The structure and function of the blood-brain barrier.
It is now clear that the phenomenon of a blood-brain barrier results from the high-resistance endothelium of cerebral vessels. The glial sheath appears to have no transport function but determines the specific characteristics of the cerebral endothelium. Among the transport mechanisms present in the endothelium is a potent sodium-potassium pump in the abluminal membrane. The endothelium probably secretes a small volume of fluid into the cerebral interstitium. Ouabain-insensitive potassium transport has been investigated in isolated cerebral capillaries. This component is very dependent on the osmolality of the medium, being markedly increased in a hypertonic medium and decreased in hypotonic conditions. This behavior may well be important in determining the net exchanges of potassium across the blood-brain barrier, which contribute to volume control of the brain in osmotic disturbances.


Safety of enteral rehabilitative therapy in rat small bowel transplantation.
To evaluate the safety of enteral rehabilitative therapy in rat small bowel transplantation. Forty-eight rat recipients of allogeneic heterotopic small bowel transplantation (SD and Wistar) were randomly divided into 4 groups according to the application or not of enteral rehabilitative therapy and cyclosporine A (CsA). The pathological changes of the graft, IL-2 receptor expression in lamina propria lymphocytes, serum IL-2 concentrations, results of spleen lymphocytes transformation test and the IL-2 secretion capacity were determined and compared. Enteral rehabilitative therapy could promote the immune function of the recipient so as to augment the acute rejection. But such effects could be blocked by the commonly used immunosuppressant CsA. Under the immunosuppression of CsA (10 mg.kg(-1).d(-1), i.m.), application of enteral rehabilitative therapy did not induce or aggravate acute rejection. Under effective immunosuppression, application of enteral rehabilitative therapy is safe in rat small bowel transplantation.


Quality of life, health, and government perception during COVID-19 times: Data from Colombia.
This analysis presents data collected through an online survey about the quality of life, health, subjective wellbeing, and government perception in four cities in Colombia during the COVID-19 crisis. Four universities and a local newspaper promoted the survey to assess how the pandemic affected the population's quality of life in a broad range of social and economic aspects. Respondents were adults (+18 years old) living in the largest Colombia cities: Bogotá, Medellín, Cali, and Barranquilla, totaling 1637 complete observations. Researchers used snowballing sampling strategy, social networks, a web page, and an advertisement in the partner newspaper for data collection. This data set helps to conduct social research and policy reports about the consequences of the pandemic. The data enclosed in this paper includes socioeconomic variables, income reduction, employment, household composition, teleworking, indebtedness, physical and mental health, physical activity behavior, subjective wellbeing, affective and communal relationships, institutional trust, and perception of government performance during COVID-19. We aim at contributing to a better understating of the consequences of the pandemic in Colombia and general in the Global South through the collection and dissemination of data for academic and policy purposes.


Host species barriers to Jaagsiekte sheep retrovirus replication and carcinogenesis.
Understanding the factors governing host species barriers to virus transmission has added significantly to our appreciation of virus pathogenesis. Jaagsiekte sheep retrovirus (JSRV) is the causative agent of ovine pulmonary adenocarcinoma (OPA), a transmissible lung cancer of sheep that has rarely been found in goats. In this study, in order to further clarify the pathogenesis of OPA, we investigated whether goats are resistant to JSRV replication and carcinogenesis. We found that JSRV induces lung tumors in goats with macroscopic and histopathological features that dramatically differ from those in sheep. However, the origins of the tumor cells in the two species are identical. Interestingly, in experimentally infected lambs and goat kids, we revealed major differences in the number of virus-infected cells at early stages of infection. These differences were not related to the number of available target cells for virus infection and cell transformation or the presence of a host-specific immune response toward JSRV. Indeed, we also found that goats possess transcriptionally active endogenous retroviruses (enJSRVs) that likely influence the host immune response toward the exogenous JSRV. Overall, these results suggest that goat cells, or at least those cells targeted for viral carcinogenesis, are not permissive to virus replication but can be transformed by JSRV.


Polyaniline functionalized reduced graphene oxide/carbon nanotube ternary nanocomposite as a supercapacitor electrode.
Aniline was chemically bonded to the surface of graphene oxide (GO) via a nucleophilic ring-opening reaction. The aniline-bonded GO was then reduced to reduced graphene oxide (RGO) in the presence of multi-walled carbon nanotubes (MWCNTs), which could prevent the RGO from aggregating. The bonded aniline was further responsible for the grafting of polyaniline (PANI) onto the surface of RGO via chemical oxidative polymerization of aniline, and the resultant PANI-RGO/MWCNT ternary nanocomposite could be used as a supercapacitor electrode with high performance.


Unraveling features of the natural MHC class II peptidome of skin-migrated dendritic cells.
Dendritic cells (DCs) migrating from peripheral tissues at steady state are considered the most efficient antigen-presenting cells (APCs) involved in the induction of peripheral T-cell tolerance via self-antigen presentation on MHC class II molecules. However, difficulties in obtaining sufficient numbers of such DCs have precluded previous analyses of their natural MHC class II peptidome in laboratory animals or humans. Here, we overcome this difficulty by collecting the large quantities of sheep DCs that migrate from the skin via the afferent lymphatics at steady state to the draining lymph node. We compared the repertoire of MHC class II-bound peptides from afferent lymph DCs with autologous APCs derived from peripheral blood. A large fraction of the MHC class II peptidome from skin DCs was derived from membrane-recycling proteins (59%) and from proteins of the antigen presentation machinery (50%), whereas these types of peptides constituted a more limited fraction in blood APCs (21 and 11%, respectively). One sheep cytokeratin peptide was identified in the skin DC peptidome indicating active processing of epithelium-derived antigens. Conversely, peptides derived from cytosolic and soluble antigens of the extracellular milieu were more represented in blood APCs than skin DCs. The biased peptidome of skin-migrated DCs indicates that these cells express a peptide repertoire for the generation of self-reactive and/or regulatory T cells mainly directed toward DC molecules from internal and external membranes and to a lesser extent toward antigens of the extracellular milieu, including some tissue-specific peptides.


[Meat and human health: excess and errors].
Many studies have examined the influence of meat consumption on human health. Meat eaters have a higher body mass index and more weight gain than vegetarians. The risk of type 2 diabetes has also been linked to high meat consumption. However, the statistical correlations with these metabolic disorders are weak. There is inconsistent evidence of a higher cardiovascular risk. A link between high meat consumption and cancer, particularly colorectal cancer, has been observed in nearly all epidemiological studies. Some studies have also shown a link with breast, prostate and lung cancer. The mode of cooking could be partly En 2 responsible for this effect, due for example to heterocyclic aromatic amines production euro during grilling and intensive cooking. Advice is given.


[Aujeszky's disease in swine in The Netherlands, A.D. 1991].
Aujeszky's disease is caused by a herpesvirus. The pig is the natural host and sole source of dissemination of the virus. After a primary infection, during which the pig excretes virus for 10-14 days, the virus remains latent in the host. Eradication has been achieved in individual herds, areas and countries with a low prevalence. The crucial question is whether it is feasible to eradicate the virus in heavily infected areas. Using so-called marker vaccines in conjunction with companion diagnostic test kits the virus was eliminated in individual herds. The preliminary results of extensive field studies to investigate whether intensive vaccination curtails the circulation of virus are encouraging.


Environmental filtering and mass effect are two important processes driving lake benthic diatoms: Results of a DNA metabarcoding study in a large lake.
Environmental filtering is often found to dominate assembly rules in diatoms. These microalgae are diverse, especially at subspecies level, and tend to exhibit a niche phylogenetic conservatism. Therefore, other rules, such as competition or mass effects, should be detectable when environmental gradients and dispersal barriers are limited. We used metabarcoding to analyse benthic littoral diatom communities in 153 sites in a large lake (Geneva) exhibiting weak geographical barriers and weak environmental gradients outside river estuaries. We assessed assembly rules using variance partitioning, phylogenetic and source tracking analyses. No phylogenetic over-dispersion of communities, indicative of exclusive competition, was detected. Instead, we found these communities to be phylogenetically over-clustered, indicating environmental filtering, which was even stronger near river estuaries where environmental gradients are stronger. Finally, using a Bayesian method (SourceTracker), we found that rivers flowing into the lake bring communities that settle, especially in sites close to estuaries. Rivers with the highest discharges are primarily responsible for immigration, explaining 27% of lake composition. Therefore, despite favourable conditions to observe other rules, our results support that diatom communities are prominently assembled by environmental filtering and immigration processes, in particular from rivers. However, this does not exclude that other assembly rules may be at play at a finer spatial, temporal and/or phylogenetic scale.


HSP72 protects cells from ER stress-induced apoptosis via enhancement of IRE1alpha-XBP1 signaling through a physical interaction.
Endoplasmic reticulum (ER) stress is a feature of secretory cells and of many diseases including cancer, neurodegeneration, and diabetes. Adaptation to ER stress depends on the activation of a signal transduction pathway known as the unfolded protein response (UPR). Enhanced expression of Hsp72 has been shown to reduce tissue injury in response to stress stimuli and improve cell survival in experimental models of stroke, sepsis, renal failure, and myocardial ischemia. Hsp72 inhibits several features of the intrinsic apoptotic pathway. However, the molecular mechanisms by which Hsp72 expression inhibits ER stress-induced apoptosis are not clearly understood. Here we show that Hsp72 enhances cell survival under ER stress conditions. The UPR signals through the sensor IRE1alpha, which controls the splicing of the mRNA encoding the transcription factor XBP1. We show that Hsp72 enhances XBP1 mRNA splicing and expression of its target genes, associated with attenuated apoptosis under ER stress conditions. Inhibition of XBP1 mRNA splicing either by dominant negative IRE1alpha or by knocking down XBP1 specifically abrogated the inhibition of ER stress-induced apoptosis by Hsp72. Regulation of the UPR was associated with the formation of a stable protein complex between Hsp72 and the cytosolic domain of IRE1alpha. Finally, Hsp72 enhanced the RNase activity of recombinant IRE1alpha in vitro, suggesting a direct regulation. Our data show that binding of Hsp72 to IRE1alpha enhances IRE1alpha/XBP1 signaling at the ER and inhibits ER stress-induced apoptosis. These results provide a physical connection between cytosolic chaperones and the ER stress response.


The processing and comprehension of wh-questions among L2 German speakers.
Using the self-paced-reading paradigm, the present study examines whether highly proficient second language (L2) speakers of German (English L1) use case-marking information during the on-line comprehension of unambiguous wh-extractions, even when task demands do not draw explicit attention to this morphosyntactic feature in German. Results support previous findings, in that both the native and the L2 German speakers exhibited an immediate subject-preference in the matrix clause, suggesting they were sensitive to case-marking information. However, only among the native speakers did this subject-preference carry over to reading times in the complement clause. The results from the present study are discussed in light of current debates regarding the ability of L2 speakers to attain native-like processing strategies in their L2.


Adrenomedullin-mediated relaxations in veins are endothelium-dependent and distinct from arteries.
In arteries, adrenomedullin (ADM) causes relaxations of rings with and without endothelium by stimulating accumulation of cyclic nucleotides resulting from activation of the ADM and calcitonin gene-related peptide (CGRP) receptors. Experiments were designed to determine the mechanism(s) of relaxation to ADM in veins. Rings of canine femoral vein with and without endothelium were suspended in organ chambers for measurement of isometric force. Rings were contracted with prostaglandin F2alpha (2 x 10(-6) M), and cumulative dose-responses to ADM (10(-11) to 10(-7) M) were obtained in the absence or presence of indomethacin (10(-5) M), indomethacin + N(G)-monomethyl-L-arginine (10(-4) M), methylene blue (10(-5) M), particulate guanylate cyclase inhibitor HS-142-1 (10(-5) M), tetraethylammonium (TEA, 10(-2) M), CGRP-receptor antagonist (CGRP 8-37, 10(-6) M), ADM-receptor antagonist (ADM 26-52, 10(-6) M), diphenhydramine (10(-6) M), 8-phenyltheophylline (3 x 10(-6) M), or superoxide dismutase (150 U/ml) plus catalase (1,200 U/ml). ADM produced concentration-dependent relaxations only in veins with endothelium. Relaxations to ADM in rings with endothelium were significantly inhibited only by methylene blue and HS-142-1. In separate experiments, incubation of rings with ADM (10(-8) M) and 3-isobutyl-1-methyl-xanthine (10(-4) M) for 3 min did not significantly affect the accumulation of cyclic guanosine monophosphate (cGMP) and cyclic adenosine monophosphate (cAMP). These data suggest that ADM-mediated relaxation in veins is endothelium dependent and is not associated with activation of CGRP receptors or currently defined ADM receptors. Further, relaxations are not mediated by nitric oxide, indomethacin-sensitive prostanoids, TEA-sensitive hyperpolarizing factors, oxygen free radicals, or accumulation of cyclic nucleotides.


Bone Loss in Revision Total Knee Arthroplasty: Evaluation and Management.
With the increasing number of total knee arthroplasty (TKA) procedures being performed annually, the burden of revision surgery is also expected to increase. Addressing bone loss during revision TKA is a considerable challenge that requires proper preoperative evaluation and surgical planning. In addition to an assessment of femoral and tibial bone loss, a systematic approach to the management of bone loss associated with TKA includes careful implant selection, adherence to the general principles of revision TKA, and an understanding of the available reconstruction options. These options include cement with screws, morcellized allograft, bulk structural allograft, modular wedges, block augments, porous metal cones or sleeves, and megaprostheses.


Hypoxia-induced oxidative stress promotes MUC4 degradation via autophagy to enhance pancreatic cancer cells survival.
Pancreatic cancer (PC) and associated pre-neoplastic lesions have been reported to be hypoxic, primarily due to hypovascular nature of PC. Though the presence of hypoxia under cancerous condition has been associated with the overexpression of oncogenic proteins (MUC1), multiple emerging reports have also indicated the growth inhibitory effects of hypoxia. In spite of being recognized as the top-most differentially expressed and established oncogenic protein in PC, MUC4 regulation in terms of micro-environmental stress has not been determined. Herein, for the first time, we are reporting that MUC4 protein stability is drastically affected in PC, under hypoxic condition in a hypoxia inducible factor 1α (HIF-1α)-independent manner. Mechanistically, we have demonstrated that hypoxia-mediated induction of reactive oxygen species (ROS) promotes autophagy by inhibiting pAkt/mTORC1 pathway, one of the central regulators of autophagy. Immunohistofluorescence analyses revealed significant negative correlation (P-value=0.017) between 8-hydroxy guanosine (8-OHG) and MUC4 in primary pancreatic tumors (n=25). Moreover, we found pronounced colocalization between MUC4 and LAMP1/LC3 (microtubule-associated protein 1A/1B-light chain 3) in PC tissues and also observed their negative relationship in their expression pattern, suggesting that areas with high autophagy rate had less MUC4 expression. We also found that hypoxia and ROS have negative impact on overall cell growth and viability, which was partially, though significantly (P<0.05), rescued in the presence of MUC4. Altogether, hypoxia-mediated oxidative stress induces autophagy in PC, leading to the MUC4 degradation to enhance survival, possibly by offering required metabolites to stressed cells.


CXCR3+ T regulatory cells selectively accumulate in human ovarian carcinomas to limit type I immunity.
Antitumor type I T-cell responses involving IFN-γ production are critical to control cancer, but the efficacy of this response is limited by a variety of immunosuppressive mechanisms that promote tumoral immune escape. One critical mechanism involves the accumulation of FOXP3(+) T regulatory cells (Treg), a class of suppressive T cells that prevent excessive tissue destruction caused by unchecked immune responses. Recent studies have revealed that FOXP3(+) Treg include distinct subsets specifically controlling over the corresponding effector subset. In particular, CXCR3(+) Treg have been described as a subset specialized in the control of type I T-cell responses in vivo. Here, we show that CXCR3(+) Treg are highly enriched in human ovarian carcinomas, particularly in solid tumor masses, where they represent the majority of Treg. Tumor-associated CXCR3(+.) Treg coexpress T-bet but do not secrete IFN-γ ex vivo and suppress proliferation and IFN-γ secretion of T effectors. In addition, they coexpress Helios, suggesting that they originate from natural Treg. Finally, we show that the proportion of CXCR3(+) Treg at tumor sites is directly correlated with that of CXCR3(+) T effectors, consistent with expression of CXCR3 ligands. Together, our findings support the concept that natural CXCR3(+) T-bet(+) Treg selectively accumulate in ovarian tumors to control type I T-cell responses, resulting in the collateral limitation of efficient antitumor immunity.


Targeting the Kynurenine Pathway for the Treatment of Cisplatin-Resistant Lung Cancer.
Cisplatin resistance is a major barrier in the effective treatment of lung cancer. Cisplatin-resistant (CR) lung cancer cells do not primarily use glucose but rather consume amino acids such as glutamine and tryptophan (Trp) for survival. CR cells activate the kynurenine (KYN) pathway (KP) to cope with excessive reactive oxygen species (ROS) and maintain homeostasis for growth and proliferation. Consequently, indoleamine 2,3-dioxygenase-1 (IDO1) becomes an essential enzyme for CR cells' survival because it initiates and regulates the first step in the KP. Increased IDO1 activities and ROS levels are found in CR cells versus cisplatin-sensitive lung cancer. Importantly, significantly greater KYN/Trp ratio (P = 0.005) is detected in serum of patients who fail cisplatin when compared with naïve treatment. Knocking down IDO1 using shRNA or IDO1 inhibitors heightens ROS levels and results in a significant growth inhibitory effect only on CR cells and not on cisplatin-sensitive cells. Exposing CR cells to antioxidant (TIRON) results in suppression of IDO1 activity and confers resistance to IDO1 inhibition, indicating an interrelationship between ROS and IDO1. Because KYN plays a critical role in reprogramming naïve T cells to the immune-suppressive regulatory T-cell (T-reg) phenotype, we observed higher expression of TGFβ, FoxP3, and CD4+CD25+ in mice bearing CR tumors compared with tumors from cisplatin-sensitive counterparts. IMPLICATIONS: Findings suggest that the enzyme-inhibitory activity and antitumor efficacy of IDO1 inhibitors rely in part on ROS levels, arguing that IDO1 expression alone may be insufficient to determine the clinical benefits for this class of experimental cancer drugs. Importantly, IDO1 inhibitors may be more suitable to treat patients with lung cancer who failed cisplatin therapy than naïve treatment patients.


Novel induction by herpes simplex virus of hybrid interferon gene transcripts driven by the strong cytomegalovirus IE94 promoter.
We have constructed stable DNA-transfected LTK+ cell lines containing two different coselected hybrid interferon (IFN) genes driven by the usually strong and constitutive promoter from the immediate-early 94K protein (IE94) gene of simian cytomegalovirus. Surprisingly, and unlike hybrid IE94-chloramphenicol acetyltransferase gene constructs, both of the IE94-IFN genes (one with and one without the complex spliced intron region) produced relatively low basal titers of biologically active human IFN in the mouse cell lines. However, IFN expression could be stimulated up to 120-fold by superinfection with herpes simplex virus (HSV), although not with cytomegalovirus. To examine the mechanism of this unexpected HSV induction process, we measured the levels of both IE94-IFN mRNA and IFN protein produced under various infection protocols. Compared with similar previously characterized cell lines containing hybrid IFN genes under the control of HSV IE or delayed-early (DE) promoters, activation of IFN expression first occurred at an intermediate time. Both IE94-IFN cell lines also produced an unusual pattern of response to infection with the HSV IE regulation-deficient mutants tsK and tsB7: stimulation of IFN synthesis occurred in the absence of a functional HSV IE175 (or ICP4) gene product, but did not occur in the absence of uncoating of virus capsids. Cycloheximide treatment (without virus infection) also gave a rapid 30-fold increase in steady-state levels of correctly initiated mRNA from both types of IE94-IFN hybrid genes, but had no effect on cells containing the IE175-IFN construct. Therefore, we conclude that the use of the IE94-IFN constructs identifies a novel HSV regulatory response that requires a previously unrecognized function of HSV and does not involve either IE175 or the pre-IE "virion factor" trans-activators that are known to stimulate transcription of HSV IE and DE genes, respectively.


A validated method for development of atovaquone as API and tablet dosage forms by UV spectroscopy.
A simple new spectrophotometric method has been developed for estimation of Atovaquone in bulk and tablet dosage form. Atovaquone is estimated to be 251 nm in methanol. The Beer's law is obeyed in the concentration range of 1-10 μg/mL of the drug. The slope and intercept values are 0.111 and 0.012, respectively. Results of analysis of this method have been validated statically and by recovery studies. The method is applied to the marketed tablet formulation. A result of the analysis of tablet formulation, given as a percentage of label claim ± standard deviation, is 99.14 ± 0.66. The precision and accuracy has been examined by performing recovery studies and found to be 100.09 ± 1.14. The developed method is simple, sensitive, and reproducible, and can be used for the routine analysis of Atovaquone in bulk and tablet dosage form.


A 10-year Audit of Penile Prosthesis Insertion.
Introduction Penile prosthesis (PP) insertion is the gold standard surgical treatment option for men with refractory Erectile Dysfunction (ED). PP insertion is considered effective but has a well-documented array of complications. Our aim was to assess outcomes following single-surgeon insertion of PP for ED within an Irish cohort. Methods Following review of the Hospital In-Patient Enquiry (HIPE) system, a retrospective chart review of all patients who underwent PP insertion over a 10-year period from 2008-2017 inclusive was performed, and an electronic database was analysed for results. Results One-hundred-and-eleven PPs were inserted in 96 patients. The most common aetiology for ED in our cohort was post-prostatectomy, affecting 25 (26%) patients. The most frequently implanted device was a 3-piece inflatable PP (3p-IPP) (AMS 700TM; American Medical Systems Inc., Minnesota, USA) and the peno-scrotal approach was utilised in the majority of patients (86, 77.5%). No intraoperative complications were recorded. Twelve (12.5%) patients developed peri-operative complications. Thirteen (13.5%) patients required device revision, the majority for device failure. Of the 71 patient satisfaction responses, 61 (85.9%) patients were satisfied with their PP. Conclusions This single-surgeon retrospective audit of PP surgery demonstrates complication rates in-line with internationally published data. Patients should be adequately counselled regarding possible complications, including device failure and erosion. PP insertion should be considered for suitable patients with refractory ED.


Albumin-based micro-composite drug carriers with dual chemo-agents for targeted breast cancer treatment.
Albumin-based drug-carrying micro-composite spheres were fabricated and studied to evaluate their potentials for breast cancer treatment. Magnetic nanoparticles and albumin were incorporated within poly(D l-lactide-co-glycolide) microspheres to increase accumulation of the microspheres at the target site. Two chemotherapeutics, cyclophosphamide and 5-fluorouracil, were encapsulated into the microspheres. The drug-release study revealed an initial burst of drug and then sustained release by diffusion. A Fourier transform infrared spectroscopy study confirmed the presence of all components of the drug delivery system. An in vitro study using fibroblast cells (3T3) and breast cancer cells (MDA-486) exhibited an effective cytotoxicity behavior when exposed to the drug delivery system in a dose- and time-dependent manner. The therapeutic influence of the drug delivery system was evaluated in vivo using a nude mouse breast cancer model. A continuous decrease in tumor size was observed in groups treated with microspheres containing the chemotherapeutics, whereas mice treated with direct chemotherapy without drug delivery system showed less efficacy and suggested tumor relapse after cessation of treatment. The enhanced therapeutic influence of the drug delivery system may be attributed to the increased uptake of the microspheres by malignant cells due to the presence of albumin and magnetic force. The bioavailability of chemotherapeutics at the target site was further increased due to the sustained release of the drugs by diffusion following the burst release. Continuous investigations will optimize the size of the drug delivery system and portions of the target driving-force components (magnetic nanoparticles and albumin) in the drug delivery system to maximize its therapeutic efficacy and minimize potential long-term side effects.


GenBank and PubMed: How connected are they?
GenBank is a public repository of all publicly available molecular sequence data from a range of sources. In addition to relevant metadata (e.g., sequence description, source organism and taxonomy), publication information is recorded in the GenBank data file. The identification of literature associated with a given molecular sequence may be an essential first step in developing research hypotheses. Although many of the publications associated with GenBank records may not be linked into or part of complementary literature databases (e.g., PubMed), GenBank records associated with literature indexed in Medline are identifiable as they contain PubMed identifiers (PMIDs). Here we show that an analysis of 87,116,501 GenBank sequence files reveals that 42% are associated with a publication or patent. Of these, 71% are associated with PMIDs, and can therefore be linked to a citation record in the PubMed database. The remaining (29%) of publication-associated GenBank entries either do not have PMIDs or cite a publication that is not currently indexed by PubMed. We also identify the journal titles that are linked through citations in the GenBank files to the largest number of sequences. Our analysis suggests that GenBank contains molecular sequences from a range of disciplines beyond biomedicine, the initial scope of PubMed. The findings thus suggest opportunities to develop mechanisms for integrating biological knowledge beyond the biomedical field.


Copper sulfide nanoparticles as high-performance cathode materials for magnesium secondary batteries.
Magnesium secondary batteries are promising candidates for large-scale energy storage systems with high safety, because of the dendrite-free electrodeposition of the magnesium anode. However, the search for available cathode materials remains a significant challenge, hindering their development. In this work, we report copper sulfide nanoparticles as high-performance cathode materials for magnesium secondary batteries, which deliver a high reversible capacity of 175 mA h g-1 at 50 mA g-1. The cathode also shows an excellent rate capability providing 90 mA h g-1 at 1000 mA g-1 and an outstanding long-term cyclability over 350 cycles. The beneficial properties are ascribed to the small-sized copper sulfide particles which facilitate the solid-state diffusion kinetics. Further investigation on the mechanism demonstrates that the reaction is a typical conversion reaction, and the excellent cycling stability is due to the CuS nanoparticles which are not facile to aggregate during cycling. This work introduces an abundant, low-cost and high-performance cathode material for magnesium secondary batteries, and provides feasibility for the practical application of magnesium secondary battery systems in large-scale energy storage devices.


Alone but feeling no pain: Effects of social exclusion on physical pain tolerance and pain threshold, affective forecasting, and interpersonal empathy.
Prior findings of emotional numbness (rather than distress) among socially excluded persons led the authors to investigate whether exclusion causes a far-reaching insensitivity to both physical and emotional pain. Experiments 1-4 showed that receiving an ostensibly diagnostic forecast of a lonesome future life reduced sensitivity to physical pain, as indicated by both (higher) thresholds and tolerance. Exclusion also caused emotional insensitivity, as indicated by reductions in affective forecasting of joy or woe over a future football outcome (Experiment 3), as well as lesser empathizing with another person's suffering from either romantic breakup (Experiment 4) or a broken leg (Experiment 5). The insensitivities to pain and emotion were highly intercorrelated.


Combining stable isotopes with contamination indicators: A method for improved investigation of nitrate sources and dynamics in aquifers with mixed nitrogen inputs.
Excessive nitrate (NO3-) concentration in groundwater raises health and environmental issues that must be addressed by all European Union (EU) member states under the Nitrates Directive and the Water Framework Directive. The identification of NO3- sources is critical to efficiently control or reverse NO3- contamination that affects many aquifers. In that respect, the use of stable isotope ratios 15N/14N and 18O/16O in NO3- (expressed as δ15N-NO3- and δ18O-NO3-, respectively) has long shown its value. However, limitations exist in complex environments where multiple nitrogen (N) sources coexist. This two-year study explores a method for improved NO3- source investigation in a shallow unconfined aquifer with mixed N inputs and a long established NO3- problem. In this tillage-dominated area of free-draining soil and subsoil, suspected NO3- sources were diffuse applications of artificial fertiliser and organic point sources (septic tanks and farmyards). Bearing in mind that artificial diffuse sources were ubiquitous, groundwater samples were first classified according to a combination of two indicators relevant of point source contamination: presence/absence of organic point sources (i.e. septic tank and/or farmyard) near sampling wells and exceedance/non-exceedance of a contamination threshold value for sodium (Na+) in groundwater. This classification identified three contamination groups: agricultural diffuse source but no point source (D+P-), agricultural diffuse and point source (D+P+) and agricultural diffuse but point source occurrence ambiguous (D+P±). Thereafter δ15N-NO3- and δ18O-NO3- data were superimposed on the classification. As δ15N-NO3- was plotted against δ18O-NO3-, comparisons were made between the different contamination groups. Overall, both δ variables were significantly and positively correlated (p < 0.0001, rs = 0.599, slope of 0.5), which was indicative of denitrification. An inspection of the contamination groups revealed that denitrification did not occur in the absence of point source contamination (group D+P-). In fact, strong significant denitrification lines occurred only in the D+P+ and D+P± groups (p < 0.0001, rs > 0.6, 0.53 ≤ slope ≤ 0.76), i.e. where point source contamination was characterised or suspected. These lines originated from the 2-6‰ range for δ15N-NO3-, which suggests that i) NO3- contamination was dominated by an agricultural diffuse N source (most likely the large organic matter pool that has incorporated 15N-depleted nitrogen from artificial fertiliser in agricultural soils and whose nitrification is stimulated by ploughing and fertilisation) rather than point sources and ii) denitrification was possibly favoured by high dissolved organic content (DOC) from point sources. Combining contamination indicators and a large stable isotope dataset collected over a large study area could therefore improve our understanding of the NO3- contamination processes in groundwater for better land use management. We hypothesise that in future research, additional contamination indicators (e.g. pharmaceutical molecules) could also be combined to disentangle NO3- contamination from animal and human wastes.


Honey bee (Apis mellifera) colonies benefit from grassland/ pasture while bumble bee (Bombus impatiens) colonies in the same landscapes benefit from non-corn/soybean cropland.
Agriculturally important commercially managed pollinators including honey bees (Apis mellifera L., 1758) and bumble bees (Bombus impatiens Cresson, 1863) rely on the surrounding landscape to fulfill their dietary needs. A previous study in Europe demonstrated that managed honey bee foragers and unmanaged native bumble bee foragers are associated with different land uses. However, it is unclear how response to land use compares between managed honey bees and a managed native bumble bee species in the United States, where honey bees are an imported species. Furthermore, to our knowledge, no such direct comparisons of bee responses to land use have been made at the colony level. To better understand how two different social bees respond to variation in land use, we monitored the weights of A. mellifera and B. impatiens colonies placed in 12 apiaries across a range of land use in Michigan, United States in 2017. Bombus impatiens colonies gained more weight and produced more drones when surrounded by diverse agricultural land (i.e., non-corn/soybean cropland such as tree fruits and grapes), while honey bee colonies gained more weight when surrounded by more grassland/pasture land. These findings add to our understanding of how different bee species respond to agricultural landscapes, highlighting the need for further species-specific land use studies to inform tailored land management.


Validating the probe-to-bone test and other tests for diagnosing chronic osteomyelitis in the diabetic foot.
To compare the diagnostic characteristics of tests used for a prompt diagnosis of chronic osteomyelitis in the diabetic foot, using bone histology as the criterion standard. The tests assessed were probe-to-bone (PTB), clinical signs of infection, radiography signs of osteomyelitis, and ulcer specimen culture. A prospective study was performed on patients with foot ulcers referred to our diabetic foot clinic. Ulcer infection was diagnosed by recording clinical signs of infection and taking specimens for culture. The presumptive diagnosis of osteomyelitis was based on these results and the findings of a plain X-ray and PTB test. All patients with a clinical suspicion of bone infection were subjected to surgical treatment of the affected bone. During surgery, bone specimens were obtained for a histological diagnosis of osteomyelitis. Over 2.5 years, 210 foot lesions were consecutively examined and 132 of these wounds with clinical suspicion of infection selected as the study sample. Of these, 105 (79.5%) lesions were diagnosed as osteomyelitis. Among the tests compared, the best results were yielded by the PTB test including an efficiency of 94%, sensitivity of 98%, specificity of 78%, positive predictive value of 95%, and negative predictive value of 91% (P < 0.001, κ 0.803); the positive likelihood ratio was 4.41, and the negative likelihood ratio was 0.02 (95% CI). In our outpatient population with a high prevalence of osteomyelitis, the PTB test was of greatest diagnostic value, especially for neuropathic ulcers, and proved to be efficient for detecting osteomyelitis in the diabetic foot.


Analyzing a stochastic time series obeying a second-order differential equation.
The stochastic properties of a Langevin-type Markov process can be extracted from a given time series by a Markov analysis. Also processes that obey a stochastically forced second-order differential equation can be analyzed this way by employing a particular embedding approach: To obtain a Markovian process in 2N dimensions from a non-Markovian signal in N dimensions, the system is described in a phase space that is extended by the temporal derivative of the signal. For a discrete time series, however, this derivative can only be calculated by a differencing scheme, which introduces an error. If the effects of this error are not accounted for, this leads to systematic errors in the estimation of the drift and diffusion functions of the process. In this paper we will analyze these errors and we will propose an approach that correctly accounts for them. This approach allows an accurate parameter estimation and, additionally, is able to cope with weak measurement noise, which may be superimposed to a given time series.


Mirror, mirror on the wall: viewing a mirror reflection of unilateral hand movements facilitates ipsilateral M1 excitability.
Primary motor cortex (M1) excitability is modulated by both ipsilateral limb movement and passive observation of movement of the contralateral limb. An interaction of these effects within M1 may account for recent research suggesting improved functional recovery of the impaired arm following stroke by viewing a mirror reflection of movements of the unimpaired arm superimposed over the (unseen) impaired arm. This hypothesis was tested in the present study using single-pulse transcranial magnetic stimulation (TMS) in eight neurologically healthy subjects. Excitability of M1 ipsilateral to a phasic, unilateral hand movement was measured while subjects performed paced (1 Hz), unilateral index finger-thumb opposition movements. Motor evoked potentials (MEPs) were obtained from the inactive first dorsal interosseous (FDI) in each of four viewing conditions: Active (viewing the active hand), Central (viewing a mark positioned between hands), Inactive (viewing the inactive hand) and Mirror (viewing a mirror-reflection of the active hand in a mirror oriented in the mid-sagittal plane) and with both hands at rest (Rest). MEPs were significantly enhanced during ipsilateral hand movement compared with the Rest condition (P<0.05). Largest MEPs were obtained in the Mirror condition, and this was significant compared with both the Inactive and Central viewing conditions (P<0.05). There was no difference between the dominant and non-dominant hand. Excitability of M1 ipsilateral to a unilateral hand movement is facilitated by viewing a mirror reflection of the moving hand. This finding provides neurophysiological evidence supporting the application of mirror therapy in stroke rehabilitation.


DataHigh: graphical user interface for visualizing and interacting with high-dimensional neural activity.
The activity of tens to hundreds of neurons can be succinctly summarized by a smaller number of latent variables extracted using dimensionality reduction methods. These latent variables define a reduced-dimensional space in which we can study how population activity varies over time, across trials, and across experimental conditions. Ideally, we would like to visualize the population activity directly in the reduced-dimensional space, whose optimal dimensionality (as determined from the data) is typically greater than 3. However, direct plotting can only provide a 2D or 3D view. To address this limitation, we developed a Matlab graphical user interface (GUI) that allows the user to quickly navigate through a continuum of different 2D projections of the reduced-dimensional space. To demonstrate the utility and versatility of this GUI, we applied it to visualize population activity recorded in premotor and motor cortices during reaching tasks. Examples include single-trial population activity recorded using a multi-electrode array, as well as trial-averaged population activity recorded sequentially using single electrodes. Because any single 2D projection may provide a misleading impression of the data, being able to see a large number of 2D projections is critical for intuition-and hypothesis-building during exploratory data analysis. The GUI includes a suite of additional interactive tools, including playing out population activity timecourses as a movie and displaying summary statistics, such as covariance ellipses and average timecourses. The use of visualization tools like the GUI developed here, in tandem with dimensionality reduction methods, has the potential to further our understanding of neural population activity.


A Study on Standard for Safety Requirements for Care Robots.
Recently, care robots are being developed that incorporate robotics into assistive products that focus on daily care for the physically disabled or elderly with reduced physical function. However, although care robots can reduce the physical burden of human intervention, they can also be dangerous depending on their situational awareness. This study describes a standardization that defines safety requirements for care robots and includes verification methods to test their safety requirements. As an example of the application of this standard, a standard for the safety and performance method of a feeding robot is shown. This standardization study is expected to contribute to the spread of care robots in the future.


Synergistic interaction between cisplatin and gemcitabine in neuroblastoma cell lines and multicellular tumor spheroids.
The efficacy and mechanism of action of cisplatin and gemcitabine were investigated in a panel of neuroblastoma cell lines and multicellular tumor spheroids. In neuroblastoma spheroids, the combination of cisplatin and gemcitabine induced a complete cytostasis at clinical relevant concentrations. A synergistic effect was observed when cells were coincubated with both drugs or preincubated with gemcitabine first. These administration sequences resulted in NASS cells in decreased ERCC1 and XPA expression, two key proteins of the NER DNA repair system, and increased platinum adduct formation in DNA. Most of these phenomena were not observed in SJNB8 cells which might explain the lack of synergy between cisplatin and gemcitabine in SJNB8 cells. Our results showed favorable interactions between cisplatin and gemcitabine in 4 out of 5 cell lines. Therefore, we feel that inclusion of gemcitabine into cisplatin-containing regiments might be a promising new strategy for the treatment of neuroblastoma.


Phase Variations and Layer Epitaxy of 2D PdSe2 Grown on 2D Monolayers by Direct Selenization of Molecular Pd Precursors.
Two-dimensional (2D) materials and van der Waals heterostructures with atomic-scale thickness provide enormous potential for advanced science and technology. However, insufficient knowledge of compatible synthesis impedes wafer-scale production. PdSe2 and Pd2Se3 are two of the noble transition-metal chalcogenides with excellent physical properties that have recently emerged as promising materials for electronics, optoelectronics, catalyst, and sensors. This research presents a feasible approach to synthesize PdSe2 and Pd2Se3 with inherently asymmetric structure on honeycomb lattice 2D monolayer substrates of graphene and MoS2. We directly deposit a molecular transition-metal precursor complex on the surface of the 2D substrates, followed by low-temperature selenization by chemical vapor flow. Parameter control leads to tuning of the material from monolayer nanocrystals with Pd2Se3 phase, to continuous few-layer PdSe2 films. Annular dark-field scanning transmission electron microscopy (ADF-STEM) reveals the structure, phase variations, and heteroepitaxy at the atomic level. PdSe2 with unconventional interlayer stacking shifts appeared as the kinetic product, whereas the bilayer PdSe2 and monolayer Pd2Se3 are the thermodynamic product. The epitaxial alignment of interlayer rotation and translation between the PdSe2 and underlying 2D substrate was also revealed by ADF-STEM. These results offer both nanoscale and atomic-level insights into direct growth of van der Waals heterostructures, as well as an innovative method for 2D synthesis by predetermined nucleation.


Inhibition of Phosphoglycerate Mutase 5 Reduces Necroptosis in Rat Hearts Following Ischemia/Reperfusion Through Suppression of Dynamin-Related Protein 1.
Necroptosis is an important form of cell death following myocardial ischemia/reperfusion (I/R) and phosphoglycerate mutase 5 (PGAM5) functions as the convergent point for multiple necrosis pathways. This study aims to investigate whether inhibition of PGAM5 could reduce I/R-induced myocardial necroptosis and the underlying mechanisms. The SD rat hearts (or H9c2 cells) were subjected to 1-h ischemia (or 10-h hypoxia) plus 3-h reperfusion (or 4-h reoxygenation) to establish the I/R (or H/R) injury model. The myocardial injury was assessed by the methods of biochemistry, H&E (hematoxylin and eosin), and PI/DAPI (propidium iodide/4',6-diamidino-2-phenylindole) staining, respectively. Drug interventions or gene knockdown was used to verify the role of PGAM5 in I/R (or H/R)-induced myocardial necroptosis and possible mechanisms. The I/R-treated heart showed the injuries (increase in infarct size and creatine kinase release), upregulation of PGAM5, dynamin-related protein 1 (Drp1), p-Drp1-S616, and necroptosis-relevant proteins (RIPK1/RIPK3, receptor-interacting protein kinase 1/3; MLKL, mixed lineage kinase domain-like); these phenomena were attenuated by inhibition of PGAM5 or RIPK1. In H9c2 cells, H/R treatment elevated the levels of PGAM5, RIPK1, RIPK3, MLKL, Drp1, and p-Drp1-S616 and induced mitochondrial dysfunctions (elevation in mitochondrial membrane potential and ROS level) and cellular necrosis (increase in LDH release and the ratio of PI+/DAPI+ cells); these effects were blocked by inhibition or knockdown of PGAM5. Inhibition of PGAM5 can reduce necroptosis in I/R-treated rat hearts through suppression of Drp1; there is a positive feedback between RIPK1 and PGAM5, and PGAM5 might serve as a novel therapeutic target for prevention of myocardial I/R injury.


Potent inhibition by ropivacaine of metastatic colon cancer SW620 cell invasion and NaV1.5 channel function.
Metastatic breast and colon cancer cells express neonatal and adult splice variants of NaV1.5 voltage-activated Na(+) channels (VASCs). Block of VASCs inhibits cell invasion. Local anaesthetics used during surgical tumour excision inhibit VASC activity on nociceptive neurones providing regional anaesthesia. Inhibition of VASCs on circulating metastatic cancer cells may also be beneficial during the perioperative period. However, ropivacaine, frequently used to provide analgesia during tumour resection, has not been tested on colon cancer cell VASC function or invasion. We used reverse transcription-polymerase chain reaction and sequencing to identify NaV1.5 variants in the SW620 metastatic colon cancer cell line. Recombinant adult and neonatal NaV1.5 variants were expressed in human embryonic kidney cells. Voltage-clamp recordings and invasion assays were used to examine the effects of ropivacaine on recombinant NaV1.5 channels and the metastatic potential of SW620 cells, respectively. SW620 cells expressed adult and neonatal NaV1.5 variants, which had similar steady-state inactivation profiles, but distinctive activation curves with the neonatal variant having a V1/2 of activation 7.8 mV more depolarized than the adult variant. Ropivacaine caused a concentration-dependent block of both NaV1.5 variants, with IC50 values of 2.5 and 3.9 µM, respectively. However, the reduction in available steady-state current was selective for neonatal NaV1.5 channels. Ropivacaine inhibited SW620 invasion, with a potency similar to that of inhibition of NaV1.5 channels (3.8 µM). Ropivacaine is a potent inhibitor of both NaV1.5 channel activity and metastatic colon cancer cell invasion, which may be beneficial during surgical colon cancer excision.


4D scanning ultrafast electron microscopy: visualization of materials surface dynamics.
The continuous electron beam of conventional scanning electron microscopes (SEM) limits the temporal resolution required for the study of ultrafast dynamics of materials surfaces. Here, we report the development of scanning ultrafast electron microscopy (S-UEM) as a time-resolved method with resolutions in both space and time. The approach is demonstrated in the investigation of the dynamics of semiconducting and metallic materials visualized using secondary-electron images and backscattering electron diffraction patterns. For probing, the electron packet was photogenerated from the sharp field-emitter tip of the microscope with a very low number of electrons in order to suppress space-charge repulsion between electrons and reach the ultrashort temporal resolution, an improvement of orders of magnitude when compared to the traditional beam-blanking method. Moreover, the spatial resolution of SEM is maintained, thus enabling spatiotemporal visualization of surface dynamics following the initiation of change by femtosecond heating or excitation. We discuss capabilities and potential applications of S-UEM in materials and biological science.


Molecular pathogenesis of intrahepatic cholangiocarcinoma.
Cholangiocarcinoma (CCA) is an orphan cancer of the hepatobiliary tract, the incidence of which has increased in the past decade. The molecular pathogenesis of this treatment-refractory disease is poorly understood. Desmoplasia is a key causal feature of CCA; however, a majority of tumors develop with no apparent etiological background. The impact of the stromal compartment on tumor progression as well as resistance to therapy is in vogue, and the epithelial-stromal crosstalk may present a target for novel treatment strategies. As such, the complexity of tumor cellularity and the molecular mechanisms underlying the diversity of growth patterns of this malignancy remain a clinical concern. It is crucial to advance our present understanding of the molecular pathogenesis of CCA to improve current clinical strategies and patient outcome. This will facilitate the delineation of patient subsets and individualization for precision therapies. Many questions persevere as to the evolutionary process and cellular origin of the initial transforming event, the context of intratumoral plasticity and the causal driver action. Next-generation sequencing has begun to underline the persistent alterations, which may be the trigger of acquired drug resistance, and the cause of metastasis and disease recurrence. A complex issue that remains is to account for the heterogeneous pool of "backseat" aberrations, which in chromosomal proximity to the causative variant are likely to influence, for example, drug response. This review explores the recent advances in defining the molecular pathways implicated in the development of this devastating disease and, which present putative clinical strategies.


[Deaths of patients with tuberculosis in combination with HIV infection from various causes, other than tuberculosis].
The structure of tuberculosis patients with HIV infection who died from any causes, other than tuberculosis, in the Sverdlovsk Region in 2000-2006 did not differ from that of patients with concurrent pathology who died from tuberculosis. The general health care facilities had revealed tuberculosis during life in most patients who died from terminal-stage HIV infection; at death these patients had endstage HIV infection which tuberculosis joined to 4 years or more after HIV infection registration. A fatal outcome in 88.1% of the patients died from the terminal stage of HIV infection was observed within the first year after tuberculosis detection, in half the cases the background disease being generalized tuberculosis at autopsy; every two patients discharged microorganisms during life, drug resistance in the causative agent of tuberculosis was found in every three patients. Half the patients who died from other causes, other than HIV infection and tuberculosis lost their life within the first year after registration of the tuberculous process. Lifetime bacterial discharge was recorded in half the patients; drug resistance of Mycobacterium tuberculosis has developed a third of the patients with comorbidity who died from other causes, other than tuberculosis and HIV infection. The volume of lifetime specialized care for HIV infection to deceased patients with comorbidity had been inadequate. The importance of the problem of notification of cases, when the autopsy background disease was tuberculosis in comorbidity patients not included into the regional tuberculosis morbidity and mortality statistics, will increase with further development of the epidemics of tuberculosis and HIV infection to the Svedlovsk Region.


Furanocoumarin Notopterol: Inhibition of Hepatocellular Carcinogenesis through Suppression of Cancer Stemness Signaling and Induction of Oxidative Stress-Associated Cell Death.
Hepatocellular carcinoma (HCC) remains an aggressive malignancy with a poor prognosis and a leading cause of cancer-related mortality globally. Cumulative evidence suggests critical roles for endoplasmic reticulum (ER) stress and unfolded protein response (UPR) in chronic liver diseases. However, the role of ER stress in HCC pathogenesis, aggressiveness and therapy response remains unclear and understudied. Against this background, the present study evaluated the therapeutic efficacy and feasibility of notopterol (NOT), a furanocoumarin and principal component of Notopterygium incisum, in the modulation of ER stress and cancer stemness, and the subsequent effect on liver oncogenicity. An array of biomolecular methods including Western blot, drug cytotoxicity, cell motility, immunofluorescence, colony and tumorsphere formation, flow-cytometric mitochondrial function, GSH/GSSG ratio, and tumor xenograft ex vivo assays were used in the study. Herein, we demonstrated that NOT significantly suppresses the viability, migration, and invasion capacity of the human HCC HepJ5 and Mahlavu cell lines by disrupting ATF4 expression, inhibiting JAK2 activation, and downregulating the GPX1 and SOD1 expression in vitro. NOT also markedly suppressed the expression of vimentin (VIM), snail, b-catenin, and N-cadherin in the HCC cells, dose-dependently. Treatment with NOT significantly attenuated cancer stem cells (CSCs)-like phenotypes, namely colony and tumorsphere formation, with the concomitant downregulation of stemness markers OCT4, SOX2, CD133, and upregulated PARP-1 cleavage, dose-dependently. We also demonstrated that NOT anticancer activity was strongly associated with increased cellular reactive oxidative stress (ROS) but, conversely, reduced mitochondrial membrane potential and function in the HepJ5 and Mahlavu cells in vitro. Our tumor xenograft studies showed that compared with sorafenib, NOT elicited greater tumor growth suppression without adverse changes in mice body weights. Compared with the untreated control and sorafenib-treated mice, NOT-treated mice exhibited markedly greater apoptosis ex vivo, and this was associated with the co-suppression of stemness and drug-resistance markers OCT4, SOX2, ALDH1, and the upregulation of endoplasmic reticulum stress and oxidative stress factors PERK and CHOP. In summary, we demonstrated for the first time that NOT exhibits strong anticancer activity via the suppression of cancer stemness, enhanced endoplasmic reticulum stress and increased oxidative stress thus projecting NOT as a potentially effective therapeutic agent against HCC.


[Experiences with scoring systems SAPS II and NEMS for registration of activities in an intensive care unit].
Intensive care is highly expensive, hence the requirement of efficient use of resources calls for evaluation of the severity of disease or injury for each individual patient, so that management may be optimised. At Ullevaal University Hospital, two different scoring systems have been used since 1996: SAPS II (Simplified Acute Physiology Score) for severity of illness and NEMS (Nine Equivalents of Nursing Manpower Use Score) for assessment of resource utilisation. A significant relationship between severity of illness and mortality was found. Length of stay in ICU correlated with increasing SAPS II score in the range 0 to 50, but at higher scores no relationship was found. Patients with trauma and cardiovascular surgical patients used most of the resources. 79% of total NEMS score consisted of vital signs monitoring, intravenous drug medication and mechanical ventilation of the patients. SAPS II and NEMS scoring systems provide useful information needed in the assessment of patient care in the ICU and are helpful in the documentation of financial and medical resource needs.


Novel omega-conotoxins from Conus catus discriminate among neuronal calcium channel subtypes.
omega-Conotoxins selective for N-type calcium channels are useful in the management of severe pain. In an attempt to expand the therapeutic potential of this class, four new omega-conotoxins (CVIA-D) have been discovered in the venom of the piscivorous cone snail, Conus catus, using assay-guided fractionation and gene cloning. Compared with other omega-conotoxins, CVID has a novel loop 4 sequence and the highest selectivity for N-type over P/Q-type calcium channels in radioligand binding assays. CVIA-D also inhibited contractions of electrically stimulated rat vas deferens. In electrophysiological studies, omega-conotoxins CVID and MVIIA had similar potencies to inhibit current through central (alpha(1B-d)) and peripheral (alpha(1B-b)) splice variants of the rat N-type calcium channels when coexpressed with rat beta(3) in Xenopus oocytes. However, the potency of CVID and MVIIA increased when alpha(1B-d) and alpha(1B-b) were expressed in the absence of rat beta(3), an effect most pronounced for CVID at alpha(1B-d) (up to 540-fold) and least pronounced for MVIIA at alpha(1B-d) (3-fold). The novel selectivity of CVID may have therapeutic implications. (1)H NMR studies reveal that CVID possesses a combination of unique structural features, including two hydrogen bonds that stabilize loop 2 and place loop 2 proximal to loop 4, creating a globular surface that is rigid and well defined.


Utility of routine surveillance blood cultures in asymptomatic allogeneic hematopoietic stem cell transplant recipients with indwelling central venous catheters at a comprehensive cancer center.
Many transplant centers obtain surveillance blood cultures (SBCs) from asymptomatic allogeneic hematopoietic stem cell transplant (allo-HCT) recipients with central venous catheters for early detection of potential blood stream infections. The aim of this study was to determine the utility of this practice. We conducted a retrospective study of all patients who underwent allo-HCT to determine the frequency, clinical significance, and costs associated with SBCs. From 776 patients, 6,801 SBCs were obtained (median, 9 per patient). Most (96.89%) were negative. Of the 211 positive SBCs, 171 (81%) had minimal clinical significance. The remaining 40 positive cultures (19%) were considered potentially significant. The frequency of potentially significant SBCs was 5.1% for the entire cohort and 0.59% of all SBCs drawn. All potentially significant cultures and some that were deemed to have minimal significance led to medical intervention, some of which were probably unnecessary. No adverse outcomes occurred in patients with positive SBCs for the first 30 days following the positive result, regardless of the pathogen isolated or the quantitative colony count. The frequency of clinically significant positive SBCs in asymptomatic adult allo-HCT recipients is very low. Routine use of this practice leads to some unnecessary medical interventions and added costs.


Evaluation of Improvements in the Separation of Monolayer and Multilayer Films via Measurements in Transflection and Application of Machine Learning Approaches.
Small plastic packaging films make up a quarter of all packaging waste generated annually in Austria. As many plastic packaging films are multilayered to give barrier properties and strength, this fraction is considered hardly recyclable and recovered thermally. Besides, they can not be separated from recyclable monolayer films using near-infrared spectroscopy in material recovery facilities. In this paper, an experimental sensor-based sorting setup is used to demonstrate the effect of adapting a near-infrared sorting rig to enable measurement in transflection. This adaptation effectively circumvents problems caused by low material thickness and improves the sorting success when separating monolayer and multilayer film materials. Additionally, machine learning approaches are discussed to separate monolayer and multilayer materials without requiring the near-infrared sorter to explicitly learn the material fingerprint of each possible combination of layered materials. Last, a fast Fourier transform is shown to reduce destructive interference overlaying the spectral information. Through this, it is possible to automatically find the Fourier component at which to place the filter to regain the most spectral information possible.


Two weeks of western diet disrupts liver molecular markers of cholesterol metabolism in rats.
The present study was designed to test the hypothesis that in the liver, excessive fat accumulation impairs cholesterol metabolism mainly by altering the low-density lipoprotein-receptor (LDL-R) pathway. Young male Wistar rats were fed standard (SD), high fat (HFD; 60% kcal) or Western (WD; 40% fat + 35% sucrose (17.5% fructose)) diets for 2 or 6 weeks. Weight gain (~ 40 g) was observed only following 6 weeks of the obesogenic diets (P < 0.01). Compared to the 2-week treatment, obesogenic diets tripled fat pad weight (~ 20 vs 7 g) after 6 weeks. Hepatic triglyceride (TG) levels were greater in response to both the WD and HFD compared to the SD (P < 0.01) at 2 and 6 weeks and their concentrations were greater (P < 0.05) in WD than HFD at 2 weeks. Plasma total cholesterol levels were higher (P < 0.05) in animals submitted to WD. After 2 and 6 weeks, liver expression of LDL-R, proprotein convertase subtilisin/kexin 9 (PCSKk9) and sterol regulatory element binding protein 2 (SREBP2), involved in LDL-cholesterol uptake, was lower in animals submitted to WD than in others treated with HFD or SD (P < 0.01). Similarly, low-density lipoprotein-receptor-related protein 1 (LRP1) and acyl-CoA cholesterol acyltransferase-2 (ACAT-2) mRNA levels were lower (P < 0.01) among WD compared to SD-fed rats. Expression of the gene coding the main regulator of endogenous cholesterol synthesis, 3-hydroxy-3-methyl-glutaryl-CoA reductase (HMGCoAR) was reduced in response to WD compared to SD and HFD at 2 (P < 0.001) and 6 (P < 0.05) weeks. Being enriched in fructose, the WD strongly promoted the expression of carbohydrate-response element binding protein (ChREBP) and acetyl-CoA carboxylase (ACC), two key regulators of de novo lipogenesis. These results show that the WD promptly increased TG levels in the liver by potentiating fat storage. This impaired the pathway of hepatic cholesterol uptake via the LDL-R axis, promoting a rapid increase in plasma total cholesterol levels. These results indicate that liver fat content is a factor involved in the regulation of plasma cholesterol.


Community analysis of ammonia oxidizer in the oxygen-limited nitritation stage of OLAND system by DGGE of PCR amplified 16S rDNA fragments and FISH.
OLAND (oxygen limited autotrophic nitrification and denitrification) nitrogen removal system was constructed by coupling with oxygen limited nitritation stage and anaerobic ammonium oxidation stage. Ammonia oxidizer, as a kind of key bacteria in N cycle, plays an important role at the oxygen limited nitritation stage of OLAND nitrogen removal system. In this study, specific amplification of 16S rDNA fragment of ammonia oxidizer by nested PCR, separation of mixed PCR samples by denaturing gradient gel electrophoresis (DGGE), and the quantification of ammonia oxidizer by fluorescence in situ hybridization (FISH) were combined to investigate the shifts of community composition and quantity of ammonia oxidizer of the oxygen limited nitritation stage in OLAND system. It showed that the community composition of ammonia oxidizer changed drastically when dissolved oxygen was decreased gradually, and the dominant ammonia oxidizer of the steady nitrite accumulation stage were completely different from that of the early stage of oxygen limited nitritation identified by DGGE. It was concluded that the Nitrosomonas may be the dominant genus of ammonia oxidizer at the oxygen limited nitritation stage of OLAND system characterized by nested PCR-DGGE and FISH, and the percentage of Nitrosomonas was 72.5% +/- 0.8% of ammonia oxidizer at the steady nitrite accumulation stage detected by FISH.


Non-conscious choice in cutaneous backward masking.
A simple but effective method has been devised to explore backward masking in cutaneous sensation. In this method a weak electrical shock to the palm (test stimulus) is followed by a stronger shock to the same site (masking stimulus). By combining this stimulation strategy with a choice reaction time paradigm, it has been possible to show that the nervous system responds to the test stimuli even though they are not consciously perceived. Further, the distribution of errors in the different types of trial suggests that, under these experimental conditions, the identification of the stimulus and the choice of response may also be made at a non-conscious level. A tentative scheme for the timing of the various neural events is proposed.


PATHWAYS FOR BIOSYNTHESIS OF A BACTERIAL CAPSULAR POLYSACCHARIDE. IV. CAPSULE RESYNTHESIS BY DECAPSULATED RESTING-CELL SUSPENSIONS.
Juni, Elliot (Emory University, Atlanta, Ga.), and Gloria A. Heym. Pathways for biosynthesis of a bacterial capsular polysaccharide. IV. Capsule resynthesis by decapsulated resting-cell suspensions. J. Bacteriol. 87:461-467. 1964.-Methods were devised for stripping capsules from encapsulated bacteria. By use of stripped resting-cell suspensions of a gram-negative capsule-forming coccus, it was shown that polysaccharide capsule resynthesis depends upon the presence of air and an oxidizable substrate. Capsule resynthesis proceeds linearly with time. For a given quantity of stripped cells, the net amount of polysaccharide capsule synthesized is a linear function of the amount of substrate oxidized. The only factor that appears to limit the extent of capsule synthesis by resting cells is the amount of substrate utilized. A series of photomicrographs of wet mounts made in India ink show the appearance of stripped cells and resynthesized capsules formed during oxidation of pyruvate and glucose by stripped resting-cell suspensions.


Recovery from Bell Palsy after Transplantation of Peripheral Blood Mononuclear Cells and Platelet-Rich Plasma.
Peripheral blood mononuclear cells (PBMCs) are multipotent, and plasma contains growth factors involving tissue regeneration. We hypothesized that transplantation of PBMC-plasma will promote the recovery of paralyzed facial muscles in Bell palsy. This case report describes the effects of PBMC-plasma transplantations in a 27-year-old female patient with right side Bell palsy. On the affected side of the face, the treatment resulted in both morphological and functional recovery including voluntary facial movements. These findings suggest that PBMC-plasma has the capacity of facial muscle regeneration and provides a promising treatment strategy for patients suffering from Bell palsy or other neuromuscular disorders.


Comparison of adriamycin (NSC-123127) and the combination of 5-fluorouracil (NSC-19893) and cyclophosphamide (NSC-26271) in advanced prostatic cancer: a preliminary report.
A preliminary report on chemotherapy for advanced prostate malignancy is presented. Nine patients have been entered in the study, of which eight are evaluable for tumor response. Of four patients initially treated with adriamycin, only one brief (5-week) objective response was noted in a patient with soft tissue disease. No responses have been seen in four patients treated with the combination of 5-fluorouracil and cyclophosphamide. No responses have been noted in five patients who failed initial treatment and were crossed over to the other treatment. Durg toxicity has been quite acceptable.


Arabidopsis CULLIN4 Forms an E3 Ubiquitin Ligase with RBX1 and the CDD Complex in Mediating Light Control of Development.
Repression of photomorphogenesis in Arabidopsis thaliana requires activity of the COP9 signalosome (CSN), CDD, and COP1 complexes, but how these three complexes work in concert to accomplish this important developmental switch has remained unknown. Here, we demonstrate that Arabidopsis CULLIN4 (CUL4) associates with the CDD complex and a common catalytic subunit to form an active E3 ubiquitin ligase both in vivo and in vitro. The partial loss of function of CUL4 resulted in a constitutive photomorphogenic phenotype with respect to morphogenesis and light-regulated gene expression. Furthermore, CUL4 exhibits a synergistic genetic interaction with COP10 and DET1. Therefore, this CUL4-based E3 ligase is essential for the repression of photomorphogenesis. This CUL4-based E3 ligase appears to associate physically with COP1 E3 ligase and positively regulates the COP1-dependent degradation of photomorphogenesis-promoting transcription factors, whereas the CSN controls the biochemical modification of CUL4 essential for E3 activity. Thus, this study suggests a biochemical activity connection between CSN and CDD complexes in their cooperation with COP1 in orchestrating the repression of photomorphogenesis.


The evolution of cranial base and face in Cercopithecoidea and Hominoidea: Modularity and morphological integration.
The evolutionary relationship between the base and face of the cranium is a major topic of interest in primatology. Such areas of the skull possibly respond to different selective pressures. Yet, they are often said to be tightly integrated. In this paper, we analyzed shape variability in the cranial base and the facial complex in Cercopithecoidea and Hominoidea. We used a landmark-based approach to single out the effects of size (evolutionary allometry), morphological integration, modularity, and phylogeny (under Brownian motion) on skull shape variability. Our results demonstrate that the cranial base and the facial complex exhibit different responses to different factors, which produces a little degree of morphological integration between them. Facial shape variation appears primarily influenced by body size and sexual dimorphism, whereas the cranial base is mostly influenced by functional factors. The different adaptations affecting the two modules suggest they are best studied as separate and independent units, and that-at least when dealing with Catarrhines-caution must be posed with the notion of strong cranial integration that is commonly invoked for the evolution of their skull shape.


Passive and active colloidal chemotaxis in a microfluidic channel: mesoscopic and stochastic models.
Chemotaxis is the response of a particle to a gradient in the chemical composition of the environment. While it was originally observed for biological organisms, it is of great interest in the context of synthetic active particles such as nanomotors. Experimental demonstration of chemotaxis for chemically-powered colloidal nanomotors was reported in the literature in the context of chemo-attraction in a still fluid or in a microfluidic channel where the gradient is sustained by a specific inlet geometry. In this work, we use mesoscopic particle-based simulations of the colloid and solvent to demonstrate chemotaxis in a microfluidic channel. On the basis of this particle-based model, we evaluate the chemical concentration profiles in the presence of passive or chemically active colloids, compute the chemotactic force acting upon them and propose a stochastic model that rationalises our findings on colloidal chemotaxis. Our model is also able to explain the results of an earlier simulation work that uses a simpler geometry and to extend its interpretation.


Purification and partial characterization of a soluble suppressor factor from papillomavirus-infected patients.
A molecular form of soluble suppressor factor was identified in serum-free supernatant fluid of cultured peripheral blood mononuclear cells (PBMC) from patients with severe forms of papillomavirus infections (epidermodysplasia verruciformis and large treatment-resistant condyloma acuminatum). The papillomavirus-induced soluble suppressor factor (SSF-H) was highly purified by gel filtration, anion-exchange chromatography, chromatofocusing, hydrophobic chromatography, and nondissociating polyacrylamide gel electrophoresis (PAGE). The factor exhibited an apparent molecular weight of 67 kDa as determined by sodium dodecylsulfate-PAGE. The isoelectric point ranged from 4.8 to 5.2 as judged by chromatofocusing. The biologic activity of the SSF-H during purification and in subsequent characterization was examined by in vitro CTLL bioassay. Purified SSF-H was stable at 56 degrees C for 1 h and in the pH range of 4.0 to 8.0. Complete inactivation was observed at 80 degrees C. Trypsin and proteinase K treatment destroyed the biologic activity associated with purified SSF-H. This factor seems to be different in its molecular weight and other physicochemical properties from the known suppressor factors that affect lymphocyte functions.


In vitro investigations with the histamine H1 receptor antagonist, epinastine (WAL 801 CL), on isolated human allergic effector cells.
Skin mast cells, basophils and eosinophils are effector cells of acute and subacute allergic responses due to their capacity to produce a large number of (pro)inflammatory mediators. Histamine H1 receptor antagonists, such as epinastine (WAL 801 CL), have been described to partially exert antiallergic and antiinflammatory effects both in vivo and in vitro in addition to their antihistaminergic properties. The aim of the present study was to investigate whether epinastine could influence the in vitro activation of isolated human skin mast cells, basophils and eosinophils induced by different secretagogues. Cells were isolated from healthy women following plastic surgery and healthy blood donors, respectively. Mast cells were isolated by enzymatic digestion of the skin. Blood cells were isolated by gradient centrifugation and negative selection with magnetic beads. A wide range of concentrations of the drug (1 nmol/l to 100 micromol/l) did not significantly inhibit histamine release from basophils induced by immunologic (anti-IgE, concanavalin A, priming factors interleukin-3 and interleukin-5) and non immunologic (A23187, ionomycin, 12-o-tetradecanoyl-phorbol-13-acetate, C5a, formyl-methionyl-leucyl-phenylalanine) stimuli. Furthermore, the drug had no effect on A23187-induced release of eosinophil cationic protein from eosinophils. However, at a concentration >0.1 nmol/l, IgE-mediated LTC4 production from basophils was significantly suppressed. Histamine release from skin mast cells due to anti-IgE or A23187 was inhibited by epinastine in a dose-dependent fashion, whereas substance P-induced activation as well as stem cell factor priming were not. Epinastine did not inhibit isolated protein kinase C from rat brain. The results confirm previous in vivo and in vitro observations obtained from animal models that epinastine exerts antiallergic and antiinflammatory effects. Whether the observed effects are due to non specific membrane interactions or by influencing intracellular signal transduction elements has to be further elucidated.


Veno-venous extra-corporeal membrane oxygenation-assisted right tracheal-sleeve pneumonectomy.
Tracheal sleeve pneumonectomy for lung cancer is an old technique, and it is reserved for exceptional cases with tracheal carina involvement. Intra-operative airways management of this operation is incredibly complex, involving thoracic surgeons, anaesthesiologists and pulmonologists. We report a case of a 38-year-old male with no clinical history, referred to our department for an adenoid-cystic carcinoma involving distal trachea, carina and main right bronchus. Tracheal sleeve pneumonectomy was performed using extra-corporeal membrane oxygenation (ECMO). A veno-venous ECMO circuit was established through a heparin-coated percutaneous cannula in the right femoral vein and a heparin-coated percutaneous cannula in the internal right jugular vein by ultrasound assistance. No major complications occurred, and the patient was discharged after 30-day bronchoscopic control, showing the absence of fistula and negativity of the methylene blue test. ECMO-assisted surgery ensures adequate respiratory support, haemodynamic stability, lower risk of bleeding complications with a clean operating field and better brain and myocardial oxygenation.


Bioequivalence of two intravenous formulations of antithrombin III: a two-way crossover study in healthy Korean subjects.
Treatment with antithrombin (AT)-III is indicated for patients with sepsis or hereditary AT deficiency. The purpose of this study was to compare the pharmacokinetic and pharmacodynamic characteristics of 2 AT-III formulations in healthy Korean volunteers to satisfy the regulatory requirements for bioequivalence for marketing purposes. A single-center, single-dose, open-label, randomized, 2-period, 2-sequence crossover study was conducted in healthy Korean volunteers. Blood samples for the drug analysis were collected for up to 216 hours after drug administration. Participants received either the test or reference formulation of AT-III 100 U/kg IV for 20 minutes in the first period and the alternative formulation in the second period. Both the AT-III activity and antigen (Ag) were measured for the analysis of pharmacokinetic properties, and the prothrombin time and the activated partial thromboplastin time were assessed for the analysis of pharmacodynamic properties. Because AT-III is an endogenous compound, the analysis used data corrected from baseline values. The tolerability of the 2 formulations was also assessed based on physical examinations including vital sign measurements, laboratory tests, and 12-lead ECG. Of the 20 subjects enrolled (mean [SD] age, height, and weight, 25.3 [2.3] years, 175.3 [4.5] cm, and 67.4 [6.3] kg, respectively), 19 completed both treatment periods; 1 subject withdrew consent for personal reasons. The observed mean (SD) Cmax, AUClast, and AUC0-∞ of AT-III activity were, respectively, 279.24% (35.92), 14,364.10 (2325.25) %·h, and 17,526.38 (3150.81) %·h with the test formulation and 249.75% (31.96), 12,962.95 (1897.52) %·h, and 15,957.67 (3189.21) %·h with the reference formulation. The observed mean (SD) Cmax, AUClast, and AUC0-∞ of AT-III Ag were 62.58 (5.66) mg/dL, 3051.94 (401.87) mg/dL·h, and 3639.80 (726.01) mg/dL·h, respectively, with the test formulation and 58.63 (5.27) mg/dL, 2805.08 (272.38) mg/dL·h, and 3340.00 (428.46) mg/dL·h with the reference formulation. The geometric mean ratios (90% CI) of the log-transformed data for AT-III activity between the 2 formulations were 1.11494 (1.08994-1.14053) for Cmax, 1.11305 (1.05435-1.17503) for AUClast, and 1.11527 (1.03754-1.19889) for AUC0-∞; corresponding values for AT-III Ag were 1.08802 (1.06258-1.11405), 1.10905 (1.05804-1.16242), and 1.11460 (1.02058-1.21726). During the study period, 8 adverse events were reported, and all were transient, mild, and resolved completely during the treatment period. The results of the present study showed that these 2 AT-III formulations met the regulatory criteria for pharmacokinetic bioequivalence with respect to AT-III activity and Ag in these healthy Korean subjects. ClinicalTrials.gov identifier: NCT00846274.


Fatty acyl desaturation in isolated hepatocytes from Atlantic salmon (Salmo salar): stimulation by dietary borage oil containing gamma-linolenic acid.
The effects of different dietary oils on the fatty acid compositions of liver phospholipids and the desaturation and elongation or [1-14C]18:3n-3 and [1-14C]18:2n-6 were investigated in isolated hepatocytes from Atlantic salmon. Atlantic salmon smolts were fed diets containing either a standard fish oil (FO) as a control diet, a 1:1 blend of Southern Hemisphere marine oil and tuna orbital oil (MO/TO), sunflower oil (SO), borage oil (BO), or olive oil (OO) for 12 wk. The SO and BO diets significantly increased the percentages of 18:2n-6, 18:3n-6, 20:2n-6, 20:3n-6, and total n-6 polyunsaturated fatty acids (PUFA) in salmon liver lipids in comparison with the FO diet. The BO diet also increased the percentage of 20:4n-6. Both the SO and BO diets significantly reduced the percentages of all n-3 PUFA in comparison with the FO diet. The OO diet significantly increased the percentages of 18:1n-3, 18:2n-6, total monoenes, and total n-6 PUFA in liver lipids compared to the FO diet, and the percentages of all n-3 PUFA were significantly reduced. With [1-14C]18:3n-3, the recovery of radioactivity in the products of delta 6 desaturation was significantly greater in the hepatocytes from salmon fed SO, BO, and OO in comparison with the FO diet. The BO diet also increased the recovery of radioactivity in the products of delta 5 desaturation. Only the BO diet significantly affected the desaturation of [1-14C]18:2n-6, increasing recovery of radioactivity in both delta 6- and delta 5-desaturation products. In conclusion, dietary BO, enriched in gamma-linolenic acid (18:3n-6), significantly increased the proportions of both 20:3n-6 and 20:4n-6 in salmon liver phospholipids and also significantly increased the desaturation of both 18:2n-6 and 18:3n-3 in salmon hepatocytes. The possible relationships between dietary fatty acid composition, tissue phospholipid fatty acid composition, and desaturation/elongation activities are discussed.


Neuroprotective potential of erythropoietin and its derivative carbamylated erythropoietin in periventricular leukomalacia.
Periventricular leukomalacia (PVL) is the predominant pathology in premature infants, characterized by prominent cerebral white matter injury, and commonly caused by hypoxia-ischemia and inflammation. Activated microglia trigger white matter damage and play a major role in the development of PVL. Erythropoietin (EPO) and its derivative carbamylated erythropoietin (CEPO) have been shown to be neuroprotective in several brain disease models. Here we investigated whether EPO and CEPO could provide protection in mouse models of PVL induced by hypoxia-ischemia or hypoxia-ischemia-inflammation. We administered EPO or CEPO to mice with PVL, and found that both EPO and CEPO treatments decreased microglia activation, oligodendrocyte damage and myelin depletion. We also noted improved performance in neurological function assays. Inhibited disease progression in PVL mice by EPO or CEPO treatment was associated with decreased poly-(ADP-ribose) polymerase-1 (PARP-1) activity. PARP-1 activity was increased dramatically in activated microglia in untreated mice with PVL. Furthermore, we demonstrated that the neuroprotective properties of EPO and CEPO were diminished after PARP-1 gene depletion. The therapeutic doses of EPO and CEPO used in this study did not interfere with normal oligodendrocyte maturation and myelination. Together, our data demonstrate that EPO and CEPO are neuroprotective in cerebral white matter injury via a novel microglial PARP-1 dependent mechanism, and hold promise as a future treatment for PVL and other hypoxic-ischemic/inflammatory white matter diseases.


[Prayers for patients with internal and cardiological diseases--an applicable therapeutic method?].
Previous intercessory prayer studies showed symptom improvement and lower rates of complications in patients who were prayed for, whether it was a direct prayer or a distant intercessory prayer. The effect of intercessory prayer was not observed in cardiology patients during two new multicentre studies (MANTRA, STEP). The STEP study actually showed an unfavourable effect when the patient knew of distant prayers made on his/her behalf. Thus, prayer as an "applicable therapeutic method" could not be empirically verified. However, active prayer within the framework of a doctor-patient relationship can strengthen the patient's optimism and activate the body's healing resources.


Monoclonal antibody radioimmunodetection of human-derived colon cancer.
This study was designed to determine whether monoclonal antibody directed against carcinoembryonic antigen could successfully be used in the scintigraphic localization of a human-derived colon carcinoma in a hamster model. An immunoglobulin G (IgG)-1 kappa monoclonal antibody, prepared in this laboratory, against carcinoembryonic antigen (CEA) was radiolabeled with iodine-131 (131I). Four Syrian hamsters bearing GW-39 human colon cancers received intracardiac injections of 50 mu Ci of 131I (14 micrograms of antibody). Gamma camera images were obtained at 24-hour intervals. Animals were sacrificed at 11 days, and the tumors and entire animals were counted. A double-label antibody experiment was conducted with 131I anti-CEA and nonspecific MOPC 21 IgG iodine-125 (125I) to assess localization specificity. The scintiphotos clearly showed the tumor at 24 hours, but there was significant background (blood-pool activity). Later images at six and 11 days showed a gradual decrease in background activity and more clear definition of the tumor. Animals sacrificed at 11 days showed 48-80% of residual whole body radioactivity to be present in the tumor. However, these tumors were large at sacrifice, weighing 8.9 to 12.4 g. Specific localization was confirmed by the double-label experiments where specific localization was twice nonspecific accretion of IgG in the tumor. This study has shown that a specific monoclonal antibody can successfully be used to scintigraphically localize a colon tumor of human origin. Although clearance of background activity is a gradual process, eventually most radioactivity left in the animal is localized in the tumor. This study illustrates that the potential radiolabeled monoclonal antibodies hold as immunodiagnostic agents.


Respiratory syncytial virus induces NRF2 degradation through a promyelocytic leukemia protein - ring finger protein 4 dependent pathway.
Respiratory syncytial virus (RSV) is the most important cause of viral acute respiratory tract infections and hospitalizations in children, for which no vaccine or specific treatments are available. RSV causes airway mucosa inflammation and cellular oxidative damage by triggering production of reactive oxygen species and by inhibiting at the same time expression of antioxidant enzymes, via degradation of the transcription factor NF-E2-related factor 2 (NRF2). RSV infection induces NRF2 deacetylation, ubiquitination, and degradation through a proteasome-dependent pathway. Although degradation via KEAP1 is the most common mechanism, silencing KEAP1 expression did not rescue NRF2 levels during RSV infection. We found that RSV-induced NRF2 degradation occurs in an SUMO-specific E3 ubiquitin ligase - RING finger protein 4 (RNF4)-dependent manner. NRF2 is progressively SUMOylated in RSV infection and either blocking SUMOylation or silencing RNF4 expression rescued both NRF2 nuclear levels and transcriptional activity. RNF4 associates with promyelocytic leukemia - nuclear bodies (PML-NBs). RSV infection induces the expression of PML and PML-NBs formation in an interferon (INF)-dependent manner and also induces NRF2 - PMN-NBs association. Inhibition of PML-NB formation by blocking IFN pathway or silencing PML expression resulted in a significant reduction of RSV-associated NRF2 degradation and increased antioxidant enzyme expression, identifying the RNF4-PML pathway as a key regulator of antioxidant defenses in the course of viral infection.


Generalized estimating equations in cluster randomized trials with a small number of clusters: Review of practice and simulation study.
Generalized estimating equations are a common modeling approach used in cluster randomized trials to account for within-cluster correlation. It is well known that the sandwich variance estimator is biased when the number of clusters is small (≤40), resulting in an inflated type I error rate. Various bias correction methods have been proposed in the statistical literature, but how adequately they are utilized in current practice for cluster randomized trials is not clear. The aim of this study is to evaluate the use of generalized estimating equation bias correction methods in recently published cluster randomized trials and demonstrate the necessity of such methods when the number of clusters is small. Review of cluster randomized trials published between August 2013 and July 2014 and using generalized estimating equations for their primary analyses. Two independent reviewers collected data from each study using a standardized, pre-piloted data extraction template. A two-arm cluster randomized trial was simulated under various scenarios to show the potential effect of a small number of clusters on type I error rate when estimating the treatment effect. The nominal level was set at 0.05 for the simulation study. Of the 51 included trials, 28 (54.9%) analyzed 40 or fewer clusters with a minimum of four total clusters. Of these 28 trials, only one trial used a bias correction method for generalized estimating equations. The simulation study showed that with four clusters, the type I error rate ranged between 0.43 and 0.47. Even though type I error rate moved closer to the nominal level as the number of clusters increases, it still ranged between 0.06 and 0.07 with 40 clusters. Our results showed that statistical issues arising from small number of clusters in generalized estimating equations is currently inadequately handled in cluster randomized trials. Potential for type I error inflation could be very high when the sandwich estimator is used without bias correction.


Clinical patterns of neurological involvement in Behçet's disease: evaluation of 200 patients. The Neuro-Behçet Study Group.
In order to define the patterns of neurological involvement in Behçet's disease and to assess prognostic factors, 558 files of the neuro-Behçet out-patient clinic were reviewed. Those patients without any evidence of objective neurological involvement as well as the patients with other possible explanations for the neurological picture, and cases not fulfilling the criteria for Behçet's disease were excluded. The remaining 200 cases (155 male, 45 female) were evaluated: 162 had parenchymal CNS involvement (brainstem or 'brainstem +' involvement in 51%, spinal cord involvement in 14%, hemispheric involvement in 15% and isolated pyramidal signs in 19%) while 38 had secondary or non-parenchymal CNS involvement. In the first group the most common findings were pyramidal signs, hemiparesis, behavioural changes and sphincter disturbance, whereas in the second group the syndrome of raised intracranial pressure due to dural sinus thrombosis was the main clinical manifestation. In 60% of the cases with parenchymal involvement, CSF was hypercellular and/or had an elevated protein level, whereas in cases with non-parenchymal involvement the CSF was usually normal except for the elevated pressure. In more than half of the patients with parenchymal involvement, MRI showed brainstem and/or basal ganglion lesions. Forty-one per cent of the cases had a course with at least one attack and remission, another 28% also had attack(s) but showed secondary progression, 10% had primary progression and 21% had silent neurological involvement. Survival analysis was performed in patients who had at least a 3-year duration of neurological disease. Parenchymal involvement, elevated protein and/or pleocytosis in the CSF, 'brainstem +' type involvement, primary or secondary progressive course and relapse during steroid tapering were all associated with a poorer prognosis.


Isthmin-1 (ISM1), a novel adipokine that reflects abdominal adipose tissue distribution in individuals with obesity.
The assessment of obesity-related health risks has traditionally relied on the Body Mass Index and waist circumference, but their limitations have propelled the need for a more comprehensive approach. The differentiation between visceral (VIS) and subcutaneous (SC) fat provides a finer-grained understanding of these risks, yet practical assessment methods are lacking. We hypothesized that combining the SC-VIS fat ratio with non-invasive biomarkers could create a valuable tool for obesity-related risk assessment. A clinical study of 125 individuals with obesity revealed significant differences in abdominal fat distribution measured by CT-scan among genders and distinct models of obesity, including visceral, subcutaneous, and the SC/VIS ratio. Stratification based on these models highlighted various metabolic changes. The SC/VIS ratio emerged as an excellent metric to differentiate metabolic status. Gene expression analysis identified candidate biomarkers, with ISM1 showing promise. Subsequent validation demonstrated a correlation between ISM1 levels in SC and plasma, reinforcing its potential as a non-invasive biomarker for fat distribution. Serum adipokine levels also correlated with the SC/VIS ratio. The Receiver Operating Characteristic analysis revealed ISM1's efficacy in discriminating individuals with favorable metabolic profiles based on adipose tissue distribution. Correlation analysis also suggested that ISM1 was involved in glucose regulation pathways. The study's results support the hypothesis that the SC-VIS fat ratio and its derived non-invasive biomarkers can comprehensively assess obesity-related health risks. ISM1 could predict abdominal fat partitioning and be a potential biomarker for evaluating obesity-related health risks.


Recollections of bullying at school and their long-term implications for lesbians, gay men, and bisexuals.
This study explores the relationship between experiences of bullying at school, adult mental health status, and symptoms of posttraumatic stress among a sample of 119 UK residents who identified as lesbian, gay, or bisexual. Participants completed a series of questionnaires that focused upon school experiences, suicide ideation at school, sexual history, relationship status and negative affect, recent positive and negative life-events, internalized homophobia, and symptoms associated with posttraumatic stress. The results suggested that posttraumatic stress was a potential issue for 17% of participants who also scored significantly higher for depression, and reported having had more casual sexual partners than their peers. However, those who were found to exhibit symptoms of post-traumatic stress were also more accepting of their sexual orientation. A small number of participants used prescription or nonprescription drugs, or alcohol to help them cope with memories of bullying. It is suggested that posttraumatic stress may be a feature of the adult lives of men and women who experienced frequent and prolonged bullying at school as a result of their actual or perceived sexual orientation.