A novel archaeal group in the phylum Crenarchaeota found unexpectedly in an eukaryotic survey in the Cariaco Basin.
Archaea have been found in many more diverse habitats than previously believed due in part to modern molecular approaches to discovering microbial diversity. We report here an unexpected expansion of the habitat diversity of the Archaea in the Cariaco Basin we found using a primer set designed for 18S eukaryotic rDNA sequence analysis. The results presented here expand the originally identified 9 archaeal clones reported in this environment using bacterial/archaeal primers to 152 archaeal clones: 67 (18 OTU) of these clones were found at a depth of 900 m of station A while 71 (9 OTU) of them were at a depth of between 300 approximately 335 m of station B&C depending upon which location the samples were taken. We used three phylogenetic analysis methods and detected 20 phylotypes belonging to a single previously unreported group distantly related to the Crenarchaeota. Also, we determined that the original nine sequences did not fall into any of the known phyla of the Archaea suggesting that they may represent a novel group within the Kingdom Archaea. Thus, from these two studies, we suggest that Archaea in the Cariaco Basin could be unique; however, further studies using archaeal-specific primers and the design of new primers as well as the systematic use of several different primer combinations may improve the chances of understanding the archeal diversity in the Cariaco Basin.


From first symptoms to diagnosis of amyotrophic lateral sclerosis: perspectives of an Irish informal caregiver cohort-a thematic analysis.
Informal caregivers play an integral part in the management of amyotrophic lateral sclerosis (ALS). The objective of this study was to explore the journey from first problem symptoms to diagnosis from the perspective of informal caregivers providing care to people with ALS. As part of a semistructured interview, information was collected on a range of caregiver demographic details, and from an open-ended question their experiences of the time of symptom onset to diagnosis. We carried out descriptive statistical analysis and thematic analysis of qualitative data. Home interviews with informal caregivers (n=74) of people with ALS attending the National ALS/Motor Neuron Disease Clinic at Beaumont Hospital, Dublin, Ireland. This was a largely female and spousal cohort of caregivers, living with the patient for whom they provided informal care. The majority of patients were men and were spinal onset. Caregivers described the time from first symptoms to diagnosis. Using a primarily inductive approach, the coding was data driven and the codes and themes derived from the content of these descriptions. Two main themes were identified (1) problem signs and symptoms (A) noticing and (B) reaction; (2) interaction with the health services. Exploring the perspectives of caregivers from first problem symptoms to diagnosis provides valuable insights into the development of the condition, impediments to its recognition, help-seeking behaviours and interactions with healthcare services. The journey from early symptoms to diagnosis is important for future decision-making, affects readiness for caregiving and could negatively impact on caregiver health and well-being. The early acknowledgement by healthcare professionals of stressors along the journey to diagnosis, and appreciation of their possible impact on caregivers is important. The separate needs of caregivers should be assessed on a regular basis.


Reconstitution of pure acetylcholine receptor in phospholipid vesicles and comparison with receptor-rich membranes by the use of a potentiometric dye.
Acetylcholine receptor, isolated in Triton X-100 on a cobra alpha-neurotoxin affinity column was incorporated into unilamellar phospholipid vesicles by a detergent depletion method using Amberlite XAD-2. Vesicles of an average diameter of 25 nm were formed, as verified by freeze-fracture electron microscopy and gel filtration. 85 to 95% of the alpha-bungarotoxin binding sites of the reconstituted acetylcholine receptor were oriented towards the outside of the vesicles. In the reconstituted receptor one molecule of residual Triton X-100 per 2.5 alpha-bungarotoxin binding sites on the receptor molecule could be assessed. The reconstituted protein was not accessible to papain digestion, whereas the pure acetylcholine receptor, solubilized by Triton X-100 was split into smaller polypeptides under the same condition. Reconstituted acetylcholine receptor and receptor-rich membranes did not exhibit the same behavior as measured by use of a potentiometric dye. This is interpreted as an irreversible alteration of at least 95% of the receptors purified in the presence of Triton X-100. Furthermore, it could be shown that the fluorescence intensity changes induced by carbamylcholine in receptor-rich membranes did not reflect ion fluxes, but conformational changes of the protein or a displacement of the dye from the protein.


Dectin-1 activation controls maturation of β-1,3-glucan-containing phagosomes.
Elimination of fungal pathogens by phagocytes requires phagosome maturation, a process that involves the recruitment and fusion of intracellular proteins. The role of Dectin-1, a β-1,3-glucan receptor, critical for fungal recognition and triggering of Th17 responses, to phagosomal maturation has not been defined. We show that GFP-Dectin-1 translocates to the fungal phagosome, but its signal decays after 2 h. Inhibition of acidification results in retention of GFP-Dectin-1 to phagosome membranes highlighting the requirement for an acidic pH. Following β-1,3-glucan recognition, GFP-Dectin-1 undergoes tyrosine phosphorylation by Src kinases with subsequent Syk activation. Our results demonstrate that Syk is activated independently of intraphagosomal pH. Inhibition of Src or Syk results in prolonged retention of GFP-Dectin-1 to the phagosome signifying a link between Syk and intraphagosomal pH. β-1,3-glucan phagosomes expressing a signaling incompetent Dectin-1 failed to mature as demonstrated by prolonged Dectin-1 retention, presence of Rab5B, failure to acquire LAMP-1 and inability to acidify. Phagosomes containing Candida albicans also require Dectin-1-dependent Syk activation for phagosomal maturation. Taken together, these results support a model where Dectin-1 not only controls internalization of β-1,3-glucan containing cargo and triggers proinflammatory cytokines, but also acts as a master regulator for subsequent phagolysosomal maturation through Syk activation.


Bilirubin helps to overcome the two-cell block in mouse oocyte cultures.
In vitro fertilization and culture of mouse oocytes, under normal atmospheric oxygen tension, subjects them to severe oxidative stress. Oocytes from some strains of mice lack the natural protective mechanism that guards them against this oxidative stress and fail to develop beyond the two-cell stage. We could overcome the toxic effects of oxygen metabolites by adding 0.2-0.4 mg/dl bilirubin in a lactate-pyruvate culture medium defined by Whitten (1971). Six- to 8-week-old ICR (Institute of Cancer Research) female mice were super ovulated by intra peritoneal injection of 5 IU PMSG (pregnant mare serum gonadotropin) followed by 10 IU hCG 48 h later. The oocytes were collected from the distended fallopian tubes and inseminated with 1-2 million sperm from 3-4-month-old ICR male mice. The eggs were scored at 24, 48, and 72 h after the hCG injection. With 0.4 mg/dl bilirubin supplement, by the end of 72 h, 82% of the eggs progressed from the two-cell stage to the four-cell stage. Routine inclusion of bilirubin can improve embryo development in vitro.


Historical overview of development in methods to estimate burden of disease due to congenital disorders.
Congenital disorders (often also called birth defects) are an important cause of mortality and disability. They encompass a wide range of disorders with differing severity that can affect any aspect of structure or function. Understanding their epidemiology is important in developing appropriate services both for their prevention and treatment. The need for epidemiological data on congenital disorders has been recognised for many decades. Here, we provide a historical overview of work that has led to the development of the Modell Global Database of Congenital Disorders (MGDb)-a tool that can be used to generate evidence-based country, regional and global estimates of the birth prevalence and outcomes of congenital disorders.


The promoter activity of human Mfn2 depends on Sp1 in vascular smooth muscle cells.
Mitofusin-2 (Mfn2) expression is dysregulated in vascular proliferative disorders and its overexpression attenuates the proliferation of vascular smooth muscle cells (VSMCs) and neointimal lesion development after balloon angioplasty. We sought to gain insight into the mechanisms that control Mfn2 expression in VSMCs. We cloned and characterized 2 kb of the 5'-flanking region of the human Mfn2 gene. Its TATA-less promoter contains a CpG island. In keeping with this, 5'-rapid amplification of cDNA ends revealed six transcriptional start sites (TSSs), of which TSS2 and TSS5 were the most frequently used. The strong CpG island was found to be non-methylated under conditions characterized by large differences in Mfn2 gene expression. The proximal Mfn2 promoter contains six putative Sp1 motifs. Sp1 binds to the Mfn2 promoter and its overexpression activates the Mfn2 promoter in VSMCs. Chemical inhibition of Sp1 reduced Mfn2 expression, and Sp1 silencing reduced transcriptional activity of the Mfn2 promoter. In keeping with this view, Sp1 and Mfn2 mRNA levels were down-regulated in the aorta early after an atherogenic diet in apolipoprotein E-knockout mice or in VSMCs cultured in the presence of low serum. Sp1 is a key factor in maintaining basal Mfn2 transcription in VSMCs. Given the anti-proliferative actions of Mfn2, Sp1-induced Mfn2 transcription may represent a mechanism for prevention of VSMC proliferation and neointimal lesion and development.


A high affinity site for sugar transport at the inner face of the human erythrocyte membrane?
A disagreement centering on a method of analysis as to the existence of a high affinity site for glucose transport at the inner face of the human red cell membrane is resolved by using direct fitting methods to confirm the original parameter estimates.


From rational design to serendipity: Discovery of novel thiosemicarbazones as potent trypanocidal compounds.
Chagas disease is a major public health problem caused by Trypanosoma cruzi, with an estimated 6-7 million people infected and 70 million at risk of infection. T. brucei gambiense and T. brucei rhodesiense are two subspecies of related parasites that cause human African trypanosomiasis, a neglected tropical disease with also millions of people at risk of infection. Pharmacotherapy for both diseases suffers from low efficacy, side effects, or drug resistance. Recently, we reported a noncovalent competitive inhibitor of cruzain (IC50 26 μM, Ki 3 μM) and TbrCatL (IC50 50 μM), two cysteine proteases considered promising drug targets for trypanosomiasis. Here, we describe the design and synthesis of derivatives of our lead compound. The new thiosemicarbazone derivatives showed potency in the nanomolar concentration range against the two enzymes, but they were later characterized as aggregators. Nevertheless, the thiosemicarbazone derivatives showed promising antiparasitic activities against T. b. brucei (EC50 13-49.7 μM) and T. cruzi (EC50 0.027-0.59 μM) under in vitro conditions. The most active thiosemicarbazone was 200-fold more potent than the current anti-chagasic drug, benznidazole, and showed a selectivity index of 370 versus myoblast cells. We have identified an excellent candidate for further optimization and in vivo studies.


Differential response of immortalized human amnion mesenchymal and epithelial cells against oxidative stress.
Cells are equipped with various antioxidant defense factors to antagonize insults from reactive oxygen species (ROS), thus the antioxidant capacity has been characterized by a variety of cellular responses during the pathophysiological processes. Amniotic cells have been extensively applied in clinical practice for burn treatment, corneal repair, and tissue regeneration. However, the antioxidative properties of amniotic cells have not yet been fully understood. Therefore, the current study was aimed to observe the response of amniotic cells against ROS stimuli, and to investigate the underlying molecular mechanisms. The immortalized human amniotic mesenchymal cells (iHAMs) and immortalized human amniotic epithelial cells (iHAEs) were used. The human skin fibroblast (HSF) was used as a control cell line. Changes in intracellular ROS generation, cell viability, and cellular morphology were investigated to reveal the response of amniotic cells against oxidative stresses induced by x-rays and hydrogen peroxide. In addition, expression of apoptosis-related proteins and response to antioxidative stress was also examined. The intracellular ROS level and cell apoptosis in iHAMs was remarkably increased. iHAEs showed relatively high resistance to ROS stimulation, which can be attributed to the high SOD2 expression and up-regulation of Nrf2, HO-1 after x-rays exposure. In contrast, iHAMs were found sensitive to oxidative damage. Expression of caspase-3, caspase-8 and BAX was increased, whereas down-regulation of Bcl-xL, Nrf2, HO-1, and TrxR-1. Taken together, findings have highlighted the characterization of response of amniotic derived epithelial and mesenchymal cells to oxidative stress. In physiological processes, iHAMs may play an important role to maintain the homeostasis of the pregnancy environment. However, under oxidative stimulations, iHAEs provides protection against oxidative damage in amnion tissue.


Maternal and foetal effects of methoxyflurane anaesthesia in the pregnant ewe.
Maternal and foetal cardiovascular, blood gas and acid-base changes were studied during 90 minutes of methoxyflurane anaesthesia. At 1.0 and 1.5 MAC anaesthesia, despite slight to moderate falls in maternal blood pressure, cardiac output and uterine blood flow, no serious foetal deterioration was seen. 2.0 MAC methoxyflurane was associated with marked falls in maternal blood pressure, cardiac output and uterine blood flow. Foetal hypoxaemia and a mixed respiratory and metabolic acidosis developed. Little foetal cardiovascular depression was seen with any level of anaesthesia. There was no direct effect of methoxyflurane on the uterine vasculature.


Cellular and molecular characterization of the impact of laboratory setup on bovine in vitro embryo production.
One of the main objectives related to performing comparative analysis of embryonic transcriptomes is to share information with other reproductive biologists or commercial service providers. Biological extracts influence performance of in vitro production systems and affect the reproducibility of results between production sites; these sources of variation could impede the potential for knowledge transfer. The objective of the present study was to assess the impact of the production site when sharing a common in vitro embryo production protocol. Biological extracts and semen were shared between production sites and thus removed as potential sources of variation. To remove the impact of blastocyst staging, all comparisons used expanded blastocysts. Although blastocyst yields and the number of Tunel positive cells per embryo differed between production sites, blastocysts were morphologically very similar in regards to cell number, their allocation to either the trophoblast or inner cell mass, or their gender ratio. These observations were also confirmed at the gene expression level, as indicated by highly similar transcript abundances. Only 36 genes out of the 16,121 expressed during bovine prehatching development were statistically differentially expressed, of which a large proportion were associated with the apoptotic process. These results highlighted the impact of laboratory set up, including personnel experience, when replicating an in vitro production system. Although inherent differences may arise, given the similarity of results between production sites, we concluded that embryo production protocols have the potential to be transferred and shared.


The luxury effect beyond cities: bats respond to socioeconomic variation across landscapes.
The luxury effect describes the positive relationship between affluence and organism diversity or activity in urban ecosystems. Driven by human activities, the luxury effect can potentially be found at a broader scale across different landscapes. Previously, the luxury effect relationship has been established within a city for two bat species, the red bat (Lasiurus borealis) and the evening bat (Nycticeius humeralis). We examined landscape-scale patterns of bat activity distribution-using empirical data for seven bat species for the luxury effect. We also identified bat-land cover associations for each species. Across North Carolina, USA, we used the mobile transect survey protocol of the North American Monitoring Program to record bat activity at 43 sites from 2015 to 2018. We collected land cover and income data at our transect locations to construct generalized linear mixed models to identify bat-land cover and bat-income relationships. We found that across landscapes, activity of the red bat and the evening bat was positively correlated to income independent of land cover, consistent with previous single-city results. We found a negative relationship between hoary bat (Lasiurus cinereus) activity and income. All seven species had specific land cover associations. Additionally, we found a positive interaction term between income and evergreen forest for the red bat and a positive interaction term between income and woody wetland for hoary bat. Our results demonstrated that the luxury effect is an ecological pattern that can be found at a broad spatial scale across different landscapes. We highlight the need for multi-scale ecology studies to identify the mechanism(s) underlying the luxury effect and that the luxury effect could cause inequity in how people receive the ecosystem services provided by bats.


An elemental diet controls inflammation in indomethacin-induced small bowel disease in rats: the role of low dietary fat and the elimination of dietary proteins.
Elemental diets (EDs) are effective in treating Crohn's disease. We hypothesize that low dietary fat and amino acids used as the sole nitrogen source are the major contributors for the success of EDs. We examined the influences of the addition of dietary fat and protein to an ED using an indomethacin-induced inflammation model in rat small intestine. In the ED-fed rats, the intestinal damage score was decreased compared with that in the standard chow group with decreasing intestinal permeability. By supplementing an ED with soybean oil (SO), intestinal permeability was increased to a level similar to that of the standard chow group. For this group, the intestinal damage score also increased compared with that of the ED group but did not reach the levels observed in the standard chow group. The addition of dietary proteins (using heat-denatured pancreatin) resulted in intestinal damage scores that were significantly higher than those of the ED+SO-fed group. The dietary protein increased the intestinal damage score. These results suggest that EDs control inflammation by decreasing intestinal permeability and the elimination of dietary proteins.


Psychosocial Intervention Use in Long-Stay Dementia Care: A Classic Grounded Theory.
The objective of this study was to develop a substantive grounded theory of staff psychosocial intervention use with residents with dementia in long-stay care. "Becoming a person again" emerged as the core category accounting for staffs' psychosocial intervention use within long-stay care. Interview data were collected from participants in nine Irish long-stay settings: 14 residents with dementia, 19 staff nurses, one clinical facilitator, seven nurse managers, 21 nursing assistants, and five relatives. Constant comparative method guided the data collection and analysis. The researcher's theoretical memos, based on unstructured observation, and applicable extant literature were also included as data. By identifying the mutuality of the participants' experiences, this classic grounded theory explains staff motivation toward psychosocial intervention use within long-stay care. It also explains how institutional factors interact with those personal factors that incline individuals toward psychosocial intervention use.


Modifiable Lifestyle Factors and Triple-negative Breast Cancer Survival: A Population-based Prospective Study.
Very little is known about the effect of modifiable lifestyle factors on outcomes of triple-negative breast cancer. We examined this association in a population-based prospective cohort study of patients with triple-negative breast cancer. A total of 518 women with confirmed triple-negative breast cancer, recruited by the Shanghai Breast Cancer Survival Study, completed 6-, 18-, 36-, and 60-month postdiagnosis surveys. We applied Cox proportional hazard models to evaluate the associations. The mean age at diagnosis was 53.4 (standard deviation = 10.6) years old. After a median follow-up of 9.1 years (range: 0.6-11.8), 128 deaths and 112 recurrences were documented. Exercise during the first 60 months postdiagnosis was inversely associated with total mortality and recurrence/disease-specific mortality with adjusted hazard ratios (HRs) of 0.67 (95% confidence interval [CI] = 0.46, 0.96) and 0.58 (95% CI = 0.39, 0.86), respectively. Women with higher exercise-metabolic equivalent scores (≥7.6 metabolic equivalent-hours/week) and longer duration of exercise (≥2.5 hours/week) had lower risk of total and recurrence/disease-specific mortality than did nonexercisers. Compared with nontea drinkers, survival was better among women who were regular tea drinkers during the first 60 months for all cause (HR = 0.57, 95% CI = 0.34, 0.93) and recurrence/disease-specific mortality (HR = 0.54, 95% CI = 0.31, 0.96). There was no dose-response pattern for tea consumption. No interactions were observed for body mass index, menopausal status, and comorbidity. These findings show that postdiagnosis exercise and tea intake were associated with improved survival among women with triple-negative breast cancer.


Development and validation of the Ascertain Dementia 8 (AD8) Thai version.
The Ascertain Dementia 8 (AD8) is a brief informant-based questionnaire which reliably distinguishes patients with neurocognitive disorder (NCD) and normal cognition. Our objective was to translate the AD8 into Thai and test its validity as a potentially useful measure to detect patients with the mild stage of major NCD due to Alzheimer disease (major NCD-AD). Evaluations of 144 informant-patient dyads were made. Participants were patients who attended the memory clinic at Ramathibodi Hospital and non-patient volunteers. The AD8-Thai version was administered separately from doctor's interviews and neuropsychological assessments. Diagnostic workups included a complete medical history, physical and neurological examinations, neuropsychological testing, the Clinical Dementia Rating Scale-Sum of Boxes (CDR-SOB), Montreal Cognitive Assessment (MoCA), blood tests and brain imaging, preferably with magnetic resonance imaging. All researchers were blinded to the AD8 results. An AD8 score ≥3 had a sensitivity of 95.5%, a specificity of 89.5%, a positive predictive value of 84%, and a negative predictive value of 97.1% for screening major NCD-AD and those with normal cognition. A subgroup analysis with participants aged ≥65 years and with ≥12 years of education revealed comparable ability to the whole group. AD8 scores had a moderate negative relationship with MoCA scores (r = -0.470) and a strong positive relationship with CDR-SOB scores (r = 0.547). The performance of AD8 scores in differentiating mild NCD from normal cognition was not as good as for major NCD-AD. AD8-Thai version is an acceptable screening tool for major NCD-AD. For patients aged 65 years or older, with at least 12 years of education, and with cognitive complaints in memory clinic, an AD8-Thai score of 3 or more would be sufficient to work on major NCD diagnosis. The performance of the AD8-Thai in mild NCD needs further investigation.


Estimation of age-specific rates of acquisition and loss of Wuchereria bancrofti infection.
This study uses a reversible catalytic model to estimate the age-specific rates of gain and loss of Wuchereria bancrofti infection from data collected during a control programme in Pondicherry, South India. The data describe the infection status in 1981 and 1986 of two cohorts of individuals, one living in an area where vector reduction had been achieved, and the other in a comparable endemic area. The rate of loss of infection in the absence of reinfection is estimated for the cohort in the control area, and the rate of gain of infection by the cohort in the endemic area estimated by substitution in the model. The mean expected life span of patent infection is estimated to be 5.4 years. The instantaneous rate of loss of infection is independent of age, while the rate of gain of infection exhibits a convex age-profile, peaking in the 16-20 year age-class. The reduced rate of gain in adults is largely attributable to the increasing proportion of potentially resistant individuals with clinical disease. The results suggest that the age-distribution of bancroftian filariasis is primarily determined by age-dependency in the rate of acquisition of infection.


Measuring the shape of the biodiversity-disease relationship across systems reveals new findings and key gaps.
Diverse host communities commonly inhibit the spread of parasites at small scales. However, the generality of this effect remains controversial. Here, we present the analysis of 205 biodiversity-disease relationships on 67 parasite species to test whether biodiversity-disease relationships are generally nonlinear, moderated by spatial scale, and sensitive to underrepresentation in the literature. Our analysis of the published literature reveals that biodiversity-disease relationships are generally hump-shaped (i.e., nonlinear) and biodiversity generally inhibits disease at local scales, but this effect weakens as spatial scale increases. Spatial scale is, however, related to study design and parasite type, highlighting the need for additional multiscale research. Few studies are unrepresentative of communities at low diversity, but missing data at low diversity from field studies could result in underreporting of amplification effects. Experiments appear to underrepresent high-diversity communities, which could result in underreporting of dilution effects. Despite context dependence, biodiversity loss at local scales appears to increase disease, suggesting that at local scales, biodiversity loss could negatively impact human and wildlife populations.


Lasp-1, a novel type of actin-binding protein accumulating in cell membrane extensions.
The Lasp-1 gene, which has been localized to the q12-q21 region of human chromosome 17, is amplified and overexpressed in human breast cancers. In addition to the previously reported LIM and SH3 domains of Lasp-1, we report here the identification of an actin-binding domain in the core of the protein. This domain is functional as we demonstrate that Lasp-1 binds actin in vivo and in vitro. In addition, confocal analysis of the Lasp-1 subcellular distribution shows that the protein is colocalized with actin at peripheral cell extensions in individual epithelial cancer cells and in transformed fibroblastic cells. Moreover, Lasp-1 is tyrosine phosphorylated in fibroblast cell lines transformed by a constitutively active form of c-Src (c-SrcY527F). Altogether, our results show that Lasp-1 defines a new type of actin-binding protein and suggest that the protein may play a role in a signaling pathway involved in the organization of the cytoskeleton.


High-risk and selected benign breast lesions diagnosed on core needle biopsy: Evidence for and against immediate surgical excision.
The vast majority of image-detected breast abnormalities are diagnosed by percutaneous core needle biopsy (CNB) in contemporary practice. For frankly malignant lesions diagnosed by CNB, the standard practice of excision and multimodality therapy have been well-defined. However, for high-risk and selected benign lesions diagnosed by CNB, there is less consensus on optimal patient management and the need for immediate surgical excision. Here we outline the arguments for and against the practice of routine surgical excision of commonly encountered high-risk and selected benign breast lesions diagnosed by CNB. The entities reviewed include atypical ductal hyperplasia, atypical lobular hyperplasia, lobular carcinoma in situ, intraductal papillomas, and radial scars. The data in the peer-reviewed literature confirm the benefits of a patient-centered, multidisciplinary approach that moves away from the reflexive "yes" or "no" for routine excision for a given pathologic diagnosis.


Surgical treatment of pNET - Experience of a "high-volume" center.
Neuroendocrine tumors of the pancreas (pNETs) are a rare disease. Grading according to the Ki67-index is the most validated risk factor. Nevertheless, controversies exist concerning other prognostic factors. The aim of this study was to evaluate published risk factors. Patients with pancreatic NETs who underwent surgery at our department from 2000 to 2014 were analyzed. The patient and tumor characteristics were evaluated. Kaplan-Meier analyses, univariate calculations as well as multivariate analyses were performed. In total, 98 patients underwent surgery due to a pNET. The final study population consisted of 88 patients. Univariate analysis demonstrated that overall survival is influenced by tumor grading, local resection margin and presence of distant metastases. However, in the multivariate analysis, only grading and the resection margin had prognostic significance. The size of the primary tumor directly correlated with the probability of metastases. Multivisceral operations had no influence on morbidity or mortality. Resection of pNETs is the only curative treatment and is safe. Since the incidence of pNETs is low, treatment should be performed at a high-volume center.


A sarcoma of myofibroblasts: an ultrastructural study.
A low-grade spindle cell sarcoma was shown by electron microscopy to be composed of myofibroblasts. It is suggested that the occurrence of myofibroblasts in spindle cell sarcomas might be of prognostic significance and that a systematic study to elucidate this point would be a valuable exercise.


The neostriatum and response selection in overt sentence production: an fMRI study.
A number of premotor and prefrontal brain areas have been recently shown to play a significant role in response selection in overt sentence production. These areas are anatomically connected to the basal ganglia, a set of subcortical structures that has been traditionally involved in response selection across behavioral domains. The putamen and the caudate, the two major inputs to the basal ganglia, have been shown to undertake motor- as well as non-motor-related selection operations in language processing. Here we investigate the role of these basal ganglia structures in sentence repetition and generation in healthy adults. Although sentence generation is known to activate prefrontal and premotor cortical areas that reciprocally connect with these two neostriatal structures, their specific contributions are not known. We present evidence suggesting that that the putamen undertakes articulation-related aspects across tasks, while the caudate selectively supports selection processes in sentence generation.


Pilot study of the completeness of notification of adult tuberculosis in Athens, Greece.
Sotiria Chest Diseases Hospital (SCDH), a referral hospital in Athens, Greece, 2012. To assess the completeness of the mandatory notification system for tuberculosis (TB) at the SCDH, and compare the observed and estimated annual incidence rates. Record linkage and the capture-recapture method were applied. Data sources were the registers from the national mandatory notification register (Hellenic Centre for Disease Control and Prevention [HCDCP]), the National Reference Laboratory for Mycobacteria (NRLM) and SCDH records. The log-linear model with the lowest Akaike information criterion was selected as the most valid statistical model. The observed and estimated TB under-reporting rates at the national level were respectively 55% (95%CI 49-60) and 75% (95%CI 71-78). The observed completeness of the HCDCP, NRLM and SCDH registers were respectively 45% (95%CI 40-51), 66% (95%CI 61-71) and 36.5% (95%CI 31-42). The estimated TB incidence rate was 15 cases per 100 000 (range 13-19/100 000), compared to the 4.9/100 000 rate officially notified. Adult TB incidence has been largely underestimated, and the TB burden is likely to be much higher than officially notified in our setting. A thorough review of the notification system should be carried out. The implementation of a network-based notification system and retraining of all relevant personnel is advised.


Diffuse gliomas exhibit whole brain impaired cerebrovascular reactivity.
Cerebral diffuse gliomas exhibit perilesional impaired cerebrovascular reactivity (CVR), yet the degree of impairment as well as its full spatial extent in the brain remains unknown. With quantitative fMRI, we studied twelve subjects with untreated brain diffuse glioma and twelve healthy controls to assess CVR impairment and determine its distribution throughout the brain. In a prospective case-control study, quantitative CVR measurements were derived from BOLD fMRI volumes during standardized iso-oxic changes in carbon dioxide. Whole brain CVR was assessed with additional detailed analyses using specific tumor and tissue masks and compared to datasets of healthy controls. Whole brain CVR was significantly impaired compared to healthy controls (0.11±0.10 versus 0.28±0.8, p<0.01). All diffuse glioma patients exhibited even more severely impaired intralesional CVR (mean 0.01±0.06). Increasing tumor volume significantly correlated with severity of intralesional CVR impairment (p<0.05, R2=0.38), and whole brain CVR impairment (p<0.05, R2=0.55). Patients with brain diffuse glioma exhibit intralesional and whole brain impaired CVR with severity correlating to tumor volume. Quantitative fMRI may be entertained to study antitumor therapy efficacy by tracking CVR changes and may have a complementary role to better interpret BOLD associated neurovascular uncoupling.


Effect of omeprazole treatment on diazepam plasma levels in slow versus normal rapid metabolizers of omeprazole.
The effect of omeprazole treatment on diazepam plasma levels was studied in four slow and six rapid metabolizers of omeprazole. Single intravenous doses of diazepam (0.1 mg/kg) were administered after 1 week of oral treatment with omeprazole (20 mg) and placebo. This was a double-blind crossover study with randomized placebo and omeprazole treatments. Blood was collected up to 120 hours after diazepam dosing (still during one-daily omeprazole and placebo administration) for measurement of diazepam and its major metabolite desmethyldiazepam. The slow metabolizers of omeprazole also metabolized diazepam slowly, exhibiting only half the diazepam plasma clearance of the others. The mean clearance of diazepam was decreased 26% after omeprazole in the rapid metabolizers, whereas the slow group showed no apparent interaction. The mean plasma concentrations of desmethyldiazepam showed a more rapid formation in the rapid compared with the slow metabolizers, which is a logical consequence of the rate of diazepam metabolism.


The role of HRCT in Tropical Pulmonary Eosinophilia.
To determine the pattern of pulmonary involvement in clinically confirmed patients of tropical pulmonary eosinophilia (TPE). An observational study on 13 patients with clinically confirmed TPE was performed to determine the CT scan appearances. The predominant CT scan finding is the presence widespread ill-defined bronchocentric nodules, which need to be differentiated from other conditions. The pattern of lung involvement on a CT scan can give a clue to the diagnosis of TPE in the correct clinical context. Radiologists in tropical countries should have a high index of suspicion for this diagnosis when reading scans showing widespread ill-defined bronchocentric nodules.


Hydrogen peroxide-mediated induction of SOD1 gene transcription is independent from Nrf2 in a cellular model of neurodegeneration.
It is still unclear whether oxidative stress (OS) is a disease consequence or is directly involved in the etiology of neurodegenerative disorders (NDs) onset and/or progression; however, many of these conditions are associated with increased levels of oxidation markers and damaged cell components. Previously we demonstrated the accumulation of reactive oxygen species (ROS) and increased SOD1 gene expression in H2O2-treated SH-SY5Y cells, recapitulating pathological features of Amyotrophic Lateral Sclerosis (ALS). Since we observed a post-transcriptional regulation of SOD1 gene in this cellular model, we investigated the transcriptional regulation of SOD1 mRNA under oxidative stress (OS). In response to H2O2 treatment, PolII increased its association to SOD1 promoter. Electrophoretic mobility shift assays and mass spectrometry analyses on SOD1 promoter highlighted the formation of a transcriptional complex bound to the ARE sequences. Western Blotting experiments showed that in our in vitro model, H2O2 exposure increases Nrf2 expression in the nuclear fraction while immunoprecipitation confirmed its phosphorylation and release from Keap1 inhibition. However, H2O2 treatment did not modify Nrf2 binding on SOD1 promoter, which seems to be regulated by different transcription factors (TFs). Although our data suggest that SOD1 is transcriptionally regulated in response to OS, Nrf2 does not appear to associate with SOD1 promoter in this cellular model of neurodegeneration. Our results open new perspectives in the comprehension of two key antioxidant pathways involved in neurodegenerative disorders.


Mental rotation within linguistic and non-linguistic domains in users of American sign language.
American sign language (ASL) uses space itself to encode spatial information. Spatial scenes are most often described from the perspective of the person signing (the 'narrator'), such that the viewer must perform what amounts to a 180 degrees mental rotation to correctly comprehend the description. But scenes can also be described, non-canonically, from the viewer's perspective, in which case no rotation is required. Is mental rotation during sign language processing difficult for ASL signers? Are there differences between linguistic and non-linguistic mental rotation? Experiment 1 required subjects to decide whether a signed description matched a room presented on videotape. Deaf ASL signers were more accurate when viewing scenes described from the narrator's perspective (even though rotation is required) than from the viewer's perspective (no rotation required). In Experiment 2, deaf signers and hearing non-signers viewed videotapes of objects appearing briefly and sequentially on a board marked with an entrance. This board either matched an identical board in front of the subject or was rotated 180 degrees. Subjects were asked to place objects on their board in the orientation and location shown on the video, making the appropriate rotation when required. All subjects were significantly less accurate when rotation was required, but ASL signers performed significantly better than hearing non-signers under rotation. ASL signers were also more accurate in remembering object orientation. Signers then viewed a video in which the same scenes were signed from the two perspectives (i.e. rotation required or no rotation required). In contrast to their performance with real objects, signers did not show the typical mental rotation effect. Males outperformed females on the rotation task with objects, but the superiority disappeared in the linguistic condition. We discuss the nature of the ASL mental rotation transformation, and we conclude that habitual use of ASL can enhance non-linguistic cognitive processes thus providing evidence for (a form of) the linguistic relativity hypothesis.


Massive Osteolysis in a Modern Total Knee Prosthesis.
An 82-year-old woman underwent right total knee replacement with a sequentially irradiated and annealed highly cross-linked polyethylene insert. At 9 years, she was found to have a massive femoral osteolysis with an impending fracture. This case demonstrates a rare occurrence of massive femoral osteolysis, requiring revision surgery, with a sequentially irradiated and annealed highly cross-linked polyethylene.


Secondary lysosomes as an integral part of the cytoskeleton: a morphological study in rat Kupffer cells.
Rat Kupffer cells contain the three major cytoskeletal components: microfilaments (MF), microtubules (MT), and intermediate filaments (IF) of the vimentin type. Previous cytomagnetometric data obtained from alveolar macrophages and rat Kupffer cells in culture provided evidence that actin filaments contribute to the movements of lysosomes. The lysosomal transport in living cells was affected, when the MFs were selectively disturbed, whereas the depolymerization of the MTs had no effect on the lysosomal movement measured by cytomagnetometric means. Immunofluorescence and ultrastructural studies of isolated and cultured rat Kupffer cells, presented in this paper, will investigate the relationship between lysosomes and the cytoskeleton. The principal filamentous structure in the peripheral cytoplasm of Kupffer cells in a dense meshwork of actin filaments. The dimension of the meshes combined with the dimensions of lysosomes implies the necessity of either (i) disintegration of the actin filament cross-links, (ii) depolarymerization and redistribution of MF's, or (iii) a displacement of actin filaments by the lysosomes during the organelle transport. The presence of microtubules in cytoplasmic protrusions and their track from the periphery to the perinuclear region during interphase might play a role in the transport mechanism of lysosomes, the more so because microtubules could often be demonstrated in closest association with lysosomes even in the first phase of endocytosis. The distribution pattern of vimentin, found as a dense interconnected framework surrounding the lysosomes like a basket, could play a role in positioning the organelles. The dynamic functions of MF's and MT's and their multifunctionality led to an adaptive and flexible organization of these filaments which may both be involved in lysosomal motion.(ABSTRACT TRUNCATED AT 250 WORDS)


Extending the coordination chemistry of 1,3,5-triaza-7-phosphaadamantane (PTA) to cobalt centers: first examples of co-PTA complexes and of a metal complex with the PTA oxide ligand.
Water-soluble Co (III) and Co (II) complexes with P- or N-coordinated PTA or PTA oxide ligands, respectively, have been prepared and fully characterized, constituting the first examples of cobalt compounds bearing PTA or any ligand with a cage-like PTA core, the latter complex providing also the first PTA oxide coordination to a metal center.


[The pyrogenal test in assessing body immune reactivity in acute diseases of the respiratory organs].
The authors studied the reaction of hypersensitivity of the delayed type to administration of pyrogenal in 94 patients with different acute diseases of the respiratory organs. Different changes of the pyrogenal test were accompanied by changes of leucocytosis, ESR, lymphocyte count, A, G, M immunoglobulins. An inverse relationship was found between the pyrogenal test and number of A and M immunoglobulins. The pyrogenal test may be used in outpatient conditions for preliminary evaluation of cellular immunity.


The implications of non-invasive prenatal testing failures: a review of an under-discussed phenomenon.
Non-invasive prenatal testing (NIPT) using cell-free DNA in maternal blood is a relatively new screening modality for the common trisomies of chromosomes 21, 18 and 13 and sex chromosome aneuploidies. For some patients, however, results are not reported because of laboratory technical issues such as low fetal fraction and sequencing failures. In this review, the clinical implications of NIPT test failures are discussed. A Medline search was performed for all studies on NIPT that include >1000 samples. The failure rates were assessed by technology. Methods based on massive parallel sequencing have been found to have the lowest failure rate (1.58%), while tests based on single-nucleotide polymorphism analysis have the highest failure rate (6.39%). Recent publications suggest that patients who receive a 'no call' result are at increased risk of aneuploidy. Some professional societies have therefore recommended that these patients undergo genetic counseling and be offered invasive diagnostic testing. NIPT technology that has a high failure rate may increase the false positive rates, decrease the positive predictive value, and increase the procedure-related pregnancy loss. © 2016 John Wiley & Sons, Ltd.


Fetal movements in hypertensive pregnancies.
The maternally perceived fetal movements counts of 273 hypertensive patients were compared to normal controls and their degree of fetal activity was related to perinatal outcome. Pregnancy induced hypertension was associated with fetal activity which was significantly lower than controls at the beginning of the third trimester and significantly higher at term. In cases of chronic hypertension patients, 8 had cessation or marked reduction of fetal movements to less than 4 in 12 hours, and all had poor fetal outcome. The remaining 265 hypertensive patients had a 92% incidence of good fetal outcome. Markedly reduced fetal movements in hypertensive patients are highly suggestive of fetal distress and following verification by additional tests the appropriate clinical measures should be undertaken.


Mechanisms of detachment in fibrillar adhesive systems.
Several creatures can climb on smooth surfaces with the help of hairy adhesive pads on their legs. A rapid change from strong attachment to effortless detachment of the leg is enabled by the asymmetric geometry of the tarsal hairs. In this study, we propose mechanisms by which the hairy pad can be easily detached, even when the hairs possess no asymmetry. Here, we examine the possible function of the tibia-tarsus leg joint and the claws. Based on a spring-based model, we consider three modes of detachment: vertically pulling the pad while maintaining either a (1) fixed or a (2) free joint, or by (3) flexing the pad about the claw. We show that in all cases, the adhesion force can be significantly reduced due to elastic forces when the hairs deform non-uniformly across the array. Our proposed model illustrates the design advantage of such fibrillar adhesive systems, that not only provide strong adhesion, but also allow easy detachment, making them suitable as organs for fast locomotion and reliable hold. The presented approaches can be potentially used to switch the adhesion state in bio-inspired fibrillar adhesives, by incorporating artificial joints and claws into their design, without the need of asymmetric or stimuli-responsive fibrillar structures.


The Neuroprotective Effect of Mesna on Cisplatin-Induced Neurotoxicity: Behavioral, Electrophysiological, and Molecular Studies.
Peripheral neuropathy and cognitive impairments following cisplatin administration may interfere with the clinical usage of the drug. Mesna is a chemoprotective agent with anti-inflammatory and anti-oxidant effects. Our study aimed to investigate the protective effects of mesna against cisplatin-induced neurotoxicity. Neurotoxicity was induced by the administration of 2.5 mg/kg cisplatin twice a week for four consecutive weeks in male Wistar rats. The neuroprotective effect of mesna (150 mg/kg/day) was evaluated through behavioral, electrophysiological, and molecular studies. Cisplatin treatment caused passive avoidance memory impairment, increased anxiety-like behaviors, altered thermal sensitivity, and decreased muscle strength in a grip strength test. Our electrophysiological studies indicated that administration of cisplatin induced peripheral sensory neuropathy and decreased the amplitudes of the compound action potential of sensory nerves. Cisplatin administration increased MDA and 4-HNE levels and decreased anti-oxidant (SOD and GPx) enzymes. Proinflammatory cytokines (IL-1β and TNF-α) and metalloproteinase-2 and 9 (MMP-2/9) were increased by cisplatin treatment. Morphological alterations were observed in the dorsal root ganglion (DRG) of cisplatin-treated rats. Cognitive impairments, anxiety, muscle strength, and thermal sensitivity changes induced by cisplatin were improved with mesna treatment. The reduced conduction velocity in sensory nerves was recovered in the cisplatin + mesna group. Mesna partially alleviated redox imbalance, reduced the proinflammatory cytokines, and MMP-2/9 levels. Mesna administration also relieved the morphological changes in DRG of cisplatin-treated rats. In conclusion, our results revealed that mesna can alleviate cisplatin-induced central and peripheral nervous system toxicity. These results support the concept that chemotherapy-induced neuropathy can be partially inhibited via mesna.


Analysis of the Relationship between Antioxidant Enzyme Gene Polymorphisms and Their Activity in Post-Traumatic Gonarthrosis.
Analysis of polymorphisms of genes encoding antioxidant enzymes SOD1 (G7958A), SOD2 (T58C), CAT (C-262T), and GSTP1 (Ile105Val) in 93 patients with post-traumatic gonarthrosis showed that GSTP1 Ile105Val polymorphism is often associated with heterozygous mutation in catalase gene CAT C-262T. In gonarthrosis, catalase activity in peripheral blood mononuclear cells in patients with CT genotype of the C-262T locus of CAT gene more than 2-fold surpassed that in CC genotype and more than 50% surpassed the normal. Changes in the balance of activity of antioxidant enzymes can affect viability of mononuclear cells.


Importance of screening for synchronous malignant neoplasms in patients with hepatocellular carcinoma: impact of FDG PET/CT.
Synchronous neoplasms (SNs) are occasionally found in hepatocellular carcinoma (HCC). We examined such cases and the efficacy of 18-fluoro-2-deoxyglucose positron-emission tomography computed tomography (PET/CT), retrospectively. We investigated 687 naïve HCC, who were admitted to our hospitals, encountered from October 2006 to December 2010 and evaluated the clinical backgrounds. All study protocols, was approved by our Institutional Ethics Committee. The usefulness of detecting SNs by PET/CT was evaluated in 234 patients who underwent PET/CT (PET group) and in 453 (non-PET group) examined in the same period. We noted the presence of SNs, defined as primary extrahepatic malignant neoplasms within 1 year of diagnosis of HCC. SNs were observed in 48 of 687 patients (54 tumours, 7.0%). SNs were detected by PET/CT in 18, which was 7.7% of PET group. The detection rate for SNs, were increased to 11.1% (26/234) in PET group by using together with upper gastrointestinal endoscopy and routine enhanced CT for HCC, which was greater than that of non-PET group (22/453, 4.9%) (P < 0.001). SNs were pointed out more frequently in PET group than non-PET group (11.1% vs. 4.9%). FDG PET/CT can enhance the detection ability for SNs in naïve HCC.


The RNF220 domain nuclear factor Teyrha-Meyrha (Tey) regulates the migration and differentiation of specific visceral and somatic muscles in Drosophila.
Development of the visceral musculature of the Drosophila midgut encompasses a closely coordinated sequence of migration events of cells from the trunk and caudal visceral mesoderm that underlies the formation of the stereotypic orthogonal pattern of circular and longitudinal midgut muscles. Our study focuses on the last step of migration and morphogenesis of longitudinal visceral muscle precursors and shows that these multinucleated precursors utilize dynamic filopodial extensions to migrate in dorsal and ventral directions over the forming midgut tube. The establishment of maximal dorsoventral distances from one another, and anteroposterior alignments, lead to the equidistant coverage of the midgut with longitudinal muscle fibers. We identify Teyrha-Meyhra (Tey), a tissue-specific nuclear factor related to the RNF220 domain protein family, as a crucial regulator of this process of muscle migration and morphogenesis that is further required for proper differentiation of longitudinal visceral muscles. In addition, Tey is expressed in a single somatic muscle founder cell in each hemisegment, regulates the migration of this founder cell, and is required for proper pathfinding of its developing myotube to specific myotendinous attachment sites.


Nucleic acid separations using superficially porous silica particles.
Ion pair reverse-phase liquid chromatography has been widely employed for nucleic acid separations. A wide range of alternative stationary phases have been utilised in conjunction with ion pair reverse-phase chromatography, including totally porous particles, non-porous particles, macroporous particles and monolithic stationary phases. In this study we have utilised superficially porous silica particles in conjunction with ion pair reverse-phase liquid chromatography for the analysis of nucleic acids. We have investigated a range of different pore-sizes and phases for the analysis of a diverse range of nucleic acids including oligonucleotides, oligoribonucleotides, phosphorothioate oligonucleotides and high molecular weight dsDNA and RNA. The pore size of the superficially porous silica particles was shown to significantly affect the resolution of the nucleic acids. Optimum separations of small oligonucleotides such as those generated in RNase mapping experiments were obtained with 80Å pore sizes and can readily be interfaced with mass spectrometry analysis. Improved resolution of larger oligonucleotides (>19mers) was observed with pore sizes of 150Å. The optimum resolution for larger dsDNA/RNA molecules was achieved using superficially porous silica particles with pore sizes of 400Å. Furthermore, we have utilised 150Å pore size solid-core particles to separate typical impurities of a fully phosphorothioated oligonucleotide, which are often generated in the synthesis of this important class of therapeutic oligonucleotide.


Secondary metabolism: regulation by phosphate and trace elements.
Secondary metabolism and cellular differentiation occur within a range of concentrations of phosphate and, in specific taxonomic groups, of zinc, manganese, and/or iron that is much narrower than that permittee for primary metabolism. Possible molecular sites of action of the four elements are reviewed.


Using insurance data to quantify the multidimensional impacts of warming temperatures on yield risk.
Previous research predicts significant negative yield impacts from warming temperatures, but estimating the effects on yield risk and disentangling the relative causes of these losses remains challenging. Here we present new evidence on these issues by leveraging a unique publicly available dataset consisting of roughly 30,000 county-by-year observations on insurance-based measures of yield risk from 1989-2014 for U.S. corn and soybeans. Our results suggest that yield risk will increase in response to warmer temperatures, with a 1 °C increase associated with yield risk increases of approximately 32% and 11% for corn and soybeans, respectively. Using cause of loss information, we also find that additional losses under warming temperatures primarily result from additional reported occurrences of drought, with reported losses due to heat stress playing a smaller role. An implication of our findings is that the cost of purchasing crop insurance will increase for producers as a result of warming temperatures.


Embryonic steroid synthesis and luteal oxytocin production: controlling mechanisms for the maternal recognition of pregnancy.
In both pigs and sheep in early pregnancy the function of the corpora lutea is maintained beyond the normal time of luteal regression. The mechanism by which this is brought about appears in both species to involve reduced uterine secretion of the uterine luteolysin, prostaglandin F2 alpha, as a result of the secretion of an embryonic antiluteolysin. In the pig this antiluteolysin is an oestrogen, and in the sheep, the protein trophoblastin. Loss of luteal oxytocin also occurs in sheep early in pregnancy, and this may contribute to reduced luteolytic drive during the period between 25 and 50 days of gestation when the embryonic antiluteolysin is absent. Concentrations of oxytocin in the corpora lutea appear to be controlled by a factor of uterine origin, since luteal oxytocin is also depleted after hysterectomy.


A novel diagnostic method for human immunodeficiency virus type-1 in plasma by near-infrared spectroscopy.
Presently, the diagnosis of virus infections is based mainly on serological assays. Although polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) have been increasingly used for the diagnosis of such viral infections, the risk of transfusion-transmitted blood-borne viruses remains. Furthermore, PCR and ELISA are expensive and time-consuming, and sometimes cause falsepositive or false-negative results. Therefore, a rapid, accurate and cost-effective diagnostic procedure is needed. We subjected plasma from individuals infected with human immunodeficiency virus type-1 (HIV-1), the causative agent of acquired immune deficiency syndrome (AIDS), as well as plasma from uninfected individuals as a control to near-infrared (NIR) spectroscopy, which may provide a rapid diagnostic method for HIV-1 infection without using any reagent. NIR spectra in the 600-1,000 nm region for plasma from pre-serologically HIV-1-infected individuals and healthy donors were subjected to partial least squares (PLS) regression analysis and leave-out cross-validation to develop a multivariate model to estimate the concentration of HIV-1. Simultaneously, the same plasma samples were examined for HIV-1 p24 by ELISA. The results obtained by the NIR spectroscopy model for HIV-1 yielded a good correlation with those obtained by the reference method (HIV-1 p24 ELISA). These results suggest that NIR spectroscopy using plasma could provide a rapid, accurate, cost-effective tool for large-scale diagnosis of HIV-1 infection.


Inhibition of cyclic AMP-dependent protein kinase by the polar head group of an insulin-sensitive glycophospholipid.
A glycophospholipid has been purified from rat liver membranes and shown to copurify with an insulin-sensitive glycophospholipid isolated from H35 hepatoma cells. The polar head group of this glycophospholipid is a phospho-oligosaccharide generated by treatment with phosphatidylinositol-specific phospholipase C from Staphylococcus aureus. It has been proposed that this phospho-oligosaccharide, which is also generated in response to insulin, may play a role in insulin action. Incubation of the catalytic subunit of cyclic AMP-dependent protein kinase with this phospho-oligosaccharide inhibited the activity of the kinase to phosphorylate histone IIA, a purified preparation of phospholipid methyltransferase and kemptide, a phosphate-accepting peptide. Inhibition of kinase activity was dose-dependent and 50% inhibition of histone phosphorylation was demonstrated with a concentration of phospho-oligosaccharide of around 2 microM. This effect was demonstrated in the presence of ATP at concentrations up to 1 mM, indicating that the phospho-oligosaccharide acts at physiological concentrations of ATP and that it does not compete with this nucleotide for the same binding site in the kinase. Inhibition by the phospho-oligosaccharide of kinase activity could be reversed by dilution or dialysis and was not reproduced by up to 50 microM myo-inositol, glucosamine, galactose, myo-inositol 1-phosphate, glucosamine 1-phosphate, galactose 1-phosphate or phosphorylcholine. The inhibitory activity was resistant to mild acid treatment but was labile to treatment with alkali, exposure to nitrous acid or incubation with sodium periodate. The phospho-oligosaccharide had no effect on the phosphorylation of lysine-rich histone by rat brain protein kinase C and on the binding of cyclic AMP to a cyclic AMP-dependent protein kinase. In conclusion, the data in this study suggested that a phospho-oligosaccharide generated from an insulin-sensitive glycophospholipid may play a role in insulin action by modulating cyclic AMP-dependent protein kinase activity.


The usefulness of phosphorylated-signal transduction and activators of transcription 3 in detecting prostate cancer from negative biopsies.
To avoid the misdiagnosis of prostate cancer (PCA), many patients receive repeated biopsies, despite receiving prior negative biopsies for PCA. Signal transduction and activators of transcription 3 (STAT3), a component of the JAK-STAT signaling pathway, can be activated by tyrosine phosphorylation as P-STAT3 and involved in the regulation of cellular growth, survival and oncogenesis. We aimed to assess the reliability of detecting PCA from the expression of P-STAT3 in prostate tissue previously designated as a negative biopsy. Prostate tissues were obtained from the biopsies of 52 patients with localized PCA as well as from the biopsies of 80 patients free of PCA. Expression of P-STAT3 in these specimens was examined by immunohistochemical staining (IHC) and used to distinguish tissue with PCA from tissue designated as benign during a biopsy procedure. P-STAT3 staining intensities in all samples (initial negative biopsies, cancer positive cores and other negative cores from the same-batch biopsies) of PCA patients was significantly higher than that of benign patients (F = 23.664, P < 0.001). Analysis of the receiver operating characteristics (ROC) curve showed that the area under curve (AUC) for P-STAT3 staining was 0.785. When positive immuno-labeling of P-STAT3 in samples from initial biopsies was used as a marker for PCA, it showed relatively high sensitivity (80.8%) and specificity (76.3%). IHC of P-STAT3 could be utilized to detect PCA patients with initial negative biopsies. As a result, it can be a potential adjunctive tool for current PCA diagnostic programs. P-STAT3 can predict the onset of PCA up to 40 months earlier than currently used diagnostic approaches.


The expanding realm of ballistosporous yeasts.
The recent progress in the systematics of ballistosporous yeasts is discussed. The extensive isolation studies carried out in the last decade resulted in a marked increase in the number of ballistosporous yeast species which now number nearly fifty. The increased number of species has expanded the complexity of taxonomic properties in the following ways: expansion of the range of mol% G+C of nuclear DNA to 39-68.5, thus overlapping that of all basidiomycetous yeasts; increased complexity of conidiogenesis with the finding of the genera Ballistosporomyces and Kockovaella. Based on partial sequencing of 18S ribosomal RNA (positions 1451-1618 in Saccharomyces cerevisiae), it is suggested that mode of conidiogenesis has little value for defining genera. Consequently, ballistosporous yeasts merely represent the "ballistosporous stage" of various taxa which cover the whole evolutionary spectrum of basidiomycetous yeasts. The importance of continuing isolation of ballistosporous yeasts is stressed, which together with molecular studies, will aid further progress in the systematics of basidiomycetous yeasts.


Chronic dermatosis and cutaneous squamous cell carcinoma in the beagle dog.
Chronic dermatosis developed in sparsely-haired, lightly pigmented, ventral body skin in 397 of 1 680 beagle dogs by the time the dogs were 5 1/2 years old. Twenty-six of 55 other beagles had developed by 12 years of age. Squamous cells carcinomas developed in the sites of dermatosis in eight of the 397 younger and five of the 26 older beagles. The lesions resembled solar keratosis (solar dermatosis, actinic dermatosis or senile keratosis) in man. They developed under circumstances suggesting that solar radiation is involved in the pathogenesis, as it is in man.


Reaction dynamics of the chimeric channelrhodopsin C1C2.
Channelrhodopsin (ChR) is a key protein of the optogenetic toolkit. C1C2, a functional chimeric protein of Chlamydomonas reinhardtii ChR1 and ChR2, is the only ChR whose crystal structure has been solved, and thus uniquely suitable for structure-based analysis. We report C1C2 photoreaction dynamics with ultrafast transient absorption and multi-pulse spectroscopy combined with target analysis and structure-based hybrid quantum mechanics/molecular mechanics calculations. Two relaxation pathways exist on the excited (S1) state through two conical intersections CI1 and CI2, that are reached via clockwise and counter-clockwise rotations: (i) the C13=C14 isomerization path with 450 fs via CI1 and (ii) a relaxation path to the initial ground state with 2.0 ps and 11 ps via CI2, depending on the hydrogen-bonding network, hence indicating active-site structural heterogeneity. The presence of the additional conical intersection CI2 rationalizes the relatively low quantum yield of photoisomerization (30 ± 3%), reported here. Furthermore, we show the photoreaction dynamics from picoseconds to seconds, characterizing the complete photocycle of C1C2.


'We have more in common as humans than difference'.
Dr Gina Awoko Higginbottom, Canada Research Chair and associate professor at the University of Alberta's faculty of nursing, recounts how her drive to tackle health inequalities has been the motivating force in her career.


Circulating MicroRNA in inflammatory bowel disease.
MicroRNAs (miRNAs) consist of a group of small noncoding RNAs that partially regulate gene expression. We investigated the expression patterns of commonly deregulated miRNAs in Crohn's disease (CD) and ulcerative colitis (UC) in peripheral blood samples of inflammatory bowel disease patients. This study consisted of 128 CD and 88 UC patients, as well as 162 healthy controls. The expression patterns of the miRNA species were quantitatively assayed using reverse transcription and real-time RT-PCR. Stem-loop complementary DNAs (cDNAs) were synthesized using looped reverse transcription primers specific for each miRNA. MiR-16, miR-23a, miR-29a, miR-106a, miR-107, miR-126, miR-191, miR-199a-5p, miR-200c, miR-362-3p and miR-532-3p were expressed at significantly higher levels in the blood from patients with CD compared with the healthy controls. No significant differences were observed when the CD patients were classified according to disease location and phenotype. In the UC cases three miRNAs (miR-16, miR-21, miR-28-5p, miR-151-5p, miR-155 and miR-199a-5p) were significantly increased compared to healthy controls. miR-155 was the most highly expressed of the UC-associated miRNA in blood samples. Our results suggest that several miRNAs could distinguish CD from UC by real-time PCR. This further highlights the putative role of miRNAs as contributors to IBD pathogenesis. They may help develop new non-invasive biomarkers to distinguish UC and CD.


Traumatic brain injury elicits similar alterations in α7 nicotinic receptor density in two different experimental models.
Traumatic brain injury (TBI) is a major cause of death and disability worldwide, especially in children and young adults. Previous studies have shown alterations in the central cholinergic neurotransmission after TBI. We therefore determined α7 nicotinic acetylcholine receptor (nAChR) densities in newborn piglets and adult rats after experimental TBI. Thirteen newborn piglets (post-TBI survival time: 6 h) underwent fluid percussion (FP) injury (n = 7) or sham operation (n = 6). Furthermore, adult rats randomized into three groups of post-TBI survival times (2, 24, 72 h) received controlled cortical impact injury (CCI, n = 8) or sham operation (n = 8). Brains were frozen, sagittally cut and incubated with the α7-specific radioligand [(125)I]α-bungarotoxin for autoradiography. In injured newborn piglets, decreased α7 receptor densities were observed in the hippocampus (-38%), the hippocampus CA1 (-40%), thalamus (-30%) and colliculus superior (-30%). In adult rats, CCI decreased the receptor densities (between -16 and -47%) in almost any brain region within 2 and 24 h. In conclusion, widespread and significantly lowered α7 nAChR densities were demonstrated in both TBI models. Our results suggest that a nearly similar TBI-induced decrease in the α7 density in the brain of immature and adult animals is found, even with the differences in species, age and experimental procedures. The alterations make the α7 nAChR a suitable target for drug development and neuroimaging after TBI.


An alpha particle measurement system using an energetic neutral helium beam in ITER (invited).
An energetic helium neutral beam is involved in the beam neutralization measurement system of alpha particles confined in a DT fusion plasma. A full size strong-focusing He(+) ion source (2 A, the beam radius of 11.3 mm, the beam energy less than 20 keV). Present strong-focusing He(+) ion source shows an emittance diagram separated for each beamlet of multiple apertures without phase space mixing, despite the space charge of a beamlet is asymmetric and the beam flow is non-laminar. The emittance of beamlets in the peripheral region was larger than that of center. The heat load to the plasma electrode was studied to estimate the duty factor for the ITER application.


Impact of Global Consensus Statement on compliance with hormonal therapy for surgical menopause.
Initiation of and compliance with menopausal hormonal therapy (MHT) have been dropping due to the 2002 Women's Health Initiative (WHI) publication. We evaluated the change in practice of MHT for surgical menopause after implementing the 2013 'Global Consensus Statement on MHT' to our institutional guideline. A retrospective study was conducted in surgically menopausal women newly registering at the Siriraj Menopause Clinic in a university hospital, Thailand, from 1995 to 2013. The patients were categorized into four groups according to periods of MHT initiation: 1995-1998 (control), 2000-2003 (WHI affected), 2005-2008 (post WHI) and 2010-2013 (Global Consensus Statement affected). Their 3-year compliance with MHT was compared using forward stepwise regression analysis. There were 288, 156, 107 and 104 cases in the 1995-1998, 2000-2003, 2005-2008 and 2010-2013 groups. Their mean age at surgery was 42.8 ± 4.7 years. After the first, second and third years, overall compliance was 82.4%, 70.9% and 61.2%, respectively. The 3-year compliance drastically dropped in the 2000-2003 group, and then improved to control level in the 2010-2013 group (51.9% vs. 77.9%, p = 0.035). The initiation of MHT continuously dropped during 2000-2013; however, compliance with MHT initiated during 2010-2013 improved after implementing the 2013 'Global Consensus Statement on MHT' to our institutional guideline. Each institute should have a strategy to encourage the initiation of and compliance with MHT for surgical menopause to achieve long-term health benefits.


'A Burden on the County': Madness, Institutions of Confinement and the Irish Patient in Victorian Lancashire.
This article explores the responses of the Poor Law authorities, asylum superintendents and Lunacy Commissioners to the huge influx of Irish patients into the Lancashire public asylum system, a system facing intense pressure in terms of numbers and costs, in the latter half of the nineteenth century. In particular, it examines the ways in which patients were passed, bartered and exchanged between two sets of institution-workhouses and asylums. In the mid-nineteenth century removal to asylums was advocated for all cases of mental disorder by asylum medical superintendents and the Lunacy Commissioners; by its end, asylum doctors were resisting the attempts of Poor Law officials to 'dump' increasing numbers of chronic cases into their wards. The article situates the Irish patient at the centre of tussles between those with a stake in lunacy provision as a group recognised as numerous, disruptive and isolated.


The dimensions of skin fragments dispersed into the air during activity.
There was a 1000-fold difference between the highest and the lowest mean rate of dispersal of bacteria-carrying particles during exercise among a group of three male and three female subjects. Differences in the numbers of skin fragments dispersed and in the proportion of these carrying bacteria were almost equally responsible for this. Since there is little difference between individuals in the rate of skin replacement the differences in the rates of dispersal of skin fragments during exercise must reflect large variations in the amount of skin surface removed in other ways. The skin fragments dispersed had a wide size range extending below 5 micrometer for the minimum projected diameter (MPD). The median MPD was about 20 micrometer with 7-10% less than 10 micrometer. Many of the particles could therefore pass freely through tightly woven fabrics with pores up to 10-15 micrometer which might seem to be impenetrable to whole corneocytes, typically larger than 30 X 40 micrometer in the hydrated state.


Dialysis of the Obese Patient: Meeting Needs for a Growing Epidemic.
Increasing population obesity and associated metabolic consequences has led to increased number of obese patients with chronic kidney disease and end-stage renal disease. While obesity is associated with increased cardiovascular morbidity and mortality, obese dialysis patients present a seeming paradox for survival benefit, given other acute intervening illnesses for those on dialysis with loss of lean body mass overtime. Challenges remain in providing adequate renal replacement options and therapy for this growing segment of obese patients on dialysis which are discussed in this article.


Activity and QSAR study of baogongteng A and its derivatives as muscarinic agonists.
Baogongteng A (BGT-A), a naturally occurring tropane muscarinic agonist isolated from Chinese medicinal plant, exhibits a bioactive effect different from those of many tropane alkaloids that are muscarinic antagonists. A series of racemic derivatives of BGT-A was synthesized to study the structure-activity relationships (SAR). To explore further the SAR in this series and to ultimately design muscarinic agonists for drug development, a Comparative Molecular Field Analysis (CoMFA) was performed. The values of the leave-one-out cross-validated correlation coefficient q2 and the conventional correlation coefficient r2 for the model are 0.613 and 0.965, respectively. The regression analysis of the data indicated that the steric effect of N-substituted group on tropane of analyzed compounds critically affected the agonistic activity to muscarinic receptors.


Treating brain diseases using systemic parenterally-administered protein therapeutics: Dysfunction of the brain barriers and potential strategies.
The parenteral administration of protein therapeutics is increasingly gaining importance for the treatment of human diseases. However, the presence of practically impermeable blood-brain barriers greatly restricts access of such pharmaceutics to the brain. Treating brain disorders with proteins thus remains a great challenge, and the slow clinical translation of these therapeutics may be largely ascribed to the lack of appropriate brain delivery system. Exploring new approaches to deliver proteins to the brain by circumventing physiological barriers is thus of great interest. Moreover, parallel advances in the molecular neurosciences are important for better characterizing blood-brain interfaces, particularly under different pathological conditions (e.g., stroke, multiple sclerosis, Parkinson's disease, and Alzheimer's disease). This review presents the current state of knowledge of the structure and the function of the main physiological barriers of the brain, the mechanisms of transport across these interfaces, as well as alterations to these concomitant with brain disorders. Further, the different strategies to promote protein delivery into the brain are presented, including the use of molecular Trojan horses, the formulation of nanosystems conjugated/loaded with proteins, protein-engineering technologies, the conjugation of proteins to polymers, and the modulation of intercellular junctions. Additionally, therapeutic approaches for brain diseases that do not involve targeting to the brain are presented (i.e., sink and scavenging mechanisms).


Surface phenotype of rat bone marrow-derived mononuclear phagocytes.
Utilizing a panel of currently available monoclonal antibodies, the surface phenotype of a pure population of resting rat bone marrow-derived mononuclear phagocytes (BMM phi) was analyzed by means of flow cytometry. The present work provides an extensive list of surface markers expressed by BMM phi and also outlines advantages and limitations of flow cytometric analysis of this cell type. The results show that the majority of surface markers considered to be expressed selectively by T lymphocytes, such as Thy-1, CD2 and CD5 antigens, leukosialin (W3/13), or an alloantigen of peripheral T cells, are not expressed by BMM phi. On the other hand, the CD8 antigen and the leukocyte common antigen recognized by MRC OX-33, considered to represent specific markers of cytotoxic T cells and/or peripheral B cells, are expressed on a variable, often considerable proportion of BMM phi. Monoclonal antibodies W3/25, MRC OX-35, and MRC OX-38, directed against epitopes on the CD4 molecule, labeled a variable proportion of BMM phi. Among the 39 monoclonal antibodies examined, none appeared to recognize an epitope which is expressed selectively by mononuclear phagocytes.


Substance P enhances wound closure in nitric oxide synthase knockout mice.
The neuropeptide, substance P (SP), up-regulates nitric oxide production (NO). The purpose of this study was to determine whether SP enhances response to cutaneous injury in nitric oxide synthase knockout (NOS null) mice. We studied mice with targeted deletions of the 3 NOS genes, neuronal NOS, inducible NOS, or endothelial NOS. Full thickness dorsal wounds were treated daily (d 0-6) with topical SP or normal saline (NaCl). Wounds were analyzed by flow cytometry for macrophage, leukocyte, endothelial, and dendritic cells. Healing time and wound epithelialization were compared using analysis of variance. Wound closure in the 3 NOS null mice was slower than the control mice (P < 0.05). SP treatment enhanced wound closure in NOS null mice (P < 0.02). NOS null wounds exhibited reduced inflammation. SP increased macrophage, leukocyte, and dendritic cell densities at d 3 and d 7 (P < 0.05) in all NOS null mice. SP increased endothelial cell number in neuronal NOS and inducible NOS null mice, but not in endothelial NOS null mice (P > 0.05). SP ameliorated the impaired wound healing response observed in NOS null mice by enhancing wound closure kinetics and epithelialization. SP increased inflammatory cell density in the wounds supporting the essential role of inflammatory cells, especially macrophages, in wound repair.


Retrosigmoid implantation of an active bone conduction stimulator in a patient with chronic otitis media.
Percutaneous bone conduction implants are widely used in patients with conductive and mixed hearing loss with no benefit from conventional air conduction hearing aids. These devices have several complications including skin reaction, wound infection, growth of skin over the abutment, and implant extrusion. We describe a case of a transcutaneous bone conduction implantation (Bonebridge, Med-el) in a patient with conductive hearing loss due to chronic otitis media. Surgical planification was performed with the software 3D slicer 4.1. According to this program, the implant transductor was positioned in the retrosigmoid area. Aided thresholds demonstrate a significant benefit, with an improvement from 68dB to 25dB. Speech discrimination scores improved 35dB. The patient is very happy and uses her device daily. The Bonebridge implant is a promising transcutaneous bone conduction implant for patients with conductive hearing loss. Retrosigmoid implantation may be useful in cases with mastoid pathology or previous surgery.


Efficiency of hematological, enzymological and oxidative stress biomarkers of Cyprinus carpio to an emerging organic compound (alphamethrin) toxicity.
Alphamethrin is one of the extensively used pyrethroids. Its non-specific mode-of-action might affect the non-target-organisms. Its toxicity data on aquatic organisms are lacking. We determined the toxicity (35 days) of alphamethrin (0.6 µg/L and 1.2 µg/L) on non-target-organisms by evaluating the efficiency of hematological, enzymological and antioxidants biomarkers of Cyprinus carpio. Compared with the control group, the efficiency of the biomarkers studied was significantly (p < 0.05) impaired in the alphamethrin treated groups. Alphamethrin-toxicity altered hematology, transaminases and the potency of LDH of fish. ACP and ALP activity and biomarkers of oxidative stress in the gills, liver and muscle tissues were affected. IBRv2 index reveals that the biomarkers were inhibited. The observed impairments were the toxicity effects of alphamethrin with respect to concentration and time. The effectiveness of biomarkers for alphamethrin toxicity was like the toxicity data available on other banned insecticides. Alphamethrin could cause multiorgan toxicity on aquatic organisms at µg/L level.


Interaction of reciprocally induced inhibition and premotor facilitation of soleus H reflexes in humans.
During contraction of the soleus muscle, H reflex magnitude is facilitated and vibration induced, late reciprocal inhibition of H reflex magnitude is reduced. The current study investigated whether reduction of electrically evoked late reciprocal inhibition also occurs in the period immediately before movement. H reflexes were facilitated immediately prior to unconditioned plantar flexion movements and conditioning stimuli to the common peroneal nerve produced long-lasting inhibition of the H reflex in 5 subjects. Conditioning stimuli applied during premotor facilitation lead to a significant inhibition of the facilitated H reflex (P < 0.05) in 3 of the 5 subjects. The level of this inhibition was not reduced from that seen with conditioning stimuli alone. These observations suggest that peripherally induced, long-lasting inhibition from the antagonist nerve can alter the excitability of the motoneuronal pool immediately before movement in humans.


A Distinct Class of Bursting Neurons with Strong Gamma Synchronization and Stimulus Selectivity in Monkey V1.
Cortical computation depends on interactions between excitatory and inhibitory neurons. The contributions of distinct neuron types to sensory processing and network synchronization in primate visual cortex remain largely undetermined. We show that in awake monkey V1, there exists a distinct cell type (››30% of neurons) that has narrow-waveform (NW) action potentials and high spontaneous discharge rates and fires in high-frequency bursts. These neurons are more stimulus selective and phase locked to 30- to 80-Hz gamma oscillations than other neuron types. Unlike other neuron types, their gamma-phase locking is highly predictive of orientation tuning. We find evidence for strong rhythmic inhibition in these neurons, suggesting that they interact with interneurons to act as excitatory pacemakers for the V1 gamma rhythm. We did not find a similar class of NW bursting neurons in L2-L4 of mouse V1. Given its properties, this class of NW bursting neurons should be pivotal for the encoding and transmission of stimulus information.


Identification of nitrite-reducing bacteria using sequential mRNA fluorescence in situ hybridization and fluorescence-assisted cell sorting.
Sequential mRNA fluorescence in situ hybridization (mRNA FISH) and fluorescence-assisted cell sorting (SmRFF) was used for the identification of nitrite-reducing bacteria in mixed microbial communities. An oligonucleotide probe labeled with horseradish peroxidase (HRP) was used to target mRNA of nirS, the gene that encodes nitrite reductase, the enzyme responsible for the dissimilatory reduction of nitrite to nitric oxide. Clones for nirS expression were constructed and used to provide proof of concept for the SmRFF method. In addition, cells from pure cultures of Pseudomonas stutzeri and denitrifying activated sludge were hybridized with the HRP probe, and tyramide signal amplification was performed, conferring a strongly fluorescent signal to cells containing nirS mRNA. Flow cytometry-assisted cell sorting was used to detect and physically separate two subgroups from a mixed microbial community: non-fluorescent cells and an enrichment of fluorescent, nitrite-reducing cells. Denaturing gradient gel electrophoresis (DGGE) and subsequent sequencing of 16S ribosomal RNA (rRNA) genes were used to compare the fragments amplified from the two sorted subgroups. Sequences from bands isolated from DGGE profiles suggested that the dominant, active nitrite reducers were closely related to Acidovorax BSB421. Furthermore, following mRNA FISH detection of nitrite-reducing bacteria, 16S rRNA FISH was used to detect ammonia-oxidizing and nitrite-oxidizing bacteria on the same activated sludge sample. We believe that the molecular approach described can be useful as a tool to help address the longstanding challenge of linking function to identity in natural and engineered habitats.


A quantitative study of intercellular heterogeneity in gold nanoparticle uptake across multiple cell lines.
Quantification of intercellular heterogeneity in nanoparticle association is of paramount interest in research investigating applications of nanoparticles in the biomedical space. In this work, gold nanoparticle association (AuNP) in cell populations was quantified using synchrotron X-ray fluorescence microscopy (XRF) for 3 different cell lines (PC-3, Caco2 and MDA-MB-231) and 2 nanoparticle co-culture times (30 min and 10% of each respective cell lines' doubling time). Heterogeneity in association between single cells in the same population was dependant on cell line as well as co-culture time. AuNP association heterogeneity increased with co-culture time for 2 out of the 3 cell lines. Regardless of mean association quantity and measured intercellular heterogeneity, all data were best described by log normal distributions. Mean association between cell lines was statistically different at 30 min, yet indistinguishable at 10% doubling time. Heterogeneity between cell lines which demonstrated statistical differences in distribution can exist despite having statistically indistinguishable means.


[A new in vitro test to simulate gastric absorption of copper, lead, tin and zinc from polluted soils].
Metals in polluted soils can be ingested by young children and cause toxicity through gastro-intestinal absorption. Very few tests are used to evaluate the gastric absorption of metals from polluted soils. The Toxicity Characteristics Leaching Procedure (TCLP) of the Environmental Protection Agency of the United States is indirectly used as usually no action is taken on a polluted soil if lead is less than 5 mg l(-1) in the test. This paper presents a new In vitro simple screening test to detect soils polluted by metals which can cause gastric absorption. The gastric juice simulation test (GJST) is based on the chemistry of the stomach. The TCLP and the GJST has been applied to six different size fractions of 5 soils. Lead carbonate was the main form of lead pollution. Lead oxide, lead silicate, lead phosphate and PbaSnbOc(CO3)d, PbaTibOc(CO3)d and PbaCrbOc were also present. Copper is mainly found as elemental copper partly carbonated while zinc is as zinc oxide or carbonate. Tin is usually associated with iron alloy or oxide or in PbaSnbOc(CO3)d. The percentage of the surface occupied by a polluting phase is higher in small particles. The proportion of particles for which pollution is located near the surface of the particle is more important for small particles. The GJST leaches more copper, lead and zinc than the TCLP, particularly on small size fractions. Inorganic tin is not solubilised by the TCLP or the GJST. The percentage of metal leached from small particles is very important with the GJST, but not with the TCLP. The pH 5 of the TCLP is too high to solubilise much metals and is not a good test to evaluate polluted soils if gastric absorption is to be considered. For two soils out of five, the TCLP didn't detect problematic soils as detected with the Integrated Exposure Uptake Biokinetic model of the USEPA. The GJST detected all the problematic soils and is better to evaluate the chemical solubility of metals in the gastric environment. The GJST better represents the stomach environment (Cl-, pH 2, T = 37 degrees C). The GJST appears to be a good test to detect metal contaminated soils.


Genotoxicity, oxidative stress and transcriptomic effects of Nitenpyram on human bone marrow mesenchymal stem cells.
Despite of the global contamination and ubiquitous exposure to nitenpyram (NIT), little knowledge is available on the adverse effects to human health, with some evidence referring to its genotoxic potency to non-target organisms and esophageal squamous papilloma in rats. Human bone marrow mesenchymal stem cells (hBMSCs) was employed as an in vitro model more relevant to humans to assess the potential genotoxicity of NIT and to understand the underlying mechanisms at cellular and molecular levels. Noncytotoxic concentrations of NIT, 50-2500 μg/mL, dose-dependently elevated micronucleus (MN) and nuclear bud (NB) frequencies to 8.7-29‰ and 15-35‰, respectively. Additional metabolism by rat liver S9 fraction decreased chromosome impairment by 27-52% on MN frequencies and 63-76% on NB frequencies. A commercial NIT product, containing 20% of NIT and 60% of pymetrozine, caused higher cytotoxicity and chromosome impairment in comparison with NIT alone. Expressions of genes responses to DNA damage, ATM, ATR, p53, p21, Bax, H2AX, and GADD45A were disturbed by NIT treatment. Reactive oxygen species (ROS) amount and superoxide dismutase (SOD) activity were enhanced by NIT. Comet assay showed that lower concentrations of NIT, 12.5-100 μg/mL, induced the DNA damage. Transcriptomic analysis identified 468 differentially expressed genes (p < 0.05, |log2(Foldchange)| ≥ 1), from which 22 pathways were enriched. Multiple affected pathways were related to cancer including viral carcinogenesis and bladder cancer. NIT may produce genotoxicity via inducing oxidative stress and deregulating PI3K/Akt, AMPK and mTOR signaling pathways, associated with carcinogenetic potency. While environmental levels of NIT alone may pose little risk to human health, attention should be paid to the health risk arose from the synergistic or additive effects that may exist among NEOs and other types of pesticides.


As fast and selective as enzymatic ligations: unpaired nucleobases increase the selectivity of DNA-controlled native chemical PNA ligation.
DNA-controlled reactions offer interesting opportunities in biological, chemical, and nanosciences. In practical applications, such as in DNA sequence analysis, the sequence fidelity of the chemical-ligation reaction is of central importance. We present a ligation reaction that is as fast as and much more selective than enzymatic T4 ligase-mediated oligonucleotide ligations. The selectivity was higher than 3000-fold in discriminating matched from singly mismatched DNA templates. It is demonstrated that this enormous selectivity is the hallmark of the particular ligation architecture, which is distinct from previous ligation architectures designed as "nick ligations". Interestingly, the fidelity of the native chemical ligation of peptide nucleic acids was increased by more than one order of magnitude when performing the ligation in such a way that an abasic-site mimic was formed opposite an unpaired template base. It is shown that the high sequence fidelity of the abasic ligation could facilitate the MALDI-TOF mass-spectrometric analysis of early cancer onset by allowing the detection of as little as 0.2 % of single-base mutant DNA in the presence of 99.8 % wild-type DNA.


An Efficient DNA Extraction for a Blue Xestospongia sp. Sponge and Its Associated Microorganisms Containing Cytotoxic Substances.
Extraction of high quantity and quality DNAs from marine sponges, which contain diverse and abundant microbial communities, is important to molecular biology techniques for the analysis of nucleic acids. Several marine sponges and their associated microorganisms have been known to produce cytotoxic natural products on several cancer cell lines via DNA damage mechanisms. These marine cytotoxic substances might be one of the factors that cause the low quantity and quality of DNAs during the DNA extraction from its living origin. Therefore, the extraction of DNA of a Thai blue marine sponge Xestospongia sp. with sufficient purity and quantity for molecular study can be challenging. In this study, we developed an efficient extraction method to prepare DNAs from a Thai blue marine sponge Xestospongia sp. which accumulated a highly potent cytotoxic alkaloid with DNA-damaging activity, named Renieramycin M (RM), as a major constituent in high quantity. We demonstrated that removal of RM from the sponge samples by a simple methanolic extraction before DNA extraction dramatically increased the yield and purity of DNAs compared to the RM-unremoved sponge samples. High molecular weight (HMW) genomic DNA was obtained from sponge samples with 8 times of RM elimination by using modified NaOAc salting-out extraction method. The quantity and quality of the prepared DNAs were comparatively determined via spectrophotometry, electrophoresis, and 16S rRNA gene amplification. Our result suggests that the removal of DNA-damaging constituents from the samples is a crucial step and must be seriously taken as the necessary consideration for the practical protocol of DNA extraction.


Self-cleaning cotton functionalized with TiO2/SiO2: focus on the role of silica.
This manuscript aims to investigate the functionalization of cotton fabrics with TiO2/SiO2. In this study, the sol-gel method was employed to prepare titania and silica sols and the functionalization was carried out using the dip-pad-dry-cure process. Titanium tetra isopropoxide (TTIP) and tetra ethyl orthosilicate (TEOS) were utilized as precursors of TiO2 and SiO2, respectively. TiO2/SiO2 composite sols were prepared in three different Ti:Si molar ratios of 1:0.43, 1:1, and 1:2.33. The self-cleaning property of cotton samples functionalized with TiO2/SiO2 was assessed based on the coffee stain removal capability and the decomposition rate of methylene blue under UV irradiation. FTIR study of the TiO2/SiO2 photocatalyst confirmed the existence of Si-O-Si and Ti-O-Si bonds. Scanning electron microscopy was employed to investigate the morphology of the functionalized cotton samples. The samples coated with TiO2/SiO2 showed greater ability of coffee stain removal and methylene blue degradation compared with samples functionalized with TiO2 demonstrating improved self-cleaning properties. The role of SiO2 in improving these properties is also discussed.


The Extractive Industries: Asserting Their Place in Global Health Pedagogy.
The extractive industries play a central role in determining the social determinants of global health yet may not be a core subject of global health studies. This article describes how an undergraduate seminar in global health takes on questions and examples related to the "causes of causes" and challenges instructors to find a place for discussion of extraction of wealth, development, and health in their curricula.


A systematic review and meta-analysis of miRNAs for the detection of cervical cancer.
Aim: This study aimed to critically appraise the evidence of the diagnostic effectiveness of miRNAs for the detection of cervical cancer. Methods & materials: A systematic review and meta-analysis was performed, searching PubMed, EMBASE and Web of Science. An umbrella meta-analysis of meta-analyses of individual biomarkers was performed. A Grading of Recommendations, Assessment, Development and Evaluations (GRADE) assessment of evidence was also performed. Results: A total of 52 miRNAs were included. Umbrella meta-analysis revealed significant heterogeneity in terms of sensitivity, specificity, receiver operating characteristic (ROC), positive predictive value and/or negative predictive value. Umbrella effects were 0.76 (95% CI: 0.73-0.78), 0.78 (95% CI: 0.75-0.81), 0.77 (95% CI: 0.75-0.80), 0.75 (95% CI: 0.71-0.79) and 0.76 (95% CI: 0.74-0.79), respectively. Conclusion: Moderate quality evidence suggested miR199a-5p, miR21-5p and miR-141a had excellent diagnostic performance.


Vortex 4.0 on chip.
The orbital angular momentum (OAM) of light has promising applications, ranging from information multiplexing andhigh-speed optical communication to computation. Dynamically tunable and switchable vortex microlasers combinedwith direct photocurrent detection of the topological charges of OAM states have paved unexplored routes for thedevelopment and integration of fourth-generation (4.0) vortex technology, potentially on chip.


Chelatable lead versus lead in human trabecular and compact bone.
In active and retired lead workers there was a close correlation between urinary excretion of lead during 6 h after intake of a single oral dose of 0.5 g penicillamine, and the excretion during 24 h. In chelation tests it is thus sufficient to collect urine for only a few hours. There was a close correlation between the amount of chelatable lead and the blood-lead level, as well as the lead level in biopsies of trabecular bone from vertebrae, but there was no association with lead in compact bone, as measured in finger-bone by in vivo X-ray fluorescence. The chelatable lead probably mainly reflects the soft tissue lead pool and a fraction of the trabecular bone lead pool, which has a relatively rapid turnover. It is not a valid indicator of the pool of lead which has slowly accumulated in the compact bone, and it is thus not useful as a time-integrated index of the exposure over a long period of time.