[The pharmacokinetic effect of haloperidol on DA receptors].
There is increasing evidence that the metabolism of haloperidol is of major significance in the action of the drug. The metabolism of haloperidol is quite simple. It keeps only typical oxidative metabolic pathway. Despite of the existing knowledge of such pathways we still do not know the exact fate of the administered compound in the serum and urine. The aim of the study was the investigation of the influence of haloperidol and its oxidative forms on D(A) and its compounds in serum. The 12,0 hour investigation demonstrated a protective effect on D(A) activity. The D(A) synthesis rate constant was increased to about the same level by GBL as well as by haloperidol. Therefore, the differences between the two regions in the reactions to the drugs to a large extent could be explained by a high and low synthesis of D(A), under controlled conditions in the olfactory tubercle and the corpus striatum.


Behavioral Interventions to Increase Condom Use Among College Students in the United States: A Systematic Review.
Background. Traditional undergraduate college students in the United States are in the age range that experiences the highest rate of sexually transmitted infections (STIs) and are vulnerable to contracting STIs. Increasing condom use among college students is a prevention strategy to reduce the spread of STIs. Aim. The purpose of this systematic review of the literature was to identify behavioral interventions that increased condom use behaviors and/or intentions among college students. Method. The Preferred Reporting Items for Systematic Review and Meta-Analyses (PRISMA) guidelines were followed in systematically searching, extracting, appraising, and synthesizing the evidence. A quality assessment was also conducted with the tool provided by the Effective Public Health Practice Project. Results. The initial search yielded 715 records. After critical appraisal, seven articles remained for review. Discussion. Four of the interventions were developed using the three constructs of the information, motivation, and behavioral skills model, and all four found significant increases in condom use or condom use intentions. Additionally, interventions that included modules to increase self-efficacy for condom use, taught participants where to get condoms and how to negotiate condom use with partners, or elicited positive associations (feels) toward condoms saw increased condom use or intention to use condoms.


The Contribution of Inflammatory Cell Infiltration to Conjunctival Inclusion Cyst Formation.
Purpose: The incidence of inclusion cysts was examined histopathologically in conjunctival disorders where inflammatory cell infiltration was seen in the subepithelial connective tissue.Method: The incidence of inclusion cysts was examined histopathologically in pterygium, vernal conjunctivitis, pyogenic granuloma and pingueculitis. The specimens of pinguecula were used as control.Results: Inclusion cysts were recognized in 5/(55) cases of pterygium, 2/(12) cases of vernal conjunctivitis, 1/(4) cases of pyogenic granuloma, and 1/(2) cases of pingueculitis. On the other hand no inclusion cyst was recognized in 20 cases of pinguecula in which no inflammatory cell infiltration was seen.Conclusions: Inflammatory cell infiltration may contribute to the formation of conjunctival inclusion cysts in pterygium, pingueculitis, vernal conjunctivitis and pyogenic granuloma.


Effects of cerebral ischemia-reperfusion on pharmacokinetic fate of paeoniflorin after intravenous administration of Paeoniae Radix extract in rats.
The objective of the present study was to investigate the effects of cerebral ischemia-reperfusion on pharmacokinetics of paeoniflorin after intravenous administration of Paeoniae Radix extract (PRE) in rats. The cerebral ischemia-reperfusion rats were induced by occluding the bilateral carotid arteries of normal rats for 2 h, followed by reperfusion. The resultant animals were immediately administrated by PRE (at a dose of 60 mg/kg of paeoniflorin) via the femoral vein, whilst the same dose was injected to the normal rats. Plasma samples were collected at different time to construct pharmacokinetic profiles by plotting drug concentration versus time. Quantification of paeoniflorin in rat plasma was achieved by using a simple and rapid high-performance liquid chromatographic method. In normal rats, the major parameters of distribution half-life (t1/2alpha), elimination half-life (t1/2beta), area under the plasma concentration-time (AUC), mean retention time (MRT), and clearance (CL), estimated by an open two-compartmental model, were 0.69, 18.77 min, 5338.71 (microg min)/ml, 18.13 min and 0.0162 mg/(kg min), respectively. However, in ischemia-reperfusion rats, the corresponding parameters were 2.04, 24.51 min, 9626.00 (microg min)/ml, 29.75 min and 0.0071 mg/(kg min), respectively. The results showed that ischemia- reperfusion significantly increased AUC values, decreased CL values, and prolonged the terminal half-life of paeoniflorin. These findings suggest that the injuries of ischemia-reperfusion could play an important role in pharmacokinetic process of paeoniflorin.


Increased intracellular concentration of an initiator protein markedly reduces the minimal sequence required for initiation of DNA synthesis.
One of the most common sites used for cloning in the filamentous phages f1, fd, and M13 lies within the phage "functional origin," a sequence of 140 nucleotides that is required for phage replication. Even small insertions (four nucleotides) at this location severely reduce origin function. Secondary trans-acting mutations in the phage genome are necessary to restore efficient replication. One of these mutations, present in one of our cloning vectors, R218, has been fully characterized. It consists of a regulatory mutation within gene V that leads to a marked increase in the intracellular level of the phage gene II protein, the "initiator" of viral replication. Increased gene II protein production is sufficient to reduce the minimal sequence required for a functional origin to only 40 nucleotides, while the remaining 100 (containing the cloning site) become entirely dispensable. The general implications of these findings are discussed.


Benefits of sphingosine-1-phosphate receptor modulators in relapsing MS estimated with a treatment sequence model.
Three sphingosine-1-phosphate receptor (S1PR) modulators are currently available as disease-modifying therapies (DMTs) for relapsing MS in the Netherlands (i.e. fingolimod, ozanimod and ponesimod). We aimed to identify which S1PR modulator yields the highest benefit from a health-economic and societal perspective during a patient's lifespan. Incorporating Dutch DMT list prices, we used the ErasmusMC/iMTA MS model to compare DMT sequences, including S1PR modulators and eight other DMT classes, for treatment-naïve patients with relapsing MS in terms of health outcomes (number of lifetime relapses, time to Expanded Disability Status Scale (EDSS) 6, lifetime quality-adjusted life years (QALYs)) and cost-effectiveness (net health benefit (NHB)). We estimated the influence of list price and EDSS progression on cost-effectiveness outcomes. In deterministic and probabilistic analysis, DMT sequences with ponesimod have lower lifetime costs and higher QALYs resulting in a higher average NHB compared to sequences with other S1PR modulators. Ponesimod remains the most cost-effective S1PR modulator when EDSS progression is class-averaged. Given the variable effects on disability progression, list price reductions could make fingolimod but not ozanimod more cost-effective than ponesimod. Our model favours ponesimod among the S1PR modulators for the treatment of relapsing MS. This implies that prioritizing ponesimod over other S1PR modulators translates into a more efficacious spending of national healthcare budget without reducing benefit for people with MS. Prioritizing cost-effective choices when counselling patients contributes to affordable and accessible MS care.


Studies in experimental autoimmune encephalomyelitis do not support developmental bisphenol a exposure as an environmental factor in increasing multiple sclerosis risk.
Multiple sclerosis (MS), a demyelinating immune-mediated central nervous system disease characterized by increasing female penetrance, is the leading cause of disability in young adults in the developed world. Epidemiological data strongly implicate an environmental factor, acting at the population level during gestation, in the increasing incidence of female MS observed over the last 50 years, yet the identity of this factor remains unknown. Gestational exposure to bisphenol A (BPA), an endocrine disruptor used in the manufacture of polycarbonate plastics since the 1950s, has been reported to alter a variety of physiological processes in adulthood. BPA has estrogenic activity, and we hypothesized that increased gestational exposure to environmental BPA may therefore contribute to the increasing female MS risk. To test this hypothesis, we utilized two different mouse models of MS, experimental autoimmune encephalomyelitis (EAE) in C57BL/6J mice (chronic progressive) and in SJL/J mice (relapsing-remitting). Dams were exposed to physiologically relevant levels of BPA in drinking water starting 2 weeks prior to mating and continuing until weaning of offspring. EAE was induced in adult offspring. No significant changes in EAE incidence, progression, or severity were observed with BPA exposure, despite changes in cytokine production by autoreactive T cells. However, endocrine disruption was evidenced by changes in testes development, and transcriptomic profiling revealed that BPA exposure altered the expression of several genes important for testes development, including Pdgfa, which was downregulated. Overall, our results do not support gestational BPA exposure as a significant contributor to the increasing female MS risk.


Effect of haloperidol on expression of dopamine D2 receptor mRNAs in rat brain.
Chronic administration of the neuroleptic drug haloperidol previously has been shown to increase the density of striatal dopamine D2 receptor, which is believed to be the underlying factor in neuroleptic-induced tardive dyskinesia. To search for the mechanism of receptor upregulation, the expression of the isoforms of dopamine D2 receptor mRNA in rat striatum was analyzed by Northern, solution, and in situ hybridizations in haloperidol-treated rats (1-35 days). Northern blot analysis of poly(A)+ RNA hybridized with a probe common for both isoforms as well as an insert-specific probe for the long isoform of the receptor revealed no significant difference in hybridization signal between the control and any of the haloperidol-treated groups of rats. The receptor density, however, was increased by 30-40% in animals receiving haloperidol for 7-35 days. Solution hybridization with an antisense riboprobe specific for a consensus sequence as well as in situ hybridization with a consensus oligonucleotide probe similarly failed to detect any increase in the expression of receptor mRNA following haloperidol treatment. The results suggest that post-transcriptional mechanisms may be responsible for regulating the haloperidol-induced increase in dopamine D2 receptors.


Understanding nanoparticle assembly: a simulation approach to SERS-active dimers.
We report herein on a model built to analyze and optimize nanoparticle (NP) dimer formation. The rationale for this work stems from our interest in building effective NP dimer-based tagging systems for surface enhanced Raman scattering (SERS)-based detection. This model takes into account the behavior of the NPs in solution and the molecules on their surface, to provide a coherent and physically constrained system. The kinetics of formation of dimers and larger assemblies are investigated on suspensions of varying concentrations through a coarse-grained ad hoc computer simulation based on a Molecular Dynamics-like approach. Several different effects are considered, including the behavior and interaction of surface molecules, the interactions between the latter and the NPs, and between NPs. The surface molecules are treated as rigid structures that can occupy specific binding sites. A Brownian model is used to both integrate the particle trajectory and provide random thermal forces. These systems show a NP concentration-dependent behavior with respect to the formation of dimers versus larger assemblies over the timescale of the simulation. The simulations also indicate that these systems form low-density aggregates as opposed to the close packed formations reported previously. A dependence on the properties and the concentration of the linkers is also demonstrated.


Bioprospecting North Atlantic microalgae with fast growth and high polyunsaturated fatty acid (PUFA) content for microalgae-based technologies.
Microalgae are considered to be an important and sustainable alternative to fish oil as a source for the polyunsaturated fatty acids (PUFAs) eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Due to their health benefits, there is an increasing interest in the commercial application of these fatty acids (FA) to health and dietary products, and to aquaculture feeds. However, FA from microalgae are still expensive to produce compared to fish or plant oils. With only a few microalgal strains being cultivated on a large scale for commercial PUFA production, prospecting for new, robust and fast-growing strains with increased PUFA content is essential in order to reduce production costs. Microalgae from northern high latitudes, exposed to cold temperatures, may be especially promising candidates as previous studies have shown increasing unsaturation of FA in response to decreasing growth temperatures in different microalgae, most likely to maintain membrane fluidity and function. We have designed a screening pipeline, targeting a focused search and selection for marine microalgal strains from extreme North Atlantic locations with high robustness and biomass production, and increased levels of EPA and DHA. The pipeline includes a rational sampling plan, isolation and cultivation of clonal strains, followed by a batch growth experiment designed to obtain information on robustness, growth characteristics, and the FA content of selected isolates during both nutrient replete exponential cultivation and nutrient limited stationary cultivation. A number of clonal cultures (N = 149) have been established, and twenty of these strains have been screened for growth and FA content and composition. Among those strains, three showed growth rates ≥ 0.7 d- 1 at temperatures of 15 °C or below, and high amounts of EPA (> 3% DW), suggesting their potential as candidates for large scale production.


Inferior meatus augmentation procedure (IMAP) normalizes nasal airflow patterns in empty nose syndrome patients via computational fluid dynamics (CFD) modeling.
Empty nose syndrome (ENS) is a controversial upper airway disorder most commonly associated with tissue loss from the inferior turbinates. The inferior meatus augmentation procedure (IMAP) has been shown to effectively reduce ENS symptoms in a durable manner, but the precise mechanisms that may govern this symptomatic improvement remain unknown. Five patients with ENS who underwent bilateral IMAP via submucosal costal cartilage implant were assessed. Pre-implant and 6 months post-implant computed tomography (CT) imaging for each ENS patient was analyzed in a blinded fashion using computational fluid dynamics (CFD) modeling to investigate intrapatient changes in airflow parameters. Following surgery, ENS patients have significantly improved symptoms as indexed by Empty Nose Syndrome 6-Item Questionnaire (ENS6Q) scoring (pre-implant: 14.00 ± 4.06 [mean ± standard deviation]; 95% confidence interval [CI], 10.44 to 17.56; post-implant: 4.8 ± 2.77; 95% CI, 2.37 to 7.23; Cohen's d = 2.64; p = 0.02). Using CFD, a significant shift in nasal airflow patterns was observed, where airflow deviates away from the middle meatus upon hitting the implant (pre-implant: 67.13% ± 11.14%; 95% CI, 60.22% to 74.04%; post-implant: 46.18% ± 12.81%; 95% CI, 38.23% to 54.12%; d = 1.74; p < 0.05) toward the inferior meatus (pre-implant: 30.55% ± 11.29%; 95% CI, 23.55% to 37.55%; post-implant: 42.59% ± 9.60%; 95% CI, 36.63 to 48.54%; d = 1.14; p < 0.05). No significant changes were found in nasal resistance (pre-implant: 0.102 ± 0.015; 95% CI, 0.092 to 0.112 Pa*s/mL; post-implant: 0.105 ± 0.041; 95% CI, 0.081 to 0.130 Pa*s/mL). In addition, the improvement of ENS6Q scoring significantly correlated with percent reduction in aberrant airflow through the middle meatus (R2 = 0.60, p = 0.04). This study supports our prior working hypothesis that disordered vectors of nasal airflow congregate in the middle meatus contribute to ENS symptoms, not nasal resistance. Moreover, these data illuminate a paradoxical, but consistent, restoration of nasal airflow to the inferior meatus following the replacement of turbinate tissue volume in the inferior meatus via IMAP surgery, potentially due to the Coandă effect.


Smart systems and personalized health: the real challenge of bridging the innovation gap.
Smart miniaturized systems, emerging from the integration of heterogeneous technologies like micro- and nano electronics, photonics, biotechnology, materials and information & communication technologies are considered today, after two decades of intensive public support, proven concepts and functional prototypes, as key enablers opening up new opportunities for healthcare and in particular personalized health. They offer an enhanced ability to sense, detect, analyze, communicate, respond, and monitor phenomena from macro (e.g. body, tissues) to nano scale (e.g. molecules, genes). For the majority of these projects, planning for the next phase of prototype validation, product design, supply chain, user targeting, clinical validation and commercial roll-out are now taking full attention. The new EU Framework Program for Research and Innovation, Horizon 2020, is focusing on technology transfer support and building ecosystems and value chains to ensure better time to market and higher impact of knowledge-based technologies. The state-of-the-art and upcoming challenges for the implementation of H2020 and new opportunities in smart systems for pHealth are discussed in the paper.


Analysis of complications in 430 consecutive pediatric patients treated with intrathecal baclofen therapy: 14-year experience.
This single-center study investigated adverse events that occurred in children and adolescent patients treated with intrathecal baclofen (ITB) therapy for spasticity and/or dystonia. In a 14-year period, 430 consecutive patients with a mean age of 13.3 ± 5.9 years received ITB over a mean follow-up period of 8.6 ± 3.8 years (range 12 months to 14 years). Eighty-nine percent of these patients had cerebral palsy. Major complications, defined as those that required a surgical intervention, were infections, CSF leaks, and device problems related to the catheter or pump. Assessing infections, the authors compared the 2 groups of patients implanted with an ITB system by either the subcutaneous or subfascial technique. The temporal distribution of events related to the catheter was also considered. At least 1 complication was present in 25% of the patients: 9.3% experienced an infection, 4.9% a CSF leak, 15.1% a problem with the catheter, and 1% a problem related to the pump. Five percent of the assessed patients suffered more than 1 complication. The rate of infections was significantly lower (p < 0.001) in patients with the pump placed subfascially compared with those with the pump placed subcutaneously. A higher rate of infection was found after pump replacement compared with the first pump implantation (10.6% vs 6%, respectively). Catheter problems were the most common complication and occurred more frequently during the 1st year after the implant. While ITB is an effective treatment to manage spasticity of different origins, adverse events may occur and need to be managed. The surgical procedure should be meticulous and different techniques may have a diverse impact on the infection rate, which is the most critical complication. Despite the adverse events that occurred in this study, the majority of patients were satisfied with the treatment received.


Lifting burden for a nursing aide during patient care in a geriatric ward.
The lifting burden of a nursing aide during work in a geriatric ward was determined by using a force-plate. Three lifting operations were performed for all patients needing lifting assistance. At the same time as the forces obtained from the force-plate were recorded, the lifts were photographed. The lifting burden during nursing often equalled or exceeded the recommendations of various authors concerning permissible maximum weight loads during different types of lifts. The lifts were often performed under unfavourable conditions and seldom with an "ideal" lifting technique.


Characterization of the mouse beta-prime adaptin gene; cDNA sequence, genomic structure, and chromosomal localization.
Adaptins are important subunits of heterotetrameric complexes called adaptors, which participate in the clathrin-coated, vesicle-mediated endocytosis and intracellular receptor transport. The gene family of adaptins is divided into three classes, alpha, beta, and gamma, with further subdivision into beta- and beta-prime components. Two beta-prime adaptins, the rat AP105a and the human BAM22, have previously been characterized. The BAM22 gene is located on human Chromosome (Chr) 22q12 and can be considered a candidate meningioma tumor suppressor gene. We report here the characterization of the mouse ortholog of the BAM22 gene, and we suggest the name adtb1 for the mouse gene. Like the BAM22 gene, the adtb1 transcript is highly and ubiquitously expressed. We provide 3885-bp cDNA sequence, which entirely covers the open reading frame of the adtb1, capable of encoding a protein of 943 amino acids. The adtb1 protein is highly conserved (>96% identity) when compared with AP105a and BAM22 proteins. We also report the genomic organization of adtb1, which is similar to the BAM22 gene. The adtb1 gene has been assigned to mouse Chr 11, band 11A2, which confirms the synteny between human Chr 22q12 and mouse Chr 11.


Genetic architecture of fusarium head blight resistance in four winter triticale populations.
Fusarium head blight (FHB) is a devastating disease that causes significant reductions in yield and quality in wheat, rye, and triticale. In triticale, knowledge of the genetic architecture of FHB resistance is missing but essential due to modern breeding requirements. In our study, four doubled-haploid triticale populations (N=120 to 200) were evaluated for resistance to FHB caused by artificial inoculation with Fusarium culmorum in four environments. DArT markers were used to genotype triticale populations. Seventeen quantitative trait loci (QTL) for FHB resistance were detected across all populations; six of them were derived from rye genome and located on chromosomes 4R, 5R, and 7R, which are here reported for the first time. The total cross-validated ratio of the explained phenotypic variance for all detected QTL in each population was 41 to 68%. In all, 17 QTL for plant height and 18 QTL for heading stage were also detected across all populations; 3 and 5 of them, respectively, were overlapping with QTL for FHB. In conclusion, FHB resistance in triticale is caused by a multitude of QTL, and pyramiding them contributes to higher resistance.


Knowledge, attitude and practices of farmers and experts about the effects of pesticide residues on agricultural product users and ecosystems: A case of Fogera District, Ethiopia.
Pesticides are chemicals used to control different types of pests. Though pesticides played a role in improving the quantity and quality of production, they have been threatening ecosystems and posed effects on humans in different parts of the world. Unfortunately, there were no studies made about the effects of pesticide residues on ecosystems and consumers in the Fogera District of Amhara Region, Ethiopia. Hence, the main objective of this study was to understand the knowledge, attitude, and practices of respondents about the effects of pesticide residues on ecosystems and consumers. A cross-sectional survey complemented by focus group discussions and field observations was used to gather the required data for the study. The close-ended data were analyzed using descriptive statistics, logistic regressions, and independent t-test, and data from open-ended questions were grouped and summarized based on their similarities. The findings of the study confirmed that there was significant knowledge, attitude, and practices difference between farmers and consumers about the effects of pesticide residues on ecosystems and humans. Farmers used highly toxic pesticides to control pests and improve the glossiness of vegetables and khat. Though they didn't use the sprayed vegetables for their home consumption, some of the farmers deliberately supplied pesticide-sprayed vegetables without worrying about the negative effects of the pesticides on the consumers. There were also fishing practices from rivers after intoxicating the fish using the pesticide sprayed feed. This, in turn, might poison individuals who consume the fish. In general, pesticide application practices and consumption of pesticide-sprayed foodstuffs and surface water might pose serious health risks to ecosystems and humans. To minimize the negative effects of pesticides, rigorous awareness-raising on the effects and management of pesticides, enforcement of laws, delineation of the pesticide free buffer zone for waters, the establishment of a clear pesticide supply chain to the end users, ecosystem assessment and food safety monitoring schemes are highly required.


Cardiomyopathy associated with Ceutzfeld-Jakob disease: a diagnosis of exclusion: a case report.
Creutzfeldt-Jakob disease (CJD), the most common prion disease in humans, is primarily known for its adverse neurological impact and inevitable mortality. Data regarding myocardial involvement in CJD are scarce. A 54-year-old female patient, presented with progressive effort dyspnoea, was diagnosed with unexplained non-ischaemic cardiomyopathy. An extensive cardiac work-up including cardiac magnetic resonance imaging (MRI) did not reveal any underlying aetiology. Simultaneously, the patient developed involuntary limb movements and progressive cognitive decline. Thalamic high-signal abnormalities on diffusion-weighted images were apparent on brain MRI. Based on these findings, she was subsequently referred to a neurology department, where she suddenly died the day after her admission. Brain autopsy demonstrated spongiform encephalopathy. A genetic analysis performed to her son revealed a mutation in the PRNP gene; all of these were consistent with CJD. This case describes the clinical association of CJD and cardiomyopathy and the diagnosis prion-induced cardiomyopathy by exclusion. It is not inconceivable that the coexistence of these two clinical entities may be related to genetic expression and contemporaneously deposition of infectious prions in myocardial muscle and brain tissue. Awareness of this possible association could be of important public-safety concern, and merits further collaborative cardiac-neurological work-up to elucidate this phenotype among patients with unexplained cardiomyopathy with neurological symptoms that resemble CJD.


Thyrotropin-releasing hormone (TRH) is not thyrotropic but somatotropic in fed and starved adult chickens.
Adult fed and starved Warren chickens, 2 yr of age, and approaching the end of the second laying year, were injected iv with 1 of the following products: 10 micrograms of thyrotropin releasing hormone (TRH); 100 micrograms of bovine thyrotropin (bTSH); 100 micrograms of ovine growth hormone (oGH); saline. The influence on plasma concentrations of thyroxine (T4), triiodothyronine (T3) or chicken GH (cGH) were followed. Prior to injection, it was clear from the control values that starvation for 3 d decreased plasma levels of T3 and increased cGH, whereas 7 d of fasting increased T4 and cGH. The plasma levels of cGH were elevated greater than 10-fold at 15 min following the TRH challenge in food-deprived chickens compared to a less than 4-fold increase in normal fed hens. This increase was followed by a rise in T3 after 1 h, which was also more pronounced in the starved animals, whereas T4 decreased or remained unaffected. Increases in T4 can, however, be obtained with 100 micrograms TSH in normal fed (2-fold) or starved animals (greater than 3-fold). Following injection of 100 micrograms oGH, a significant increase in T3 levels was observed which in fed animals was already present at 30 min, but the higher levels persisted for 1 and 2 h in fed and starved hens. At the same time, a decrease in T4 was observed in both groups of GH-treated chickens. It is concluded that TRH at the dose used is not thyrotropic but has a somatotropic effect and is responsible for the peripheral conversion of T4 into T3.


Growth retardation and tumour inhibition by BRCA1.
Inherited mutations in BRCA1 predispose to breast and ovarian cancer, but the role of BRCA1 in sporadic breast and ovarian cancer has previously been elusive. Here, we show that retroviral transfer of the wild-type BRCA1 gene inhibits growth in vitro of all breast and ovarian cancer cell lines tested, but not colon or lung cancer cells or fibroblasts. Mutant BRCA1 has no effect on growth of breast cancer cells; ovarian cancer cell growth is not affected by BRCA1 mutations in the 5' portion of the gene, but is inhibited by 3' BRCA1 mutations. Development of MCF-7 tumours in nude mice is inhibited when MCF-7 cells are transfected with wild-type, but not mutant, BRCA1. Most importantly, among mice with established MCF-7 tumours, peritoneal treatment with a retroviral vector expressing wild-type BRCA1 significantly inhibits tumour growth and increased survival.


The transition from HK to LK phenotype in the red cells of newborn genetically LK lambs.
Red cells from newborn lambs were separated into different age populations by centrifugation, and cells with fetal hemoglobin (Hb) were distinguished from those with adult Hb by an acid elution technique. Changes were followed during development in rates of K+ transport (active and passive), numbers of Na+/K+ pump sites per cell, cell volumes, and numbers of Lp and L1 antigen sites per cell. These changes were correlated with the percentage of cells with adult hemoglobin. (The Lp and L1 antigens are associated with K+ transport in that specific alloantibody against Lp, anti-Lp, stimulates active transport, and anti-L1 inhibits passive transport.) Active K+ transport decreased during development because of a decline in number of Na+/K+ pumps (from measurements of ouabain binding) and because of an alteration in the affinity of the pumps for intracellular K+ (from kinetic studies in which the intracellular K+ concentration was varied). Cells with fetal Hb had fewer Lp sites and were larger than cells with adult Hb. As transport properties changed, the number of Lp sites increased and continued to increase after all the cells had adult Hb Cells with fetal Hb had as many L1 sites as lamb cells with adult Hb, but the number of L1 sites was less than those found previously for adult sheep. A population of small cells with intermediate K+ concentration and intermediate numbers of Lp sites appeared soon after birth. The various points of evidence suggested that the developmental process leading to cells with adult transport properties was a gradual one and did not coincide precisely with the switch from fetal to adult Hb.


3D printed microfluidic devices with integrated versatile and reusable electrodes.
We report two 3D printed devices that can be used for electrochemical detection. In both cases, the electrode is housed in commercially available, polymer-based fittings so that the various electrode materials (platinum, platinum black, carbon, gold, silver) can be easily added to a threaded receiving port printed on the device; this enables a module-like approach to the experimental design, where the electrodes are removable and can be easily repolished for reuse after exposure to biological samples. The first printed device represents a microfluidic platform with a 500 × 500 μm channel and a threaded receiving port to allow integration of either polyetheretherketone (PEEK) nut-encased glassy carbon or platinum black (Pt-black) electrodes for dopamine and nitric oxide (NO) detection, respectively. The embedded 1 mm glassy carbon electrode had a limit of detection (LOD) of 500 nM for dopamine and a linear response (R(2) = 0.99) for concentrations between 25-500 μM. When the glassy carbon electrode was coated with 0.05% Nafion, significant exclusion of nitrite was observed when compared to signal obtained from equimolar injections of dopamine. When using flow injection analysis with a Pt/Pt-black electrode and standards derived from NO gas, a linear correlation (R(2) = 0.99) over a wide range of concentrations (7.6-190 μM) was obtained, with the LOD for NO being 1 μM. The second application showcases a 3D printed fluidic device that allows collection of the biologically relevant analyte adenosine triphosphate (ATP) while simultaneously measuring the release stimulus (reduced oxygen concentration). The hypoxic sample (4.8 ± 0.5 ppm oxygen) released 2.4 ± 0.4 times more ATP than the normoxic sample (8.4 ± 0.6 ppm oxygen). Importantly, the results reported here verify the reproducible and transferable nature of using 3D printing as a fabrication technique, as devices and electrodes were moved between labs multiple times during completion of the study.


Transvenous catheter pulmonary embolectomy.
Acute cardiovascular collapse in the hospitalized patient is associated with a high mortality rate and remains a therapeutic dilemma. Survival could be improved in the subgroup of patients with massive pulmonary thromboembolism if prompt surgical intervention is undertaken. This report presents the cases of two patients with cardiovascular collapse who survived transvenous catheter pulmonary embolectomy (herein described in detail). This procedure can be performed in any hospital with angiographic facilities and personnel trained in the technique. In our opinion it is the procedure of choice in patients with refractory cardiovascular collapse from massive pulmonary thromboembolism.


Signals of selection in outlier loci in a widely dispersing species across an environmental mosaic.
Local adaptation reflects a balance between natural selection and gene flow and is classically thought to require the retention of locally adapted alleles. However, organisms with high dispersal potential across a spatially or temporally heterogeneous landscape pose an interesting challenge to this view requiring local selection every generation or when environmental conditions change to generate adaptation in adults. Here, we test for geographical and sequence-based signals of selection in five putatively adaptive and two putatively neutral genes identified in a previous genome scan of the highly dispersing purple sea urchin, Strongylocentrotus purpuratus. Comparing six populations spanning the species' wide latitudinal range from Canada to Baja California, Mexico, we find positive tests for selection in the putative adaptive genes and not in the putative neutral genes. Specifically, we find an excess of low-frequency and nonsynonymous polymorphisms in two transcription factors and a transporter protein, and an excess of common amino acid polymorphisms in the two transcription factors, suggestive of spatially balancing selection. We test for a genetic correlation with temperature, a dominant environmental variable in this coastal ecosystem. We find mild clines and a stronger association of genetic variation with temperature than latitude in four of the five putative adaptive loci and a signal of local adaptation in the Southern California Bight. Overall, patterns of genetic variation match predictions based on spatially or temporally balancing selection in a heterogeneous landscape and illustrate the value of geographical and coalescent tests on candidate loci identified in a genome-wide scan for selection.


Comparison of the kinetics and thermodynamics for methyl radical addition to C=C, C=O, and C=S double bonds.
The barriers, enthalpies, and rate constants for the addition of methyl radical to the double bonds of a selection of alkene, carbonyl, and thiocarbonyl species (CH(2)=Z, CH(3)CH=Z, and (CH(3))(2)C=Z, where Z = CH(2), O, or S) and for the reverse beta-scission reactions have been investigated using high-level ab inito calculations. The results are rationalized with the aid of the curve-crossing model. The addition reactions proceed via early transition structures in all cases. The barriers for addition of methyl radical to C=C bonds are largely determined by the reaction exothermicities. Addition to the unsubstituted carbon center of C=C double bonds is favored over addition to the substituted carbon center, both kinetically (lower barriers) and thermodynamically (greater exothermicities). The barriers for addition to C=O bonds are influenced by both the reaction exothermicity and the singlet-triplet gap of the substrate. Addition to the carbon center is favored over addition to the oxygen, also both thermodynamically and kinetically. For the thiocarbonyl systems, addition to the carbon center is thermodynamically favored over addition to sulfur. However, in this case, the reaction is contrathermodynamic, addition to the sulfur center having a lower barrier due to spin density considerations. Entropic differences among corresponding addition and beta-scission reactions are relatively minor, and the differences in reaction rates are thus dominated by differences in the respective reaction barriers.


Ploidy-Seq: inferring mutational chronology by sequencing polyploid tumor subpopulations.
Human cancers are frequently polyploid, containing multiple aneuploid subpopulations that differ in total DNA content. In this study we exploit this property to reconstruct evolutionary histories, by assuming that mutational complexity increases with time. We developed an experimental method called Ploidy-Seq that uses flow-sorting to isolate and enrich subpopulations with different ploidy prior to next-generation genome sequencing. We applied Ploidy-Seq to a patient with a triple-negative (ER-/PR-/HER2-) ductal carcinoma and performed whole-genome sequencing to trace the evolution of point mutations, indels, copy number aberrations, and structural variants in three clonal subpopulations during tumor growth. Our data show that few mutations (8% to 22%) were shared between all three subpopulations, and that the most aggressive clones comprised a minority of the tumor mass. We expect that Ploidy-Seq will have broad applications for delineating clonal diversity and investigating genome evolution in many human cancers.


Clinical and economic burden of revision knee arthroplasty.
Surgery is indicated for symptomatic knee osteoarthritis (OA) when conservative measures are unsuccessful. High tibial osteotomy (HTO), unicompartmental knee arthroplasty (UKA), and total knee arthroplasty (TKA) are surgical options intended to relieve knee OA pain and dysfunction. The choice of surgical intervention is dependent on several factors such as disease location, patient age, comorbidities, and activity levels. Regardless of surgical treatment, complications such as infection, loosening or lysis, periprosthetic fracture, and postoperative pain are known risks and are indications for revision surgery. The clinical and economic implications for revision surgery are underappreciated. Over 55,000 revision surgeries were performed in 2010 in the US, with 48% of these revisions in patients under 65 years. Total costs associated with each revision TKA surgery have been estimated to be in excess of $49,000. The current annual economic burden of revision knee OA surgery is $2.7 billion for hospital charges alone. By 2030, assuming a 5-fold increase in the number of revision procedures, this economic burden will exceed $13 billion annually. It is appealing to envision a therapy that could delay or obviate the need for arthroplasty. From an actuarial standpoint, this would have the theoretical downstream effect of substantially reducing the number of revision procedures. Although no known therapies currently meet these criteria, such a breakthrough would have a tremendous impact in lessening the clinical and economic burden of knee OA revision surgery.


Hepatocellular Carcinoma Incidence Threshold for Surveillance in Virologically Cured Hepatitis C Individuals.
Guidelines recommend biannual surveillance for hepatocellular carcinoma (HCC) in hepatitis C individuals with cirrhosis if the HCC incidence rate is above 1.5 per 100 person-years (PY). However, the incidence threshold for surveillance in individuals who achieve a virologic cure is unknown. We estimated the HCC incidence rate above which routine HCC surveillance is cost-effective in this growing population of virologically cured hepatitis C individuals with cirrhosis or advanced fibrosis. We developed a Markov-based microsimulation model of the natural history of HCC in individuals with hepatitis C who achieved virologic cure with oral direct-acting antivirals. We used published data on the natural history of hepatitis C, competing risk post virologic cure, HCC tumor progression, real-world HCC surveillance adherence, contemporary HCC treatment options and associated costs, and utilities of different health states. We estimated the HCC incidence above which biannual HCC surveillance using ultrasound and alpha-fetoprotein would be cost-effective. In virologically cured hepatitis C individuals with cirrhosis or advanced fibrosis, HCC surveillance is cost-effective if HCC incidence exceeds 0.7 per 100 PY using $100,000 per quality-adjusted life year willingness-to-pay. At this HCC incidence, routine HCC surveillance would result in 2650 and 5700 additional life years per 100,000 cirrhosis and advanced fibrosis persons, respectively, compared with no surveillance. At $150,000 willingness-to-pay, surveillance is cost-effective if HCC incidence exceeds 0.4 per 100 PY. Sensitivity analysis showed that the threshold mostly remained below 1.5 per 100 PY. The contemporary HCC incidence threshold is much lower than the previous 1.5% incidence value used to guide HCC surveillance decisions. Updating clinical guidelines could improve the early diagnosis of HCC.


Safety of Same-Day Discharge Posterior Lumbar Decompression and/or Discectomy: An Observational Study Using Administrative Data From England.
Retrospective cohort study. Same-day discharge is widely used in many surgical specialities. If carefully planned, it can improve patient outcomes whilst using resources efficiently. We aimed to investigate the safety of same-day discharge following a posterior lumbar decompression and/or discectomy (PLDD). This was a retrospective analysis of administrative data. We extracted data from the Hospital Episodes Statistics database for the 5 years from 1st April 2014 to 31st March 2019. Patients undergoing an elective one or two level PLDD aged 19-54 years during the index stay were included. The primary exposure variable was same-day discharge or post-surgery overnight stay and the primary outcome was emergency hospital readmission within 90 days post-discharge. Data were available for 45,814 PLDD performed across 103 hospital trusts of which 7914 (17.3%) were performed as same-day discharge. Same-day discharge rates varied from 87.7% to 0% across the 90 hospital trusts that operated on more than 50 patients during the study period. Fourteen (15.6%) trusts had same-day discharge rates above 30% and 57 (63.3%) trusts had same-day discharge rates below 10%. The odds of emergency hospital readmission within 90 days were lower for same-day discharge patients (odds ratio .72 (95% confidence interval .61 to .85). There was no difference in outcomes for patients seen at trusts with a same-day discharge rate of ≥30% compared to trusts with a same-day discharge rate of ≤10%. Same-day discharge low-complexity elective PLDD is safe in adult patients below the age of 55 years. There is potential for many providers to substantially increase their rates of same-day discharge.


Serum beta-human chorionic gonadotropin levels relate poorly with the size of a tubal pregnancy.
Ectopic implantation usually begins with relatively normal growth of trophoblast and serum beta-hCG progression. However, the trophoblast eventually erodes into vessels and a variable degree of bleeding and hematoma compromises its growth. The serum beta-hCG level then usually begins to demonstrate some degree of abnormal progression. For our patients, the length of the ectopic pregnancy varied widely, depending mainly on when the individual patient chose to seek medical treatment. Infertility patients followed from the moment of conception would thus be expected to show a better correlation between early beta-hCG levels and the size of the tubal pregnancy. Preconditions for the operative laparoscopic management of a tubal pregnancy usually include that the tube be unruptured, less than 3 cm in diameter, and readily accessible via the laparoscopic approach. Ackerman et al. suggested there was a general correlation between the serum beta-hCG level and tubal rupture, and the present data demonstrate an overall positive correlation between the size of the tubal pregnancy and the serum level. However, the range of levels is so broad for any given size of mass or tubal status that this correlation is not meaningful clinically. A reliable method to determine the size and status of a tubal pregnancy before laparoscopy would be valuable. Unfortunately, we did not find the preoperative serum beta-hCG level to be useful for making this prediction.


Structural analysis of human beta-defensin-1 and its significance in urinary tract infection.
Human beta-defensin-1 (hBD-1) is a 36-amino-acid antimicrobial peptide that functions in the host innate defense. We developed a highly sensitive radioimmunoassay for hBD-1 and identified several hBD-1 peptides in human kidney, urine, and plasma by amino acid sequencing and mass spectrometry. Large quantities of hBD-1 peptides are produced in the kidney, are released into the tubular lumen as 47-amino-acid pro-hBD1, and then undergo proteolytic processing and generate multiple truncated forms. The respective urine and plasma concentrations of hBD-1 in patients with pyelonephritis are 48.1 +/- (SEM) 15.7 pmol/mg creatinine and 2.66 +/- 0.41 pmol/ml, 3.1-fold and 1.8-fold those of normal individuals. hBD-1 is thought to contribute to mucosal defense in the urinary tract. Our findings provide a better understanding of the biosynthesis of this peptide and its pathophysiological significance in infectious diseases.


In vitro evaluation of milk-based, soy-based, and amino acid-based infant formulas on Streptococcus mutans biofilm formation.
Infant formulas are based on milk, and the addition of simple carbohydrates as a caloric source, for infants. The carbohydrates added in infant formulas can cause a significant increase of Streptococcus mutans in the oral cavity of infants adding to their cariogenicity. The aim of the study was to assess and compare the biofilm formation in three commercially available infant formulas;which are based on milk, soy and amino acid. In vitro microbiological assay of Streptococcus mutans biofilm formation in milk based, soy based and amino acid based infant formulas. Twenty-four hour-cultured S. mutans and microtiter plates were used for analysis. At microtiter plate, 190 μL of modified TSB broth containing SBF, MBF, amino acid-based infant formulas, and dairy whitener as a positive control in five dilutions (1:05, 1:10, 1:20, 1:40, and 1:80) was added into respective wells. 10 μL of cultured S. mutans was inoculated into the wells and incubated at 37°C for 24 h. Biofilm was washed, fixed, and stained with crystal violet. The absorbance was measured to evaluate biofilm growth, which was read as optical densities in a spectrophotometer at 490 nm and was tabulated. Three infant formulas tested showed S. mutans biofilm growth. Minimal biofilm growth was observed in amino acid-based formula at 1:80 dilution, followed by MBF at 1:10 dilution and SBF at 1:80 dilution. Commercially available infant formulas favor S. mutans biofilm growth and can be cariogenic. Amino acid-based infant formula was found to have less S. mutans biofilm growth than MBF and SBF.


Down-regulation of NKD1 increases the invasive potential of non-small-cell lung cancer and correlates with a poor prognosis.
As a negative modulator of the canonical Wnt signaling pathway, Naked1 (NKD1) is widely expressed in many normal tissues. However, the expression pattern and clinicopathological significance of NKD1 in patients with non-small-cell lung cancer (NSCLC) is still unclear. Immunohistochemical studies were performed on 35 cases of normal lung tissues and 100 cases of NSCLC, including 66 cases with complete follow-up records. The NKD1 protein and mRNA expressions were detected by western blot and Real-time PCR, respectively. To examine the effect of NKD1 on the invasiveness of lung cancer cells, NKD1 was down-regulated by siRNA in lung cancer cell lines and the invasive ability was then evaluated by the Matrigel invasion assay. In addition, the expressions of Dishevelled-1 and β-catenin proteins, as well as MMP mRNA were also examined in NKD1 knockdown cells. In 35 fresh lung cancer tissues examined, 27(79%) of them exhibited lower levels of NKD1 protein in comparison with their corresponding normal tissue (P = 0.009). However, the NKD1 mRNA level was significantly higher in cancerous lung tissues, compared with the adjacent normal tissues. In 100 NSCLC tissues, NKD1 was significantly lower in 78 cases (78%) than in the normal specimens, determined by immunohistochemical staining. The reduced NKD1 expression was correlated with histological type (P = 0.003), poor differentiation (P = 0.004), lymph node metastasis (P = 0.013), TNM stage (P = 0.002) and poor survival (62.88 ± 3.23 versus 23.61 ± 2.18 months, P = 0.03). In addition, NKD1 knockdown could up-regulate Dishevelled-1 and β-catenin protein levels, as well as increased MMP-7 transcription and the invasive ability of lung cancer cells. Furthermore, when the NKD1-knockdown cells were treated with Dishevelled-1 antibody, their invasive potential was significantly reduced. NKD1 protein is reduced but NKD1 mRNA is elevated in NSCLC. Reduced NKD1 protein expression correlates with a poor prognosis in NSCLC. NKD1 might inhibit the activity of the canonical Wnt pathway through Dishevelled-1.


Residues required for activity in Escherichia coli o-succinylbenzoate synthase (OSBS) are not conserved in all OSBS enzymes.
Understanding how enzyme specificity evolves will provide guiding principles for protein engineering and function prediction. The o-succinylbenzoate synthase (OSBS) family is an excellent model system for elucidating these principles because it has many highly divergent amino acid sequences that are <20% identical, and some members have evolved a second function. The OSBS family belongs to the enolase superfamily, members of which use a set of conserved residues to catalyze a wide variety of reactions. These residues are the only conserved residues in the OSBS family, so they are not sufficient to determine reaction specificity. Some enzymes in the OSBS family catalyze another reaction, N-succinylamino acid racemization (NSAR). NSARs cannot be segregated into a separate family because their sequences are highly similar to those of known OSBSs, and many of them have both OSBS and NSAR activities. To determine how such divergent enzymes can catalyze the same reaction and how NSAR activity evolved, we divided the OSBS family into subfamilies and compared the divergence of their active site residues. Correlating sequence conservation with the effects of mutations in Escherichia coli OSBS identified two nonconserved residues (R159 and G288) at which mutations decrease efficiency ≥200-fold. These residues are not conserved in the subfamily that includes NSAR enzymes. The OSBS/NSAR subfamily binds the substrate in a different orientation, eliminating selective pressure to retain arginine and glycine at these positions. This supports the hypothesis that specificity-determining residues have diverged in the OSBS family and provides insight into the sequence changes required for the evolution of NSAR activity.


Phosphorylase synthesis in diabetic hepatocytes and cardiomyocytes.
Whereas total cardiac glycogen phosphorylase activity appears to be unaffected by severe insulin deficiency, a diabetes-induced decreased in hepatic glycogen phosphorylase activity has been demonstrated by our laboratory and others using liver extracts, isolated perfused liver, and cultured hepatocytes. The loss of activity in diabetic liver can be correlated with a drop in protein levels. Using primary cultures of cells from normal and diabetic rats and phosphorylase specific antibodies, we found a corresponding decrease in phosphorylase synthesis in diabetic hepatocytes cultured for 2 days in a serum-free, chemically defined medium. When hepatocytes are cultured in the presence of insulin, triiodothyronine, and cortisol, there is a significant recovery in the rate of phosphorylase synthesis after 3 days. Over the 3-day time period, there is no significant difference in the rate of phosphorylase degradation in normal compared with diabetic hepatocytes. Total protein synthesis in both hepatocytes and cardiomyocytes is unaffected by diabetes, as is phosphorylase synthesis in cultured cardiomyocytes.


Extraordinary lithium ion storage capability achieved by SnO2 nanocrystals with exposed {221} facets.
Rational design of SnO2 nanomaterials with superior architectures and excellent electrochemical properties is highly desirable for lithium ion storage. Here, several SnO2 nanoparticles with different exposed crystal planes, such as {101}, {110} and {221} facets, are developed and further embedded into graphene/carbon nanotube (G/CNT) networks, achieving highly conductive carbon/SnO2 films (C/SnO2) with homogeneous dispersion of SnO2 nanoparticles. Three-dimensional (3D) G/CNT networks with highly porous structures and electronic contacts with imbedded SnO2 nanoparticles provide excellent pathways for transfer of electrons and ions and further buffer structural changes of SnO2 nanocrystals during lithium-ion insertion/extraction processes. Close contact between G/CNT matrix and embedded SnO2 nanoparticles ensures that all high-energy {221} facets of SnO2 are exploited during rapid electrochemical reactions. The high electrical conductivity of G/CNT networks can further prevent pulverization of nanostructured SnO2. As a result, C/SnO2 film with 90% content of octahedral SnO2 nanocrystals (C/SnO2-O (90%)) exhibits superior reversible specific capacity of 1008 mA h g-1 at 0.1 A g-1, excellent rate capability, low internal resistance and long-term cycling stability for 1000 cycles. These results further confirm that SnO2 nanocrystals with high-energy {221} facets can provide numerous active sites for lithium ion storage than other SnO2 nanomaterials.


The UCSC Ebola Genome Portal.
With the Ebola epidemic raging out of control in West Africa, there has been a flurry of research into the Ebola virus, resulting in the generation of much genomic data. In response to the clear need for tools that integrate multiple strands of research around molecular sequences, we have created the University of California Santa Cruz (UCSC) Ebola Genome Browser, an adaptation of our popular UCSC Genome Browser web tool, which can be used to view the Ebola virus genome sequence from GenBank and nearly 30 annotation tracks generated by mapping external data to the reference sequence. Significant annotations include a multiple alignment comprising 102 Ebola genomes from the current outbreak, 56 from previous outbreaks, and 2 Marburg genomes as an outgroup; a gene track curated by NCBI; protein annotations curated by UniProt and antibody-binding epitopes curated by IEDB. We have extended the Genome Browser's multiple alignment color-coding scheme to distinguish mutations resulting from non-synonymous coding changes, synonymous changes, or changes in untranslated regions. Our Ebola Genome portal at http://genome.ucsc.edu/ebolaPortal/ links to the Ebola virus Genome Browser and an aggregate of useful information, including a collection of Ebola antibodies we are curating.


Measurements of methane and nitrous oxide in human breath and the development of UK scale emissions.
Exhaled human breath can contain small, elevated concentrations of methane (CH4) and nitrous oxide (N2O), both of which contribute to global warming. These emissions from humans are not well understood and are rarely quantified in global greenhouse gas inventories. This study investigated emissions of CH4 and N2O in human breath from 104 volunteers in the UK population, to better understand what drives these emissions and to quantify national-scale estimates. A total of 328 breath samples were collected, and age, sex, dietary preference, and smoking habits were recorded for every participant. The percentage of methane producers (MPs) identified in this study was 31%. The percentage of MPs was higher in older age groups with 25% of people under the age of 30 classified as MPs compared to 40% in the 30+ age group. Females (38%) were more likely to be MPs than males (25%), though overall concentrations emitted from both MP groups were similar. All participants were found to emit N2O in breath, though none of the factors investigated explained the differences in emissions. Dietary preference was not found to affect CH4 or N2O emissions from breath in this study. We estimate a total emission of 1.04 (0.86-1.40) Gg of CH4 and 0.069 (0.066-0.072) Gg of N2O in human breath annually in the UK, the equivalent of 53.9 (47.8-60.0) Gg of CO2. In terms of magnitude, these values are approximately 0.05% and 0.1% of the total emissions of CH4 and N2O reported in the UK national greenhouse gas inventories.


Exercised-induced hematuria.
Bloody or discolored urine after exercise can have various etiologies. Exercise-induced hematuria is a common, benign cause of discolored urine following strenuous exercise. Evaluation of patients with this complaint begins with a thorough history, a focused physical examination and a microscopic examination of the urine. Using a systematic approach, the physician can detect serious, treatable problems, and testing can be limited. Patients with exercise-induced hematuria can be reassured that the condition is benign and that they can return to full activity.


The endocycle controls nurse cell polytene chromosome structure during Drosophila oogenesis.
Polytene chromosomes exhibit intricate higher order chromatin structure that is easily visualized due to their precisely aligned component strands. However, it remains unclear if the same factors determine chromatin organization in polyploid and diploid cells. We have analyzed one such factor, the cell cycle, by studying changes in Drosophila nurse cell chromosomes throughout the 10 to 12 endocycles of oogenesis. We find that nurse cells undergo three distinct types of endocycle whose parameters are correlated with chromosome behavior. The first four endocycles support complete DNA replication; poorly banded polytene euchromatin progressively condenses during the late S phases to produce blob-like chromosomes. During the unique fifth endocycle, an incomplete late S phase is followed by a mitosis-like state during which the 64C chromosomes dissociate into 32 chromatid pairs held together by unreplicated regions. All the subsequent endocycles lack any late S phase; during these cycles a new polytene chromosome grows from each 2C chromatid pair to generate 32-ploid polytene nuclei. These observations suggest that euchromatin begins to condense during late S phase and that nurse cell polytene chromosome structure is controlled by regulating whether events characteristic of late S and M phase are incorporated or skipped within a given endocycle.


Predicting vaginal birth after cesarean section: a cohort study.
To develop a model to predict vaginal birth after cesarean (VBAC) in our population and to compare the accuracy of this model to the accuracy of a previously published widely used model. Women attempting trial of labor after cesarean delivery (TOLAC) at our institution from January 1, 2000 through May 30, 2010 were evaluated for inclusion. Demographic and clinical data were collected. Associations of these characteristics with VBAC were evaluated with univariate and multivariate logistic regression. We critically compared the accuracy of the resulting model to a previously published widely utilized model for predicting VBAC. A total of 2,635 deliveries with at least 1 prior cesarean delivery were identified. TOLAC was attempted in 599 (22.7%) and resulted in 456 VBACs (76.0%) and 143 repeat cesareans (24.0%). VBAC success was independently associated with age <30 years, a body mass index <30, prior vaginal delivery, prior VBAC, and absence of a recurrent indication for cesarean. This model provided a range of successful probability of VBAC (38-98%) with an area under the receiver operating characteristic curve of 0.723. This study provides an accurate and simple model that can be utilized to guide decisions related to TOLAC.


Butyrate ameliorated ferroptosis in ulcerative colitis through modulating Nrf2/GPX4 signal pathway and improving intestinal barrier.
Oxidative stress and intestinal inflammation are main pathological features of ulcerative colitis (UC). Ferroptosis, characterized by iron accumulation and lipid peroxidation, is closely related to the pathologic process of UC. 16S rRNA sequencing for intestinal microbiota analysis and gas chromatography-mass spectrometry (GC-MS) for short-chain fatty acid (SCFA) contents clearly demonstrated lower amounts of butyrate-producing bacteria and butyrate in colitis mice. However, the precise mechanisms of sodium butyrate (NaB) in treating UC remain largely unclear. We found that ferroptosis occurred in colitis models, as evidenced by the inflammatory response, intracellular iron level, mitochondria ultrastructural observations and associated protein expression. NaB inhibited ferroptosis in colitis, significantly rescued weight loss and colon shortening in mice and reduced inflammatory lesions and mitochondrial damage. Furthermore, NaB improved intestinal barrier integrity and markedly suppressed the expression of pro-ferroptosis proteins. Conversely, the protein expression of anti-ferroptosis markers including nuclear factor erythroid-related Factor 2 (Nrf2) and glutathione peroxidase 4 (GPX4), was significantly upregulated with NaB treatment. Moreover, the knockdown of Nrf2 reversed the anti-colitis effect of NaB. Taken together, NaB exhibited a protective effect by ameliorating ferroptosis in experimental colitis through Nrf2/GPX4 signaling and improving intestinal barrier integrity, which provides a novel mechanism for NaB prevention of UC.


Two dimensional Doppler echocardiographic/M mode echocardiographic and phonocardiographic method for study of extracardiac heterograft valved conduits in the right ventricular outflow tract position.
Significant concern exists over the long-term results of right ventricular outflow tract repair using heterograft valved conduits. Because these conduits and valves are difficult to image using ultrasound, a serially applicable two dimensional Doppler echocardiographic, M mode echocardiographic and phonocardiographic method for noninvasive investigation was developed and applied in 15 children. The method provides two dimensional echocardiographic imaging of valve contour and motion, as well as M mode and phonocardiographic analysis and quantitative range-gated Doppler information about the timing of flow through the conduit. Conduit diameter in two dimensional echocardiographic images correlated well with known conduit size (r = +0.96). A thickened and stenosed heterograft valve was predicted in two patients before hemodynamic investigation. This new method provides serially obtainable information to aid in the management of children and infants with a valved conduit placed for repair of congenital heart malformations and aids in planning the timing of hemodynamic follow-up studies.


Well-being, school climate, and the social identity process: a latent growth model study of bullying perpetration and peer victimization.
The present study concerns longitudinal research on bullying perpetration and peer victimization. A focus is on school factors of school climate (academic support, group support) and school identification (connectedness or belonging), which are conceptualized as related but distinct constructs. Analysis of change on these factors as well as individual well-being across time contributes to understanding bullying behavior. Latent growth modeling was employed to examine the predictors of anxiety, depression, 2 school climate factors and school identification in understanding change in physical and verbal bullying behavior. The sample included 492 Australian school students (means age 15 years, 53.5% male) in Grades 7 to 10 who completed measures over 3 years. Academic support and group support were the strongest predictors of change in bullying and victimization. Positive change in school identification also predicted a decrease in bullying behavior over time. An increase in depression or anxiety across time predicted an increase in rates of both bullying and victimization over time. Future research should continue to examine the complex relationship between individual-psychological and social-psychological variables in impacting on incidence of school-based bullying. On a practical note, school-based intervention programs may benefit from an approach that aims to target the school climate, social identity with the school, and promote individual psychological well-being.


Brain- and heart-type fatty acid-binding proteins in the brain: tissue distribution and clinical utility.
Detection of brain injury by serum markers is not a standard procedure in clinical practice, although several proteins, such as S100B, neuron-specific enolase (NSE), myelin basic protein, and glial fibrillary acidic protein, show promising results. We investigated the tissue distribution of brain- and heart-type fatty acid-binding proteins (B-FABP and H-FABP) in segments of the human brain and the potential of either protein to serve as plasma marker for diagnosis of brain injury. B-FABP and H-FABP were measured immunochemically in autopsy samples of the brain (n = 6) and in serum samples from (a) patients with mild traumatic brain injury (MTBI; n = 130) and (b) depressed patients undergoing bilateral electroconvulsive therapy (ECT; n = 14). The protein markers S100B and NSE were measured for comparison. Reference values of B-FABP and H-FABP were established in healthy individuals (n = 92). The frontal, temporal, and occipital lobes, the striatum, the pons, and the cerebellum had different tissue concentrations of B-FABP and of H-FABP. B-FABP ranged from 0.8 microg/g wet weight in striatum tissue to 3.1 microg/g in frontal lobe. H-FABP was markedly higher, ranging from 16.2 microg/g wet weight in cerebellum tissue to 39.5 microg/g in pons. No B-FABP was detected in serum from healthy donors. H-FABP serum reference value was 6 microg/L. In the MTBI study, serum B-FABP was increased in 68% and H-FABP in 70% of patients compared with S100B (increased in 45%) and NSE (increased in 51% of patients). In ECT, serum B-FABP was increased in 6% of all samples (2 of 14 patients), whereas H-FABP was above its upper reference limit (6 microg/L) in 17% of all samples (8 of 14 patients), and S100B was above its upper reference limit (0.3 microg/L) in 0.4% of all samples. B-FABP and H-FABP patterns differ among brain tissues, with the highest concentrations in the frontal lobe and pons, respectively. However, in each part of the brain, the H-FABP concentration was at least 10 times higher than that of B-FABP. Patient studies indicate that B-FABP and H-FABP are more sensitive markers for minor brain injury than the currently used markers S100B and NSE.


Purification and characterization of a receptor for the 987P pilus of Escherichia coli.
A receptor-containing fraction that caused visible aggregation of piliated Escherichia coli strain 987 bacteria was released into solution when brush borders from adult rabbit small intestines were stored. The receptor-containing fraction was separated into more than 50 bands by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, 4 of which contained carbohydrate as demonstrated by staining with periodic acid-Schiff. Receptor activity in the receptor-containing fraction was associated with a low-molecular-weight (less than 14,000) glycoprotein as measured by Western blots. Extraction of brush borders with chloroform-methanol-water (4:8:3) resulted in increased pilus 987P receptor activity in the aqueous extract. Gel filtration chromatography of sodium dodecyl sulfate-solubilized aqueous extract yielded a fraction with 987P receptor activity that contained only the low-molecular-weight glycoprotein. The purified 987P receptor contained 81% carbohydrate and 19% amino acids by weight. Isoelectric focusing separated the 987P receptor receptor into 3 peaks corresponding to isoelectric points of 2.2, 3.8, and 4.1. Each of the peaks contained 987P receptor activity. The 987P receptor activity was sensitive to periodate oxidation and to digestion with pronase. The strain 987-agglutinating activity of the receptor was inhibited by amino sugars and their N-acetylated derivatives, by compounds having a free amino group, by high concentrations of salt, and by lectins that recognize D-galactose or L-fucose.


Responses of ocean circulation and carbon cycle to changes in the position of the Southern Hemisphere westerlies at Last Glacial Maximum.
We explore the impact of a latitudinal shift in the westerly wind belt over the Southern Ocean on the Atlantic meridional overturning circulation (AMOC) and on the carbon cycle for Last Glacial Maximum background conditions using a state-of-the-art ocean general circulation model. We find that a southward (northward) shift in the westerly winds leads to an intensification (weakening) of no more than 10% of the AMOC. This response of the ocean physics to shifting winds agrees with other studies starting from preindustrial background climate, but the responsible processes are different. In our setup changes in AMOC seemed to be more pulled by upwelling in the south than pushed by downwelling in the north, opposite to what previous studies with different background climate are suggesting. The net effects of the changes in ocean circulation lead to a rise in atmospheric pCO2 of less than 10 μatm for both northward and southward shift in the winds. For northward shifted winds the zone of upwelling of carbon- and nutrient-rich waters in the Southern Ocean is expanded, leading to more CO2outgassing to the atmosphere but also to an enhanced biological pump in the subpolar region. For southward shifted winds the upwelling region contracts around Antarctica, leading to less nutrient export northward and thus a weakening of the biological pump. These model results do not support the idea that shifts in the westerly wind belt play a dominant role in coupling atmospheric CO2 rise and Antarctic temperature during deglaciation suggested by the ice core data.


3D Printable Drug Delivery Systems: Next-generation Healthcare Technology and Regulatory Aspects.
A revolutionary shift in healthcare has been sparked by the development of 3D printing, propelling us into an era replete with boundless opportunities for personalized DDS (Drug Delivery Systems). Precise control of the kinetics of drug release can be achieved through 3D printing, improving treatment efficacy and patient compliance. Additionally, 3D printing facilitates the co-administration of multiple drugs, simplifying treatment regimens. The technology offers rapid prototyping and manufacturing capabilities, reducing development timelines and costs. The seamless integration of advanced algorithms and artificial neural networks (ANN) augments the precision and efficacy of 3D printing, propelling us toward the forefront of personalized medicine. This comprehensive review delves into the regulatory frontiers governing 3D printable drug delivery systems, with an emphasis on adhering to rigorous safety protocols to ensure the well-being of patients by leveraging the latest advancements in 3D printing technologies powered by artificial intelligence. The paradigm promises superior therapeutic outcomes and optimized medication experiences and sets the stage for an immersive future within the Metaverse, wherein healthcare seamlessly converges with virtual environments to unlock unparalleled possibilities for personalized treatments.


Novel Mutation Sites in the Development of Vancomycin- Intermediate Resistance in Staphylococcus aureus.
Increased use of vancomycin has led to the emergence of vancomycin-intermediate Staphylococcus aureus (VISA). To investigate the mechanism of VISA development, 39 methicillin-susceptible strains and 3 MRSA strains were treated with vancomycin to induce non-susceptibility, and mutations in six genes were analyzed. All the strains were treated with vancomycin in vitro for 60 days. MICs were determined by the agar dilution and E-test methods. Vancomycin was then removed to assess the stability of VISA strains and mutations. Following 60 days of vancomycin treatment in vitro, 29/42 VISA strains were generated. The complete sequences of rpoB, vraS, graR, graS, walK, and walR were compared with those in the parental strains. Seven missense mutations including four novel mutations (L466S in rpoB, R232K in graS, I594M in walk, and A111T in walR) were detected frequently in strains with vancomycin MIC ≥ 12 μg/mL. Jonckheere-Terpstra trend test indicated these mutations might play an important role during VISA evolution. After the vancomycin treatment, strains were passaged to vancomycin-free medium for another 60 days, and the MICs of all strains decreased. Our results suggest that rpoB, graS, walk, and walR are more important than vraS and graR in VISA development.


Role of the Admiral Nurse in supporting a person with dementia and their family carer.
Distress is commonly experienced by those caring for someone with dementia, and can occur whether the carer is living with a person with dementia, or supporting a person with dementia who lives alone to maintain their independence. It is essential for health and social care professionals to support family carers to balance their needs with those of the person they care for. However, this might be challenging because many influencing factors can affect these needs, and identifying the needs of the family carer and balancing them with the needs of the person with dementia is often complex. This article uses a case study to explore the role of one Admiral Nurse (KG) in supporting the main family carer of a person with dementia, and the model of reflection used by the Admiral Nurse to undertake critical thinking and analysis in relation to their role.


Identification and Characterization of Small-Molecule IRF3-Dependent Immune Activators for Pharmaceutical Development.
We sought to develop a small-molecule activator of interferon regulatory factor 3 (IRF3), an essential innate immune transcription factor, which could potentially be used therapeutically in multiple disease settings. Using a high-throughput screen, we identified small-molecule entities that activate a type I interferon response, with minimal off-target NFκB activation. We identified 399 compounds at a hit rate of 0.24% from singlicate primary screening. Secondary screening included the primary hits and additional compounds with similar chemical structures obtained from other library sources and resulted in 142 candidate compounds. The hit compounds were sorted and ranked to identify compound groups with activity in both human and mouse backgrounds to facilitate animal model engagement for translational development. Chemical modifications within two groups of small molecules produced leads with improved activity over original hits. Furthermore, these leads demonstrated activity in ex vivo cytokine release assays from human blood- and mouse bone marrow-derived macrophages. Dependence on IRF3 was demonstrated using bone marrow-derived macrophages from IRF3-deficient mice, which were not responsive to the molecules. To identify the upstream pathway leading to IRF3 activation, we used a library of CRISPR knockout cell lines to test the key innate immune adaptor and receptor molecules. These studies indicated a surprising toll-interleukin-1 receptor-domain-containing-adapter-inducing interferon-β-dependent but TLR3/4-independent mechanism of IRF3 activation.


Influenza A virus infection of intestinal epithelial cells enhances the adhesion ability of Crohn's disease associated Escherichia coli strains.
Modifications of intestinal glycoreceptors expression, in particular CEACAM6, typically found in ileal Crohn's disease (CD), favor, among the commensal species of microbiota, the enrichment in Escherichia coli. Removal of protein glycosidic residues by neuraminidase, a sialidase typical of influenza virus, increases adhesion ability of Escherichia coli to Caco-2 intestinal cells. In this study we investigated whether influenza virus infection of human intestinal epithelial cells could influence the adhesiveness of different Escherichia coli strains isolated from CD patients by altering surface glycoreceptors. Influenza virus infection of intestinal cells increased exposure of galactose and mannose residues on the cell surface. In particular, glycoreceptors Thomsen-Friedenreich and CEACAM6 were over-expressed in influenza virus infected cells. In the same experimental conditions, a significant increase in bacterial adhesiveness was observed, independently of their own adhesive ability. The increase was reverted by treatment with anti-TF and anti-CEACAM6 antibodies. Interestingly, influenza virus was able to efficiently replicate in human primary intestinal cells leading to TF exposure. Finally, intestinal infected cells produced high levels of pro-inflammatory cytokines compared to control. Overall these data suggest that influenza virus infection, could constitute an additional risk factor in CD patients.


Measuring the modulus of the spatial coherence function using an error tolerant phase shifting algorithm and a continuous lateral shearing interferometer.
The modulus of the degree of coherence can be derived from interference patterns either by using fringes and next neighbour operations or by using several interferograms produced through phase shifting. Here the latter approach will be followed by using a lateral shearing interferometer exploiting a diffractive grating wedge providing a linearly progressive shear. Phase shifting methods offer pixel-oriented evaluations but suffer from instabilities and drifts which is the reason for the derivation of an error immune algorithm. This algorithm will use five π/2-steps of the reference phase also for the calculation of the modulus of the coherence function.


Associations of fat-soluble micronutrients and redox biomarkers with frailty status in the FRAILOMIC initiative.
A poor fat-soluble micronutrient (FMN) and a high oxidative stress status are associated with frailty. Our aim was to determine the cross-sectional association of FMNs and oxidative stress biomarkers [protein carbonyls (PrCarb) and 3-nitrotyrosine] with the frailty status in participants older than 65 years. Plasma levels of vitamins A (retinol), D3 , E (α-tocopherol and γ-tocopherol) and carotenoids (α-carotene and β-carotene, lycopene, lutein/zeaxanthin, and β-cryptoxanthin), PrCarb, and 3-nitrotyrosine were measured in 1450 individuals of the FRAILOMIC initiative. Participants were classified into robust, pre-frail, and frail using Fried's frailty criteria. Associations between biomarkers and frailty status were assessed by general linear and logistic regression models, both adjusted for cohort, season of blood sampling, gender, age, height, weight, and smoking. Robust participants had significantly higher vitamin D3 and lutein/zeaxanthin concentrations than pre-frail and frail subjects; had significantly higher γ-tocopherol, α-carotene, β-carotene, lycopene, and β-cryptoxanthin concentrations than frail subjects, and had significantly lower PrCarb concentrations than frail participants in multivariate linear models. Frail subjects were more likely to be in the lowest than in the highest tertile for vitamin D3 (adjusted odds ratio: 2.15; 95% confidence interval: 1.42-3.26), α-tocopherol (2.12; 1.39-3.24), α-carotene (1.69; 1.00-2.88), β-carotene (1.84; 1.13-2.99), lycopene (1.94; 1.24-3.05), lutein/zeaxanthin (3.60; 2.34-5.53), and β-cryptoxanthin (3.02; 1.95-4.69) and were more likely to be in the highest than in the lowest tertile for PrCarb (2.86; 1.82-4.49) than robust subjects in multivariate regression models. Our study indicates that both low FMN and high PrCarb concentrations are associated with pre-frailty and frailty.


Degradation of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans by the white rot fungus Phanerochaete sordida YK-624.
A method for the degradation of dioxins by white rot fungi was developed. Degradation of a mixture of 10 kinds of tetra- to octachlorodibenzo-p-dioxins (polychlorinated dibenzo-p-dioxins [PCDDs]) and tetra- to octachlorodibenzofurans (polychlorinated dibenzofurans [PCDFs]), which were chlorinated at 2-, 3-, 7-, and 8-positions of the molecules, by the white rot fungus Phanerochaete sordida YK-624 was studied in a stationary low-nitrogen medium. The percent degradation values of PCDDs and PCDFs were approximately 40 (tetra-chloro-) to 76% (hexachloro-) and 45 (tetrachloro-) to 70% (hexachloro-), respectively. Metabolites of 2,3,7,8-tetra- and octaCDD formed by P. sordida YK-624 included 4,5-dichlorocatechol and tetrachlorocatechol, respectively. These results suggest that white rot fungus is able to substantially degrade both PCDDs and PCDFs. This is the first report of the degradation of highly chlorinated PCDDs and PCDFs by a microorganism.


Continence and complications rates after male slings as primary surgery for post-prostatectomy incontinence: a systematic review.
to analyze continence and complications rates after male slings as first line surgical treatment, in order to improve patient counseling for the management of SUI postprostatectomy. A MedLine search using specified search terms was done on January 23, 2012. This research rendered 160 records. No controlled trial was available for analysis. The majority of papers dealing with out- come and complications came from a few centres. At a median follow-up of 15 months the pooled cure rates for all kinds of slings was 77.4; in the AdVance group the pooled cure rates was 72.5%; in the InVance group it was 74.2% while in the Remeex group it was 84.3%. Only a few number observational studies addressed review selection criteria. The pooled overall cure rates is high but there are no data concerning reliable pre- and postoperative prognostic factors affecting treatment failure and complications rates, thus it is not possible to have suitable criteria for a better patient selection. The statistically pooled results obtained should be interpreted with caution because of several limitations due to several study selection limitations: observational study design, few number of analysed studies, heterogeneity, lack of outcome definition and standardisation, between-study variability, high risk of bias.


Squamous cell carcinoma arising from a recurrent ischial pressure ulcer: a case report.
Marjolin's ulcer is the malignant transformation of long-standing chronic pressure ulcers and requires prompt diagnosis and treatment. A 46-year-old man with an 8-year history of traumatic spinal injury with paraplegia presented with a recurrent ischial pressure ulcer. The initial ulcer, which developed 6 years earlier, was a Stage IV sacral ulcer. The wound was debrided and pathology showed epithelial hyperplasia, acanthosis, hyperkatosis accompanied by mild inflammation, and fibrosis without any malignant transformation. The lesion was covered with a fasciocutaneous bipedicled flap. Four years later, the patient presented with a similar ulcer in the same location. Histology showed the presence of a well-differentiated squamous cell carcinoma (SCC). Following a wide excision, the lesion was covered with a gluteal maximal V-Y musculocutaneous advancement flap. At last follow-up 14 months postoperatively, there was no evidence of recurrence or metastatic disease. Clinicians must be aware of known risk factors for the development of SCC.


Using role play to develop positive attitudes toward people with disabilities among dental students: An exploratory pilot study.
Comprehensive and interprofessional undergraduate teaching and training in special care dentistry (SCD) could influence future dentists' attitudes towards patients with disabilities. The aim of the study was to assess the influence of role play as part of SCD training on the attitudes of undergraduates towards patients with disabilities. The students attending the SCD module during the autumn semester of 2019 comprised our target population. A quasi-experimental design was used to compare the attitudinal scores between three groups of students exposed to different modalities of SCD teaching: group 1 - didactic teaching (N = 92), group 2 - didactic teaching and clinical training in the dental treatment of children or elderly patients with disabilities (N = 142), group 3 - theoretical teaching and clinical training, accompanied by role play sessions featuring persons with disabilities (N = 20). The contact rate was 100%. The outcomes of the intervention were measured using the altruism scale, which forms part of the International Association for Disability and Oral Health (iADH) Toolbox for measuring attitudes and intended behaviors towards persons with disabilities. The analysis revealed the two-factor structure of the nine-item altruism scale. The first factor was described as the acceptance of oral health care as a universal human right, while the second was the perception of the contribution of marginalised groups, including people with disabilities, to the community. The students participating in the role play scored higher on both factors in comparison to those attending other modes of SCD training. The presented findings suggested role play as a powerful tool in fostering positive attitudes towards persons with disabilities in the education of dental students and highlighted the importance of multidisciplinary training.


Misleading 18FDG-PET/CT finding caused by chronic HIV infection in a patient with gastric carcinoma: a case report.
The aim of this study was to describe the influence of HIV infection on the staging of gastric cancer by using positron emission tomography/computed tomography (PET/CT) imaging. We treated a patient with local advanced gastric cancer. PET/CT showed an unusual hypermetabolic activity of lymph nodes, and the patient was later found to be HIV-positive. PET/CT scans in cancer staging of patients with chronic HIV infection may show advanced stage disease. Spread of primary tumor to unusual sites should be a warning sign for the possible concurrent diseases.


SQUAT: A web tool to mine human, murine and avian SAGE data.
There is an increasing need in transcriptome research for gene expression data and pattern warehouses. It is of importance to integrate in these warehouses both raw transcriptomic data, as well as some properties encoded in these data, like local patterns. We have developed an application called SQUAT (SAGE Querying and Analysis Tools) which is available at: http://bsmc.insa-lyon.fr/squat/. This database gives access to both raw SAGE data and patterns mined from these data, for three species (human, mouse and chicken). This database allows to make simple queries like "In which biological situations is my favorite gene expressed?" as well as much more complex queries like: <<what are the genes that are frequently co-over-expressed with my gene of interest in given biological situations?>>. Connections with external web databases enrich biological interpretations, and enable sophisticated queries. To illustrate the power of SQUAT, we show and analyze the results of three different queries, one of which led to a biological hypothesis that was experimentally validated. SQUAT is a user-friendly information retrieval platform, which aims at bringing some of the state-of-the-art mining tools to biologists.


Synthesis and Photodimerization of 2- and 2,3-Disubstituted Anthracenes: Influence of Steric Interactions and London Dispersion on Diastereoselectivity.
There is increased evidence that the effect of bulky groups in organic, organometallic, and inorganic chemistry is not only repulsive but can be attractive because of London dispersion interactions. The influence of the size of primary alkyl substituents in 2- and 2,3-positions of anthracenes on the diastereoselectivity (anti vs syn dimer) of the [π4s + π4s] photoinduced dimerization is investigated. The synthesis of the anthracene derivatives was achieved by Suzuki-Miyaura reaction of 2,3-dibromoanthracene with alkylboronic acids as well as by reduction of anthraquinones that were obtained from 2,3-disubstituted 1,3-butadienes and naphthoquinone followed by dehydrogenation. The mixtures of dianthracene isomers were analyzed with respect to the anti/syn-ratio of the products by X-ray crystallography and nuclear Overhauser effect spectroscopy. While for the 2,3-dimethylanthracene the anti and syn isomers were formed in equal amounts, the anti dimers are the major products in all other cases. A linear correlation (R2 = 0.98) between the steric size (Charton parameter) and the isomeric ratio suggests that the selectivity is dominated by classical repulsive steric effects. An exception is the iso-butyl substituent that produces an increased amount of the syn isomer. It is suggested that this is due to an exalted effect of London dispersion interactions.


Lipid-protein correlations in nanoscale phospholipid bilayers determined by solid-state nuclear magnetic resonance.
Nanodiscs are examples of discoidal nanoscale lipid-protein particles that have been extremely useful for the biochemical and biophysical characterization of membrane proteins. They are discoidal lipid bilayer fragments encircled and stabilized by two amphipathic helical proteins named membrane scaffolding protein (MSP), ~10 nm in size. Nanodiscs are homogeneous, easily prepared with reproducible success, amenable to preparations with a variety of lipids, and stable over a range of temperatures. Here we present solid-state nuclear magnetic resonance (SSNMR) studies on lyophilized, rehydrated POPC Nanodiscs prepared with uniformly (13)C-, (15)N-labeled MSP1D1 (Δ1-11 truncated MSP). Under these conditions, by SSNMR we directly determine the gel-to-liquid crystal lipid phase transition to be at 3 ± 2 °C. Above this phase transition, the lipid (1)H signals have slow transverse relaxation, enabling filtering experiments as previously demonstrated for lipid vesicles. We incorporate this approach into two- and three-dimensional heteronuclear SSNMR experiments to examine the MSP1D1 residues interfacing with the lipid bilayer. These (1)H-(13)C and (1)H-(13)C-(13)C correlation spectra are used to identify and quantify the number of lipid-correlated and solvent-exposed residues by amino acid type, which furthermore is compared with molecular dynamics studies of MSP1D1 in Nanodiscs. This study demonstrates the utility of SSNMR experiments with Nanodiscs for examining lipid-protein interfaces and has important applications for future structural studies of membrane proteins in physiologically relevant formulations.


Characterisation of a truncated Toxoplasma gondii surface antigen 2 (SAG2) secreted by the methylotrophic yeast Pichia pastoris.
A truncated form of surface antigen 2 (SAG2) of the protozoan parasite Toxoplasma gondii was cloned and expressed in the methylotrophic yeast Pichia pastoris. This recombinant antigen, designated as recSAG2-N, contained only the N-terminal half of the native SAG2. The recSAG2-N was secreted by the Pichia pastoris into the culture supernatant, and it was harvested by using the trichloroacetic acid precipitation method. Specificity of recSAG2-N was evaluated in western blot assays. Fifty human serum samples, including 32 from confirmed cases of toxoplasmosis, were tested. Results from the assays showed that recSAG2-N reacted with sera from the toxoplasmosis cases only. In vivo experiments showed that serum from mice which received recSAG2-N reacted with the native SAG2 of T. gondii.


Doubling the resolution of a confocal spinning-disk microscope using image scanning microscopy.
Fluorescence microscopy has become an indispensable tool for cell biology. Recently, super-resolution methods have been developed to overcome the diffraction limit of light and have shown living cells in unprecedented detail. Often, these methods come at a high cost and with complexity in terms of instrumentation and sample preparation, thus calling for the development of low-cost, more accessible methods. We previously developed image scanning microscopy (ISM), which uses structured illumination to double the resolution and quadruple the contrast of a confocal microscope. Implementing this technique into a confocal spinning-disk (CSD) microscope allows recording ISM images with up to ~1 frame per second, making it ideal for imaging dynamic biological processes. Here we present a step-by-step protocol describing how to convert any existing commercial CSD microscope into a CSD-ISM, with only moderate changes to the hardware and at low cost. Operation of the CSD-ISM is realized with a field programmable gate array using the software environment Micro-Manager, a popular open-source platform for microscopy. The provided software ( https://projects.gwdg.de/projects/csdism-2020 ) takes care of all algorithmic complexities and numerical workload of the CSD-ISM, including hardware synchronization and image reconstruction. The hardware modifications described here result in a theoretical maximum increase in resolution of √2 ≈ 1.41, which can be further improved by deconvolution to obtain a theoretical maximum twofold increase. An existing CSD setup can be upgraded in ~3 d by anyone with basic knowledge in optics, electronics and microscopy software.


Vasoactive agents and production of thrombosis during intravascular coagulation. 2. alpha-Adrenergic stimulation: effects and mechanisms.
The mechanism whereby norepinephrine elicits thrombosis during intravascular coagulation was investigated further in rabbits given a 4 h infusion of thrombin (1 NIH unit/kg/min). Norepinephrine (3 micrograms/kg/min) combined with thrombin produced glomerular capillary thrombosis in all animals as compared to 4.3% with thrombin alone. Alpha-adrenergic receptors mediated this effect, as indicated (a) by prevention of glomerular thrombosis by dibenzyline but not by methysergide, and (b) by failure of histamine or acetylcholine combined with thrombin to induce the phenomenon. However, in combination with thrombin, these two agents induced duodenal mucosal microthrombosis. Study of the glomerular circulation with colloidal carbon showed that norepinephrine elicits severe glomerular capillary stasis in thrombin treated rabbits; the vasomotor reaction precedes increased fibrinogen consumption and focal deposition of fibrin in the glomeruli. Pretreatment with dibenzyline prevented glomerular stasis and reduced fibrinogen consumption. The phagocytic activity of the reticulo-endothelial system was increased 7 times by thrombin infusions, with or without norepinephrine. We conclude that stimulation of the alpha 1-adrenergic receptors triggers glomerular thrombosis by production of severe glomerular stasis which localizes formation of thrombi in the dilated vessels. These results provide a rational explanation for the role of alpha-adrenergic stimulation in the endotoxin-induced generalized Shwartzman reaction and outline some of the mechanisms and agents implied in the selection of the target organs for thrombosis during intravascular coagulation.


Opportunities for improved risk assessments of exotic species in Canada using bioclimatic modeling.
This paper briefly reviews the process of exotic pest risk assessments and presents some examples of emerging opportunities for spatial bioclimatic modeling of exotic species in Canada. This type of analysis can support risk assessments but does not replace the need for on-going high quality field-based observations to validate and update models. Bioclimatic analysis of several exotic pests is provided to illustrate both opportunities and limits. A link is demonstrated to the National Forest Inventory to characterize timber volumes at risk for one exotic species. 'Challenges' are both scientific and administrative. More accessible and current field survey data are required to improve models. Our experience is that for many exotic species, historical, and even current, data are not always digital or quality controlled for taxonomic identity and accurate geo-referencing. This inhibits their use for integrated spatial modeling applications.


Sustainable Health Informatics: Health Informaticians as Alchemists.
The digital transformation of health care delivery remains an elusive work in progress. Contextual variation continues to be a significant barrier to the development of sustainable health information systems. In this paper we characterize health informaticians as modern alchemists and use this characterization to describe informatics progress in addressing four key healthcare challenges. We highlight the need for informaticians to be diligent and loyal to basic methodological principles while also appreciating the role that contextual variation plays in informatics research. We also emphasize that meaningful health systems transformation takes time. The insight presented in this paper helps informaticians in our quest to develop sustainable health information systems.