ketoacid) production. Although blood ketones were stable, ketoacid excretion, reflecting net ketoacid production, was decreased by 59% with acid and increased by 66% with base compared with NaCl (control) ingestion. Thus, in this state of chronic ketoacidosis, challenges to acid-base balance were countered by a rapidly occurring, sustained, reversible, and quantitatively significant modification of net acid production which acted as an effective mechanism for acid-base regulation.


NMR evidence for a two-step phase separation in Nd1.85Ce0.15CuO4-delta.
By Cu NMR we studied the spin and charge structure in Nd(2-x)Ce(x)CuO(4-delta). For x=0.15, starting from a superconducting sample, the low temperature magnetic order in the sample reoxygenated under 1 bar oxygen at 900 degrees C reveals a peculiar modulation of the internal field, indicative of a phase characterized by large charge droplets ("blob" phase). By prolonged reoxygenation at 4 bars the blobs break up and the spin structure changes to that of an ordered antiferromagnet. We conclude that the superconductivity in the n-type systems competes with a genuine type I Mott-insulating state.


Synthesis and Thermal Properties of Solid-State Structural Isomers: Ordered Intergrowths of SnSe and MoSe2.
A family of structural isomers [(SnSe)1.05]m(MoSe2)n were prepared using the modulated elemental reactant method by varying the layer sequence and layer thicknesses in the precursor. By varying the sequence of Sn-Se and Mo-Se layer pairs deposited and annealing the precursors to self-assemble the targeted compound, all six possible isomers [(SnSe)1.05]4(MoSe2)4, [(SnSe)1.05]3(MoSe2)3[(SnSe)1.05]1(MoSe2)1, [(SnSe)1.05]3(MoSe2)2[(SnSe)1.05]1(MoSe2)2, [(SnSe)1.05]2(MoSe2)3[(SnSe)1.05]2(MoSe2)1, [(SnSe)1.05]2(MoSe2)1[(SnSe)1.05]1(MoSe2)2[(SnSe)1.05]1(MoSe2)1, and [(SnSe)1.05]2(MoSe2)2[(SnSe)1.05]1(MoSe2)1[(SnSe)1.05]1(MoSe2)1 were prepared. The structures were characterized by X-ray diffraction and electron microscopy which showed that all of the compounds have very similar c-axis lattice parameters and in-plane constituent lattice parameters yet distinct isomeric structures. These studies confirm that the structure, order, and thickness of the constituent layers match that of the precursors. The cross-plane thermal conductivity is found to be very low (∼0.08 Wm(-1) K(-1)) and independent of the number of SnSe-MoSe2 interfaces within uncertainty. The poor thermal transport in these layered isomers is attributed to a large cross-plane thermal resistance created by SnSe-MoSe2 and MoSe2-MoSe2 turbostratically disordered van der Waals interfaces, the density of which has less variation among the different compounds than the SnSe-MoSe2 interface density alone.


PATHWAYS OF MEDICAL PROGRESS.
During the three decades that have passed, medical science has ascended to a high plateau of achievement. The climb has involved several pathways; among them: (1) the physiological approach toward disease as experiments which nature performs on organisms, (2) the more intelligent interpretation of the functional reactions of the body in disease in accordance with latest discoveries in physiology, (3) the supplementation of observable phenomena through use of laboratory instruments, (4) the assumption of active investigation both on patients and experimental animals by clinicians themselves, (5) the shuttling of problems between clinical and experimental laboratories and (6) correlated research in clinical and physiological departments. As we look down from the heights we have reached, we have reason to be pleased with our progress; but when we look ahead we become aware that there are still high mountain ranges to be climbed. We realize that their ascent can not be accomplished by employing merely the methods, equipment and strategy that have proved successful so far; we must improve the application of principles that are old and well established, and evolve others that are new. Above all, we from laboratories and clinics must join hands to help each other climb; and through correlated team-work overcome the great obstacles that jealous nature places in our way. I have ventured to suggest a few directions which such mutual help may take. They include (1) means by which new fundamental discoveries can be utilized more quickly by clinicians and practitioners of medicine; (2) plans by which younger clinical investigators can be given approximately the same opportunity for training in research technique as their colleagues entering experimental sciences; (3) pleas that the shuttling of problems between hospitals and laboratories of fundamental science may continue in order that the ultimate significance of clinical results may be better understood and that the applicability of fundamental discoveries to nature's experiments may be tested; (4) judicious combination of talents of laboratory and clinical workers, whenever this leads to greater economy of effort and does not infringe upon the primary duties of each participant to his calling. The spirit of correlation which is involved in all these plans of advance is a silent force which grows not only through mutual interest in each other's problems but also through frank respectful criticism of each other's trends. With such a spirit of correlated effort science marches on.


Event-related fast optical signal in a rapid object recognition task: improving detection by the independent component analysis.
Noninvasive recording of fast optical signals presumably reflecting neuronal activity is a challenging task because of a relatively low signal-to-noise ratio. To improve detection of those signals in rapid object recognition tasks, we used the independent component analysis (ICA) to reduce "global interference" (heartbeat and contribution of superficial layers). We recorded optical signals from the left prefrontal cortex in 10 right-handed participants with a continuous-wave instrument (DYNOT, NIRx, Brooklyn, NY). Visual stimuli were pictures of urban, landscape and seashore scenes with various vehicles as targets (target-to-non-target ratio 1:6) presented at ISI=166 ms or 250 ms. Subjects mentally counted targets. Data were filtered at 2-30 Hz and artifactual components were identified visually (for heartbeat) and using the ICA weight matrix (for superficial layers). Optical signals were restored from the ICA components with artifactual components removed and then averaged over target and non-target epochs. After ICA processing, the event-related response was detected in 70%-100% of subjects. The refined signal showed a significant decrease from baseline within 200-300 ms after targets and a slight increase after non-targets. The temporal profile of the optical signal corresponded well to the profile of a "differential ERP response", the difference between targets and non-targets which peaks at 200 ms in similar object detection tasks. These results demonstrate that the detection of fast optical responses with continuous-wave instruments can be improved through the ICA method capable to remove noise, global interference and the activity of superficial layers. Fast optical signals may provide further information on brain processing during higher-order cognitive tasks such as rapid categorization of objects.


[Cold pressed oils].
In recent years the interest in cold pressed oils has increased. In the Polish market, besides a wide assortment of imported products, Polish oils, mainly rapeseed oil, are also ever more frequently available. Cold pressed oils are used in food and pharmaceutical industry and also in the production of cosmetics. The production of cold pressed oils is simple and the investments for it are low. A limitation of the use of this method is low efficiency of the process (6-15% of oil remains in the pressed residues). For obtaining of proper oil it is indispensable to use ripe, clean, non-damaged and correctly stored seeds. Moreover, the conditions and parameters of the production and storage are important, especially the temperature and duration of pressing. The quality of oil is reduced at higher temperatures. At the temperatures below 40 degrees C the process yields oil of good quality permitted for consumption. The quality and the durability of the obtained oil depend on many factors, among them on the composition of fatty acids, the conditions of production and storage, on the presence of prooxidants and antioxidants. The analogue of the Polish cold pressed oil may be "cold pressed oil" and "virgin oil" the definitions and properties of which were proposed at the 21st Session of the Committees for Codex Alimentarius for Fats and Oils of the FAO/WHO (London, 1993).


Early acquisition of anogenital human papillomavirus among teenage men who have sex with men.
Anogenital human papillomavirus (HPV) is common among men who have sex with men (MSM) and causes anal cancer. This study examined the determinants of initial anogenital HPV infection among teenage MSM. Two hundred MSM aged 16 to 20 years were recruited via community and other sources. Men were tested for HPV DNA from the anus and penis. The proportion of men with anal HPV of any type increased from 10.0% in men reporting no prior receptive anal sex to 47.3% in men reporting ≥ 4 receptive anal sex partners (P < .001).A similar pattern was also seen with HPV type 16 (P = .044). The proportion of men with penile HPV increased from 3.7% in men reporting no prior insertive anal sex to 14.8% in men reporting ≥ 4 insertive anal sex partners (P = .014). Overall, 39.0% (95% confidence interval (CI), 32.2%-46.1%) of men had at least 1 HPV type: 23.0% (95% CI, 17.4%-29.5%) had a vaccine-preventable type (6, 11, 16 or 18). Early and high per partner transmission of HPV occurred between men soon after their first sexual experiences. HPV vaccination needs to commence early for maximal prevention of HPV among MSM.


The effect of the type of dietary protein on the development of ovarian cancer.
We evaluated whether different dietary protein qualities (isocaloric diets involving animal (casein) or plant protein (soy protein) could inhibit the ovarian cancer growth in mice and improve their prognosis and whether chemotherapy had different tumor reducing effects on these mice. In the mice of the 20% plant protein group, the ovarian cancer growth at 5 weeks after tumor implantation was clearly reduced in comparison to the mice in the 20% animal protein group (p< 0.001). The serum levels of insulin and IGF-1 levels were both lower in the mice of the 20% plant protein group than in the mice of the 20% animal protein group (p<0.001 and p<0.01, respectively). Immunohistochemistry revealed that the level of eukaryotic initiation factor 4E-binding protein 1 (p-4EBP1) activity-one of the major downstream effectors of the mTOR pathway -of the plant protein group was significantly weaker than that of the animal protein group (p<0.001). The prognosis of the 20% plant protein group was better than that of the 20% animal protein group (log-rank test, p=0.0062). The ovarian cancer growth in the 20% plant protein plus cisplatin treatment group was not significantly reduced in comparison to the 20% animal protein plus cisplatin treatment group. Our findings suggest that a diet high in plant protein reduces the growth of human ovarian cancer cells in mice compared to a diet high in animal protein, -possibly through the lack of activation of the IGF/Akt/mTOR pathway, and leads to a better prognosis with or without cisplatin treatment.


Assessing the Genotoxicity of Cellulose Nanomaterials in a Co-Culture of Human Lung Epithelial Cells and Monocyte-Derived Macrophages.
Cellulose micro/nanomaterials (CMNMs) are innovative materials with a wide spectrum of industrial and biomedical applications. Although cellulose has been recognized as a safe material, the unique properties of its nanosized forms have raised concerns about their safety for human health. Genotoxicity is an endpoint that must be assessed to ensure that no carcinogenic risks are associated with exposure to nanomaterials. In this study, we evaluated the genotoxicity of two types of cellulose micro/nanofibrils (CMF and CNF) and one sample of cellulose nanocrystals (CNC), obtained from industrial bleached Eucalyptus globulus kraft pulp. For that, we exposed co-cultures of human alveolar epithelial A549 cells and THP-1 monocyte-derived macrophages to a concentration range of each CMNM and used the micronucleus (MN) and comet assays. Our results showed that only the lowest concentrations of the CMF sample were able to induce DNA strand breaks (FPG-comet assay). However, none of the three CMNMs produced significant chromosomal alterations (MN assay). These findings, together with results from previous in vitro studies using monocultures of A549 cells, indicate that the tested CNF and CNC are not genotoxic under the conditions tested, while the CMF display a low genotoxic potential.


Ischaemic preconditioning preferentially increases protein S-nitrosylation in subsarcolemmal mitochondria.
Nitric oxide (NO) and protein S-nitrosylation (SNO) have been shown to play important roles in ischaemic preconditioning (IPC)-induced acute cardioprotection. The majority of proteins that show increased SNO following IPC are localized to the mitochondria, and our recent studies suggest that caveolae transduce acute NO/SNO cardioprotective signalling in IPC hearts. Due to the close association between subsarcolemmal mitochondria (SSM) and the sarcolemma/caveolae, we tested the hypothesis that SSM, rather than the interfibrillar mitochondria (IFM), are major targets for NO/SNO signalling derived from caveolae-associated eNOS. Following either control perfusion or IPC, SSM and IFM were isolated from Langendorff perfused mouse hearts, and SNO was analysed using a modified biotin switch method with fluorescent maleimide fluors. In perfusion control hearts, the SNO content was higher in SSM compared with IFM (1.33 ± 0.19, ratio of SNO content Perf-SSM vs. Perf-IFM), and following IPC SNO content significantly increased preferentially in SSM, but not in IFM (1.72 ± 0.17 and 1.07 ± 0.04, ratio of SNO content IPC-SSM vs. Perf-IFM, and IPC-IFM vs. Perf-IFM, respectively). Consistent with these findings, eNOS, caveolin-3, and connexin-43 were detected in SSM, but not in IFM, and IPC resulted in a further significant increase in eNOS/caveolin-3 levels in SSM. Interestingly, we did not observe an IPC-induced increase in SNO or eNOS/caveolin-3 in SSM isolated from caveolin-3(-/-) mouse hearts, which could not be protected with IPC. In conclusion, these results suggest that SSM may be the preferential target of sarcolemmal signalling-derived post-translational protein modification (caveolae-derived eNOS/NO/SNO), thus providing an important role in IPC-induced cardioprotection.


Meconium increases type 1 angiotensin II receptor expression and alveolar cell death.
The pulmonary renin-angiotensin system (RAS) contributes to inflammation and epithelial apoptosis in meconium aspiration. It is unclear if both angiotensin II receptors (ATR) contribute, where they are expressed and if meconium modifies subtype expression. We examined ATR subtypes in 2 wk rabbit pup lungs before and after meconium exposure and with and without captopril pretreatment or type 1 receptor (AT1R) inhibition with losartan, determining expression and cellular localization with immunoblots, RT-PCR and immunohistochemistry, respectively. Responses of cultured rat alveolar type II pneumocytes were also examined. Type 2 ATR were undetected in newborn lung before and after meconium instillation. AT1R were expressed in pulmonary vascular and bronchial smooth muscle and alveolar and bronchial epithelium. Meconium increased total lung AT1R protein approximately 3-fold (p = 0.006), mRNA 29% (p = 0.006) and immunostaining in bronchial and alveolar epithelium and smooth muscle, which were unaffected by captopril and losartan. Meconium also increased AT1R expression >3-fold in cultured type II pneumocytes and caused concentration-dependent cell death inhibited by losartan. Meconium increases AT1R expression in newborn rabbit lung and cultured type II pneumocytes and induces AT1R-mediated cell death. The pulmonary RAS contributes to the pathogenesis of meconium aspiration through increased receptor expression.


Exercise as Medicine for Mental and Substance Use Disorders: A Meta-review of the Benefits for Neuropsychiatric and Cognitive Outcomes.
Exercise may improve neuropsychiatric and cognitive symptoms in people with mental disorders, but the totality of the evidence is unclear. We conducted a meta-review of exercise in (1) serious mental illness (schizophrenia spectrum, bipolar disorder and major depression (MDD)); (2) anxiety and stress disorders; (3) alcohol and substance use disorders; (4) eating disorders (anorexia nervosa bulimia nervosa, binge eating disorders, and (5) other mental disorders (including ADHD, pre/post-natal depression). Systematic searches of major databases from inception until 1/10/2018 were undertaken to identify meta-analyses of randomised controlled trials (RCTs) of exercise in people with clinically diagnosed mental disorders. In the absence of available meta-analyses for a mental disorder, we identified systematic reviews of exercise interventions in people with elevated mental health symptoms that included non-RCTs. Meta-analysis quality was assessed with the AMSTAR/+. Overall, we identified 27 systematic reviews (including 16 meta-analyses representing 152 RCTs). Among those with MDD, we found consistent evidence (meta-analyses = 8) that exercise reduced depression in children, adults and older adults. Evidence also indicates that exercise was more effective than control conditions in reducing anxiety symptoms (meta-analyses = 3), and as an adjunctive treatment for reducing positive and negative symptoms of schizophrenia (meta-analyses = 2). Regarding neurocognitive effects, exercise improved global cognition in schizophrenia (meta-analyses = 1), children with ADHD (meta-analyses = 1), but not in MDD (meta-analyses = 1). Among those with elevated symptoms, positive mental health benefits were observed for exercise in people with pre/post-natal depression, anorexia nervosa/bulimia nervosa, binge eating disorder, post-traumatic stress disorder and alcohol use disorders/substance use disorders. Adverse events were sparsely reported. Our panoramic meta-overview suggests that exercise can be an effective adjunctive treatment for improving symptoms across a broad range of mental disorders.


Genetic evidence that SMAD2 is not required for gonadal tumor development in inhibin-deficient mice.
Inhibin is a tumor-suppressor and activin antagonist. Inhibin-deficient mice develop gonadal tumors and a cachexia wasting syndrome due to enhanced activin signaling. Because activins signal through SMAD2 and SMAD3 in vitro and loss of SMAD3 attenuates ovarian tumor development in inhibin-deficient females, we sought to determine the role of SMAD2 in the development of ovarian tumors originating from the granulosa cell lineage. Using an inhibin alpha null mouse model and a conditional knockout strategy, double conditional knockout mice of Smad2 and inhibin alpha were generated in the current study. The survival rate and development of gonadal tumors and the accompanying cachexia wasting syndrome were monitored. Nearly identical to the controls, the Smad2 and inhibin alpha double knockout mice succumbed to weight loss, aggressive tumor progression, and death. Furthermore, elevated activin levels and activin-induced pathologies in the liver and stomach characteristic of inhibin deficiency were also observed in these mice. Our results indicate that SMAD2 ablation does not protect inhibin-deficient females from the development of ovarian tumors or the cachexia wasting syndrome. SMAD2 is not required for mediating tumorigenic signals of activin in ovarian tumor development caused by loss of inhibin.


Educational Gaming for Pharmacy Students - Design and Evaluation of a Diabetes-themed Escape Room.
Objective. To design an educational game that will increase third-year professional pharmacy students' knowledge of diabetes mellitus disease management and to evaluate their perceived value of the game. Methods. Faculty members created an innovative educational game, the diabetes escape room. An authentic escape room gaming environment was established through the use of a locked room, an escape time limit, and game rules within which student teams completed complex puzzles focused on diabetes disease management. To evaluate the impact, students completed a pre-test and post-test to measure the knowledge they've gained and a perception survey to identify moderating factors that could help instructors improve the game's effectiveness and utility. Results. Students showed statistically significant increases in knowledge after completion of the game. A one-sample t-test indicated that students' mean perception was statistically significantly higher than the mean value of the evaluation scale. This statically significant result proved that this gaming act offers a potential instructional benefit beyond its novelty. Conclusion. The diabetes escape room proved to be a valuable educational game that increased students' knowledge of diabetes mellitus disease management and showed a positive perceived overall value by student participants.


Results of a randomized study of 3 schedules of low-dose decitabine in higher-risk myelodysplastic syndrome and chronic myelomonocytic leukemia.
Epigenetic therapy with hypomethylating drugs is now the standard of care in myelodysplastic syndrome (MDS). Response rates remain low, and mechanism-based dose optimization has not been reported. We investigated the clinical and pharmacodynamic results of different dose schedules of decitabine. Adults with advanced MDS or chronic myelomonocytic leukemia (CMML) were randomized to 1 of 3 decitabine schedules: (1) 20 mg/m2 intravenously daily for 5 days; (2) 20 mg/m2 subcutaneously daily for 5 days; and (3) 10 mg/m2 intravenously daily for 10 days. Randomization followed a Bayesian adaptive design. Ninety-five patients were treated (77 with MDS, and 18 with CMML). Overall, 32 patients (34%) achieved a complete response (CR), and 69 (73%) had an objective response by the new modified International Working Group criteria. The 5-day intravenous schedule, which had the highest dose-intensity, was selected as optimal; the CR rate in that arm was 39%, compared with 21% in the 5-day subcutaneous arm and 24% in the 10-day intravenous arm (P < .05). The high dose-intensity arm was also superior at inducing hypomethylation at day 5 and at activating P15 expression at days 12 or 28 after therapy. We conclude that a low-dose, dose-intensity schedule of decitabine optimizes epigenetic modulation and clinical responses in MDS.


Low-stress and high-reflectance Mo/Si multilayers for extreme ultraviolet lithography by magnetron sputtering deposition with bias assistance.
To explore the potential of achieving low-stress and high-reflectance Mo/Si multilayers deposited by conventional magnetron sputtering with bias assistance, we investigated the effects of varying Ar gas pressure, substrate bias voltage, and a bias-assisted Si ratio on the stress and extreme ultraviolet (EUV) reflectance of Mo/Si multilayers. To reduce the damage of ion bombardments on an Si-on-Mo interface, only the final part of the Si layer was deposited with bias assistance. Bias voltage has strong influence on the stress. The compressive stress of Mo/Si multilayers can be reduced remarkably by increasing bias voltage due to the increase of Mo-on-Si interdiffusion and postponement of Mo crystallization transition. Properly choosing gas pressure and a bias-assisted Si ratio is critical to obtain high EUV reflectance. Appropriately decreasing gas pressure can reduce the interface roughness without increasing interdiffusion. Too much bias assistance can seriously reduce the optical contrast between Mo and Si layers and lead to a remarkable decrease of EUV reflectance. Thus, by appropriately choosing gas pressure, bias voltage, and a bias-assisted Si ratio, the stress values of Mo/Si multilayers can be reduced to the order of -100 MPa with an EUV reflectance loss of about 1%.


DNMT cooperativity--the developing links between methylation, chromatin structure and cancer.
Controversy has reigned for some time over the biological connection between DNA methylation and cancer. For this reason, the methylation mechanism responsible for increased cancer risk has received greater attention in recent years. Tumor suppressor genes are often hypermethylated resulting in gene silencing. Although some have questioned this interpretation of the link between methylation and cancer, it appears that both hypermethylation and hypomethylation events can create epigenetic changes that can contribute to cancer development. Recent studies have shown that the methyltransferases DNMT1 and DNMT3b cooperatively maintain DNA methylation and gene silencing in human cancer cells. Disruption of the human DNMT3b only slightly reduces the overall global DNA methylation; however, demethylation was markedly potentiated when both DNMT1 and DNMT3b were simultaneously deleted. The results to these experiments provide compelling evidence towards a role for DNA methylation in cancer. This review discusses the current understanding of cancer-epigenetic information and highlights recent studies that connect the methylation machinery and chromatin remodelling with cancer susceptibility.


Reflex control of intestinal gas dynamics and tolerance in humans.
Intestinal transit of gas is normally adapted to the luminal gas load, but in some patients impaired transit may lead to gas retention and symptoms. We hypothesized that intestinal gas transit is regulated by reflex mechanisms released by segmental distension at various gut levels. In 24 healthy subjects, we measured gas evacuation and perception of jejunal gas infusion (12 ml/min) during simultaneous infusion of duodenal lipids mimicking the postprandial caloric load (Intralipid, 1 kcal/min). We evaluated the effects of proximal (duodenal) distension (n = 8), distal (rectal) distension (n = 8), and sham distension, as control (n = 8). Duodenal lipid infusion produced gas retention (366 +/- 106 ml) with low abdominal perception (1.5 +/- 0.8 score). Distension of either the duodenum or rectum during lipid infusion expedited gas transit and prevented retention (-120 +/- 164 and -124 +/- 162 ml retention, respectively; P < 0.05 vs. control). However, the tolerance to the intestinal gas load differed markedly, depending on the site of distension; perception remained low during rectal distension (2.6 +/- 0.7 score; not significant vs. control) but increased during duodenal distension (4.4 +/- 0.7 score; P < 0.05 vs. control). We conclude that focal gut distension, either at proximal or distal sites, accelerates gas transit, but the symptomatic response depends on the site of stimulation.


Transcriptional profiling reveals gene expression changes associated with inflammation and cell proliferation following short-term inhalation exposure to copper oxide nanoparticles.
Our recent studies revealed a dose-dependent proinflammatory response to copper oxide nanoparticles (CuO NPs) in rats following short-term inhalation exposure for five consecutive days. Here transcriptomics approaches were applied using the same model to assess global gene expression in lung tissues obtained 1 day post-exposure and after a recovery period of 22 days from rats exposed to clean air or 6 hour equivalent doses of 3.3 mg m-3 (low dose) and 13.2 mg m-3 (high dose). Microarray analyses yielded about 1000 differentially expressed genes in the high-dose group and 200 in low-dose compared to the clean air control group, and less than 20 after the recovery period. Pathway analysis indicated cell proliferation/survival and inflammation as the main processes triggered by exposure to CuO NPs. We did not find significant perturbations of pathways related to oxidative stress. Upregulation of epithelial cell transforming protein 2 (Ect2), a known oncogene, was noted and ECT2 protein was upregulated in the lungs of exposed animals. Proliferation of alveolar epithelial cells was demonstrated based on Ki67 expression. The gene encoding monocyte chemoattractant protein 1 (or CCL2) was also upregulated and this was confirmed by immunohistochemistry. However, no aberrant DNA methylation of inflammation-associated genes was observed. In conclusion, we have found that inhalation of CuO NPs in rats causes upregulation of the oncoprotein ECT2 and the chemokine CCL2 and other proinflammatory markers as well as proliferation in bronchoalveolar epithelium after a short-term inhalation exposure. Thus, pathways known to be associated with neoplastic processes and inflammation were affected in this model.


dynr.mi: An R Program for Multiple Imputation in Dynamic Modeling.
Assessing several individuals intensively over time yields intensive longitudinal data (ILD). Even though ILD provide rich information, they also bring other data analytic challenges. One of these is the increased occurrence of missingness with increased study length, possibly under non-ignorable missingness scenarios. Multiple imputation (MI) handles missing data by creating several imputed data sets, and pooling the estimation results across imputed data sets to yield final estimates for inferential purposes. In this article, we introduce dynr.mi(), a function in the R package, Dynamic Modeling in R (dynr). The package dynr provides a suite of fast and accessible functions for estimating and visualizing the results from fitting linear and nonlinear dynamic systems models in discrete as well as continuous time. By integrating the estimation functions in dynr and the MI procedures available from the R package, Multivariate Imputation by Chained Equations (MICE), the dynr.mi() routine is designed to handle possibly non-ignorable missingness in the dependent variables and/or covariates in a user-specified dynamic systems model via MI, with convergence diagnostic check. We utilized dynr.mi() to examine, in the context of a vector autoregressive model, the relationships among individuals' ambulatory physiological measures, and self-report affect valence and arousal. The results from MI were compared to those from listwise deletion of entries with missingness in the covariates. When we determined the number of iterations based on the convergence diagnostics available from dynr.mi(), differences in the statistical significance of the covariate parameters were observed between the listwise deletion and MI approaches. These results underscore the importance of considering diagnostic information in the implementation of MI procedures.


Clinical Outcome of Arthroscopic Bankart Repair Combined With Simultaneous Capsular Repair.
A capsular tear and humeral avulsion of the glenohumeral ligament lesion are not uncommon findings in association with a Bankart lesion. However, there have been few reports regarding the prevalence of such capsular lesions and the postoperative recurrence after capsular repair. Purpose/Hypothesis: This study investigated the prevalence of capsular lesions and clarified their influence on the postoperative recurrence of instability. In addition, factors were identified that were associated with the occurrence of capsular lesions and the postoperative recurrence of instability. We hypothesized that clinical outcomes would be improved by combining arthroscopic Bankart repair with simultaneous capsular repair. Cohort study; Level of evidence, 3. Capsular lesions were retrospectively examined through operative records, still pictures, and videos in 172 shoulders with traumatic anterior instability. First, the prevalence of capsular lesions and their severity were investigated. Then, postoperative recurrence was determined in shoulders observed for a minimum of 2 years. Finally, factors were assessed that were associated with the occurrence of capsular lesions and the postoperative recurrence of instability. A capsular lesion was recognized in 37 shoulders (21.5%), being severe and mild in 20 and 17, respectively. All were repaired simultaneously with the arthroscopic Bankart procedure. After follow-up for at least 2 years, recurrence of instability was detected in 10 of 34 shoulders (29.4%), including 6 (31.6%) with severe capsular lesions and 4 (26.7%) with mild lesions. The recurrence rate was significantly higher in shoulders with a capsular lesion than in shoulders without a capsular lesion (18 of 120, 15%; P = .013), but there was no significant difference between severe and mild lesions. Regardless of the sport played, capsular lesions were significantly more frequent in patients ≥30 years old, patients with complete dislocation, and patients with a coexisting Hill-Sachs lesion. Postoperative recurrence of instability was significantly more frequent in patients <30 years and competitive athletes. In shoulders undergoing arthroscopic Bankart repair, capsular lesions were often present and were associated with higher postoperative recurrence of instability. While these lesions were more frequent in older patients, postoperative recurrence of instability was more likely in young competitive athletes.


A box H/ACA small nucleolar RNA-like domain at the human telomerase RNA 3' end.
Simple sequence repeat telomeric DNA is maintained by a specialized reverse transcriptase, telomerase. The integral RNA subunit of telomerase contains a template region that determines the sequence added to chromosome ends. Aside from providing the template, little is known about the role of the telomerase RNA. In addition, no hypotheses have been suggested to account for the striking evolutionary divergence in size and sequence between telomerase RNAs of ciliates, yeasts, and mammals. We show that the two- to threefold increase in size of the mammalian telomerase RNAs relative to ciliate telomerase RNAs is due to the presence of an extra domain resembling a box H/ACA small nucleolar RNA (snoRNA). The human telomerase RNA (hTR) H/ACA domain is essential in vivo for hTR accumulation, hTR 3' end processing, and telomerase activity. By substituting the U64 box H/ACA snoRNA for the hTR H/ACA domain, we demonstrate that a heterologous snoRNA can function to promote chimeric RNA accumulation and 3' end processing but not telomerase activity. In addition, we show that maturation of full-length hTR and its assembly into active telomerase occur from an mRNA promoter-driven RNA polymerase II transcript but not from a U6 snRNA promoter-driven RNA polymerase III transcript. Finally, we show that a small percentage of hTR is associated with nucleoli. These results have implications for the biogenesis and structure of hTR and the human telomerase ribonucleoprotein complex. They also expand the structural and functional diversity of the box H/ACA snoRNA motif.


Effects of 2-bromoterguride, a dopamine D2 receptor partial agonist, on cognitive dysfunction and social aversion in rats.
2-Bromoterguride, a dopamine D2 receptor partial agonist with antagonist properties at serotonin 5-HT2A receptors and α2C-adrenoceptors, meets the prerequisites of a putative atypical antipsychotic drug (APD). We recently showed that 2-bromoterguride is effective in tests of positive symptoms of schizophrenia in rats without inducing extrapyramidal side effects or metabolic changes. In continuation of our recent work, we now investigated the effect of 2-bromoterguride on apomorphine and phencyclidine (PCP)-induced disruptions of prepulse inhibition (PPI) of the acoustic startle response, a measure of sensory gating. In addition, we used subchronic PCP treatment to produce cognitive deficits and social aversion, and assessed the effect of 2-bromoterguride on the performance in the novel object recognition (NOR) task (model for studying cognitive deficit symptoms of schizophrenia) and the social interaction test (model for studying negative symptoms of schizophrenia). Finally, we extended the side effect profile of 2-bromoterguride by measuring the prolactin response to systemic administration of the drug in rats. Treatment with 2-bromoterguride (0.1 and 0.3 mg/kg) reversed PPI deficits induced by apomorphine and PCP, respectively. Subchronic PCP induced impairments in object memory and social interaction behavior which were ameliorated by 2-bromoterguride but not by clozapine and aripiprazole, respectively. Prolactin concentration in blood serum was not elevated at 1, 2, or 4 h post-2-bromoterguride treatment, which further supports the safe and effective use of this drug. Our data support 2-bromoterguride as a promising APD candidate due to its beneficial effect on cognitive impairments and negative symptoms of schizophrenia.


Common genetic variants in the 9p21 region and their associations with multiple tumours.
The chromosome 9p21.3 region has been implicated in the pathogenesis of multiple cancers. We systematically examined up to 203 tagging SNPs of 22 genes on 9p21.3 (19.9-32.8 Mb) in eight case-control studies: thyroid cancer, endometrial cancer (EC), renal cell carcinoma, colorectal cancer (CRC), colorectal adenoma (CA), oesophageal squamous cell carcinoma (ESCC), gastric cardia adenocarcinoma and osteosarcoma (OS). We used logistic regression to perform single SNP analyses for each study separately, adjusting for study-specific covariates. We combined SNP results across studies by fixed-effect meta-analyses and a newly developed subset-based statistical approach (ASSET). Gene-based P-values were obtained by the minP method using the Adaptive Rank Truncated Product program. We adjusted for multiple comparisons by Bonferroni correction. Rs3731239 in cyclin-dependent kinase inhibitors 2A (CDKN2A) was significantly associated with ESCC (P=7 × 10(-6)). The CDKN2A-ESCC association was further supported by gene-based analyses (Pgene=0.0001). In the meta-analyses by ASSET, four SNPs (rs3731239 in CDKN2A, rs615552 and rs573687 in CDKN2B and rs564398 in CDKN2BAS) showed significant associations with ESCC and EC (P<2.46 × 10(-4)). One SNP in MTAP (methylthioadenosine phosphorylase) (rs7023329) that was previously associated with melanoma and nevi in multiple genome-wide association studies was associated with CRC, CA and OS by ASSET (P=0.007). Our data indicate that genetic variants in CDKN2A, and possibly nearby genes, may be associated with ESCC and several other tumours, further highlighting the importance of 9p21.3 genetic variants in carcinogenesis.


Training a Hyperdimensional Computing Classifier Using a Threshold on Its Confidence.
Hyperdimensional computing (HDC) has become popular for light-weight and energy-efficient machine learning, suitable for wearable Internet-of-Things devices and near-sensor or on-device processing. HDC is computationally less complex than traditional deep learning algorithms and achieves moderate to good classification performance. This letter proposes to extend the training procedure in HDC by taking into account not only wrongly classified samples but also samples that are correctly classified by the HDC model but with low confidence. We introduce a confidence threshold that can be tuned for each data set to achieve the best classification accuracy. The proposed training procedure is tested on UCIHAR, CTG, ISOLET, and HAND data sets for which the performance consistently improves compared to the baseline across a range of confidence threshold values. The extended training procedure also results in a shift toward higher confidence values of the correctly classified samples, making the classifier not only more accurate but also more confident about its predictions.


Measles vaccination before the measles-mumps-rubella vaccine.
At the beginning of the 1960s, it was clear that a vaccine against measles would soon be available. Although measles was (and remains) a killer disease in the developing world, in the United States and Western Europe this was no longer so. Many parents and many medical practitioners considered measles an inevitable stage of a child's development. Debating the desirability of measles immunization, public health experts reasoned differently. In the United States, introduction of the vaccine fit well with Kennedy's and Johnson's administrations' political commitments. European policymakers proceeded cautiously, concerned about the acceptability of existing vaccination programs. In Sweden and the Netherlands, recent experience in controlling polio led researchers to prefer an inactivated virus vaccine. Although in the early 1970s attempts to develop a sufficiently potent inactivated vaccine were abandoned, we have argued that the debates and initiatives of the time during the vaccine's early history merit reflection in today's era of standardization and global markets.


Light and electron microscope observations of the lymphatic drainage units of the peritoneal cavity of rodents.
Fluid, particles, and cells are taken up from the peritoneal cavity by lymphatic drainage units, which, in the mouse and rat, are located along the peritoneal surface of the muscular portion of the diaphragm. The drainage units are composed of three specifically differentiated components: a lymphatic lacuna, a covering of lacunar mesothelium, and intervening submesothelial connective tissue. The units are drained by connecting lymphatic vessels that cross the diaphragm to empty into collecting lymphatic vessels running along the pleural surface of the diaphragm. The collecting lymphatics empty into parasternal lymphatic trunks. In this report, we briefly review critical features of the drainage apparatus and describe new observations, summarized below, about their structure. Around the rim of stomata, the mesothelial openings that lead into the lymphatic lacunae, plasma membranes of lacunar mesothelial cells and of lacunar endothelial cells abut but are not linked to one another by recognizable junctional specializations. Lacunar endothelial cells often extend valve-like processes that bridge the distal end of the channel beneath the stoma. The configuration of the endothelial processes may be complex. Occasionally, processes from fibroblasts in the submesothelial connective tissue adjacent to stomata make contact with the interstitial surface of lacunar endothelial cells. A discontinuous elastic layer in the submesothelial connective tissue spans the roof of each lacuna. Connecting and collecting lymphatics, which drain lymphatic lacunae, possess endothelial valves. Possible functions for each of these newly described structural features are discussed.


Interventions for emergency contraception.
Emergency contraception (EC) is using a drug or copper intrauterine device (Cu-IUD) to prevent pregnancy shortly after unprotected intercourse. Several interventions are available for EC. Information on the comparative effectiveness, safety and convenience of these methods is crucial for reproductive healthcare providers and the women they serve. This is an update of a review previously published in 2009 and 2012. To determine which EC method following unprotected intercourse is the most effective, safe and convenient to prevent pregnancy. In February 2017 we searched CENTRAL, MEDLINE, Embase, PsycINFO, CINAHL, Popline and PubMed, The Chinese biomedical databases and UNDP/UNFPA/WHO/World Bank Special Programme on Human Reproduction (HRP) emergency contraception database. We also searched ICTRP and ClinicalTrials.gov as well as contacting content experts and pharmaceutical companies, and searching reference lists of appropriate papers. Randomised controlled trials including women attending services for EC following a single act of unprotected intercourse were eligible. We used standard methodological procedures recommended by Cochrane. The primary review outcome was observed number of pregnancies. Side effects and changes of menses were secondary outcomes. We included 115 trials with 60,479 women in this review. The quality of the evidence for the primary outcome ranged from moderate to high, and for other outcomes ranged from very low to high. The main limitations were risk of bias (associated with poor reporting of methods), imprecision and inconsistency. Comparative effectiveness of different emergency contraceptive pills (ECP)Levonorgestrel was associated with fewer pregnancies than Yuzpe (estradiol-levonorgestrel combination) (RR 0.57, 95% CI 0.39 to 0.84, 6 RCTs, n = 4750, I2 = 23%, high-quality evidence). This suggests that if the chance of pregnancy using Yuzpe is assumed to be 29 women per 1000, the chance of pregnancy using levonorgestrel would be between 11 and 24 women per 1000.Mifepristone (all doses) was associated with fewer pregnancies than Yuzpe (RR 0.14, 95% CI 0.05 to 0.41, 3 RCTs, n = 2144, I2 = 0%, high-quality evidence). This suggests that if the chance of pregnancy following Yuzpe is assumed to be 25 women per 1000 women, the chance following mifepristone would be between 1 and 10 women per 1000.Both low-dose mifepristone (less than 25 mg) and mid-dose mifepristone (25 mg to 50 mg) were probably associated with fewer pregnancies than levonorgestrel (RR 0.72, 95% CI 0.52 to 0.99, 14 RCTs, n = 8752, I2 = 0%, high-quality evidence; RR 0.61, 95% CI 0.45 to 0.83, 27 RCTs, n = 6052, I2 = 0%, moderate-quality evidence; respectively). This suggests that if the chance of pregnancy following levonorgestrel is assumed to be 20 women per 1000, the chance of pregnancy following low-dose mifepristone would be between 10 and 20 women per 1000; and that if the chance of pregnancy following levonorgestrel is assumed to be 35 women per 1000, the chance of pregnancy following mid-dose mifepristone would be between 16 and 29 women per 1000.Ulipristal acetate (UPA) was associated with fewer pregnancies than levonorgestrel (RR 0.59; 95% CI 0.35 to 0.99, 2 RCTs, n = 3448, I2 = 0%, high-quality evidence). Comparative effectiveness of different ECP dosesIt was unclear whether there was any difference in pregnancy rate between single-dose levonorgestrel (1.5 mg) and the standard two-dose regimen (0.75 mg 12 hours apart) (RR 0.84, 95% CI 0.53 to 1.33, 3 RCTs, n = 6653, I2 = 0%, moderate-quality evidence).Mid-dose mifepristone was associated with fewer pregnancies than low-dose mifepristone (RR 0.73; 95% CI 0.55 to 0.97, 25 RCTs, n = 11,914, I2 = 0%, high-quality evidence). Comparative effectiveness of Cu-IUD versus mifepristoneThere was no conclusive evidence of a difference in the risk of pregnancy between the Cu-IUD and mifepristone (RR 0.33, 95% CI 0.04 to 2.74, 2 RCTs, n = 395, low-quality evidence). Adverse effectsNausea and vomiting were the main adverse effects associated with emergency contraception. There is probably a lower risk of nausea (RR 0.63, 95% CI 0.53 to 0.76, 3 RCTs, n = 2186 , I2 = 59%, moderate-quality evidence) or vomiting (RR 0.12, 95% CI 0.07 to 0.20, 3 RCTs, n = 2186, I2 = 0%, high-quality evidence) associated with mifepristone than with Yuzpe. levonorgestrel is probably associated with a lower risk of nausea (RR 0.40, 95% CI 0.36 to 0.44, 6 RCTs, n = 4750, I2 = 82%, moderate-quality evidence), or vomiting (RR 0.29, 95% CI 0.24 to 0.35, 5 RCTs, n = 3640, I2 = 78%, moderate-quality evidence) than Yuzpe. Levonorgestrel users were less likely to have any side effects than Yuzpe users (RR 0.80, 95% CI 0.75 to 0.86; 1 RCT, n = 1955, high-quality evidence). UPA users were more likely than levonorgestrel users to have resumption of menstruation after the expected date (RR 1.65, 95% CI 1.42 to 1.92, 2 RCTs, n = 3593, I2 = 0%, high-quality evidence). Menstrual delay was more common with mifepristone than with any other intervention and appeared to be dose-related. Cu-IUD may be associated with higher risks of abdominal pain than mifepristone (18 events in 95 women using Cu-IUD versus no events in 190 women using mifepristone, low-quality evidence). Levonorgestrel and mid-dose mifepristone (25 mg to 50 mg) were more effective than Yuzpe regimen. Both mid-dose (25 mg to 50 mg) and low-dose mifepristone(less than 25 mg) were probably more effective than levonorgestrel (1.5 mg). Mifepristone low dose (less than 25 mg) was less effective than mid-dose mifepristone. UPA was more effective than levonorgestrel.Levonorgestrel users had fewer side effects than Yuzpe users, and appeared to be more likely to have a menstrual return before the expected date. UPA users were probably more likely to have a menstrual return after the expected date. Menstrual delay was probably the main adverse effect of mifepristone and seemed to be dose-related. Cu-IUD may be associated with higher risks of abdominal pain than ECPs.


Role of LONELY GUY genes in indeterminate nodulation on Medicago truncatula.
LONELY GUY (LOG) genes encode cytokinin riboside 5'-monophosphate phosphoribohydrolases and are directly involved in the activation of cytokinins. To assess whether LOG proteins affect the influence of cytokinin on nodulation, we studied two LOG genes of Medicago truncatula. Expression analysis showed that MtLOG1 and MtLOG2 were upregulated during nodulation in a CRE1-dependent manner. Expression was mainly localized in the dividing cells of the nodule primordium. In addition, RNA interference revealed that MtLOG1 is involved in nodule development and that the gene plays a negative role in lateral root development. Ectopic expression of MtLOG1 resulted in a change in cytokinin homeostasis, triggered cytokinin-inducible genes and produced roots with enlarged vascular tissues and shortened primary roots. In addition, those 35S:LOG1 roots also displayed fewer nodules than the wild-type. This inhibition in nodule formation was local, independent of the SUPER NUMERIC NODULES gene, but coincided with an upregulation of the MtCLE13 gene, encoding a CLAVATA3/EMBRYO SURROUNDING REGION peptide. In conclusion, we demonstrate that in M. truncatula LOG proteins might be implicated in nodule primordium development and lateral root formation.


Smoking, Binge Drinking, and Drug Use Among Childhood Cancer Survivors: A Meta-Analysis.
Childhood cancer survivors are at risk for late effects of therapy, some of which may be exacerbated by smoking, alcohol, or drug use. We undertook a meta-analysis of the literature to determine whether survivors engage in risk-taking behaviors at rates different from their peers/siblings. Studies comparing current engagement in risk-taking behaviors between cancer survivors and siblings or matched peers were identified in MEDLINE (1946-), EMBASE (1947-), PsychINFO (1806-), and the Cochrane Controlled Trials Register. Two reviewers assessed publications for inclusion and extracted data independently. Studies were combined using inverse variance weighting to determine odds ratios (OR) and prevalence rates of risk-taking behaviors in survivors compared to controls. Fourteen of 1,713 studies satisfied inclusion criteria. Twelve assessed smoking, six binge drinking, and seven drug use. Among survivors, 22% (95% confidence interval 0.19, 0.26) smoked, 20% (0.08, 0.51) were binge drinkers, and 15% (0.10, 0.23) used drugs. Survivors were less likely than siblings to smoke (OR 0.68 [0.49, 0.96]) or binge drink (OR 0.77 [0.68, 0.88]), but similarly likely to use drugs (OR 0.33 [0.03, 3.28]). Survivors were less likely than matched peers to smoke (OR 0.54 [0.42, 0.70]) or use drugs (OR 0.57 [0.40, 0.82]), but equally likely to binge drink (OR 0.97 [0.38, 2.49]). Childhood cancer survivors engage in similar or lower rates of risk taking than their siblings/peers. Future studies should identify survivors most likely to benefit from focused interventions, and determine the impact of risk-taking behaviors on the risk for late effects of cancer therapy.


Pheochromocytoma and paraganglioma.
Pheochromocytoma is a very special kind of tumor full of duplicity. On the one hand it represents its own microworld with unique clinical, biochemical and pathological features, while on the other it constitutes a tremendously significant part of whole body system, playing a vital role for practically every organ system. It has a very special character - sometimes like a child it can be sweet and predictable, while at times it can behave like a deadly wild beast, crashing and tearing everything on its path in a fierce rage. It also consists of the amazingly intelligent neuroendocrine cells that possess a magical ability to make miraculous substances of many kinds. But most of all, it is a system that is able to drive our curiosity and the itch of "Cogito, ergo sum" to limitless depths and year by year it still amazes us with new and unexpected discoveries that move our understanding of multiple pathways and metabolic events closer to the ultimate truth. Recent discoveries of succinate dehydrogenase (SHD) and prolyl hydroxylase (PHD) mutations, for example, propelled our understanding of neuroendocrine tumorigenesis as a whole, as well as physiology of mitochondrial respiratory chain and phenomenon of pseudohypoxia in particular. Good old discoveries make their way from dusty repositories to shine with new meaning, appropriate for the current level of knowledge. This acquired wisdom makes us better physicians - knowing the specific expression makeup of catecholamine transporters, GLUTs and SRIFs allows for better tailored imaging and therapeutic manipulations. There are still long ways to go, keeping in mind that pheochromocytoma is but so very special, and we are optimistic and expect many great things to come.


Chronic daily headache.
Headache is a relatively neglected neurological disorder. Indeed, many neurologists find outpatient headache management--particularly of chronic daily headache--one of the least engaging parts of their job. The neglect of headache as a research problem has been reversed by the relatively recent emergence of strong programmes in centres such as Copenhagen, London, Philadelphia, New York, Liege and Leiden. Partly as a result of this, the pejorative attitude to headache as a clinical problem is less than it was, but many neurologists are still bemused by the intrusion of headache, both at a local level when headache patients occupy scarce specialist beds, and at a global level where the World Health Organization ranks migraine in the top 20 causes of global disability. This article reviews one of the commonest headache syndromes encountered by neurologists--chronic daily headache.


Passive air sampling: the use of the index of microbial air contamination.
Bioaerosol plays an important role in human life with potentially infectious, allergic and toxic effects. Active and passive methods can be used to assess microbial air contamination, but so far there is not a unanimous consensus regarding the indications about methods to be used and how to interpret the results. The passive method has been standardized by the Index of Microbial Air contamination (IMA). Classes of contamination and maximum acceptable levels of IMA have been proposed, related to different infection or contamination risks. The aim of this study was to provide information about the use of the passive sampling method, with reference to the IMA standard. We searched PubMed and Scopus for articles published until January 2020 reporting the citation of the article by Pasquarella et al. "The index of microbial air contamination. J Hosp Infect 2000". Only studies in English language where the IMA standard was applied were considered. Studies regarding healthcare settings were excluded. 27 studies were analyzed; 12 were performed in Europe, 8 in Asia, 5 in Africa, 2 in America. Cultural heritage sites, educational buildings and food industries were the most common indoor monitored environments; in 8 studies outdoor air was monitored. This review has provided a picture of the application of standard IMA in different geographic areas and different environments at risk of airborne infection/contamination. The analysis of the results obtained, together with a wider collection of data, will provide a useful contribution towards the definition of reference limits for the various types of environments to implement targeted preventive measures.


Optimal cure rate by personalized HCV regimens in real-life: a proof-of-concept study.
Pretreatment evaluation of HCV-infected patients is a complex interplay between multiple clinical and viral parameters, leading to a tailored approach that may bring real-life sustained virological response (SVR) rates close to 98%-99%. As proof-of-concept, we evaluated the efficacy of all-oral direct-acting antiviral (DAA) regimens in patients whose personalization included pre-therapy evaluation of natural resistance-associated substitutions (RASs), in addition to international guideline recommendations. One hundred and thirty-one patients who started a first-line all-oral DAA regimen between April 2015 and December 2016 were tested for baseline NS3 and NS5A RASs by Sanger sequencing. SVR12 was defined as HCV-RNA undetectability 12 weeks after treatment discontinuation. Compatibly with a real-life context, 74.0% (97 of 131) of patients presented ≥2 pretreatment risk factors for failure to achieve SVR12 (infection by GT-1a/GT-3a; cirrhosis; previous treatment experience; HCV-RNA ≥800 000 IU/mL) and 33.6% had ≥3 risk factors. Natural RASs were frequently detected (32.1% prevalence), with substantial prevalence of NS5A RASs (15.3%), mostly represented by Y93H in GT-1b (3 of 36, 8.3%) and GT-3a (3 of 25, 12.0%) and F28C in GT-2c (2 of 11, 18.2%). Overall, personalized treatment led to 100% SVR12, even in those patients for whom treatment strategy was either strengthened (by ribavirin inclusion and/or duration increase) or simplified (by ribavirin exclusion and/or duration reduction), thanks to baseline RAS evaluation. Even with newer DAA regimens, an integrated interpretation of clinical and virological pretreatment parameters, including natural RASs, may play a relevant role in bringing SVR rates close to the highest achievable. Treatment tailoring can be foreseen in 'hard-to-treat' patients, but also in 'easy' patients with favourable indicators, whereby a simplification/shortening of recommended regimens can be indicated.


All-fiber all-normal dispersion laser with a fiber-based Lyot filter.
We propose the use of a short section of polarization-maintaining fiber as a birefringent medium to construct an all-fiber Lyot filter inside the cavity of a fiber laser. This allows mode-locked operation of an all-fiber all-normal dispersion Yb-fiber oscillator without the use of a bulk bandpass filter and using standard components. Moreover, filter bandwidth and modulation depth is easily controlled by changing the length and splice angle of the polarization-maintaining-fiber section, leading to an adjustable filter. At mode-locked operation, the 30% output fiber port delivers 1 nJ pulses that are dechirped to 230 fs duration.


CircaCompare: a method to estimate and statistically support differences in mesor, amplitude and phase, between circadian rhythms.
A fundamental interest in chronobiology is to compare patterns between groups of rhythmic data. However, many existing methods are ill-equipped to derive statements concerning the statistical significance of differences between rhythms that may be visually apparent. This is attributed to both the form of data used (longitudinal versus cross-sectional) and the limitations of the statistical tests used to draw conclusions. To address this problem, we propose that a cosinusoidal curve with a particular parametrization be used to model and compare data of two sets of observations collected over a 24-h period. The novelty of our test is in the parametrization, which allows the explicit estimation of rhythmic parameters [mesor (the rhythm-adjusted mean level of a response variable around which a wave function oscillates), amplitude and phase], and simultaneously testing for statistical significance in all three parameters between two or more groups of datasets. A statistically significant difference between two groups, regarding each of these rhythmic parameters, is indicated by a P-value. The method is evaluated by applying the model to publicly available datasets, and is further exemplified by comparison to the currently recommended method, DODR. The results suggest that the method proposed may be highly sensitive to detect rhythmic differences between groups in phase, amplitude and mesor. https://github.com/RWParsons/circacompare/.


Adrenal steroid responses to naloxone in polycystic ovarian disease.
In order to investigate the role of the adrenal gland in the pathogenesis of polycystic ovarian disease (PCOD), we evaluated the adrenal steroid response to an opiate receptor blockade. Six healthy menstruating volunteers and 6 patients with PCOD were given a saline or naloxone (4 mg i.v.) injection in the early follicular phase. Blood samples were taken prior to the injections and every 15-30 minutes in the following 2 hours. Cortisol, androstenedione (A) and dehydroepiandrosterone (DHA) plasma levels were determined by RIA after extraction (cortisol) and celite chromatography (A and DHA). While in controls naloxone increased only cortisol concentrations, in PCOD patients DHA plasma levels also were stimulated by the opiate receptor antagonist. In PCOD patients the increase of cortisol (p less than 0.05) and of DHA (p less than 0.001) levels resulted significantly higher than in controls. In both groups A plasma levels remained unchanged after naloxone administration. These data confirm that endogenous opioids exert an inhibitory control on the pituitary-adrenal axis. In PCOD patients the response to naloxone led to a hypersecretion of adrenal delta 5-androgens, which could account for the development of the syndrome.


Open Source Tools for Biological Image Analysis.
Visiting the Bio Imaging Search Engine (BISE) (Bio, BISE, Engine, http://biii.eu/, Imaging, Search) website at the time of writing this article, almost 1200 open source assets (components, workflows, collections) were found. This overwhelming range of offer difficults the fact of making a reasonable choice, especially to newcomers. In the following chapter, we briefly sketch the advantages of the open source software (OSS) particularly used for image analysis in the field of life sciences. We introduce both the general OSS idea as well as some programs used for image analysis. Even more, we outline the history of ImageJ as it has served as a role model for the development of more recent software packages. We focus on the programs that are, to our knowledge, the most relevant and widely used in the field of light microscopy, as well as the most commonly used within our facility. In addition, we briefly discuss recent efforts and approaches aimed to share and compare algorithms and introduce software and data sharing good practices as a promising strategy to facilitate reproducibility, software understanding, and optimal software choice for a given scientific problem in the future.


Equine lymphocytic-plasmacytic enterocolitis: a retrospective study of 14 cases.
Lymphocytic-plasmacytic enteritis (LPE) is a morphological diagnosis given to a type of infiltrative intestinal disease classified within the complex of idiopathic inflammatory bowel disease (IBD). The purpose of this retrospective study was to describe the details of breed, age and sex, clinical and clinicopathological findings and outcome of horses diagnosed with LPE. Data were reported from 14 horses that had a histopathological diagnosis of LPE; the median age was 12 years, and there was no breed or sex predilection. Common clinical signs were weight loss (100%), diarrhoea (50%) and lethargy (50%). Hypoproteinaemia and hypoalbuminaemia were present in 6/13 (46%) and 9/12 (75%) horses, respectively. Results of the oral glucose tolerance tests or D-xylose absorption tests were abnormal in 9/12 (75%) horses and rectal mucosal biopsies were abnormal in 3/7 (43%) horses. Corticosteroid treatment was used unsuccessfully in 4 horses. All horses were subjected to euthanasia and LPE was diagnosed at necropsy. Diffuse thickening of the small intestine was apparent grossly in 10/13 (77%) horses examined. LPE is an uncommon equine intestinal disease that is difficult to diagnose antemortem and has a poor prognosis.


Synthesis, Properties, and Mechanism of Action of New Generation of Polycyclic Glycopeptide Antibiotics.
The increased resistance of glycopeptide based antibiotics has become a serious problem for the chemotherapy of infections triggered by resistant Gram-positive bacteria. This has motivated the urgent sincere efforts to develop potent glycopeptide-based antibiotics in both academy and industry research laboratories. Understanding of the mechanism of action of natural and modified glycopeptides is considered as the basis for the rational design of compounds with valuable properties to achieve the fundamental results. Several hydrophobic glycopeptide analogues active against resistant strains were developed during the last two decades. Three drugs, namely, oritavancin, telavancin and dalbavancin were approved by FDA in 2013-2014. It was found that hydrophobic derivatives act through different mechanisms without binding with the modified target of resistant bacteria. Types: Different types of chemical modifications led to several glycopeptide analogues active against Gram-negative bacteria as advocated by in vitro studies or demonstrating potent antiviral activity in the cell models. A new class of glycopeptide antibiotics with potent activity against sensitive and resistant bacterial strains has been recently reported with the aim to overcome the resistance, however, there are a lot of obscure problems in the complete understanding of their mechanisms of actions. In this review, we summarized the achievements of synthetic methods devoted to the construction of new polycyclic glycopeptide antibiotics and described the studies related to their mechanism of actions.


Mapping leptin-interacting sites in recombinant leptin-binding domain (LBD) subcloned from chicken leptin receptor.
The binding domain of the chicken leptin receptor [chLBD (chicken leptin-binding domain)], subcloned from the full-size chicken leptin receptor and prepared in an Escherichia coli system, was subjected to site-directed mutagenesis to identify the amino acids involved in leptin binding. A total of 22 electrophoretically pure, >90% monomer-containing mutants were expressed, refolded and purified. The effects of the mutations were tested by the ability to form complexes with ovine leptin, and the kinetic parameters of interaction were determined by surface plasmon resonance. Six mutants were used to determine whether mutations of several amino acids that differ between chLBD and mammalian LBDs will affect affinity: none showed any such effect, except the mutant A105D (Ala(105)-->Asp), which exhibited some decrease in affinity. Surface plasmon resonance analysis identified six mutants in which binding activity was totally abolished (F73A, Y14A/F73A, V76A/F77A, L78A/L79A, V76A/F77A/L78A/L79A and A105D/D106V) and six mutants (Y14A, R41A, R41A/S42A/K43A, V103A, V135A/F136A and F136A) in which affinity for the hormone was reduced, mainly by increased dissociation rates. Gel-filtration experiments indicated the formation of a 1:1 ovine or human leptin-chLBD complex with a molecular mass of approx. 41 kDa. Gel-filtration experiments yielded 1:1 complexes with those mutants in which affinity had decreased, but not with the six mutants, which had totally lost their binding capacity. Modelling the leptin-chLBD complex indicated that the binding domain of the latter is located mainly in the L3 loop, which contributes nine amino acid residues interacting with leptin. Contact-surface analysis identified the residues having the highest contribution to the recognition site to be Phe73, Phe77 and Leu79.


Clonal dissemination of vancomycin-resistant Enterococcus faecium ST412 in a Brazilian region.
Vancomycin-resistant Enterococcus faecium (VREfm) has emerged as an important global nosocomial pathogen, and this trend is associated with the spread of high-risk clones. Here, we determined the genetic and phenotypic features of 93 VREfm isolates that were obtained from patients in 13 hospitals in Vitória, Espírito Santo, Brazil, during 2012-2013. All the isolates were vancomycin-resistant and harbored the vanA gene. Only 6 (6.5%) of the VREfm isolates showed the ability to form biofilm. The 93 isolates analyzed belong to a single pulsed-field gel electrophoresis lineage and presented six subtypes. MLST genotyping showed that all VREfm belonged to ST412 (the high-risk clone, hospital-adapted). The present study describes the dissemination of ST412 clone in the local hospitals. The clonal spread of these ST412 isolates in the area we analyzed as well as other hospitals in southeastern Brazil supports the importance of identifying and controlling the presence of these microorganisms in health care-related services.


spaP gene of Streptococcus mutans in dental plaque and its relationship with early childhood caries.
Streptococcus mutans and Streptococcus sobrinus are the main pathogens associated with the development of dental caries in humans. Recently, the real-time polymerase chain reaction (qPCR-TR) has been used for fast and exact quantification of these bacteria species. This molecular biology method has made the detection of these bacteria in saliva and dental plaque possible; additionally, it aids the development of illness risk prediction. The purpose of this prospective, analytic, transversal, observational and unicenter study was to quantify the spaP gene of the Streptococcus mutans and its correlation with caries in a group of children using isolated DNA from plaque samples processed through qPCR-TR, using specific oligonucleotides for this gene detection. The cariogenic potential of Streptococcus mutans in the dental plaque was analysed in a group of patients aged 12 to 46 months. A descriptive statistical analysis was performed. The Spearman's correlation coefficient was used to establish the correlation between caries (dmft) index (decayed/missing/filled primary teeth), spaP gene and age group. The Wilcoxon test was used to compare MSB cultivation technique and qPCR-TR. In the molecular trials, a close association between caries prevalence in childhood and the presence and high proportion of the spaP gene of S. mutans was found. The average caries prevalence was 3.71, and it increased as age range increased. The highest caries prevalence was observed in female patients and in the oldest age range studied (40 46 months) which contrasts with the 12-18 months age that had a caries (dmft) index of zero. The amplification using as initiator the gene spaP of the nucleic acids extracted from the S. mutans resulted positive in 91.3% of the cases. Every child with caries was positive for the spaP and only 8.75% were negative, this group included children without caries. In conclusion, there was a correlation with infant caries prevalence and S. mutans.


Fangchinoline inhibits SARS-CoV-2 and MERS-CoV entry.
The COVID-19 pandemic caused by SARS-CoV-2, lead to mild to severe respiratory illness and resulted in 6.9 million deaths worldwide. Although vaccines are effective in preventing COVID-19, they may not be sufficient to protect immunocompromised individuals from this respiratory illness. Moreover, novel emerging variants of SARS-CoV-2 pose a risk of new COVID-19 waves. Therefore, identification of effective antivirals is critical in controlling SARS and other coronaviruses, such as MERS-CoV. We show that Fangchinoline (Fcn), a bisbenzylisoquinoline alkaloid, inhibits replication of SARS-CoV, SARS-CoV-2, and MERS-CoV in a range of in vitro assays, by blocking entry. Therapeutic use of Fcn inhibited viral loads in the lungs, and suppressed associated airway inflammation in hACE2. Tg mice and Syrian hamster infected with SARS-CoV-2. Combination of Fcn with remdesivir (RDV) or an anti-leprosy drug, Clofazimine, exhibited synergistic antiviral activity. Compared to Fcn, its synthetic derivative, MK-04-003, more effectively inhibited SARS-CoV-2 and its variants B.1.617.2 and BA.5 in mice. Taken together these data demonstrate that Fcn is a pan beta coronavirus inhibitor, which possibly can be used to combat novel emerging coronavirus diseases.


Cancer incidence in a cohort of infertile women.
Among 2,496 infertile Israeli women treated between 1964 and 1974, 143 cancer cases were observed as compared with 116.1 expected (standardized incidence ratio (SIR) = 1.2, 95% confidence interval (CI) 1.0-1.5) through 1991. Site-specific analysis revealed 12 ovarian cancers versus 7.2 expected (SIR = 1.6, 95% CI 0.8-2.9), 21 endometrial cancers versus 4.3 expected (SIR = 4.85, 95% CI 3.0-7.4), and 59 breast cancers versus 46.6 expected (SIR = 1.3, 95% CI 0.96-1.6). Sensitivity analysis revealed that confounding was unlikely to explain the raised risk of endometrial cancer, but nulliparity might explain the increased risk of ovarian cancer. The excess of endometrial cancer was prominent among patients with normal estrogen production but progesterone deficiency (SIR = 9.4, 95% CI 5.0-16.0). The risk for ovarian cancer was similar among the total groups of treated and untreated patients (SIR = 1.7 vs. 1.6). The standardized incidence ratio for endometrial cancer was higher among the treated group than the untreated group, although not significantly. Treatment with ovulation-inducing drugs does not appear to increase the risk for ovarian cancer, but its role cannot be completely excluded.


A novel polar surface polysaccharide from Rhizobium leguminosarum binds host plant lectin.
Rhizobium bacteria produce different surface polysaccharides which are either secreted in the growth medium or contribute to a capsule surrounding the cell. Here, we describe isolation and partial characterization of a novel high molecular weight surface polysaccharide from a strain of Rhizobium leguminosarum that nodulates Pisum sativum (pea) and Vicia sativa (vetch) roots. Carbohydrate analysis showed that the polysaccharide consists for 95% of mannose and glucose, with minor amounts of galactose and rhamnose. Lectin precipitation analysis revealed high binding affinity of pea and vetch lectin for this polysaccharide, in contrast to the other known capsular and extracellular polysaccharides of this strain. Expression of the polysaccharide was independent of the presence of a Sym plasmid or the nod gene inducer naringenin. Incubation of R. leguminosarum with labelled pea lectin showed that this polysaccharide is exclusively localized on one of the poles of the bacterial cell. Vetch roots incubated with rhizobia and labelled pea lectin revealed that this bacterial pole is involved in attachment to the root surface. A mutant strain deficient in the production of this polysaccharide was impaired in attachment and root hair infection under slightly acidic conditions, in contrast to the situation at slightly alkaline conditions. Our data are consistent with the hypothesis that rhizobia can use (at least) two mechanisms for docking at the root surface, with use of a lectin-glycan mechanism under slightly acidic conditions.


Temporal summation of heat pain in humans: Evidence supporting thalamocortical modulation.
Noxious cutaneous contact heat stimuli (48 degrees C) are perceived as increasingly painful when the stimulus duration is extended from 5 to 10s, reflecting the temporal summation of central neuronal activity mediating heat pain. However, the sensation of increasing heat pain disappears, reaching a plateau as stimulus duration increases from 10 to 20s. We used functional magnetic resonance imaging (fMRI) in 10 healthy subjects to determine if active central mechanisms could contribute to this psychophysical plateau. During heat pain durations ranging from 5 to 20s, activation intensities in the bilateral orbitofrontal cortices and the activation volume in the left primary (S1) somatosensory cortex correlated only with perceived stimulus intensity and not with stimulus duration. Activation volumes increased with both stimulus duration and perceived intensity in the left lateral thalamus, posterior insula, inferior parietal cortex, and hippocampus. In contrast, during the psychophysical plateau, both the intensity and volume of thalamic and cortical activations in the right medial thalamus, right posterior insula, and left secondary (S2) somatosensory cortex continued to increase with stimulus duration but not with perceived stimulus intensity. Activation volumes in the left medial and right lateral thalamus, and the bilateral mid-anterior cingulate, left orbitofrontal, and right S2 cortices also increased only with stimulus duration. The increased activity of specific thalamic and cortical structures as stimulus duration, but not perceived intensity, increases is consistent with the recruitment of a thalamocortical mechanism that participates in the modulation of pain-related cortical responses and the temporal summation of heat pain.


NOD2-C2 - a novel NOD2 isoform activating NF-kappaB in a muramyl dipeptide-independent manner.
The innate immune system employs several receptor families that form the basis of sensing pathogen-associated molecular patterns. NOD (nucleotide-binding and oligomerization domain) like receptors (NLRs) comprise a group of cytosolic proteins that trigger protective responses upon recognition of intracellular danger signals. NOD2 displays a tandem caspase recruitment domain (CARD) architecture, which is unique within the NLR family. Here, we report a novel alternative transcript of the NOD2 gene, which codes for a truncated tandem CARD only protein, called NOD2-C2. The transcript isoform is highest expressed in leucocytes, a natural barrier against pathogen invasion, and is strictly linked to promoter usage as well as predominantly to one allele of the single nucleotide polymorphism rs2067085. Contrary to a previously identified truncated single CARD NOD2 isoform, NOD2-S, NOD2-C2 is able to activate NF-kappaB in a dose dependent manner independently of muramyl dipeptide (MDP). On the other hand NOD2-C2 competes with MDPs ability to activate the NOD2-driven NF-kappaB signaling cascade. NOD2 transcripts having included an alternative exon downstream of exon 3 (exon 3a) are the endogenous equivalents of a previously described in vitro construct with the putative protein composed of only the two N-terminal CARDs. This protein form (NOD2-C2) activates NF-kappaB independent of an MDP stimulus and is a potential regulator of NOD2 signaling.


Does sleep-dependent consolidation favour weak memories?
Sleep stabilizes newly acquired memories, a process referred to as memory consolidation. According to recent studies, sleep-dependent consolidation processes might be deployed to different extents for different types of memories. In particular, weaker memories might benefit greater from post-learning sleep than stronger memories. However, under standard testing conditions, sleep-dependent consolidation effects for stronger memories might be obscured by ceiling effects. To test this possibility, we devised a new memory paradigm (Memory Arena) in which participants learned temporospatial arrangements of objects. Prior to a delay period spent either awake or asleep, training thresholds were controlled to yield relatively weak or relatively strong memories. After the delay period, retrieval difficulty was controlled via the presence or absence of a retroactive interference task. Under standard testing conditions (no interference), a sleep-dependent consolidation effect was indeed observed for weaker memories only. Critically though, with increased retrieval demands, sleep-dependent consolidation effects were seen for both weaker and stronger memories. These results suggest that all memories are consolidated during sleep, but that memories of different strengths require different testing conditions to unveil their benefit from post-learning sleep.


Socio-Demographic Characteristics and Patterns of Substance Use Disorder in Oman: A retrospective study of the National Registry Surveillance Programme between 2004 and 2018.
Substance use disorder is a global challenge. Therefore, this study aimed to provide an updated view of socio-demographic characteristics and patterns of substance use in Oman. This retrospective descriptive study was conducted between 2004 and 2018. Data were retrieved from Oman's National Drug Addict Registry. The data collected included the socio-demographic characteristics of registered cases, the proportion of various psychoactive substances' consumption and their routes of administration, the associated sociodemographic determinants as well as comorbid conditions. A total of 6,453 cases were registered during the study's timeframe. The majority of which were Omani (97.9%), male (98.7%), single (57.9%), unemployed (50.2%), had an education level below university (81.0%) and were adolescents and young adults (77.0%). Opiates were the most common substance used (66.6%) and more than half of the sample were polydrug users (51.0%). Injecting-drug users constituted 53.4% of the total registered cases. The proportion of people with hepatitis virus C, hepatits virus B and HIV among the registered cases were 46.9%, 5.1% and 3.7%, respectively. The findings are in favour of rapidly escalating the introduction of a substance use preventive programme at all school levels as well as making opioid substitution therapy and other harm reduction programmes available in Oman.


Coronectomy - An alternative approach to remove impacted teeth in oncological patients.
Coronectomy involves the section of the tooth crown leaving the roots in the socket. Possibility of inferior alveolar nerve injury and mandibular fracture are the main indications for this approach. Herein, we describe a case series of coronectomy to highlight its indication in normal and oncological patients. A total of 9 patients were submitted to coronectomy, 6 of them were oncological. Three patients were evaluated before head and neck radiotherapy (HNRT), 2 after HNRT and 1 before bone marrow transplantation. Mean age of the patients was 49 years, most of them male (n=7). Lower third molars were the main teeth that received this procedure, and all cases presented intimate anatomic relationship between the roots and the mandibular canal. Moreover, three cases also presented evident mandibular fracture risk in removing the tooth. During the follow-up period, none complications were observed related to coronectomy and oncological treatment. Coronectomy is a safe approach including for cancer patients and it should be considered in high-risk impacted teeth extractions. Key words:Coronectomy, impacted teeth, oncological patients, postoperative complications.


Dynamic risk assessment in patients with differentiated thyroid cancer.
The stratification of patients with differentiated thyroid cancer based on their initial risk of recurrence, according to specific clinical, histopathological and perioperative data, is an important starting point for tailoring the follow-up during the first 1-2 years after initial therapy (surgery with or without radioiodine ablation). However, risk of recurrence re-stratification based on new clinical data that becomes available after considering the response to treatment (dynamic risk assessment) provides a more accurate prediction of the status at final follow-up and a more individualized approach. In this review, we summarized the available data regarding dynamic risk of recurrence and the suggested management of differentiated thyroid cancer patients according to the response to treatment. The use of this strategy is crucial to avoid overtreatment and intensive follow-up of the vast majority of patients who will have a very good prognosis and, on the other hand, focus therapeutic efforts on those patients who will have a worse prognosis. In the future, molecular biology analysis of the tumors and well-designed prospective studies will probably refine the risk of recurrence prediction.


Long-term juxtafoveal retinal telangiectasia.
To determine the long-term natural history of idiopathic juxtafoveal telangiectasia (IJRT). Record review in 2 university-based and 2 private vitreoretinal practices sought patients with IJRT documented by color photographs and fluorescein angiograms (FAs) during the period January 1, 1980, to December 31, 1993. Patients then had repeated examinations and FAs. Twenty patients with IJRT in 32 eyes had follow-up examinations. Fifteen patients had color photographs and FAs, and one had color photographs alone. Follow-up varied from 10 years to 21 years (average, 15 years). Six eyes were treated by laser photocoagulation at onset. Twenty-four of the 26 untreated eyes lost vision as measured by Snellen testing. Visual loss and morphologic progression depended on IJRT type. Six of 8 untreated eyes with type IA IJRT lost vision by >/ or = 3 lines (Snellen), 4 to 20/70 or worse. Vision loss was caused by progressive telangiectatic changes and intraretinal edema. Fifteen of 20 initially untreated eyes with type IIA IJRT developed either central retinal pigment epithelium membranes or subretinal neovascularization with loss of vision to 20/80 or less. IJRT prognosis depends on type and clinical features. Long-term prognosis for central vision is poor.


Individual and combined effects of low dissolved oxygen and low pH on survival of early stage larval blue crabs, Callinectes sapidus.
A large number of coastal ecosystems globally are subjected to concurrent hypoxic and acidified conditions that will likely intensify and expand with continued climate change. In temperate regions, the spawning of many important organisms including the Atlantic blue crab Callinectes sapidus occurs during the summer months when the severity of coastal hypoxia and acidification is the greatest. While the blue crab earliest larval stage can be exposed to co-occurring hypoxia and acidification observed in many coastal ecosystems, the effects of these concurrent stressors on larval blue crab survival is unknown. This study investigated the individual and combined consequences of low dissolved oxygen (DO) and low pH on blue crab larvae survival through a series of short-term experiments. During 14-day experiments with moderately hypoxic conditions (117-127 μM O2 or 3.74-4.06 mg L-1) and acidified conditions (pH on total scale of 7.16-7.33), low DO and low pH individually and significantly reduced larval survival by 60% and 49%, respectively, with the combination of stressors reducing survival by 87% compared to the control treatment (210-269 μM O2 or 6.72-8.61 mg L-1, 7.91-7.94 DO and pH, respectively). During 4-day experiments with lower DO levels (68-83 μM O2 or 2.18-2.62 mg L-1) and comparable pH levels of 7.29-7.39, low DO individually reduced survival by >90% compared to the control (261-267 μM O2 or 8.35-8.54 mg L-1, 7.92-7.97 DO and pH, respectively), whereas low pH had no effect and there was no interaction between stressors. Over a 4-day period, the DO threshold at which 50% of the larval blue crab population died (LC50) was 121 μM O2 (3.86 mgL-1). In 14-day experiments, the DO and pH effects were additive, yielding survival rates lower than the individual treatments, and significantly correlated with DO and pH concentrations. Collectively, these findings indicate that blue crab sensitivity to both low DO and low pH are acute within the larval stage, depend on the intensity and duration of exposure, and leads to mortality, thereby potentially contributing to the interannual variability and possible regional declines of this fishery.


Immediate and delayed reactions to cosmetic ingredients.
The purpose of this study was to investigate the incidence and etiology of cutaneous reactions caused by cosmetics, with an emphasis on perfume sensitivity. 19 control subjects and 31 patch test clinic patients (16 with a history of adverse cosmetic reactions) were examined for sensitivity by history, open and patch testing using the North American Contact Dermatitis Group (NACDG) fragrance screening series and 11 other common allergens found in cosmetics. Contact urticaria was very frequent to certain chemicals; however, patients with a history of cosmetic sensitivity were not found to have a significant increase in positive reactions when compared to controls or patients with eczematous skin. 12 subjects had positive patch test reactions, most of which were not clinically relevant. 3 patients with a history of cosmetic sensitivity had positive reactions, only 1 of which was in the fragrance screening series (cinnamic alcohol). There were 6 reactions in patients with eczematous skin, 4 of which were to preservatives. 3 controls had positive reactions, each to thimerosal. A history of cosmetic sensitivity was not confirmed by open and closed skin testing in our subjects.


Paenibacillus koreensis sp. nov., a new species that produces an iturin-like antifungal compound.
A bacterial strain, YC300T, that produces an iturin-like antifungal antibiotic was isolated from compost and identified as member of the genus Paenibacillus. Gram reaction of the strain was variable depending upon growth stages and culture media. Three different types of colonies were developed on tryptic soy agar. The organism was facultatively anaerobic and grew at 50 degrees C. The DNA G+C content was 54 mol % and anteiso-C15:0 was the major fatty acid. A 0.9 kb fragment was produced by PCR amplification of strain YC300T DNA using primers PAEN515F and 1377R. Levels of 16S rDNA similarity between strain YC300T and other Paenibacillus species were between 89.8 and 94.8%. Phylogenetically, strain YC300T formed a significant monophyletic clade with Paenibacillus validus. It is clear from polyphasic evidence that the isolate should be classified as Paenibacillus koreensis sp. nov., the type strain of which is YC300T (= KCTC 2393T, KCCM 40903T).


Clusterin increases post-ischemic damages in organotypic hippocampal slice cultures.
Clusterin or apolipoprotein J is a heterodimeric glycoprotein which is known to be increased during tissue involution in response to hormonal changes or injury and under circumstances leading to apoptosis. Previous studies in wild-type (WT) and clusterin-null (Clu-/-) mice indicated a protective role of clusterin over-expression in astrocytes lasting up to 90 days post-ischemia. However, in in vitro and in vivo models of neonatal hypoxia-ischemia, clusterin exacerbates necrotic cell death. We developed recombinant forms of clusterin and examined their effect on propidium iodide uptake, neuronal and synaptic markers as well as electrophysiological recordings in hippocampal slice cultures from Clu-/- and WT mice subjected to oxygen-glucose deprivation (OGD). WT mice displayed a marked up-regulation of clusterin associated with electrophysiological deficits and dramatic increase of propidium iodide uptake 5 days post-OGD. Immunocytochemical and western blot analyses revealed a substantial decrease of neuronal nuclei and synaptophysin immunoreactivity that predominated in WT mice. These findings contrasted with the relative post-OGD resistance of Clu-/- mice. The addition of biologically active recombinant forms of human clusterin for 24 h post-OGD led to the abolishment of the ischemic tolerance in Clu-/- slices. This deleterious effect of clusterin was reverted by the concomitant administration of the NMDA receptor antagonist, d-2-amino-5-phosphonopentanoate. The present data indicate that in an in vitro model of ischemia characterized by the predominance of NMDA-mediated cell death, clusterin exerts a negative effect on the structural integrity and functionality of hippocampal neurons.


The plastidic 2-cysteine peroxiredoxin is a target for a thioredoxin involved in the protection of the photosynthetic apparatus against oxidative damage.
The chloroplastic drought-induced stress protein of 32 kD (CDSP32) is composed of two thioredoxin modules and is induced by environmental and oxidative stress conditions. We investigated whether the plastidic protein BAS1, which is related to eubacterial 2-Cys peroxiredoxin, is a target for CDSP32. Using a CDSP32 active-site mutant, we showed that the BAS1 and CDSP32 proteins form a mixed disulfide complex in vitro. Moreover, affinity chromatography indicated that BAS1 is a major target for CDSP32 in chloroplasts. CDSP32 was able to reduce BAS1 in vitro, and BAS1 displayed CDSP32-dependent peroxidase activity. The function of CDSP32 was investigated in transgenic potato lines without detectable levels of the protein as a result of cosuppression. Under conditions of photooxidative stress induced by incubation with either methyl viologen or t-butyl hydroperoxide or by exposure to low temperature under high light, plants lacking CDSP32 exhibited decreased maximal photosystem II photochemical efficiencies compared with the wild type and transgenic controls. In addition, plants without CDSP32 retained much less chlorophyll than controls under stress, indicating increased damage to photosynthetic membranes. We conclude that CDSP32 is a thioredoxin with a critical role in plastid defense against oxidative damage and that this role is related to its function as a physiological electron donor to the BAS1 peroxiredoxin.