Expression of two differentiation antigens on normal and cultured human T cells.
An antiserum specific for human T lymphocytes (AMT) was used to examine patterns of T cell surface antigen expression and to isolate their reactive membrane antigens. By a quantitative adsorption assay, different plateaus of AMT reactivity with blood T cells were observed after serial adsorptions with individual T cell lines. MOLT-3 cells removed 95% of AMT activity to blood T cells whereas MOLT-4 removed 70% and HSB-2 removed only 30%. A cross-adsorption analysis demonstrated that each of the three cell lines differed in their adsorbing efficiency to remove AMT antibodies reactive with the reciprocal cell lines. Radiolabeled membrane proteins were solubilized with either sodium deoxycholate (DOC) or NP-40 detergents, precipitated with AMT, and Staphylococcus aureus Cowan strain I, and analyzed by sodium dodecyl sulfate-polyacrylamide disc gel electrophoresis (SDS-PAGE). Two distinct T lymphocyte antigens of approximately 25,000 daltons (p25) and 16,000 (p16) were identified on MOLT-3 cells. Similar relative quantities of p25 and p16 were detected on human peripheral T cells and thymocytes. MOLT-4 cells contained less of the p25 peak than did MOLT-3. HSB-2 cells gave a small peak in the same general location as the MOLT-3 p25 peak, and a relatively large p16 peak. Adsorption of AMT with HSB-2 removed the capacity of the antiserum to precipitate p16 from MOLT-3, but did not eliminate reactivity with p25.


Processing of syndecan-2 by matrix metalloproteinase-14 and effect of its cleavage on VEGF-induced tube formation of HUVECs.
Syndecans (SDCs) are transmembrane proteoglycans that are involved in cell adhesion and cell communication. Specifically, SDC2 plays a key role in tumorigenesis, metastasis, and angiogenesis. Previously, we found that rat SDC2 is shed by matrix metalloproteinase-7 (MMP-7) in colon cancer cells. Here, we analyzed the susceptibility of rat SDC2 to various MMPs. We found that the rat SDC2 ectodomain (ECD) fused to the C-terminal Fc region, which was expressed in mammalian cells, was cleaved more efficiently by MMP-14 than MMP-7. Likewise, when anchored on the surface of HeLa cells, rat SDC2 was cleaved more efficiently by the treatment of MMP-14 than MMP-7 and was shed more readily by membrane-anchored MMP-14 than soluble MMP-14. Furthermore, MMP-14 cleaved recombinant SDC2-ECD expressed in Escherichia coli into multiple fragments. Using N-terminal amino acid sequencing and the top-down proteomics approach, we determined that the major cleavage sites were S88↓L89, T98↓M99, T100↓L101, D132↓P133, and N148↓L149 for rat SDC2-ECD and S55↓G56, S65↓P66, P75↓K76, N92↓I93 D122↓P123, and S138↓L139 for human SDC2-ECD. Finally, the rat and human SDC2-ECD lost the ability to suppress vascular endothelial growth factor-induced formation of capillary-like tubes by human umbilical vein endothelial cells following cleavage by MMP-14, but its major cleavage-site mutant of rat SDC2-ECD did not. These results suggest that MMP-14 is a novel enzyme responsible for degrading SDC2 and impairing its physiological roles including angiogenesis.


[Dependence of oligomerization of influenza virus nucleoprotein on the species affiliation of the virus].
Comparison of human and avian influenza virus nucleoprotein (NP) oligomerization showed that the efficiency of NP oligomerization is different in influenza viruses of different origin. NP oligomerization is virtually complete in avian influenza viruses, while in human influenza viruses only part of monomeric NP is oligomerized. The authors discuss the utilization of NP oligomerization efficiency as a sign for identification of the origin of influenza virus.


Intracoronary administration of FGF-2: a computational model of myocardial deposition and retention.
This study uses a computational model to characterize the myocardial deposition and retention of basic fibroblast growth factor (FGF-2) at the cellular level after intracoronary (IC) administration of exogenous FGF-2. The model is applied to the in situ conditions present within the myocardium of a dog for which the plasma pharmacokinetics resulting from IC injection of FGF-2 were recorded. Our estimates show that the processes involved in FGF-2 signaling are not diffusion limited; rather, the response time is determined by the reaction time of FGF-2 binding to cell surface receptors. Additionally, the processes of receptor secretion and internalization are found to play crucial roles in the FGF-2 dynamics; future experiments are required to quantify these processes. The model predictions obtained in this study suggest that IC administration of FGF-2 via either a single bolus or repetitive injections causes a transient increase (time scale of hours) in myocardial FGF-2 concentration if the endogenous level of free interstitial FGF-2 is low enough to allow permeation of FGF-2 molecules from the microvascular to the interstitial spaces. The model shows that the majority (64%) of the extracellular FGF-2 ligands are located within the interstitium, and similar fractions are found in the basement membrane and extracellular matrix. Among the FGF-2 molecules found within the interstitium, 2% are free and 98% are bound to interstitial heparan sulfate proteoglycans. These results support the theory of extracellular control of the bioavailability of FGF-2 via dynamic storage of FGF-2 within the basement membrane and extracellular matrix.


Single dissociation findings of ADHD deficits in vigilance but not anterior or posterior attention systems.
Following a distributed network model of visuospatial attention, the authors used an A-X version of the Continuous Performance Test and a covert orienting paradigm to examine the vigilance, anterior, and posterior attention systems. Compared with control participants without attention-deficit/hyperactivity disorder (ADHD), children with the predominantly inattentive (ADHD-I) and combined (ADHD-C) subtypes had lower sensitivity (d') to detect targets from nontargets. Children with ADHD-C, but not ADHD-I, additionally had a highly activated response style (lnbeta). Performance for both subtypes decreased to a greater extent over time in a manner consistent with problems in sustained attention. Together, these results suggest the presence of vigilance system deficits in participants with both ADHD subtypes. However, consistent with previous meta-analytic work, there was no evidence for anterior or posterior system orienting dysfunctions in either subtype.


Hepatitis B and C Virus Infections Among Human Immunodeficiency Virus-Infected People Who Inject Drugs in Lahore, Pakistan.
Hepatitis B virus (HBV) and hepatitis C virus (HCV) are the major cause of the global burden of hepatitis. One of the main routes of transmission for both viruses is through exposure to infected blood, which includes sharing blood-contaminated syringes and needles. Human immunodeficiency virus (HIV) attacks the immune system and results in acquired immune deficiency syndrome and opportunistic infections. The objective of this study was to assess the epidemiology of HBV and HCV infections among HIV-infected people who inject drugs (PWID). The study enrolled 100 PWID from different addiction centers of the city of Lahore in Pakistan. All subjects were HIV-infected males and were above 16 years of age. Screening of HBV and HCV infections was performed through immunochromatography tests and enzyme-linked immunosorbent assays. The prevalence of HCV and HBV infections among the 100 HIV-infected PWID was 55% and 6%, respectively. HIV monoinfection was found in 37% of the subjects, while triple infection was detected in 2% of the subjects. Majority of the HIV-infected PWID were using heroin and Avil injections (65%). Half of the subjects had used injection drugs for 1-5 years, while 32% had used injection drugs for 6-10 years. HCV infection was more common than HBV infection among the enrolled subjects. Most of the PWID were practicing heroin and Avil injections.


Pediatric Brain Tumors: Narrating Suffering and End-of-Life Decisionmaking.
When talking about decisionmaking for children with a life-threatening condition, the death of children with brain tumors deserves special attention. The last days of the lives of these children can be particularly harsh for bystanders, and raise questions about the suffering of these children themselves. In the Netherlands, these children are part of the group for whom a wide range of end-of-life decisions are discussed, and questions raised. What does the end-of-life for these children look like, and what motivates physicians and parents to make decisions that may affect the life and death of these children? This article highlights the story of the parents of the sisters Roos and Noor. When both their daughters were diagnosed with a hereditary brain tumor, they had to make similar decisions twice. Their story sheds light on the suffering of children in the terminal phase, and how this suffering may motivate parents and physicians to make decisions that influence the end of life of these children's lives.We argue that complete knowledge about suffering in the terminal phase of children with brain tumors is impossible. However, by collecting experiences like those of Roos and Noor, we can move toward an experienced-based understanding and better guide parents and physicians through these hardest of decisions.


Risk factors for developing atopic dermatitis.
The aim of this thesis was to investigate possible risk factors affecting the development of AD. AD is a frequent disease among children and has a substantial impact on the lives of both the child and its family. A better understanding of the disease would enable better treatment, prevention and information to the families involved. Previous risk factor studies have been hampered by an unsuitable study design and/or difficulties in standardization when diagnosing AD, which limit their conclusions. In paper I, we conducted a traditional cross-sectional analysis testing 40 possible risk factors for developing AD at 3 years of age. Our data suggested a strong heredity of AD and confirmed the risk associated with the non-functional FLG allele mutations after adjustments for confounders. Besides this mother's dermatitis and father's allergic rhinitis were found to increase the risk of AD. Perinatal exposure to dog was the only environmental exposure that significantly reduced the disease manifestation, suggesting other, yet unknown environmental factors affecting the increasing prevalence of AD in children. Length at birth was shown to be inversely associated with the risk of later developing AD. This traditional risk factor analysis led to two borderline significant results: duration of exclusive breastfeeding and mother's alcohol intake during the 3rd trimester. Since these possible two risk factors could neither be rejected nor accepted, we decided to do two in-depth studies, further investigating these, using longitudinal data information and data analysis instead of the traditional cross-sectional approach (paper II & III). In paper II, we investigated the risk of developing AD and wheezy symptoms until age 2 years depending on duration of breastfeeding. We found an increased risk of AD, but a protective effect on wheezy disorders in infancy from exclusive breastfeeding. The effect of exclusive breastfeeding on the risk of development of AD was significant after adjustment for demographics, FLG variants R501X and 2282del4 status, parent's AD and pets at home (RR 2.09, 95% CI 1.15-3.80, p=0.016). In addition, there was a significant effect of duration of exclusive breastfeeding (p=0.043), as the relative risk of AD was increased in proportion to increased duration of breastfeeding. The risk associated with exclusive breastfeeding was not explained by the fatty acid composition of mother's milk, though a trend showed higher risk of AD if mother's milk had low concentrations of n-3 fatty acids. In paper III, we found that alcohol intake during pregnancy was associated with a significantly higher risk of developing AD in the offspring, with the effect persisting throughout the whole 7 years follow-up period (HR 1.44, 95% CI 1.05-1.99, p=0.024). The increased risk was still significant after confounder adjustment for mother's education, AD and smoking habits during the 3rd trimester. There was no association between alcohol intake during pregnancy and other atopic endpoints (wheeze episodes, asthma, allergic rhinitis, blood eosinophil count, total IgE, sensitization, cord blood IgE and nasal eosinophilia). However, the underlying explanation was not clear. The thesis is based on data collected as part of the ongoing COPSAC cohort. The cohort is a longitudinal, prospective birth cohort following 411 children born to mothers with asthma. This selection of high-risk children restricts the interpretation of the results and they cannot necessarily be expanded to apply to the general population.


The concept of 'intent' within Australian coronial data: factors affecting the National Coronial Information System's classification of mortality attributable to intentional self-harm.
Within Australia all unexpected deaths are investigated by the Coroners Court; specifically, the coroner investigates the identity of the deceased and the cause and circumstances of death. This 'unexpected death' category inevitably includes cases of self-harm and suicide. Concerns regarding the accurate reporting of national suicide statistics resulted in a review of the coding process undertaken by the Australian Bureau of Statistics (ABS), which produces the national statistics, and a formal Commonwealth Government Senate Inquiry in 2009. This article reflects data and opinions collected prior to the Senate Inquiry or the adjustment of the ABS coding processes, and explores the role of the Coroner in determining the intent of the deceased person and the role the National Coronial Information System (NCIS) 1 database plays in the provision of this information. At the Case Notification and Case Closure stages of the coronial process, administrative coders abstract from the coronial file the 'intent' of the deceased and enter the data into relevant administrative systems (which upload to the NCIS). The relevant intent code in the NCIS is 'Intentional Self-Harm', which incorporates deliberate actions of self-harm and suicide. A mixed-method study was employed to investigate anecdotal reports of a problematic coronial coding process surrounding this category of cases. A sample of Australian coroners (n=16), and of the national population of NCIS coders (n=36), were surveyed using separate instruments, and an unobtrusive case review of sampled NCIS cases (n=127) reflecting nine key mechanisms-of-death, was undertaken. Each Australian state and territory has its own Coroners Act, none of which provides legislative direction regarding the determination of intent by the coroner. Neither the coroner-respondents nor the coders favoured a standard proforma to record 'intent'. In order to inform their classificatory decision-making regarding the deceased's 'intent', the coders need to abstract extensively from the entire case file, scrutinising documentary materials from different investigators. They rely primarily on the police report at Case Notification and the coroner's finding at Case Completion. Coders do not generally perceive the classification of 'intent' to be problematic; however, despite NCIS-provided coder (technical) support materials, there exist inconsistent coder work practices and, sometimes, absent documentary evidence reflecting lack of information for ascertainment and interpretation by the coroner, investigators, and forensic experts on the 'intent' of the deceased. The gap between what a coroner is legally required to document regarding 'intent' and what society needs to know for statistical and preventive purposes, seems problematic to bridge.


The consumption of viruses returns energy to food chains.
Viruses impact host cells and have indirect effects on ecosystem processes. Plankton such as ciliates can reduce the abundance of virions in water, but whether virus consumption translates into demographic consequences for the grazers is unknown. Here, we show that small protists not only can consume viruses they also can grow and divide given only viruses to eat. Moreover, the ciliate Halteria sp. foraging on chloroviruses displays dynamics and interaction parameters that are similar to other microbial trophic interactions. These results suggest that the effect of viruses on ecosystems extends beyond (and in contrast to) the viral shunt by redirecting energy up food chains.


Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM).
We have developed a high-resolution fluorescence microscopy method based on high-accuracy localization of photoswitchable fluorophores. In each imaging cycle, only a fraction of the fluorophores were turned on, allowing their positions to be determined with nanometer accuracy. The fluorophore positions obtained from a series of imaging cycles were used to reconstruct the overall image. We demonstrated an imaging resolution of 20 nm. This technique can, in principle, reach molecular-scale resolution.


The antioxidant systems in Toxoplasma gondii and the role of cytosolic catalase in defence against oxidative injury.
Superoxide dismutase, catalase, glutathione peroxidase and peroxiredoxins form an antioxidant network protecting cells against reactive oxygen species (ROS). Catalase is a potent H2O2-detoxifying enzyme, which is unexpectedly absent in some members of the Kinetoplastida and Apicomplexa, but present in Toxoplasma gondii. In T. gondii, catalase appears to be cytosolic. In addition, T. gondii also possesses genes coding for other types of peroxidases, including glutathione/thioredoxin-like peroxidases and peroxiredoxins. This study presents a detailed analysis of the role of catalase in the parasite and reports the existence of antioxidant enzymes localized in the cytosol and the mitochondrion of T. gondii. The catalase gene was disrupted and, in addition, T. gondii cell lines overexpressing either catalase or a cytosolic 1-cys peroxiredoxin, TgPrx2, under the control of a strong promoter were created. Analysis of these mutants confirmed that the catalase activity is cytosolic and is encoded by a unique gene in T. gondii. Furthermore, the catalase confers protection against H2O2 exposure and contributes to virulence in mice. The overexpression of Prx2 also increases protection against H2O2 treatment, suggesting that catalase and other peroxidases function as a defence mechanism against endogenously produced reactive oxygen intermediates and the oxidative stress imposed by the host.


Superoxide dismutase polyethylene glycol improves survival in hemorrhagic shock.
Oxygen free radicals are known to form after reperfusion of ischemic tissue. To test the role and importance of oxygen free radicals in hemorrhagic shock, an animal model of hemorrhagic shock and resuscitation was utilized. Sprague-Dawley rats were anesthetized with halothane and then subjected to approximately 50 per cent blood volume hemorrhage (30 cc/kg), followed by a 60 min shock period. Resuscitation was performed over 1 hour with lactated ringers (LR) at a volume of two times blood loss (60 cc/kg). This model results in a survival rate of 25 per cent over 72 hrs. Using this model, animals were randomized to receive either LR, Superoxide Dismutase-Polyethylene Glycol (SOD-PEG) (15,000 units/kg) with LR or Catalase-Polyethylene Glycol (CAT-PEG) (175,000 units/kg) with LR. The group treated with SOD-PEG demonstrated significantly increased survival rates vs the group treated with LR (67% vs 25%, P = 0.02). The group treated with CAT-PEG demonstrated no significant improvement in survival when compared to the LR-treated group (20% vs 24%). These data suggest that treatment directed toward oxygen free radicals and reperfusion injury may play an important role in hemorrhagic shock resuscitation.


Status of genome projects for nonpathogenic bacteria and archaea.
Since the first microbial genome was sequenced in 1995, 30 others have been completed and an additional 99 are known to be in progress. Although the early emphasis of microbial genomics was on human pathogens for obvious reasons, a significant number of sequencing projects have focused on nonpathogenic organisms, beginning with the release of the complete genome sequence of the archaeon Methanococcus jannaschii in 1996. The past 18 months have seen the completion of the genomes of several unusual organisms, including Thermotoga maritima, whose genome reveals extensive potential lateral transfer with archaea; Deinococcus radiodurans, the most radiation-resistant microorganism known; and Aeropyrum pernix, the first Crenarchaeota to be completely sequenced. Although the functional characterization of genomic data is still in its initial stages, it is likely that microbial genomics will have a significant impact on environmental, food, and industrial biotechnology as well as on genomic medicine.


Postoperative care following single-stage laryngotracheoplasty.
Single-stage laryngotracheoplasty (SSLTP) provides a method of correcting mild-to-moderate laryngotracheal stenosis while avoiding the risks of prolonged laryngeal stenting. Pediatric patients are orally intubated for 5 to 7 days postoperatively to ensure an adequate airway while edema resolves and healing begins. During this period, continuous neuromuscular blockade has been advocated in infants and young children to avoid endotracheal tube trauma to the fresh graft and potentially life-threatening accidental decannulation. Pulmonary atelectasis is the most common morbidity associated with prolonged neuromuscular blockade. Neuromuscular weakness also may follow prolonged paralysis and prolong hospitalization. This paper compares the postoperative course of 17 patients who underwent 18 SSLTP procedures by the senior author. The first 8 patients received continuous neuromuscular blockade in the early postoperative period. To reduce perceived morbidity, the last 9 patients were managed with a protocol that incorporated daily 4- to 8-hour "interruptions" of paralysis. Seven patients tolerated this protocol modification. As a group, these patients had less postoperative pulmonary atelectasis prior to extubation (p < .05) and were extubated sooner than patients receiving continuous neuromuscular blockade (p<.05) without compromising the surgical success of the procedure. Intermittent paralysis permitted for more accurate assessment of pain control and protected against accidental drug accumulation. Although self-extubation did not occur, diligent nursing care with adequate sedation and analgesia is necessary to avoid the risk of accidental extubation.


Cerebral Diffusivity Changes in Fetuses with Chiari II Malformation.
Chiari II malformation is a congenital abnormality characterized by a small posterior fossa with downward displacement of the hindbrain into the foramen magnum. Diffusion-weighted imaging (DWI) can be used to quantitatively characterize brain injury and physiological maturation. We aim to evaluate DWI parameters of the infra- and supratentorial brain in fetuses with Chiari II malformation. MRI and US studies of 26 fetuses with Chiari II malformation and 15 controls were evaluated for the presence/severity of hydrocephalus, myelomeningocele level, and brain apparent diffusion coefficient (ADC) values. Cerebral flow impedance parameters (resistance and pulsatile indexes, resistance index [RI], and pulsatility index [PI] respectively) from concurrently preformed fetal US were also evaluated. Of the Chiari II group, 16 fetuses had hydrocephalus. There was increasing severity of lateral ventriculomegaly in the high compared to low spinal dysraphism subgroups. There were significant lower ADC values in the frontal (p = 0.01) and temporal lobes (p = 0.05) in Chiari II group compared with normal, regardless of the presence or severity of hydrocephalus. Fetuses with Chiari II showed significantly lower RI and PI values. Abnormal ADC values indicate supratentorial microstructural changes in fetuses with Chiari II malformation. Further investigation of the role of diffusion imaging metrics in evaluating abnormal brain development, parenchymal damage and efficacy of fetal surgery is needed.


Nitric oxide mediates selective degeneration of hypothalamic orexin neurons through dysfunction of protein disulfide isomerase.
We addressed the role of nitric oxide (NO) in orexin neuron degeneration that has been observed under various pathological conditions. Administration of an NO donor NOC18 (50 nmol) into the third ventricle of mice resulted in a significant decrease of orexin-immunoreactive (-IR) neurons, in contrast to a modest change in melanin-concentrating hormone-IR neurons. In addition, NOC18 promoted formation of orexin-A-IR aggregates within orexin neurons. An endoplasmic reticulum stress inducer tunicamycin replicated the effect of NOC18 with regard to decrease of orexin-IR neurons and formation of aggregates. We also found that NOC18 caused an increase in S-nitrosation of protein disulfide isomerase (PDI) and a decrease in PDI activity in hypothalamic tissues. Moreover, PDI inhibitors, such as cystamine and securinine, caused a selective decrease of orexin neurons and promoted formation of orexin-A-IR aggregates. Aggregate formation in orexin-IR neurons was also induced by local injection of small interfering RNA targeting PDI. Interestingly, sleep deprivation for 7 consecutive days induced a selective decrease of orexin-IR neurons, which was preceded by aggregate formation in orexin-IR neurons and an increase in S-nitrosated PDI in the hypothalamus. Activity of neuronal NO synthase (nNOS)-positive neurons in the lateral hypothalamus as assessed by c-Fos expression was elevated in response to sleep deprivation. Finally, sleep deprivation-induced decrease of orexin-IR neurons, formation of aggregates, and S-nitrosation of PDI were not observed in nNOS knock-out mice. These results indicate that nNOS-derived NO may mediate specific pathological events in orexin neurons, including neuropeptide misfolding via S-nitrosation and inactivation of PDI.


Effects of prenatal nicotine exposure on biochemical development of rat brain regions: maternal drug infusions via osmotic minipumps.
The effects of a continuous 16-day gestational exposure to nicotine on brain development were examined in the offspring of dams who received a minipump implant on the 4th day of gestation. Maternal viability was unaffected and weight gain was only reduced slightly, but nearly half the dams failed to give birth; dams delivering pups had normal litter sizes. Examination of fetal macromolecules on the 18th day of gestation revealed specific deficits in cell number (DNA) in developing brain tissue as opposed to the rest of the fetus, accompanied by parallel shortfalls in other macromolecules (RNA, protein). After birth, brain development in the nicotine-exposed animals showed persistent abnormalities in the timing of maturational events, with elevated levels of ornithine decarboxylase (an enzymatic marker related to cellular maturation) detectable in all brain regions. Subsequent effects on macromolecules were highly selective regionally, with clear distinctions between areas in which neuronal replication occurs relatively late (cerebellum) compared to early-developing regions (midbrain plus brainstem). Differences apparent between the effects of infused maternal nicotine and those noted previously in studies with nicotine injections indicate that the drug does exert primary effects on developing neural tissues, but that other factors associated with the injection route (such as hypoxia and ischemia consequent to acute effects of nicotine) can interact with the drug to influence brain cell maturation.


Inhibition of respiratory neural discharges by clonidine and 5-hydroxytryptophan.
Activation of receptors for norepinephrine or serotonin in the central nervous system by i.v. injection of clonidine (10-50 micrograms/kg) or 5-hydroxytryptophan (20-40 mg/kg) inhibits phrenic neural discharges in anesthetized, artificially ventilated cats. Clonidine induces a rapid and complete inhibition of phrenic nerve activity which lasts for 1 to 3.2 hr. The inhibition is prevented by prior administration of phenoxybenzamine (10 mg/kg) or tolazoline (3 mg/kg). 5-Hydroxytryptophan, injected after inhibition of peripheral amino acid decarboxylase (carbidopa, 30-50 mg/kg), elicits a gradual but complete inhibition of phrenic nerve discharges which persists for 1 to 10 hr and is unaltered by alpha or beta adrenoceptor blocking agents. The inhibitions produced by clonidine and 5-hydroxytryptophan are overcome transiently during hypercapnia. Stimulation of carotid body chemoreceptors by i.a. injections of lobeline, doxapram or 0.015 N HCl in saline also briefly reinstates phrenic nerve discharges after inhibition by clonidine. Inhibition is also overcome during electrical stimulation of the carotid sinus nerve.


Skp2-dependent ubiquitination and activation of LKB1 is essential for cancer cell survival under energy stress.
LKB1 is activated by forming a heterotrimeric complex with STRAD and MO25. Recent studies suggest that LKB1 has pro-oncogenic functions, besides acting as a tumor suppressor. How the LKB1 activity is maintained and how LKB1 regulates cancer development are largely unclear. Here we show that K63-linked LKB1 polyubiquitination by Skp2-SCF ubiquitin ligase is critical for LKB1 activation by maintaining LKB1-STRAD-MO25 complex integrity. We further demonstrate that oncogenic Ras acts upstream of Skp2 to promote LKB1 polyubiquitination by activating Skp2-SCF ubiquitin ligase. Moreover, Skp2-mediated LKB1 polyubiquitination is required for energy-stress-induced cell survival. We also detected overexpression of Skp2 and LKB1 in late-stage hepatocellular carcinoma (HCC), and their overexpression predicts poor survival outcomes. Finally, we show that Skp2-mediated LKB1 polyubiquitination is important for HCC tumor growth in vivo. Our study provides new insights into the upstream regulation of LKB1 activation and suggests a potential target, the Ras/Skp2/LKB1 axis, for cancer therapy.


Leadership development practices and hospital financial outcomes.
Affordable Care Act legislation is requiring leaders in US health systems to adapt to new and very different approaches to improving operating performance. Research from other industries suggests leadership development can be a helpful component of organizational change strategies; however, there is currently very little healthcare-specific research available to guide design and deployment. The goal of this exploratory study is to examine potential relationships between specific leadership development practices and health system financial outcomes. Results from the National Center for Healthcare Leadership survey of leadership development practices were correlated with hospital and health system financial performance data from the 2013 Medicare Cost Reports. A general linear regression model, controlling for payer mix, case-mix index, and bed size, was used to assess possible relationships between leadership practices and three financial performance metrics: operating margin, days cash on hand, and debt to capitalization. Statistically significant associations were found between hospital-level operating margins and 5 of the 11 leadership practices as well as the composite score. Relationships at the health system level, however, were not statistically significant. Results provide preliminary evidence of an association between hospital financial performance and investments made in developing their leaders.


Influence of implant-abutment interface design on bone and soft tissue levels around immediately placed and restored single-tooth implants: a randomized controlled clinical trial.
The purpose of the present study was to compare the clinical and radiographic outcomes of single implants immediately placed and restored with two different implant-abutment connections. Forty subjects requiring single maxillary premolar replacement were consecutively included in this study and prospectively followed for 12 months. One implant was placed at the time of tooth extraction and immediately restored in each patient. Subjects were randomly selected to receive either prosthetic abutments with a Morse taper connection and a platform switch (test group) or conventional abutments with an internal connection and a matching diameter (control group). A provisional screw-retained crown was positioned and adjusted for nonfunctional loading within 24 hours. Four months later, the definitive crowns were delivered. Periodontal parameters and clinical and radiographic measurements of soft and hard tissue levels were recorded at the moment of prosthesis placement and at 4 and 12 months afterward. Means of the two groups were compared using paired and independent-sample t tests (P = .05). Of the 40 patients recruited, 38 (24 women and 14 men) completed the study. No implants were lost in the control group, whereas one implant failed in the test group. At the 12-month examination, no statistically significant differences were seen between the two groups for periodontal parameters, marginal soft tissue level change, or papilla height (P > .05), but greater marginal bone loss was observed at the control sites (0.51 ± 0.24 mm) compared to the test sites (0.2 ± 0.17 mm) (P = .0004). Although the control group demonstrated a slight increase in marginal bone loss compared to the test group, the peri-implant soft tissues were very stable with both types of implant-abutment connection after 12 months of loading.


Genomic structure of the mouse gene (Lmnb1) encoding nuclear lamin B1.
The mouse gene (Lmnb1) that encodes nuclear lamin B1 was isolated. Structural analyses revealed that the lamin B1 gene spans about 43 kb of the genome and consists of 11 exons and 10 introns. Exon/intron structure of the B1 gene clearly showed the conserved organization shared among the intermediate filament protein family genes. The presumptive promoter region has high GC content and contains a CAAT box and multiple SP1 sites but no classical TATA box, suggesting that the lamin B1 gene has a typical housekeeping gene promoter with a CpG island. These data reveal the gene structure of the only remaining unanalyzed mouse somatic lamin gene. Gene structures of all the mouse somatic lamins are compared.


A systematic review and meta-analysis of the randomized controlled trials on adjuvant intraperitoneal chemotherapy for resectable gastric cancer.
The purpose of this systematic review and meta-analysis was to determine the effectiveness and safety of adjuvant intraperitoneal chemotherapy for patients with locally advanced resectable gastric cancer. Studies eligible for this systematic review included those in which patients with gastric cancer were randomly assigned to receive surgery combined with intraperitoneal chemotherapy versus surgery without intraperitoneal chemotherapy. There were no language restrictions. After independent quality assessment and data extraction, data were pooled for meta-analysis. Thirteen reports of randomized controlled trials (RCTs) were included for quality appraisal and data extraction. Ten reports were judged to be of fair quality and subjected to meta-analysis. A significant improvement in survival was associated with hyperthermic intraoperative intraperitoneal chemotherapy (HIIC) alone (hazard ratio [HR] = 0.60; 95% CI = 0.43 to 0.83; p = 0.002) or HIIC combined with early postoperative intraperitoneal chemotherapy (EPIC) (HR = 0.45; 95% CI = 0.29 to 0.68; p = 0.0002). There was a trend towards survival improvement with normothermic intraoperative intraperitoneal chemotherapy (p = 0.06), but this was not significant with either EPIC alone or delayed postoperative intraperitoneal chemotherapy. Intraperitoneal chemotherapy was also found to be associated with higher risks of intra-abdominal abscess (RR = 2.37; 95% CI = 1.32 to 4.26; p = 0.003) and neutropenia (RR = 4.33; 95% CI = 1.49 to 12.61; p = 0.007). The present meta-analysis indicates that HIIC with or without EPIC after resection of advanced gastric primary cancer is associated with improved overall survival. However, increased risk of intra-abdominal abscess and neutropenia are also demonstrated.


[Progressive necrotizing otitis externa in diabetics with disorders of the cerebral nerves (author's transl)].
The disease occurs without exception in elderly diabetics. Pseudomonas aeruginosa (pyocyaneus) is always found in the secretion. In our reported case of a 71-year old man appeared in addition to a fetid, festering, granulating otitis externa, disturbances of swallowing, hoarseness and severe occipital headaches appear after five months. Neurologically, the N. glossopharyngeus, the N. recurrens and the N. hypoglossus were shown to be paretic. The inflammatory process has pushed forward via the bone of the auditory meatus on to the base of the skull to the jugular foramen and the foramen lacerum. Surgical removal of the inflammatory changes and subsequent treatment with carbenicillin is recommended as the treatment of choice.


Functional analysis of NLP genes from Botrytis elliptica.
SUMMARY We functionally analysed two Nep1-like protein (NLP) genes from Botrytis elliptica (a specialized pathogen of lily), encoding proteins homologous to the necrosis and ethylene-inducing protein (NEP1) from Fusarium oxysporum. Single gene replacement mutants were made for BeNEP1 and BeNEP2, providing the first example of transformation and successful targeted mutagenesis in this fungus. The virulence of both mutants on lily leaves was not affected. BeNEP1 and BeNEP2 were individually expressed in the yeast Pichia pastoris, and the necrosis-inducing activity was tested by infiltration of both proteins into leaves of several monocots and eudicots. Necrotic symptoms developed on the eudicots tobacco, Nicotiana benthamiana and Arabidopsis thaliana, and cell death was induced in tomato cell suspensions. No necrotic symptoms developed on leaves of the monocots rice, maize and lily. These results support the hypothesis that the necrosis-inducing activity of NLPs is limited to eudicots. We conclude that NLPs are not essential virulence factors and they do not function as host-selective toxins for B. elliptica.


Construction of miRNA-miRNA networks revealing the complexity of miRNA-mediated mechanisms in trastuzumab treated breast cancer cell lines.
Trastuzumab is a monoclonal antibody frequently used to prevent the progression of HER2+ breast cancers, which constitute approximately 20% of invasive breast cancers. microRNAs (miRNAs) are small, non-coding RNA molecules that are known to be involved in gene regulation. With their emerging roles in cancer, they are recently promoted as potential candidates to mediate therapeutic actions by targeting genes associated with drug response. In this study we explored miRNA-mediated regulation of trastuzumab mechanisms by identifying the important miRNAs responsible for the drug response via homogenous network analysis. Our network model enabled us to simplify the complexity of miRNA interactions by connecting them through their common pathways. We outlined the functionally relevant miRNAs by constructing pathway-based miRNA-miRNA networks in SKBR3 and BT474 cells, respectively. Identification of the most targeted genes revealed that trastuzumab responsive miRNAs favourably regulate the repression of targets with longer 3'UTR than average considered to be key elements, while the miRNA-miRNA networks highlighted central miRNAs such as hsa-miR-3976 and hsa-miR-3671 that showed strong interactions with the remaining members of the network. Furthermore, the clusters of the miRNA-miRNA networks showed that trastuzumab response was mostly established through cancer related and metabolic pathways. hsa-miR-216b was found to be the part of the most powerful interactions of metabolic pathways, which was defined in the largest clusters in both cell lines. The network based representation of miRNA-miRNA interactions through their shared pathways provided a better understanding of miRNA-mediated drug response and could be suggested for further characterization of miRNA functions.


A high-density graphene-sulfur assembly: a promising cathode for compact Li-S batteries.
This work reports a high-density graphene/sulfur assembly for compact Li-S batteries with high volumetric capacity, which retains good structural stability and conductivity. This dense assembly was prepared by a reduction-triggered self-assembly of graphene oxide with simultaneous deposition of sulfur, followed by unique evaporation-induced spatial volume shrinkage. This assembly has an ultrahigh density, delivering an unprecedented volumetric capacity that is much higher than common carbon/sulfur cathodes. In particular, the unique spatial confinement derived from the shrinkage of the graphene/sulfur assembly is favorable for stabilizing sulfur cathodes.


Cutaneous Melanocytic Tumor with CRTC1::TRIM11 Fusion: Review of the Literature of a Potentially Novel Entity.
"Cutaneous melanocytic tumor with CRTC1::TRIM11 fusion" (CMTCT) is a recently described entity belonging to the family of superficial tumors displaying melanocytic differentiation. Thirteen cases have been reported so far, on the head and neck, extremities, and trunk of adults of all ages (12 cases) and one in an 11-year-old child. Histopathologically, it is a nodular or multilobulated tumor composed of spindle and epithelioid cells arranged in nests, fascicles, or bundles that are surrounded by thin collagenous septa. By immunohistochemistry, the tumor shows variable immunoreactivity for S100-protein, SOX10, and MITF, as well as specific melanocytic markers such as MelanA and HMB-45. The neoplasm's biologic behavior remains uncertain since the reported cases are limited and the follow-up is short (median 12 months). However, local recurrence and synchronous distant metastasis after 13 years of initial resection has been described in one case. Herein, we present a comprehensive literature review of CMTCT hoping to raise awareness among the dermatopathologists of this potentially novel entity.


Etiological factors and temporomandibular treatment outcomes: the effects of trauma and psychological dysfunction.
This paper examines the effect of trauma and psychological dysfunction as etiological factors in temporomandibular disorder (TMD). It employs a thoroughly validated measurement system, the TMJ Scale, to determine the effects of traumatic temporomandibular joint injury as well as pre-treatment stress and psychological dysfunction levels upon presenting symptom levels. It also addresses these parameters for the eventual treatment outcome. During the course of the study, 754 patients were evaluated at the author's practice, which is limited to the diagnosis and Phase I treatment of temporomandibular dysfunction. Of those individuals, 693 (91.9%) were found to have clinically treatable temporomandibular disorders. At the time of this study, 201 consecutive patients (29%) have completed treatment and were deemed to have reached Maximum Medical Improvement (MMI). The validated measurement system of the TMJ Scale was readministered to this post treatment population. Data analysis revealed that trauma patients did not differ from non-trauma patients in initial symptom levels, nor in levels of symptom improvement (with the exception of a higher palpation pain level reported by the trauma patients). Stress and psychological dysfunction were predictive of higher initial symptom perception levels, but were not significantly related to treatment outcomes. These findings have important implications for practitioners in the field of temporomandibular studies. If it can be confirmed that psychological variables have no impact on treatment outcome, it would be difficult to justify the now frequently employed "dual axis" classifications and major emphasis placed on psychological treatment for temporomandibular patients.


Use of Services among Female Sex Workers Receiving a Comprehensive HIV Enhanced Prevention Intervention in Kampala, Uganda: A Cross-sectional Study.
We estimated the prevalence and factors associated with the use of a dedicated HIV prevention, care, and treatment service for female sex workers (FSWs) in Kampala, Uganda. Between October 2017 to January 2018, we conducted a cross-sectional study among FSWs at a dedicated clinic. We defined use as the use of the HIV prevention, care, and treatment services by FSWs at least once within the past six months. We used the log-binomial model to determine the factors associated with use of clinic services. Eight hundred and seventy-four women were included in the analysis. The overall prevalence of use of clinic services was 81%. At adjusted analysis, use of clinic services was independently associated with being HIV positive and being treated for STIs in the past three months. The prevalence of use of clinic services was high. Dedicated services for FSWs are required to support their use of HIV and STI care.


Nucleus accumbens mediates the pronociceptive effect of sleep deprivation: the role of adenosine A2A and dopamine D2 receptors.
Sleep disorders increase pain sensitivity and the risk of developing painful conditions; however, the underlying mechanisms are poorly understood. It has been suggested that nucleus accumbens (NAc) influences sleep-wake cycle by means of a balance between adenosine activity at A2A receptors and dopamine activity at D2 receptors. Because the NAc also plays an important role in pain modulation, we hypothesized that the NAc and its A2A and D2 receptors mediate the pronociceptive effect of rapid eye movement (REM) sleep deprivation (SD). We found that 24 hours of REM-SD induced an intense pronociceptive effect in Wistar rats, which decreases progressively over a sleep rebound period. Although the level of fecal glucocorticoid metabolites increased with SD within group, it did not differ between sleep-deprived group and control group, indicating a stress response with similar magnitude between groups. The pronociceptive effect of REM-SD was prevented by excitotoxic lesion (N-Methyl-D-aspartate, 5.5 μg) of NAc and reverted by its acute blockade (Qx-314, 2%). The administration of an A2A receptor antagonist (SCH-58261, 7 ng) or a D2 receptor agonist (piribedil, 6 μg) into the NAc increased home cage activity and blocked the pronociceptive effect of REM-SD. Complementarily, an A2A receptor agonist (CGS-21680, 24 ng) impaired the reversal of the pronociceptive effect and decreased home cage activity, as it did a D2 receptor antagonist (raclopride, 5 μg). Rapid eye movement SD did not affect the expression of c-Fos protein in NAc. These data suggest that SD increases pain by increasing NAc adenosinergic A2A activity and by decreasing NAc dopaminergic D2 activity.


Evaluation of a method for determining concentrations of isoeugenol, an AQUI-S residue, in fillet tissue from freshwater fish species.
AQUI-S is a fish anesthetic/sedative that is approved for use in a number of countries throughout the world and has the potential for use in the United States. The active ingredient in AQUI-S is isoeugenol. A method for determining isoeugenol concentrations in edible fillet tissue is needed for regulatory purposes, including surveillance and potential use in studies fulfilling human food safety data requirements if U.S. Food and Drug Administration approval is pursued. A method was developed and evaluated for determining isoeugenol concentrations in fillet tissue using relatively common procedures and equipment. The method produced accurate and precise results with fillet tissue from 10 freshwater fish species. The percentage of isoeugenol recovered from samples fortified with isoeugenol at nominal concentrations of 1, 50, and 100 microg/g for all species was always >80 and <97%. Within-day precision for samples fortified at those same concentrations was < or =10%, and day-to-day precision was < or =4.0%. Method precision with fillet tissue containing biologically incurred isoeugenol was < or =8.1%. There were no or minimal chromatographic interferences in control fillet tissue extracts from 9 of the 10 species. The method detection limits for all but one species ranged from 0.004 to 0.014 microg/g, and the quantitation limits ranged from 0.012 to 0.048 microg/g.


Injectable Biomedical Devices for Sensing and Stimulating Internal Body Organs.
The rapid pace of progress in implantable electronics driven by novel technology has created devices with unconventional designs and features to reduce invasiveness and establish new sensing and stimulating techniques. Among the designs, injectable forms of biomedical electronics are explored for accurate and safe targeting of deep-seated body organs. Here, the classes of biomedical electronics and tools that have high aspect ratio structures designed to be injected or inserted into internal organs for minimally invasive monitoring and therapy are reviewed. Compared with devices in bulky or planar formats, the long shaft-like forms of implantable devices are easily placed in the organs with minimized outward protrusions via injection or insertion processes. Adding flexibility to the devices also enables effortless insertions through complex biological cavities, such as the cochlea, and enhances chronic reliability by complying with natural body movements, such as the heartbeat. Diverse types of such injectable implants developed for different organs are reviewed and the electronic, optoelectronic, piezoelectric, and microfluidic devices that enable stimulations and measurements of site-specific regions in the body are discussed. Noninvasive penetration strategies to deliver the miniscule devices are also considered. Finally, the challenges and future directions associated with deep body biomedical electronics are explained.


Factors affecting commencement and cessation of smoking behaviour in Malaysian adults.
Tobacco consumption peak in developed countries has passed, however, it is on the increase in many developing countries. Apart from cigarettes, consumption of local hand-rolled cigarettes such as bidi and rokok daun are prevalent in specific communities. Although factors associated with smoking initiation and cessation has been investigated elsewhere, the only available data for Malaysia is on prevalence. This study aims to investigate factors associated with smoking initiation and cessation which is imperative in designing intervention programs. Data were collected from 11,697 adults by trained recording clerks on sociodemographic characteristics, practice of other risk habit and details of smoking such as type, duration and frequency. Smoking commencement and cessation were analyzed using the Kaplan-Meier estimates and log-rank tests. Univariate and multivariate Cox proportional hazard regression models were used to calculate the hazard rate ratios. Males had a much higher prevalence of the habit (61.7%) as compared to females (5.8%). Cessation was found to be most common among the Chinese and those regularly consuming alcoholic beverages. Kaplan-Meier plot shows that although males are more likely to start smoking, females are found to be less likely to stop. History of betel quid chewing and alcohol consumption significantly increase the likelihood of commencement (p < 0.0001), while cessation was least likely among Indians, current quid chewers and kretek users (p < 0.01). Gender, ethnicity, history of quid chewing and alcohol consumption have been found to be important factors in smoking commencement; while ethnicity, betel quid chewing and type of tobacco smoked influences cessation.


Medication adherence among community-dwelling patients with heart failure.
To determine medication use and adherence among community-dwelling patients with heart failure (HF). Residents of Olmsted County, Minnesota, with HF were recruited from October 10, 2007, through February 25, 2009. Pharmacy records were obtained for the 6 months after enrollment. Medication adherence was measured by the proportion of days covered (PDC). A PDC of less than 80% was classified as poor adherence. Factors associated with medication adherence were investigated. Among the 209 study patients with HF, 123 (59%) were male, and the mean ± SD age was 73.7 ± 13.5 years. The median (interquartile range) number of unique medications filled during the 6-month study period was 11 (8-17). Patients with a documented medication allergy were excluded from eligibility for medication use within that medication class. Most patients received conventional HF therapy: 70% (147/209) were treated with β-blockers and 75% (149/200) with angiotensin-converting enzyme inhibitors or angiotensin II receptor blockers. Most patients (62%; 127/205) also took statins. After exclusion of patients with missing dosage information, the proportion of those with poor adherence was 19% (27/140), 19% (28/144), and 13% (16/121) for β-blockers, angiotensin-converting enzyme inhibitors or angiotensin II receptor blockers, and statins, respectively. Self-reported data indicated that those with poor adherence experienced more cost-related medication issues. For example, those who adhered poorly to statin therapy more frequently reported stopping a prescription because of cost than those with good adherence (46% vs 6%; P < .001), skipping doses to save money (23% vs 3%; P = .03), and not filling a new prescription because of cost (46% vs 6%; P < .001). Community-dwelling patients with HF take a large number of medications. Medication adherence was suboptimal in many patients, often because of cost.


Clustering of the self-organizing map.
The self-organizing map (SOM) is an excellent tool in exploratory phase of data mining. It projects input space on prototypes of a low-dimensional regular grid that can be effectively utilized to visualize and explore properties of the data. When the number of SOM units is large, to facilitate quantitative analysis of the map and the data, similar units need to be grouped, i.e., clustered. In this paper, different approaches to clustering of the SOM are considered. In particular, the use of hierarchical agglomerative clustering and partitive clustering using k-means are investigated. The two-stage procedure--first using SOM to produce the prototypes that are then clustered in the second stage--is found to perform well when compared with direct clustering of the data and to reduce the computation time.


Voltage-Gated Proton Channels in the Tree of Life.
With a single gene encoding HV1 channel, proton channel diversity is particularly low in mammals compared to other members of the superfamily of voltage-gated ion channels. Nonetheless, mammalian HV1 channels are expressed in many different tissues and cell types where they exert various functions. In the first part of this review, we regard novel aspects of the functional expression of HV1 channels in mammals by differentially comparing their involvement in (1) close conjunction with the NADPH oxidase complex responsible for the respiratory burst of phagocytes, and (2) in respiratory burst independent functions such as pH homeostasis or acid extrusion. In the second part, we dissect expression of HV channels within the eukaryotic tree of life, revealing the immense diversity of the channel in other phylae, such as mollusks or dinoflagellates, where several genes encoding HV channels can be found within a single species. In the last part, a comprehensive overview of the biophysical properties of a set of twenty different HV channels characterized electrophysiologically, from Mammalia to unicellular protists, is given.


Acute EEG findings in HIV-infected Zambian adults with new-onset seizure.
To describe acute EEG findings in HIV-infected adults with new-onset seizure, assess baseline clinical characteristics associated with EEG abnormalities, and evaluate the relationship between EEG abnormalities and recurrent seizure. Eighty-one HIV-infected adults with new-onset seizure had EEG recordings during their index admission. Baseline characteristics assessed included HIV stage, seizure semiology, serum and CSF studies, neuroimaging, cognitive function based on the Zambian Mini-Mental State Examination and International HIV Dementia Scale, and psychiatric symptoms using the Shona Symptom Questionnaire. We evaluated the relationship between baseline characteristics and EEG abnormalities. Patients were followed for seizure recurrence, and the association between acute EEG abnormalities and seizure recurrence was assessed. Death was a secondary outcome. Fifty-five patients had abnormal EEGs (68%): 18 (22%) had interictal spikes (12) or a recorded seizure (6). Among baseline clinical characteristics, more advanced HIV disease (p = 0.039) and any imaging abnormality (p = 0.027) were associated with abnormal EEGs. Cortical (p = 0.008) and white matter (p = 0.004) abnormalities were associated with slow posterior dominant rhythm. Patients were followed for a median of 303 days (interquartile range 103-560). Twenty-four (30%) died and 23 (28%) had recurrent seizures. EEG abnormalities were not associated with recurrent seizure. There was a nonsignificant association between seizures recorded during EEG and death (67% vs 26%, p = 0.051). EEG abnormalities are common in this population, particularly in patients with imaging abnormalities and advanced HIV. Acute EEG abnormalities were not associated with recurrent seizure, but high mortality rates during follow-up limited this analysis.


Origins of the Greenland shark (Somniosus microcephalus): Impacts of ice-olation and introgression.
Herein, we use genetic data from 277 sleeper sharks to perform coalescent-based modeling to test the hypothesis of early Quaternary emergence of the Greenland shark (Somniosus microcephalus) from ancestral sleeper sharks in the Canadian Arctic-Subarctic region. Our results show that morphologically cryptic somniosids S. microcephalus and Somniosus pacificus can be genetically distinguished using combined mitochondrial and nuclear DNA markers. Our data confirm the presence of genetically admixed individuals in the Canadian Arctic and sub-Arctic, and temperate Eastern Atlantic regions, suggesting introgressive hybridization upon secondary contact following the initial species divergence. Conservative substitution rates fitted to an Isolation with Migration (IM) model indicate a likely species divergence time of 2.34 Ma, using the mitochondrial sequence DNA, which in conjunction with the geographic distribution of admixtures and Pacific signatures likely indicates speciation associated with processes other than the closing of the Isthmus of Panama. This time span coincides with further planetary cooling in the early Quaternary period followed by the onset of oscillating glacial-interglacial cycles. We propose that the initial S. microcephalus-S. pacificus split, and subsequent hybridization events, were likely associated with the onset of Pleistocene glacial oscillations, whereby fluctuating sea levels constrained connectivity among Arctic oceanic basins, Arctic marginal seas, and the North Atlantic Ocean. Our data demonstrates support for the evolutionary consequences of oscillatory vicariance via transient oceanic isolation with subsequent secondary contact associated with fluctuating sea levels throughout the Quaternary period-which may serve as a model for the origins of Arctic marine fauna on a broad taxonomic scale.


A novel high-risk aortic root anatomy for right coronary artery occlusion during TAVI.
Acute coronary occlusion during transcatheter aortic valve implantation (TAVI) is a rare but potentially lethal complication. Main mechanisms are sinus insufficiency or sinus sequestration with well-described risk factors. We present two cases of acute right coronary artery occlusion during TAVI with a self-expanding valve in the absence of classical risk factors and propose a novel mechanism.


Determinants of Morbidity and Mortality Associated With Isolated Tricuspid Valve Surgery.
Background Whether the poor outcomes of isolated tricuspid valve surgery are related to the operation itself or to certain patient characteristics including late referral is unknown. Methods and Results Adult patients who underwent isolated tricuspid valve surgery were identified in the Nationwide Readmissions Database (2016-2017). Patients who had redo tricuspid valve surgery, endocarditis, or congenital heart disease were excluded. Multivariable logistic regression was performed to identify contributors to postoperative mortality. A total of 1513 patients were included (mean age 55.7±16.6 years, 49.6% women). Surrogates of late referral were frequent: 41% of patients were admitted with decompensated heart failure, 44.3% had a nonelective surgery status, 16.8% had advanced liver disease, and 31% had an unplanned hospitalization in the prior 90 days. The operation was performed on day 0 to 1 of the hospitalization in only 50% of patients, and beyond day 10 in 22% of patients. In-hospital mortality occurred in 8.7% of patients. Median length of stay was 14 days (7-35 days), and median cost was $87 223 ($43 122-$200 872). In multivariable logistic regression analysis, surrogates for late referrals (acute heart failure decompensation, nonelective surgery status, or advanced liver disease) were the strongest predictors of in-hospital mortality (odds ratio [OR], 4.75; 95% CI, 2.74-8.25 [P<0.001]). This was also consistent in a second model incorporating unplanned hospitalizations in the 90 days before surgery as a surrogate for late referral (OR, 5.50; 95% CI, 2.28-10.71 [P<0.001]). Conclusions The poor outcomes of isolated tricuspid valve surgery may be largely explained by the late referral for intervention. Studies are needed to determine the role of early intervention for severe isolated tricuspid regurgitation.


Hirano bodies and other cytoskeletal abnormalities develop in fetal rat CNS grafts isolated for long periods in peripheral nerve.
Dissociated cells from the telencephalic region of 12-day-old rat embryos were cultured in vitro for 3-5 days and transplanted into segments of the sciatic nerve of adult inbred rats. Transplanted progenitor cells survived, differentiated, and expressed various morphological and molecular features characteristic of neurons and glia. Six to twelve months after grafting, many neurons underwent changes compatible with an alteration of their cytoskeleton. These included: (1) a strong perikaryal immunoreactivity to the monoclonal antibody RT97, directed against the 200-kDa phosphorylated neurofilament subunit and (2) the formation of Hirano bodies within dendrites. Similar cytoskeletal abnormalities are seen as part of the spectrum of changes that occur in some human neurodegenerative diseases and in aging. The approach we have used may provide new possibilities for the study of the pathogenesis of these lesions under controlled laboratory conditions.


Phagocyte-generated oxygen metabolites and cellular injury.
Phagocytic leukocytes are motile cells capable of inducing damage and lysis of a wide variety of biologic targets. Recent insights into the mechanisms of phagocyte-mediated destruction have derived from the observation that these cells can consume and metabolize oxygen to generate an impressive array of reactive oxygen intermediates. The role of oxygen metabolites in antimicrobial defense mechanisms has been the subject of intensive study, but only recently has attention focused on the potential importance of oxygen in phagocyte-mammalian cell interactions. In this review we will examine evidence obtained in a variety of in vitro model systems demonstrating the ability of intact phagocytes to generate oxygen metabolites capable of destroying normal or neoplastic cells. A basic understanding of the biochemistry of phagocyte-mediated oxygen-dependent events should allow us to elucidate and potentially modulate immunologic defenses against neoplastic invasion, the destruction of normal tissue in pathogenic states, and the course of the inflammatory response. The role of phagocyte-derived oxygen metabolites in microbicidal activity will not be the subject of this report, and the reader is referred to recent comprehensive reviews.


Patterns of engrailed protein in early Drosophila embryos.
By the onset of gastrulation during nuclear cycle 14 of Drosophila embryogenesis, the engrailed gene is expressed in fourteen one-cell-wide stripes. Each stripe defines the anlagen of the posterior compartment of a metameric segment. We report here several observations relating to the role and disposition of the engrailed protein during the embryonic stages that precede cellularization. We demonstrate that in embryos mutant for the engrailed gene, there were characteristic morphological abnormalities as early as the 6th cleavage cycle. In addition, the engrailed protein was detected in pre-cycle-9 embryos by Western blot analysis. When localization of engrailed protein begins during cycle 14, engrailed expression was first present in broad anterior and posterior regions before the fourteen-stripe pattern appeared.


The Effects of Music-Based Interventions for Pain and Anxiety Management during Vaginal Labour and Caesarean Delivery: A Systematic Review and Narrative Synthesis of Randomised Controlled Trials.
Music-based interventions are not physically invasive, they usually have minimal side effects, and they are increasingly being implemented during the birthing process for pain and anxiety relief. The aim of this systematic review is to summarise and evaluate published, randomised controlled trials (RCTs) assessing the effects of music-based interventions for pain and anxiety management during vaginal labour and caesarean delivery. Following the PRISMA guidelines, a systematic search of the literature was conducted using: PsychInfo (Ovid), PubMed, and Web of Science. Studies were included in the review if they were RCTs that assessed the effects of music on pain and anxiety during vaginal and caesarean delivery by human mothers. A narrative synthesis was conducted on 28 identified studies with a total of 2835 participants. Most, but not all, of the included studies assessing music-based interventions resulted in reduced anxiety and pain during vaginal and caesarean delivery. Music as part of a comprehensive treatment strategy, participant-selected music, music coupled with another therapy, and relaxing/instrumental music was specifically useful for reducing light to moderate pain and anxiety. Music-based interventions show promising effects in mitigating pain and anxiety in women during labour. However, the long-term effects of these interventions are unclear.


[Study of immunological status in women with post-term pregnancy].
The aim of the work was to study immunological homeostasis by comparing the immune responses during physiological, post-term and prolonged pregnancy. 51 women were divided into 3 groups. In the first group there were 13 pregnant women with post-term pregnancy, in the second group--8 pregnant women with prolonged pregnancy and in the third group--30 pregnant women with term pregnancy. All of them were practically healthy pregnant women. We could not find reliable differences between full-term and prolonged pregnancies, but in latter cases there was a trend towards deficiency of immunologic factors. Post-term pregnancy was characterized as an immunophatologic condition. It may be concluded that post-term newborns must be included into the high-risk group and be permanently observed by pediatricians.


Exploring the similarity of single-layer covalent organic frameworks using electronic structure calculations.
Two-dimensional Covalent Organic Frameworks (2D COFs) have attracted considerable interest because of their potential for a broad range of applications. Different combinations of the monomeric units can lead to potentially novel materials with varying physico-chemical properties. In this study, we investigate the electronic properties of various 2D COFs with square lattice topology based on a tight-binding density functional theory approach. We first classify the 2D COFs into different classes according to the degree of π-conjugation. Interestingly, this classification is recovered by using a similarity measure based on specific features of the electronic band-structure of the COFs. Further, we study the effect of aromaticity on the electronic structure of fully-conjugated COFs. Our results show that the conjugation and aromaticity are keys in the electronic band-structure of COFs.


D-xylose absorption in the adult bovine.
A D-xylose absorption curve was determined on 10 apparently healthy adult cows. The D-xylose was placed directly into the abomasum by abomasocentesis. Various doses of D-xylose were placed in the abomasum of cows with rumen fistula to determine the most useful dose. By giving 0.5 gm of D-xylose per dg of body weight, a blood level was produced that peaked around 90 minutes at 13.8 +/- 3 mg/di. The technique for abomasal centesis was developed by first using embalmed cows and then cows with rumen fistulae.


Characterization of Sfp, a Bacillus subtilis phosphopantetheinyl transferase for peptidyl carrier protein domains in peptide synthetases.
The Bacillus subtilis enzyme Sfp, required for production of the lipoheptapeptide antibiotic surfactin, posttranslationally phosphopantetheinylates a serine residue in each of the seven peptidyl carrier protein domains of the first three subunits (SrfABC) of surfactin synthetase to yield docking sites for amino acid loading and peptide bond formation. With recombinant Sfp and 16-17-kDa peptidyl carrier protein (PCP) domains excised from the SrfB1 and SrfB2 modules as apo substrates, kcat values of 56-104 min-1 and K(m) values of 1.3-1.8 microM were determined, indicating equivalent recognition of the adjacent PCP domains by Sfp. In contrast to other phosphopantetheinyl transferases (PPTases) previously examined, Sfp will modify the apo forms of heterologous recombinant proteins, including the PCP domain of Saccharomyces cerevisiae Lys2 (involved in lysine biosynthesis), the aryl carrier protein (ArCP) domain of Escherichia coli EntB (involved in enterobactin biosynthesis), and the E. coli acyl carrier protein (ACP) subunit, suggesting Sfp as a good candidate for heterologous coexpression with peptide and polyketide synthase genes to overproduce holo-synthase enzymes. Cosubstrate coenzyme A (CoA), the phosphopantetheinyl group donor, has a K(m) of 0.7 microM. Desulfo-CoA and homocysteamine-CoA are also substrates of Sfp, and benzoyl-CoA and phenylacetyl-CoA are also utilized by Sfp, resulting in direct transfer of acyl phosphopantetheinyl moieties into the carrier protein substrate. Mutagenesis in Sfp of five residues conserved across the PPTase family was assessed for in vivo effects on surfactin production and in vitro effects on PPTase activity.


Citrulline and intestinal fatty acid-binding protein: longitudinal markers of postweaning small intestinal function in pigs?
The objective of the current study was to investigate whether plasma citrulline or intestinal fatty acid-binding protein (I-FABP) concentrations might be used as longitudinal markers for small intestinal function in piglets after weaning. Plasma citrulline and I-FABP concentrations were measured longitudinally in weaned and unweaned piglets, and related to intestinal absorption values (i.e., plasma mannitol and 3-xylose concentrations in a sugar absorption test). Within each litter (n = 10), 2 piglets with a close-to-litter-average BW were selected. At 20.8 +/- 0.4 d of age, the selected piglets per litter were either weaned conventionally (CW) or remained with the sow (UNW). One day before, and 0.5, 2, 4, and 7 d after weaning of the CW piglets, the selected piglets of both groups were subjected to a sugar absorption test. After a 2-h fast, piglets were administered an oral dose of 2 mL/kg of sugar solution, containing 50 mg/kg of mannitol and 100 mg/kg of 3-xylose. One hour after administration, a blood sample was collected from a jugular vein for determination of plasma I-FABP, citrulline, mannitol, and 3-xylose concentrations. Plasma I-FABP concentration showed great variation within treatments, and no difference was observed in plasma I-FABP concentrations between the CW and UNW treatments (P = 0.63). The absorption of 3-xylose was not different between treatments (P = 0.83). Mannitol absorption, however, was less in the weaned CW piglets compared with the UNW piglets (P = 0.003), with the nadir on d 4 postweaning. Weaning also reduced plasma citrulline concentrations in the CW treatment compared with the UNW treatment (P < 0.001). On d 4 and 7 postweaning, plasma citrulline concentrations of CW piglets were less (P < 0.001 and P = 0.0013) than preweaning values. Furthermore, in the CW treatment, plasma citrulline concentrations correlated with plasma mannitol concentrations at d 4 postweaning (r = 0.89, P = 0.008) and overall (r = 0.76, P = 0.001). Based on these results, plasma citrulline concentration seems to be a possible marker for monitoring intestinal function in pigs after weaning.


Settlement patterns of the coral Acropora millepora on sediment-laden surfaces.
Successful recruitment in corals is important for the sustenance of coral reefs, and is considered a demographic bottleneck in the recovery of reef populations following disturbance events. Yet several factors influence larval settlement behaviour, and here we quantified thresholds associated with light attenuation and accumulated sediments on settlement substrates. Sediments deposited on calcareous red algae (CRA) directly and indirectly impacted coral settlement patterns. Although not avoiding direct contact, Acropora millepora larvae were very reluctant to settle on surfaces layered with sediments, progressively shifting their settlement preference from upward to downward facing (sediment-free) surfaces under increasing levels of deposited sediment. When only upward-facing surfaces were presented, 10% of settlement was inhibited at thresholds from 0.9 to 16mgcm-2 (EC10), regardless of sediment type (carbonate and siliciclastic) or particle size (fine and coarse silt). These levels equate to a very thin (<150μm) veneer of sediment that occurs within background levels on reefs. Grooves within settlement surfaces slightly improved options for settlement on sediment-coated surfaces (EC10: 29mgcm-2), but were quickly infilled at higher deposited sediment levels. CRA that was temporarily smothered by sediment for 6d became bleached (53% surface area), and inhibited settlement at ~7mgcm-2 (EC10). A minor decrease in settlement was observed at high and very low light intensities when using suboptimal concentrations of a settlement inducer (CRA extract); however, no inhibition was observed when natural CRA surfaces along with more realistic diel-light patterns were applied. The low deposited sediment thresholds indicate that even a thin veneer of sediment can have consequences for larval settlement due to a reduction of optimal substrate. And while grooves and overhangs provide more settlement options in high deposition areas, recruits settling at these locations may be subject to ongoing stress from shading, competition, and sediment infilling.


Monozygotic twins with discordant karyotypes following preimplantation genetic screening and single embryo transfer: case report.
to report a case of monozygotic monochorial diamniotic twins with discordant karyotypes. the pregnancy was achieved following a treatment cycle with intracytoplasmic sperm injection (ICSI) and preimplantation genetic screening (PGS) for chromosomes X, Y, 13, 16, 18, 21, 22. One embryo euploid for studied chromosomes was transferred. Prenatal ultrasonography revealed monozygotic twins. One fetus had growth retardation, multiple organ abnormalities and polyhydramnion. The other twin had normal ultrasound appearance. Delivery on week 29 of gestation resulted in the birth of two females, a stillborn twin with karyotype 45,XX,-13[12]/46,XX,r(13)[3] and a healthy twin with normal karyotype. the discordance in the twins' karyotypes originated from a mosaic embryo. Structural chromosomal abnormality of the affected twin could not be revealed using standard PGS investigation. Embryo splitting occurred probably due to apoptotic process in an early stage of embryo development. Apoptosis represents one of the possible mechanisms which can explain the embryo twinning process globally.


Circulating concentrations of bovine pregnancy-associated glycoproteins and late embryonic mortality in lactating dairy herds.
The objectives of these experiments were as follows: (1) to determine the association between circulating concentrations of pregnancy-associated glycoproteins (PAG) and late embryonic mortality (EM) in lactating dairy cattle following fixed-time artificial insemination (TAI) on d 0 or timed embryo transfer (TET) on d 7, (2) to identify a circulating concentration of PAG on d 31 below which late EM would be likely to occur, and (3) to identify when during gestation (d 31-59) late EM is occurring. Cows were diagnosed pregnant on d 31 of gestation based on presence of a fetal heartbeat and reconfirmed to be pregnant on d 59 of gestation. Late EM occurred when a cow had a viable embryo on d 31 of gestation but not on d 59 following TAI or TET. Only pregnant cows on d 31 were included in the analysis (TAI-maintained, n=413; TAI-EM, n=77; TET-maintained, n=238; TET-EM, n=47). Cows that were pregnant at d 31 of gestation and maintained the pregnancy until d 59 had significantly higher circulating concentrations of PAG at d 31 of gestation compared with cows that experienced late EM between d 31 and 59 of gestation in both TAI and TET. To conduct a more stringent test of the effectiveness of a single circulating PAG concentration (d 31) to predict EM, a receiver-operating characteristic curve was generated to identify a PAG concentration on d 31 that would predict EM with ≥95% accuracy in cows that received TAI or TET. Based on positive and negative predicative value analysis, a circulating concentration of PAG below 1.4 ng/mL (TAI; minimal detectable level 0.28 ng/mL) and 1.85 ng/mL (TET) was 95% accurate in predicting EM (between d 31 and 59) at d 31 of gestation, respectively. Following TET, embryonic loss was tracked by Doppler ultrasound, progesterone, and PAG from d 24 to 59 of gestation, with more than 50% of the loss occurring between d 31 and 38 of gestation. In summary, circulating concentrations of PAG on d 31 of gestation may provide a good marker for predicting EM between d 31 and 59 of gestation, and the data suggest that this model could help predict which cows will undergo late EM.


Valproic acid and immune thrombocytopenia.
During the course of a prospective serial study over a period of one year, findings compatible with immune-mediated thrombocytolysis accompanied the administration of valproic acid in about half of 45 children with epilepsy. Thrombocytopenia occurred in 15 and neutropenia in 12 patients, but was transient and self-limiting in each. The association is probably one of cause and effect, and it may reflect the structural similarity between valproic acid and constituents of cell membranes.


[Effect of motor activity on aerobic performance of healthy subjects].
Bicycle ergometry studies in which 120 healthy male volunteers, aged 20--40, look part, showed that aerobic performance depended on their normal motor activity. In the test subjects, leading an active mode of life, maximum oxygen uptake and general work capacity were significantly higher than in those leading a sedentary mode of life. The data obtained give evidence that the people with diminished muscle activity should be recommended to do regular exercises in order to maintain aerobic performance.


The Size of Labia Minora and Perception of Genital Appearance: A Cross-Sectional Study.
The aims of the study were to describe and to assess the size of labia minora in normal women, including the women's perception of their genitals to offer better counseling to the women seeking for labiaplasty. This cross-sectional study was undertaken in the outpatient clinic at the Department of Obstetrics and Gynaecology at Kolding Medical Center between May 2012 and October 2013, when study participants were recruited in connection with their routine visits. Main outcomes of interest were visibility and width of the patients' labia minora. In addition, women were asked whether they perceived their genitals as being normal. Information about age, body mass index, and parity was also collected. We used descriptive statistics and paired t test to compare the clinical measurements. In total, 244 women were included in the study. The median width of labia was 15 mm, and 90% of labia were less than 26.5 mm. Fifty-six percent of the women had visible labia minora. Regarding perception, 86.5% considered their labia as being normal. Within the group of women who perceived their genitals to be abnormal, 73.3% had visible labia minora. Among women with the largest labia (>26.5 mm), two thirds considered themselves normal. There was no statistically significant association between the measurements of labia minora and age, body mass index, or parity. One of 10 women had a measurement of labia minora of at least 26.5 mm in width. It is just as common to have visible labia minora as hidden labia minora. Women who considered their genitals to be abnormal more often had visible labia minora than women who considered their genitals to be normal.


Kainate receptor agonists, antagonists and allosteric modulators.
Interest in kainate receptors has increased over the past few years. Our understanding of their physiology and pharmacology has improved markedly since their original cloning and expression in the early 1990s. For example, agonist profiles at recombinant kainate receptors have been used to identify and distinguish kainate receptors in neurons. Furthermore, the development of selective antagonists for kainate receptor subtypes has increased our understanding of the functional roles of kainate receptors in neurons and synaptic transmission. In this review we described the activity of agonists and antagonists at kainate receptors and their selectivity profiles at NMDA and non-NMDA receptors.


Effect of the Timing of Surgery on Neurological Recovery for Patients with Incomplete Paraplegia Caused by Metastatic Spinal Cord Compression.
This study aimed to investigate the effect of timing of surgery on neurological recovery for patients with metastatic spinal cord compression (MSCC). According to the timing of surgery, 75 patients with incomplete paraplegia caused by MSCC were assigned to 3 groups: within 3 days (group A), between 4 days and 7 days (group B), and after 7 days (group C). T-test, one-way ANOVA, Mann-Whitney U-test, and Chi-square test were used to evaluate the difference in the improvement of American Spinal Injury Association Impairment Scale (AIS) and ambulatory status, the incidence of perioperative complications, surgical site infection, and the length of hospital stay between 3 groups. Patients with incomplete paraplegia treated in our department had an average of 17.4±1.8 days delayed and most occurred before hospitalization (4.0±0.4 vs 13.2±1.8, P<0.001). There was no significant difference in the AIS improvement between patients with different pre-op AIS. The timing of surgery was significantly correlated with AIS improvement (correlation coefficient=-0.257, P=0.019). Sub-analysis showed that patients who underwent surgery within 7 days (group A and group B) had significantly better AIS improvement compared with group C (improved at least 1 grade, P=0.043; improved more than 1 grade, P=0.039) and the surgery timing was more important for patients with AIS B and C. The timing of surgery was significantly correlated with the length of hospital stay (correlation coefficient=0.335, P=0.003). Patients of group C had the longest length of hospital stay (P=0.002). The incidence of perioperative complications and surgical site infection did not differ significantly between the 3 groups. Delay surgery was common in incomplete paraplegia patients with MSCC. Patients with AIS B and C who underwent surgery within 7 days had better AIS improvement.


Effects of nonsteroidal anti-inflammatory drugs on post-operative renal function in adults.
Nonsteroidal anti-inflammatory drugs (NSAIDs) can play a major role in the management of acute pain in the peri-operative period. However, there are conflicting views on whether NSAIDs are associated with adverse renal effects. The primary objective of this review was to determine the effects of NSAIDs on post-operative renal function in adults. Electronic searches for relevant randomised and quasi-randomised controlled trials in Cochrane Controlled Trials Register, MEDLINE and EMBASE were performed. Attempts were also made to identify trials from citation lists of relevant trials, review articles and clinical practice guidelines. Hand-searching of conference abstracts published in major anaesthetic journals was also performed. The inclusion criteria were randomised or quasi-randomised comparisons of individual NSAIDs with either each other or placebo for treatment of post-operative pain, with relevant post-operative renal outcome measures, in adult surgical patients. Of the 14 trials that fulfilled the selection criteria for this review, eight trials were relevant with sufficient data for meta-analysis. The data was extracted independently by two reviewers. The primary outcome measure was creatinine clearance within the first two days after surgery. Secondary outcome measures included serum creatinine, urine volume, urinary sodium level, urinary potassium level, fractional excretion of sodium, fractional excretion of potassium, need for dialysis and need for diuretic or dopamine treatment for renal insufficiency. Weighted mean differences for continuous outcomes and relative risk for dichotomous outcomes were estimated. As a group, NSAIDs reduced creatinine clearance by 18ml/min (95%CI: 6 to 31) and potassium output by 38mmol/day (95%CI: 19 to 56) on the first day after surgery compared to placebo. Serum creatinine clearance increased on the second day after surgery by 15umol/L (95%CI: 2 to 28) compared to placebo. No significant reduction in urine volume during the early post-operative period was found. There was no significant difference in serum creatinine in the early post-operative period between patients receiving ketorolac and diclofenac in one trial. No cases of post-operative renal failure requiring dialysis were described. NSAIDs caused a clinically unimportant transient reduction in renal function in the early post-operative period. NSAIDs should not be withheld from adults with normal pre-operative renal function because of concerns about post-operative renal impairment.


Translating guidelines into clinical practice: benefits of an acute heart failure unit .
Heart failure (HF) is a very prevalent condition, with high mortality, morbidity and costs, despite the development of effective drug therapy. Many patients still do not have access to appropriate treatment. Heart failure clinics have shown success in improving delivery of appropriate HF management, helping to reduce morbidity and mortality and costs. To compare the quality of diagnosis and drug therapy at discharge in an unselected population of patients admitted to a medical ward of an urban central teaching hospital before and after the creation of a heart failure unit (HFU) and to evaluate its impact on long-term mortality and morbidity from all causes and from HF. We compared a population of 153 patients, aged 75 +/- 12.8 years, 50.3% male, with a diagnosis of HF according to the guidelines of the European Society of Cardiology, NYHA classes II-IV (74% III or IV), admitted consecutively to a medical ward (MW) before the creation of an HFU (Group 1), with a population of 82 patients, aged 73 +/- 12.4 years, 62.2% male, admitted consecutively to the HFU for acute decompensated HF (Group 2). The prescription of diuretics, spironolactone, angiotensin-converting enzyme (ACE) inhibitors, beta-blockers, angiotensin receptor antagonists (ARAs) and warfarin in the two groups at discharge was analyzed. The endpoints of the study were: in-hospital mortality (IM), mortality and hospitalization due to all causes (P1), mortality and hospitalization due to HF (P2), all-cause mortality (P3), and mortality due to HF (P4). The follow-up duration was 18 months. The two populations were not statistically different in gender, age or type of HF. HF diagnosis was correct in 67.7% of cases in Group 1, vs. 86% in Group 2. The increase in the prescription of beta-blockers in Group 2 compared with Group 1 was highly significant (4.9% vs. 36.0%, p = 0.000 in all-cause HF patients and 28.9% vs. 56.8%, p = 0.000, in HF due to systolic dysfunction), as was that of warfarin (16.7% vs. 32.6%, p = 0.003 in all-cause HF and 17.6% vs. 44.4%, p = 0.001, in HF due to systolic dysfunction). IM in the MW vs. HFU was 7.7% vs. 8.5% (p = 0.42). The probabilities for the different end-points in MW/HFU at 6, 12 and 18 months were: P1: 55/35% (p = 0.013), 68/46% (p = 0.0005) and 73/51% (p = 0.0004); P2: 26/19% (p = 0.1), 34/20% (p = 0.008) and 39/23% (p = 0.004); P3: 25.5/21% (p = 0.22), 31/30% (p = 0.44) and 38/35% (p = 0.32); P4: 16/19% (p = 0.28), 23/20% (p = 0.3) and 27/33% (p = 0.25). These results are in agreement with reported morbidity and mortality from the syndrome after hospitalization. There is a tendency for better outcome in all the studied endpoints among the population treated in the HFU, despite greater severity of the disease, compared to those treated in the MW. A statistically significant improvement was found in the composite endpoints of mortality and hospitalization from all causes and due to HF, at 12 and 18 months. The results of the study indicate that the HFU had a better performance in reducing morbidity than mortality compared to the MW. As hospitalization accounts for 60% of HF costs, HFUs are expected to be cost effective.


Gene Editing: A New Tool for Viral Disease.
The emergence of the CRISPR/Cas system of antiviral adaptive immunity in bacteria as a facile system for gene editing in mammalian cells may well lead to gene editing becoming a novel treatment for a range of human diseases, especially those caused by deleterious germline mutations. Another potential target for gene editing are DNA viruses that cause chronic pathogenic diseases that cannot be cured by using currently available drugs. We review the current state of this field and discuss the potential advantages and problems with using a gene editing approach as a treatment for diseases caused by DNA viruses.


Weighting regressions by propensity scores.
Regressions can be weighted by propensity scores in order to reduce bias. However, weighting is likely to increase random error in the estimates, and to bias the estimated standard errors downward, even when selection mechanisms are well understood. Moreover, in some cases, weighting will increase the bias in estimated causal parameters. If investigators have a good causal model, it seems better just to fit the model without weights. If the causal model is improperly specified, there can be significant problems in retrieving the situation by weighting, although weighting may help under some circumstances.


Antiinflammatory and immunomodulating properties of fungal metabolites.
We discuss current information on the ability of extracts and isolated metabolites from mushrooms to modulate immune responses. This can result in a more enhanced innate and acquired disease resistance. The major immunomodulating effects of these active substances derived from mushrooms include mitogenicity and activation of immune effector cells, such as lymphocytes, macrophages, and natural killer cells, resulting in the production of cytokines, including interleukins (ILs), tumor necrosis factor alpha (TNF)-alpha, and interferon gamma (INF)-gamma. In particular, the ability of selective mushroom extracts to modulate the differentiation capacity of CD4(+) T cells to mature into T(H)1 and/or T(H)2 subsets will be discussed. As a consequence these extracts will have profound effects in particular diseases, like chronic autoimmune T(H)1-mediated or allergic T(H)2-mediated diseases. Immunosuppressive effects by mushroom components have also been observed. The therapeutic effects of mushrooms, such as anticancer activity, suppression of autoimmune diseases, and allergy have been associated with their immunomodulating effects. However, further studies are needed to determine the molecular mechanisms of the immunomodulating effects of mushrooms metabolites both individually and in complex mixtures, for example, extracts.


ERRF sensitizes ERBB2-positive breast cancer cells to lapatinib treatment likely by attenuating MCL1 and ERBB2 expression.
Previously we found that the estrogen receptor (ER) related factor ERRF regulates cell proliferation and tumor growth, and its expression is positively associated with ER status and better survival but inversely associated with ERBB2 (also named HER2) status in breast cancer. Here we report that ERRF also plays an important role in the response of ERBB2-positive breast cancer cells to lapatinib, a dual tyrosine kinase inhibitor that interrupts the ERBB2 and EGFR pathway. In ERBB2-positive breast cancer cell lines, lower levels of ERRF expression correlated with lapatinib resistance, restoration of ERRF expression in lapatinib-resistant cell lines JIMT-1 and MDA-MB-453 enhanced their lapatinib responses, and knockdown of ERRF in lapatinib sensitive cell lines BT-474 and SK-BR-3 caused lapatinib resistance. ERRF-enhanced lapatinib sensitivity was also confirmed in xenograft tumors of JIMT-1 cells. In patients with ERBB2-positive breast cancer, higher level of ERRF expression correlated with both pathologic complete response (pCR) to lapatinib and better survival. Mechanistically, ERRF expression in resistant cells promoted lapatinib-induced apoptosis by attenuating MCL1 and ERBB2 expression. These results suggest that ERRF plays an important role in lapatinib response of ERBB2-positive breast cancer, and further study of ERRF could lead to improved prediction and sensitivity of lapatinib response.


Time-invariant biological networks with feedback loops: structural equation models and structural identifiability.
Quantitative analyses of biological networks such as key biological parameter estimation necessarily call for the use of graphical models. While biological networks with feedback loops are common in reality, the development of graphical model methods and tools that are capable of dealing with feedback loops is still in its infancy. Particularly, inadequate attention has been paid to the parameter identifiability problem for biological networks with feedback loops such that unreliable or even misleading parameter estimates may be obtained. In this study, the structural identifiability analysis problem of time-invariant linear structural equation models (SEMs) with feedback loops is addressed, resulting in a general and efficient solution. The key idea is to combine Mason's gain with Wright's path coefficient method to generate identifiability equations, from which identifiability matrices are then derived to examine the structural identifiability of every single unknown parameter. The proposed method does not involve symbolic or expensive numerical computations, and is applicable to a broad range of time-invariant linear SEMs with or without explicit latent variables, presenting a remarkable breakthrough in terms of generality. Finally, a subnetwork structure of the C. elegans neural network is used to illustrate the application of the authors' method in practice.


Swapped-domain constructs of the glycoprotein-41 ectodomain are potent inhibitors of HIV infection.
The conformational rearrangement of N- and C-heptad repeats (NHR, CHR) of the HIV-1 glycoprotein-41 (gp41) ectodomain into a trimer of hairpins triggers virus-cell fusion by bringing together membrane-spanning N- and C-terminal domains. Peptides derived from the NHR and CHR inhibit fusion by targeting a prehairpin intermediate state of gp41. Typically, peptides derived from the CHR are low nanomolar inhibitors, whereas peptides derived from the NHR are low micromolar inhibitors. Here, we describe the inhibitory activity of swapped-domain gp41 mimics of the form CHR-loop-NHR, which were designed to form reverse hairpin trimers exposing NHR grooves. We observed low nanomolar inhibition of HIV fusion in constructs that possessed the following properties: an extended NHR C-terminus, an exposed conserved hydrophobic pocket on the NHR, high helical content, and trimer stability. Low nanomolar activity was independent of CHR length. CD studies in membrane mimetic dodecylphosphocholine micelles suggested that bioactivity could be related to the ability of the inhibitors to interact with a membrane-associated prehairpin intermediate. The swapped-domain design resolves the problem of unstable and weakly active NHR peptides and suggests a different mechanism of action from that of CHR peptides in inhibition of HIV-1 fusion.


The versatility of presculptured homograft incus prostheses.
A twelve-year experience with banked homograft ossicles has developed into a system of presculptured incus allografts adapted for the correction of many ossicular defects encountered in tympanoplasty and revision stapes surgery. The standardized preparation of a number of variations of modeled incus prostheses for an operating room bank by a well-trained ear homograft laboratory technician has been an effective and time-saving technique.