Development and characterization of an oral microbiome transplant among Australians for the treatment of dental caries and periodontal disease: A study protocol.
Oral microbiome transplantation (OMT) is a novel concept of introducing health-associated oral microbiota into the oral cavity of a diseased patient. The premise is to reverse the state of oral dysbiosis, and restore the ecological balance to maintain a stable homeostasis with the host immune system. This study will assess the effectiveness, feasibility, and safety of OMT using an interdisciplinary approach. To find donors suitable for microbial transplantation, supragingival plaque samples will be collected from 600 healthy participants. Each sample (200μL) will subsequently be examined in two ways: 1) 100μL of the sample will undergo high-throughput 16S rRNA gene amplicon sequencing and shotgun sequencing to identify the composition and characterisation of a healthy supragingival microbiome, 2) the remaining 100μL of the plaque sample will be mixed with 25% artificial saliva medium and inoculated into a specialised in-vitro flow cell model containing a hydroxyapatite disk. To obtain sufficient donor plaque, the samples would be grown for 14 days and further analysed microscopically and sequenced to examine and confirm the growth and survival of the microbiota. Samples with the healthiest microbiota would then be incorporated in a hydrogel delivery vehicle to enable transplantation of the donor oral microbiota. The third step would be to test the effectiveness of OMT in caries and periodontitis animal models for efficacy and safety for the treatment of oral diseases. If OMTs are found to be successful, it can form a new treatment method for common oral diseases such as dental caries and periodontitis. OMTs may have the potential to modulate the oral microbiota and shift the ecological imbalances to a healthier state.


A plasmid-based Escherichia coli gene expression system with cell-to-cell variation below the extrinsic noise limit.
Experiments in synthetic biology and microbiology can benefit from protein expression systems with low cell-to-cell variability (noise) and expression levels precisely tunable across a useful dynamic range. Despite advances in understanding the molecular biology of microbial gene regulation, many experiments employ protein-expression systems exhibiting high noise and nearly all-or-none responses to induction. I present an expression system that incorporates elements known to reduce gene expression noise: negative autoregulation and bicistronic transcription. I show by stochastic simulation that while negative autoregulation can produce a more gradual response to induction, bicistronic expression of a repressor and gene of interest can be necessary to reduce noise below the extrinsic limit. I synthesized a plasmid-based system incorporating these principles and studied its properties in Escherichia coli cells, using flow cytometry and fluorescence microscopy to characterize induction dose-response, induction/repression kinetics and gene expression noise. By varying ribosome binding site strengths, expression levels from 55-10,740 molecules/cell were achieved with noise below the extrinsic limit. Individual strains are inducible across a dynamic range greater than 20-fold. Experimental comparison of different regulatory networks confirmed that bicistronic autoregulation reduces noise, and revealed unexpectedly high noise for a conventional expression system with a constitutively expressed transcriptional repressor. I suggest a hybrid, low-noise expression system to increase the dynamic range.


The cost of hospital-acquired complications for older people with and without dementia; a retrospective cohort study.
Increased length of stay and high rates of adverse clinical events in hospitalised patients with dementia is stimulating interest and debate about which costs may be associated and potentially avoided within this population. A retrospective cohort study was designed to identify and compare estimated costs for older people in relation to hospital-acquired complications and dementia. Australia's most populous state provided a census sample of 426,276 discharged overnight public hospital episodes for patients aged 50+ in the 2006-07 financial year. Four common hospital-acquired complications (urinary tract infections, pressure areas, pneumonia, and delirium) were risk-adjusted at the episode level. Extra costs were attributed to patient length of stay above the average for each patient's Diagnosis Related Group, with separate identification of fixed and variable costs (all in Australian dollars). These four complications were found to be associated with 6.4% of the total estimated cost of hospital episodes for people over 50 (A$226million/A$3.5billion), and 24.7% of the estimated extra cost of above-average length of stay spent in hospital for older patients (A$226million/A$914million). Dementia patients were more likely than non-dementia patients to have complications (RR 2.5, p <0.001) and these complications comprised 22.0% of the extra costs (A$49million/A$226million), despite only accounting for 10.4% of the hospital episodes (44,488/426,276). For both dementia and non-dementia patients, the complications were associated with an eightfold increase in length of stay (813%, or 3.6 days/0.4 days) and doubled the increased estimated mean episode cost (199%, or A$16,403/A$8,240). Urinary tract infections, pressure areas, pneumonia and delirium are potentially preventable hospital-acquired complications. This study shows that they produce a burdensome financial cost and reveals that they are very important in understanding length of stay and costs in older and complex patients. Once a complication occurs, the cost is similar for people with and without dementia. However, they occur more often among dementia patients. Advances in models of care, nurse skill-mix and healthy work environments show promise in prevention of these complications for dementia and non-dementia patients.


New pharmaceuticals reduce cost of illness.
The cost of illness includes not only the funds required to treat illness, but also the effect on the patient's quality of life. Recent concern about rising health costs have focused on the direct expenditures without noting that the cost of illness in terms of mortality and morbidity has declined significantly. Pharmaceuticals have played a major role in reducing the total cost of illness. Several studies of the cost-effectiveness of past introductions of vaccines and pharmaceuticals reveal large cost savings. Although the focus of most studies has been on major advances, the continuing process of less dramatic therapeutic improvements has significantly trimmed the cost of illness. Cost-benefit studies of new drugs or changes in drug use, while more difficult to perform, make it possible to influence the selection of therapy. Since pharmaceuticals represent less than 10% of total treatment costs, reduction in the cost of pharmaceutical products can only have a minor impact on the total cost of illness. Pharmaceuticals can reduce the cost of illness by providing alternative therapies that reduce direct treatment cost or improve the public health.


The effect of the Meniett device in patients with Ménière's disease: long-term results.
To review our long-term experience with the Meniett device in patients who failed medical management to analyze its efficacy in controlling vertigo and its impact on the functional level of the patient. Retrospective case review with long-term follow-up of patients receiving the Meniett device at our institution between May 2002 and April 2005. Tertiary referral center. Patients who failed medical therapy for Ménière's disease and opted to receive the Meniett over chemical labyrinthectomy, surgical labyrinthectomy, endolymphatic shunt surgery, or vestibular nerve section. The Meniett is a small, portable, low-pressure pulse generator meant to be self-administered, requiring 5-minute sessions 3 times a day. Patient's current perception of usefulness of Meniett device for controlling vertigo and current level of Meniett treatment. Of 14 patients meeting the study criteria, 12 were able to be contacted, giving an average follow-up of 4 years. Three patients (25%) showed no benefit; 9 patients (75%) did respond to the Meniett and thought it had reduced the frequency and severity of their vertigo attacks. Seven patients (58%) continued to use the device on a daily basis; the other 2 patients (17%) went into remission 2 and 3 years after initiating treatment. The Meniett device is a minimally invasive, nondestructive treatment that may be used to provide longer-term reduction of vestibular symptoms in patients with Ménière's disease.


Everolimus with or without bevacizumab in advanced pNET: CALGB 80701 (Alliance).
Treatment with the MTOR inhibitor everolimus improves progression-free survival (PFS) in pancreatic neuroendocrine tumors (pNETs), but it is not known if the addition of a VEGF pathway inhibitor to an MTOR inhibitor enhances antitumor activity. We performed a randomized phase II study evaluating everolimus with or without bevacizumab in patients with advanced pNETs. One hundred and fifty patients were randomized to receive everolimus 10 mg daily with or without bevacizumab 10 mg/kg i.v. every 2 weeks. Patients also received standard dose of octreotide in both arms. The primary endpoint was PFS, based on local investigator review. Treatment with the combination of everolimus and bevacizumab resulted in improved progression-free survival compared to everolimus (16.7 months compared to 14.0 months; one-sided stratified log-rank P = 0.1028; hazard ratio (HR) 0.80 (95% CI 0.56-1.13)), meeting the predefined primary endpoint. Confirmed tumor responses were observed in 31% (95% CI 20%, 41%) of patients receiving combination therapy, as compared to only 12% (95% CI 5%, 19%) of patients receiving treatment with everolimus (P = 0.0053). Median overall survival duration was similar in the everolimus and combination arm (42.5 and 42.1 months, respectively). Treatment-related toxicities were more common in the combination arm. In summary, treatment with everolimus and bevacizumab led to superior PFS and higher response rates compared to everolimus in patients with advanced pNETs. Although the higher rate of treatment-related adverse events may limit the use of this combination, our results support the continued evaluation of VEGF pathway inhibitors in pNETs.


Motor Cortex Function in Fibromyalgia: A Study by Functional Near-Infrared Spectroscopy.
Previous studies indicated changes of motor cortex excitability in fibromyalgia (FM) patients and the positive results of transcranial stimulation techniques. The present study aimed to explore the metabolism of motor cortex in FM patients, in resting state and during slow and fast finger tapping, using functional Near-Infrared Spectroscopy (fNIRS), an optical method which detects in real time the metabolism changes in the cortical tissue. We studied 24 FM patients and 24 healthy subjects. We found a significant slowness of motor speed in FM patients compared to controls. During resting state and slow movement conditions, the metabolism of the motor areas was similar between groups. The oxyhemoglobin concentrations were significantly lower in patients than in control group during the fast movement task. This abnormality was independent from FM severity and duration. The activation of motor cortex areas is dysfunctional in FM patients, thus supporting the rationale for the therapeutic role of motor cortex modulation in this disabling disorder.


Insights into congenital stationary night blindness based on the structure of G90D rhodopsin.
We present active-state structures of the G protein-coupled receptor (GPCRs) rhodopsin carrying the disease-causing mutation G90D. Mutations of G90 cause either retinitis pigmentosa (RP) or congenital stationary night blindness (CSNB), a milder, non-progressive form of RP. Our analysis shows that the CSNB-causing G90D mutation introduces a salt bridge with K296. The mutant thus interferes with the E113Q-K296 activation switch and the covalent binding of the inverse agonist 11-cis-retinal, two interactions that are crucial for the deactivation of rhodopsin. Other mutations, including G90V causing RP, cannot promote similar interactions. We discuss our findings in context of a model in which CSNB is caused by constitutive activation of the visual signalling cascade.


Multiplex polymerase chain reaction on FTA cards vs. flow cytometry for B-lymphocyte clonality.
Two-colour flow cytometry was compared with multiplex PCR with capillary electrophoresis for clonality determination in specific categories of B-cell lymphoma. FTA cards were evaluated for preserving DNA from node imprints and expediting molecular analysis. A single-tube multiplex PCR targeted IGH and lymphoma-specific translocations in DNA extracted from 180 frozen lymphoid tissues and DNA bound to FTA cards from 192 fresh tissues and 137 aspirates. PCR results were compared with flow cytometry in the extracted and aspirated samples. Overall, single-tube multiplex PCR sensitivity was equivalent in the sample groups (intergroup range 79%-91%). False negatives were associated with tumour origin in the follicle centre. Multiplex PCR and flow cytometry were equally sensitive and together detected 98% of B-cell lymphomas. Additional two-tube targeting of IGK suggested an overall molecular sensitivity >90%. False positive (pseudoclonal) single-tube multiplex PCR was associated with necrosis and sparse lymphocytes. Multiplex PCR using template DNA bound to an FTA card effectively detects B-lymphocyte clonality, obviates DNA extraction and refrigeration, and can be used without diminished sensitivity in fine needle aspirates or node imprints as a replacement for or complement to flow cytometry at any point in the diagnostic work-up.


A new look at measles.
Since the measles vaccine was licensed in the United States in 1963, the number of measles cases has declined by 98%. Nevertheless, measles has not been eliminated as had been hoped, and, in fact, has started to increase in incidence. The increase in the number of cases has been accompanied by a change in measles epidemiology; the highest attack rate now occurs in preschool-age children and in older school-age and college students. The latter is the basis for the adoption of a two-dose measles immunization schedule by the ACIP and AAP. In preschool-age children, however, the problem is more disturbing and reflects low rates of immunization, particularly among inner-city populations. A major public health effort must be directed to achieve and maintain high vaccination rates if measles elimination is to be accomplished. Otherwise, measles outbreaks, with their accompanying morbidity and mortality, will continue to occur.


Tracking and visualization of space-time activities for a micro-scale flu transmission study.
Infectious diseases pose increasing threats to public health with increasing population density and more and more sophisticated social networks. While efforts continue in studying the large scale dissemination of contagious diseases, individual-based activity and behaviour study benefits not only disease transmission modelling but also the control, containment, and prevention decision making at the local scale. The potential for using tracking technologies to capture detailed space-time trajectories and model individual behaviour is increasing rapidly, as technological advances enable the manufacture of small, lightweight, highly sensitive, and affordable receivers and the routine use of location-aware devices has become widespread (e.g., smart cellular phones). The use of low-cost tracking devices in medical research has also been proved effective by more and more studies. This study describes the use of tracking devices to collect data of space-time trajectories and the spatiotemporal processing of such data to facilitate micro-scale flu transmission study. We also reports preliminary findings on activity patterns related to chances of influenza infection in a pilot study. Specifically, this study employed A-GPS tracking devices to collect data on a university campus. Spatiotemporal processing was conducted for data cleaning and segmentation. Processed data was validated with traditional activity diaries. The A-GPS data set was then used for visual explorations including density surface visualization and connection analysis to examine space-time activity patterns in relation to chances of influenza infection. When compared to diary data, the segmented tracking data demonstrated to be an effective alternative and showed greater accuracies in time as well as the details of routes taken by participants. A comparison of space-time activity patterns between participants who caught seasonal influenza and those who did not revealed interesting patterns. This study proved that tracking technology an effective technique for obtaining data for micro-scale influenza transmission research. The findings revealed micro-scale transmission hotspots on a university campus and provided insights for local control and prevention strategies.


Changes in health care spending and quality 4 years into global payment.
Spending and quality under global budgets remain unknown beyond 2 years. We evaluated spending and quality measures during the first 4 years of the Blue Cross Blue Shield of Massachusetts Alternative Quality Contract (AQC). We compared spending and quality among enrollees whose physician organizations entered the AQC from 2009 through 2012 with those among persons in control states. We studied spending changes according to year, category of service, site of care, experience managing risk contracts, and price versus utilization. We evaluated process and outcome quality. In the 2009 AQC cohort, medical spending on claims grew an average of $62.21 per enrollee per quarter less than it did in the control cohort over the 4-year period (P<0.001). This amount is equivalent to a 6.8% savings when calculated as a proportion of the average post-AQC spending level in the 2009 AQC cohort. Analogously, the 2010, 2011, and 2012 cohorts had average savings of 8.8% (P<0.001), 9.1% (P<0.001), and 5.8% (P=0.04), respectively, by the end of 2012. Claims savings were concentrated in the outpatient-facility setting and in procedures, imaging, and tests, explained by both reduced prices and reduced utilization. Claims savings were exceeded by incentive payments to providers during the period from 2009 through 2011 but exceeded incentive payments in 2012, generating net savings. Improvements in quality among AQC cohorts generally exceeded those seen elsewhere in New England and nationally. As compared with similar populations in other states, Massachusetts AQC enrollees had lower spending growth and generally greater quality improvements after 4 years. Although other factors in Massachusetts may have contributed, particularly in the later part of the study period, global budget contracts with quality incentives may encourage changes in practice patterns that help reduce spending and improve quality. (Funded by the Commonwealth Fund and others.).


Characterization of Porous Materials by Fluorescence Correlation Spectroscopy Super-resolution Optical Fluctuation Imaging.
Porous materials such as cellular cytosol, hydrogels, and block copolymers have nanoscale features that determine macroscale properties. Characterizing the structure of nanopores is difficult with current techniques due to imaging, sample preparation, and computational challenges. We produce a super-resolution optical image that simultaneously characterizes the nanometer dimensions of and diffusion dynamics within porous structures by correlating stochastic fluctuations from diffusing fluorescent probes in the pores of the sample, dubbed here as "fluorescence correlation spectroscopy super-resolution optical fluctuation imaging" or "fcsSOFI". Simulations demonstrate that structural features and diffusion properties can be accurately obtained at sub-diffraction-limited resolution. We apply our technique to image agarose hydrogels and aqueous lyotropic liquid crystal gels. The heterogeneous pore resolution is improved by up to a factor of 2, and diffusion coefficients are accurately obtained through our method compared to diffraction-limited fluorescence imaging and single-particle tracking. Moreover, fcsSOFI allows for rapid and high-throughput characterization of porous materials. fcsSOFI could be applied to soft porous environments such hydrogels, polymers, and membranes in addition to hard materials such as zeolites and mesoporous silica.


[Solvent extraction-flow injection analysis for determination of trimethoprim in compound preparations].
Trimethoprim (TMP) in four pharmaceutical preparations (compound sulfamethoxazol tablets, compound tetracycline tablets, compound trimethoprim and sulfamethoxazol tablets; and compound berberine injection) is determined by solvent extraction-flow injection spectrophotometry. It can be extracted into chloroform directly, and the absorbance at a wavelength of 280 nm of the organic phase is measured after phase separation. The manifold comprises two streams. The sample is injected into a 0.2 mol/L NaOH carrier stream, and extracted with chloroform in a 200-cm coil (ID 0.7 mm) after a 50 cm reaction tube (ID 1.0 mm). Calibration graph is linear in the range of 25-150 micrograms/ml. The average recovery is 101.4% with a relative standard deviation of 1.1%. The proposed system permits the analysis of about 50 samples per hour. Precise results in agreement with those obtained with official methods are achieved.


Human rights abuses, transparency, impunity and the Web.
This paper reviews how human rights advocates during the "war-on-terror" have found new ways to use the World Wide Web (Web) to combat human rights abuses. These include posting of human rights reports; creating large, open-access and updated archives of government documents and other data, tracking CIA rendition flights and maintaining blogs, e-zines, list-serves and news services that rapidly distribute information between journalists, scholars and human rights advocates. The Web is a powerful communication tool for human rights advocates. It is international, instantaneous, and accessible for uploading, archiving, locating and downloading information. For its human rights potential to be fully realized, international law must be strengthened to promote the declassification of government documents, as is done by various freedom of information acts. It is too early to assess the final impact of the Web on human rights abuses in the "war-on-terror". Wide dissemination of government documents and human rights advocates' reports has put the United States government on the defensive and some of its policies have changed in response to public pressure. Even so, the essential elements of secret prisons, detention without charges or trials, and illegal rendition remain intact.


A (31)P-NMR study of the H(+)-MoO(4)(2-)-(HP)O(3)(2-)-HPO(4)(2-)-(C(6)H(5)P)O(3)(2-)-(CH(3)P)O(3)(2-) system at low Mo(tot)/P(tot) ratio - Formation of mixed-hetero X2M5-type polyanions.
Formation of X2M5-type heteropolyanions in the aqueous mixed-hetero H(+)-MoO(4)(2-)-(HP)O(3)(2-)-HPO(4)(2-)-(C(6)H(5)P)O(3)(2-)-(CH(3)P)O(3)(2-) system has been studied by (31)P-NMR at [Mo](tot)/[P(all)](tot) = 1.5 at 298(1) K in 0.600 M Na(Cl) ionic medium. The -log[H(+)] range 1.4-6.2 has been covered. The pK(a) values of methylphosphonate were determined from (31)P-NMR chemical shift data in the range of 0 < -log[H(+)] < 13.2 as H(CH(3)P)O(3)(-) 7.30 and H(2)(CH(3)P)O(3) 2.05. All four-component H(+)-MoO(4)(2-)-P1-P2 subsystems (P1 and P2 are two of the four phosphorus-containing components), except H(+)-MoO(4)(2-)-(HP)O(3)(2-)-HPO(4)(2-), which has been studied previously, were examined instead of the full six-component system in order to simplify the evaluation of the NMR spectra. The concentrations were adjusted to [Mo](tot) = 60 mM and [P1](tot) = [P2](tot) = 20 mM in the measurements. All conceivable P2M5 and (P1)(P2)M5 heteropolyanions were identified. The following novel X2M5-type polyanions were found and their formation constants are given in logarithmic values with 3sigma in parentheses: (CH(3)P)(2)Mo(5)O(21)(4-) 69.68(7), (HP)(C(6)H(5)P)Mo(5)O(21)(4-) 67.09(4), (C(6)H(5)P)PMo(5)O(22)(5-) 63.23(5), H(C(6)H(5)P)PMo(5)O(22)(4-) 68.11(4), (HP)(CH(3)P)Mo(5)O(21)(4-) 67.85(4), (CH(3)P)PMo(5)O(22)(5-) 64.14(7), H(CH(3)P)PMo(5)O(22)(4-) 68.89(4) and (C(6)H(5)P)(CH(3)P)Mo(5)O(21)(4-) 69.18(4). The equilibrium conditions are illustrated in distribution diagrams. Phosphite proved to bind more weakly in X2M5-type polyoxometallates than phosphate or phosphonates, which are almost equally favoured in complexation.


Clinical Outcomes and Reduced Pulmonary Artery Pressure With Intra-Aortic Balloon Pump During Central Extracorporeal Life Support.
Patients supported with extracorporeal life support (ECLS) can experience severe complications from increased left ventricular afterload. The intra-aortic balloon pump (IABP) is thought to unload the left ventricle (LV) and is routinely used with ECLS despite conflicting evidence of its clinical benefit. This retrospective, single-center study examined the effect of the simultaneous use of IABP and centrally cannulated ECLS on patient outcomes and provides new insights into IABP-mediated LV unloading. Thirty patients supported with central ECLS and IABP (extracorporeal life support-IABP group, ECLS-I) were compared with 30 patients with central ECLS alone (ECLS) for cardiogenic shock. Rates of survival to 30 days (p = 0.06) and intensive care unit (ICU) discharge (p = 0.17), and clinical outcomes were not significantly different between the two groups. In patients with pulmonary artery pressure monitoring, mean pulmonary artery (PA) pressure was significantly reduced after 24 (p = 0.007) and 48 hours (p = 0.002) in the ECLS-I group. No significant difference in PA pressure was observed in the ECLS group after 24 or 48 hours. The IABP has the ability to reduce pulmonary artery pressure in patients supported by central ECLS. However, this did not translate into improved survival or clinical outcomes in our population.


A participatory assessment of ecosystem services and human wellbeing in rural Costa Rica using photo-voice.
Human well-being is intricately connected to ecosystem services. A keystone contribution to the ecosystem service literature has been the Millennium Ecosystem Assessment, MA, (Ecosystems and human well-being: a framework for assessment, Island Press, Washington, DC; 2003, 2005). Much of the work on ecosystem services to date has focused on the assessment and classification of environmental functions. The need for inclusion of community perspectives in ecosystem assessments has been widely recognized in order to better understand the distribution of impacts and benefits resulting from natural resource use. Communities can offer a direct route to understanding the complex relationships between ecosystems and human well-being and how environmental management affects their livelihoods. Photovoice has been made popular as a tool for participatory needs assessment but it has had limited use in ecosystem assessments to date. The purpose of this paper is twofold: (1) to present the results of a community-level assessment of environmental services in a watershed dominated by pineapple monoculture in Costa Rica; and (2) to evaluate the strengths and the limitations of photovoice as a tool for mapping the relationship between ecosystems and people. I argue that photovoice is an underutilized methodology that has the potential to complement biophysical ecosystem service assessments in the context of impoverished and resource-dependent communities, particularly, since assessing ecosystem services and acting upon that information requires integrating the knowledges of diverse stakeholders, recognizing power imbalances, and grappling with the complexity of social-ecological systems. Processes such as photovoice have the potential to catalyze community self-organization, which is a critical component for empowerment.


Clonal Evolution of a High-Grade Pediatric Glioma With Distant Metastatic Spread.
High-grade glioma (HGG) rarely spreads outside the CNS. To test the hypothesis that the lesions were metastases originating from an HGG, we sequenced the relapsing HGG and distant extraneural lesions. We performed whole-exome sequencing of an HGG lesion, its local relapse, and distant lesions in bone and lymph nodes. Phylogenetic reconstruction and histopathologic analysis confirmed the common glioma origin of the secondary lesions. The mutational profile revealed an IDH1/2 wild-type HGG with an activating mutation in EGFR and biallelic focal loss of CDKN2A (9p21). In the metastatic samples and the local relapse, we found an activating PIK3CA mutation, further copy number gains in chromosome 7 (EGFR), and a putative pathogenic driver mutation in a canonical splice site of FLNA. Our findings demonstrate tumor spread outside the CNS and identify potential genetic drivers of metastatic dissemination outside the CNS, which could be leveraged as therapeutic targets or potential biomarkers.


Interleukin-1beta induction of NFkappaB is partially regulated by H2O2-mediated activation of NFkappaB-inducing kinase.
Reactive oxygen species (ROS) have been demonstrated to act as second messengers in a number of signal transduction pathways, including NFkappaB. However, the mechanism(s) by which ROS regulate NFkappaB remain unclear and controversial. In the present report, we describe a mechanism whereby interleukin-1beta (IL-1beta) stimulation of NFkappaB is partially regulated by H2O2-mediated activation of NIK and subsequent NIK-mediated phosphorylation of IKKalpha. IL-1beta induced H2O2 production in MCF-7 cells and clearance of this ROS through the expression of GPx-1 reduced NFkappaB transcriptional activation by inhibiting NIK-mediated phosphorylation of IKKalpha. Although IKKalpha and IKKbeta were both involved in IL-1beta-mediated activation of NFkappaB, only the IKKalpha-dependent component was modulated by changes in H2O2 levels. Interestingly, in vitro reconstitution experiments demonstrated that NIK was activated by a very narrow range of H2O2 (1-10 microM), whereas higher concentrations (100 microM to 1 mM) inhibited NIK activity. Treatment of cells with the general Ser/Thr phosphatase inhibitor (okadaic acid) lead to activation of NFkappaB and enhanced NIK activity as a IKKalpha kinase, suggesting that ROS may directly regulate NIK through the inhibition of phosphatases. Recruitment of NIK to TRAF6 following IL-1beta stimulation was inhibited by H2O2 clearance and Rac1 siRNA, suggesting that Rac-dependent NADPH oxidase may be a source of ROS required for NIK activation. In summary, our studies have demonstrated that redox regulation of NIK by H2O2 is mechanistically important in IL-1beta induction of NFkappaB activation.


[Status of the mucous membrane of the jejunum of the rat after desympathization].
Desympathization of the rat jejunum has been performed by means of periarterial sympathectomy of the cranial mesenteric artery. In total preparations of the jejunum wall, by means of water-aldehyde method, catecholamines have been revealed for controlling desympathization effectiveness. Signs of reinnervation appear in 180 days after the operation. In paraffin sections, stained by general histological methods, morphometric analysis of the most important parameters of the mucous membrane is performed (height of the villi, depth of the crypts, villus/crypt index, height of the epithelium, amount of goblet cells and interepithelial lymphocytes) Height of the villi and villus/crypt index increase in 1, 3 and 14 days, while in 7 days after the operation depth of the crypts increases, and the index mentioned decreases. Height of epithelium in the villi decreases in 14, 30 and 90 days. In 3-30 days after the operation amount of interepithelial lymphocytes increases.


Oral iron prophylaxis in pregnancy: not too little and not too much!
An adequate supply of iron is essential for normal development of the fetus and newborn child. Iron deficiency and iron deficiency anemia (IDA) during pregnancy increase the risk of preterm birth and low birth weight. Iron is important for development of the fetal brain and cognitive abilities of the newborn. Children born to iron-deficient mothers will start their lives suffering from iron deficiency or even IDA. Oral iron prophylaxis to pregnant women improves iron status and prevents development of IDA. The Danish National Board of Health has since 1992 recommended prophylactic oral iron supplements to all pregnant women and the currently advocated dose is 40-50 mg ferrous iron taken between meals from 10 weeks gestation to delivery. However, 30-40 mg ferrous iron is probably an adequate dose in most affluent societies. In developed countries, individual iron prophylaxis guided by iron status (serum ferritin) has physiological advantages compared to general iron prophylaxis. In contrast, in most developing countries, general iron prophylaxis is indicated, and higher doses of oral iron, for example, 60 mg ferrous iron or even more should be recommended, according to the present iron status situation in the specific populations of women of fertile age and pregnant women.


Is the Neutrophil-to-Lymphocyte Ratio Associated With Increased Morbidity After Colorectal Surgery?
We aimed to evaluate the association between preoperative and postoperative neutrophil-to-lymphocyte ratio (NLR) and 30-day postoperative complications after colorectal surgery. Patients undergoing elective colorectal surgery between January, 2010 and December, 2014 were identified. Patients who had preoperative and postoperative days 1 and 2 (Postoperative day [POD]-1, POD-2) NLR were included in the study. Primary study outcomes were optimal NLR cutoff values at preop, POD-1, and POD-2. A total of 1328 patients met the inclusion criteria. Of those, 518 (39%) patients experienced at least one postoperative complication. Sex (P<0.001), diabetes mellitus (DM) (P<0.001), diagnosis (P=0.001), operation type (P=0.03), and open surgery (P<0.001) were statistically associated with higher NLR (POD-1, P=0.02; POD2, P=0.01). DM (OR, 1.97; 95%CI, 1.27-3.08; P=0.003] and NLR on POD-2≥9.2 (OR, 1.43; 95%CI, 1.03-1.98; P=0.02) were significantly related to postoperative complications. NLR may provide clinicians with an additional tool for identifying patients at high risk for postoperative complications after elective colorectal surgery. Routine use of NLR may lead to early intervention and potentially improve the management of complications after colorectal surgery.


Kolliphor® HS 15-cyclodextrin Complex for the Delivery of Voriconazole: Preparation, Characterization, and Antifungal Activity.
This study aimed to reduce the amount of sulfobutylether-β-cyclodextrin (SBECD) used in the marketed voriconazole injections to meet the clinical needs of patients with moderate-to-severe renal impairment (creatinine clearance rate <50 mL/min). This study found that the surfactant Kolliphor® HS 15 (HS 15) and SBECD had significant synergistic effects on solubilizing voriconazole, and a novel voriconazole complex delivery system (VRC-CD/HS 15) was established. The complex system was characterized, and its antifungal activity was studied by dynamic light scattering, dialysis bag method, disk diffusion, and broth microdilution. Compared with the control, its encapsulation efficiency (90.07±0.48%), drug loading (7.37±0.25%) and zeta potential (-4.36±1.37 mV) were increased by 1.54%, 41.19%, and 296.36%, respectively; its average particle size (13.92±0.00 nm) was reduced by 15.69%, so the complex system had better stability. Simultaneously, its drug release behavior was similar to that of the control, and it was a first-order kinetic model. Antifungal studies indicated that the complex system had noticeable antifungal effects. With the increase of drug concentration, the inhibition zone increased. The minimum inhibitory concentrations of the complex system against Cryptococcus neoformans, Aspergillus niger and Candida albicans were 0.0313 μg/mL, 1 μg/mL and 128 μg/mL, respectively. It showed a significant inhibitory effect on C. neoformans and had a visible therapeutic effect on Kunming mice infected with C. neoformans. Consequently, VRC-CD/HS 15 had better physicochemical properties and still had an apparent antifungal effect, and was promising as a potential alternative drug for clinical application.


Regioselective benzannulation of allylic sulfur ylides with ynones: a rapid access to substituted thioanisoles.
The efficiency and selectivity of annulation reactions are often difficult to control in the presence of multiple potential reactive centers, like in the case of allylic sulfur ylides (ASY). Here, we describe a novel base mediated [3+3] benzannulation of ASY and readily available alkynones, which accomplishes the regioselective formation of multisubstituted thioanisoles, highly sought after chemical scaffolds. A new reactivity pattern of ASY has been unearthed, where it acted as both a 3C component and sulfur source in benzannulation. Use of a widely available base, operational simplicity and broad substrate scope are the additional salient features of the conversion.


Expert consensus on endoscopic papillectomy using a Delphi process.
Consensus regarding an optimal algorithm for endoscopic treatment of papillary adenomas has not been established. We aimed to assess the existing degree of consensus among international experts and develop further concordance by means of a Delphi process. Fifty-two international experts in the field of endoscopic papillectomy were invited to participate. Data were collected between August and December 2019 using an online survey platform. Three rounds were conducted. Consensus was defined as ≥70% agreement. Sixteen experts (31%) completed the full process, and consensus was achieved on 47 of the final 79 statements (59%). Diagnostic workup should include at least an upper endoscopy using a duodenoscope (100%) and biopsy sampling (94%). There should be selected use of additional abdominal imaging (75%-81%). Patients with (suspected) papillary malignancy or over 1 cm intraductal extension should be referred for surgical resection (76%). To prevent pancreatitis, rectal nonsteroidal anti-inflammatory drugs should be administered before resection (82%) and a pancreatic stent should be placed (100%). A biliary stent is indicated in case of ongoing bleeding from the papillary region (76%) or concerns for a (micro)perforation after resection (88%). Follow-up should be started 3 to 6 months after initial papillectomy and repeated every 6 to 12 months for at least 5 years (75%). This is the first step in developing an international consensus-based algorithm for endoscopic management of papillary adenomas. Surprisingly, in many areas consensus could not be achieved. These aspects should be the focus of future studies.


Proteomics of cauda epididymal fluid from mature Holstein bulls.
The proteome of cauda epididymal fluid (CEF) from Holstein bulls was defined. Fluid was collected from the vas deferens, subjected to 2-D SDS-PAGE and spots identified by CapLC-MS/MS and MALDI-ToF/ToF. Because albumin accounted for 21.1% of all spot intensities in the gels examined by PDQuest, samples were subjected to albumin depletion and then analyzed again as before. Original CEF gels had 114 ± 3 spots, including as the most abundant: albumin, epididymal secretory protein E1, prostaglandin d-synthase and gelsolin. Epididymal fluid also expressed: clusterin, transferrin, N-acetyl-β-glucosaminidase, cauxin, glutathione peroxidase, acidic seminal fluid protein (aSFP), aldehyde reductase, α-l-fucosidase, α-1-β-glycoprotein, apolipoprotein A-1, β actin, calmodulin, cathepsin D, cystatin E/M, enolase, galectin 3-binding protein, leucine amino-peptidase and nucleobindin. Albumin depletion decreased that very spot to 10% of its original intensity and the resulting gels had, on average, 137 ± 4 spots. Spots identified as dipeptidyl-peptidase 7, angiotensin-converting enzyme, arylsulfatase A, aspartylglucosaminidase, serine protease inhibitors, new isoforms of calmodulin, cystatin E/M and a 17-kDa nucleobindin appeared only in depleted maps. This study is the first to report nucleobindin and aSFP as epididymal components. We suggest that CEF proteins act to facilitate membrane remodeling, transport of lipophilic substances, protect sperm and prevent premature acrosome reaction.


H2A.Z and H2B.Z double-variant nucleosomes define intergenic regions and dynamically occupy var gene promoters in the malaria parasite Plasmodium falciparum.
Histone variants are important components of eukaryotic chromatin and can alter chromatin structure to confer specialized functions. H2B variant histones are rare in nature but have evolved independently in the phyla Apicomplexa and Trypanasomatida. Here, we investigate the apicomplexan-specific Plasmodium falciparum histone variant Pf H2B.Z and show that within nucleosomes Pf H2B.Z dimerizes with the H2A variant Pf H2A.Z and that Pf H2B.Z and Pf H2A.Z occupancy correlates in the subset of genes examined. These double-variant nucleosomes also carry common markers of euchromatin like H3K4me3 and histone acetylation. Pf H2B.Z levels are elevated in intergenic regions across the genome, except in the var multigene family, where Pf H2A.Z/Pf H2B.Z double-variant nucleosomes are only enriched in the promoter of the single active var copy and this enrichment is developmentally regulated. Importantly, this pattern seems to be specific for var genes and does not apply to other heterochromatic gene families involved in red blood cell invasion which are also subject to clonal expression. Thus, Pf H2A.Z/Pf H2B.Z double-variant nucleosomes appear to have a highly specific function in the regulation of P. falciparum virulence.


High frequency of p53 gene mutations in primary breast cancers in Japanese women, a low-incidence population.
The pattern of acquired mutations in the p53 tumour-suppressor gene is potentially useful for determining factors contributing to carcinogenesis in diverse populations differing in incidence and/or mortality from the disease. We previously reported differences in mutational patterns of the p53 gene in primary breast cancers from Midwest US Caucasian, African-American and Austrian women. Herein, we report 16 mutations in 27 primary breast cancers from Japanese women from Hirosaki, a population with a low incidence of breast cancer. The frequency of 59.3% of p53 mutations is the highest reported in breast cancers from a particular ethnic group thus far. A relatively high number of mutations (7/16) were heterozygous in at least some tumour cell clusters. Intergroup comparisons of the mutational pattern between this population and several other US, European and Japanese populations do not show any statistically significant differences. There were recurrent mutations at two sites, codon 273 (R --> H; three mutations), a common hotspot of mutations in breast and other cancers, and codon 183 (S --> Stop; two mutations), a very rare location for p53 mutations. These mutations were shown to be independent and presumably not in the germ line. The highest frequency of p53 mutations raises the possibility that p53 mutagenesis is a predominant factor for breast cancer development in this low-risk Japanese group, whereas in other cohorts different mechanisms are likely to account for the higher proportion of breast cancer. Further studies are needed to confirm the present observations.


Outcomes after salvage procedures for the painful dislocated hip in cerebral palsy.
The painful dislocated hip in the setting of cerebral palsy is a challenging problem. Many surgical procedures have been reported to treat this condition with varying success rates. The purpose of this study is to retrospectively evaluate and compare the outcomes of 3 different surgical procedures performed at our institution for pain relief in patients with spastic quadriplegic cerebral palsy and painful dislocated hips. A retrospective chart review of the surgical procedures performed by 5 surgeons for spastic, painful dislocated hips from 1997 to 2010 was performed. The procedures identified were (1) proximal femoral resection arthroplasty (PFRA); (2) subtrochanteric valgus osteotomy (SVO) with femoral head resection; and (3) proximal femur prosthetic interposition arthroplasty (PFIA) using a humeral prosthesis. Outcomes based on pain and range of motion were determined to be excellent, good, fair, or poor by predetermined criteria. Forty-four index surgeries and 14 revision surgeries in 33 patients with an average follow-up of 49 months met the inclusion criteria. Of the index surgeries, 12 hips were treated with a PFRA, 21 with a SVO, and 11 with a PFIA. An excellent or good result was noted in 67% of PFRAs, 67% of SVOs, and 73% of PFIAs. No statistical significance between these procedures was achieved. The 14 revisions were performed because of a poor result from previous surgery, demonstrating a 24% reoperation rate overall. No patients classified as having a fair result underwent revision surgery. All patients receiving revision surgery were eventually classified as having an excellent or good result. Surgical treatment for the painful, dislocated hip in the setting of spastic quadriplegic cerebral palsy remains unsettled. There continue to be a large percentage of failures despite the variety of surgical techniques designed to treat this problem. These failures can be managed, however, and eventually resulted in a good outcome. We demonstrated a trend toward better outcomes with a PFIA, but further study should be conducted to prove statistical significance. III.


Endothelin mediates renal vasodilation and hyperfiltration during pregnancy in chronically instrumented conscious rats.
Profound vasodilation of the kidneys and other nonreproductive organs transpires during early pregnancy. Because nitric oxide (NO) was found to mediate renal vasodilation and hyperfiltration in conscious pregnant rats, and endogenous endothelin (ET) was suggested to be vasodilatory in the renal circulation of nonpregnant rats, we tested whether endothelin mediates the NO-dependent changes in the renal circulation during pregnancy. Glomerular filtration rate (GFR) and effective renal plasma flow (ERPF) were measured in conscious pregnant and virgin rats before and during infusion of 30 micrograms/min RES-701-1 (a selective ETB receptor subtype antagonist). Baseline GFR and ERPF were significantly increased by 35% in gravid rats relative to virgin controls. During infusion of RES-701-1, the pregnant rats responded more robustly, showing a greater decline in both GFR and ERPF such that renal function converged in the two groups of rats. ERPF also converged in pregnant and virgin rats during infusion of SB-209760, a nonselective ETA/B receptor subtype antagonist. Combined infusion of Nomega-nitro-L-arginine methyl ester [L-NAME, an NO synthase (NOS) inhibitor] and RES-701-1 reduced GFR and ERPF to levels comparable to those reached with either agent given alone, suggesting inhibition of a common vasodilatory pathway. RES-701-1 and SB-209670 significantly lowered the cGMP content of small renal arteries from gravid and virgin rats in vitro, strengthening the link between the renal endothelial ETB receptor subtype and NO. Importantly, we showed that RES-701-1 is not a direct inhibitor of NOS. We conclude that endothelin mediates the NO-dependent changes in the renal circulation of conscious rats during pregnancy.


Polar self-assembly: steric effects leading to polar mixed-ligand coordination cages.
We present a highly unusual example of self-assembly, specifically a polar, mixed-ligand cage which forms in preference to symmetrical homo-ligand products, and which suggests that steric effects can be exploited to obtain novel non-uniform polyhedral cages. In particular, reaction between the bulky tripodal triphosphine 2,4,6-tris(diphenylphosphino)triazine, L1, the non-bulky tripodal trinitrile 2,4,6-tris(cyanomethyl)trimethylbenzene, L2 and silver hexafluoroantimonate, AgSbF6, in a 3:1:4 ratio gives the mixed-ligand aggregate [Ag4(L1)3(L2)(SbF6)]3+, 1-SbF6, instantly as the only product in quantitative yield. The X-ray crystal structure of complex 1-SbF6 is consistent with the suspected solution-state structure. The cage derives from trigonal-pyramidal geometry, the basal vertices of which are defined by three bulky triphosphines, L1, and the apical vertex by the non-bulky trinitrile, L2. There is apical elongation amounting to 19% in comparison to the ideal uniform tetrahedron. The cage also encapsulates an SbF6 anion. 19F NMR spectra in solution for the analogous PF6 complex [Ag4(L1)3(L2)(PF6)]3+, 1-PF6, confirm that one anion is also encapsulated in solution. The synthesis of the analogous CF3SO3(-) complex, [Ag4(L1)3(L2)(OTf)]3+, 1-OTf, in solution is also described, although 1-PF6 and 1-OTf could not be isolated due to slow decomposition in solution. The selective formation of these mixed-ligand cages is discussed in terms of ligand-ligand and ligand-included anion steric repulsions, which we propose prevent the formation of the competing hypothetical homo-ligand tetrahedral structure [Ag4(L1)4(SbF6)]3+, and thus favour the mixed ligand cage. "Cage cone angles" for L1 and L2 are estimated at 115 degrees and 101 degrees, respectively. Variable-temperature 31P NMR spectroscopy shows that complex 1-SbF6 and the related previously reported partial tetrahedral complex [Ag4(L1)3(anion)]3+ undergo dynamic twisting processes in solution between enantiomeric C3-symmetry conformations.


A city looks at its problem of drug abuse by injection.
A retrospective, multi-source screening study has detected 148 users (110 males, 38 females) of intravenous drugs who presented to all relevant agencies between 1971 and 1973 in Bristol, a city where maintenance therapy is a very infrequent practice. The peak prevalence was found in the age group 20-24 years, and it appeared that the chief sources of drugs were burglaries of chemists' shops and general practitioner prescriptions. Sixty-four per cent of our users had not been notified to the Home Office. Suggestions are made for the future monitoring and development of services.


Computer assisted spine surgery.
When inserting screws into a vertebral pedicle, the surgeon usually exposes the back part of the vertebra and uses his or her anatomic knowledge to align the drill in the proper direction. A slight error in direction may result in an important error in the position of the tip of the screw. This is done with no direct visibility of crucial structures (spinal cord, pleura, vessels). Statistical analysis of a series of surgical procedures has shown that 10% to 40% of the screws are not installed correctly. To reduce the risk of complication, a computer assisted method is proposed that enables the surgeon to place a screw at a position preoperatively defined in 3 dimensions using computed tomography images. This allows the surgeon to align a standard surgical drill with the optimal position and direction. The depth of the pilot hole during drilling also is monitored by the system to prevent penetration of the anterior cortex of the vertebral body. Using this procedure, in vitro tests were performed and showed that an accuracy of less than 1 mm can be obtained. Clinical trials were done in 10 patients who suffered severe scoliosis or spondylolisthesis. The trajectory of the holes drilled in L2, L3, L4, and L5 vertebrae were checked for all clinical tests. Postoperative radiographs and computed tomography scans showed that the screws were well inserted in each plane for each pedicle. This technique also can be used to perform osteosynthesis at the thoracic and cervical levels.


Nitric oxide metabolites and oxidative stress in mild essential hypertension.
Many papers have showed non univocal data about oxidative stress status and nitric oxide metabolites in essential hypertension. Considering this preamble we examined the total antioxidant status (TAS), the lipid peroxidation (LP), expressed as thiobarbituric acid-reactive substances (TBARS), the stable end products of nitric oxide (NOx) and LP/NOx ratio in 25 subjects with untreated mild essential hypertension. The obtained data show a significant increase in TBARS (p < 0.001) and NOx (p < 0.001) in hypertensives and no variation in TAS and in TBARS/NOx ratio. None of these parameters was statistically related to the metabolic parameters or to the blood pressure values. The high level of lipid peroxidation observed in this group of hypertensives suggests the timely and specific employment of antihypertensive and antioxidant agents while the NOx increase seems to confirm the inflammatory status accompanying this clinical condition.


Endoscopic Ultrasound-Guided Fine-Needle Aspiration Cytology Combined With Automated Quantitative DNA Cytometry Can Improve the Value in the Detection of Pancreatic Malignancy.
Quantitative DNA-image cytometry (ICM) is used to diagnose malignancy via detecting changes in DNA content. We aimed to estimate the value of cytology, DNA-ICM, and their combination in diagnosing pancreatic malignancy. One hundred twenty-one endoscopic ultrasound-guided fine-needle aspiration samples from 116 patients suspected for pancreatic malignancy were examined by cytology and DNA-ICM. Their results and the final diagnoses (malignancy or not) were collected. Diagnostic values were compared among cytology, DNA-ICM, and their combination. The DNA-ICM had a lower sensitivity and accuracy than cytology (64.2% vs 81.1%; 71.9% vs 85.1%). The combination of the techniques significantly improved the diagnostic accuracy compared with that of cytology or DNA-ICM alone (0.932 vs 0.905, P = 0.02; 0.932 vs 0.821, P < 0.0001). Using the Youden index, we determined that one cell with DI (DNA index) ≥2.5 is the optimal cutoff value for DNA-ICM to diagnose pancreatic malignancy. After adoption of this criterion, the sensitivity and accuracy were improved to 74.7% and 80.2% with DNA-ICM and 90.5% and 92.6% with the combined method. The DNA-ICM is an effective complementary method to cytology in diagnosing pancreatic malignancy. Although the diagnostic value for DNA-ICM is lower than that of cytology, an improved value was obtained after their combination.


Does melissa officinalis cause withdrawal or dependence?
Melissa officinalis is a medical and aromatic plant that is used for its hypnotic, sedative, and spasmolytic effects. This report presents a case study of30-year-old patient who was admitted to an emergency department with restlessness, tremor, distractibility, and sweating following a discontinuation of Melissa officinalis consumption. In this case, withdrawal symptoms may be related to the dependence effect caused by long-term use of Melissa officinalis. Although Melissa officinalis, a plant, is preferred by many patients as an alternative to pharmaceutical drugs, patients should be made aware that it may have a risk of dependency and can lead to withdrawal symptoms.


Interpersonal touch enhances cognitive control: A neurophysiological investigation.
Touch is central to mammalian communication, socialization, and wellbeing. Despite this prominence, interpersonal touch is relatively understudied. In this preregistered investigation, we assessed the influence of interpersonal touch on the subjective, neural, and behavioral correlates of cognitive control. Forty-five romantic couples were recruited (N = 90; dating >6 months), and one partner performed an inhibitory control task while electroencephalography was recorded to assess neural performance monitoring. Interpersonal touch was provided by the second partner and was manipulated between experimental blocks. A within-subject repeated-measures design was used to maximize statistical power, with our sample size providing 80% power for even small effect sizes (ds > .25). Results indicated that participants were not only happier when receiving touch, but also showed increased neural processing of mistakes. Further exploratory cognitive modeling using indirect effects tests and drift diffusion models of decision making revealed that touch was indirectly associated with both improved inhibitory control and increased rates of evidence accumulation (drift rate) through its influence on neural monitoring. Thus, beyond regulating emotion and stress, interpersonal touch appears to enhance the neurocognitive processes underling flexible goal-directed behavior. (PsycINFO Database Record


Comparison of Perioperative Outcomes between the Supraclavicular Artery Island Flap and Fasciocutaneous Free Flap.
Outcomes of the supraclavicular artery island flap (SCAIF) have not been extensively studied in comparison with free tissue transfer (FTT) flaps for head and neck reconstruction. We hypothesize that the pedicled SCAIF has decreased operating room time, length of stay, time to wound healing of recipient site, complications, and hospital charges as compared with FTT. Case series with chart review. Tertiary care teaching hospital. Medical records were reviewed for patients who underwent SCAIF (n = 45) or FTT (n = 28) reconstruction between 2011 and 2013. Total operating room time was significantly lower for the SCAIF group vs the FTT group (6.7 vs 8.1 hours, P = .002). Procedural time was 5.7 hours for the SCAIF group, as compared with 7.2 hours for FTT group (P = .0015). Mean area for SCAIF donor site was 63.89 cm(2) vs 81.8 cm(2) for the radial forearm free flap group (P = .015). There was no significant difference in mean length of stay between SCAIF (8.8 days) and FTT (11 days, P = .12). Mean length of time to wound healing of the recipient site was similar in the SCAIF group vs the FTT group (17.3 vs 22.1 days, P = .071). Ratio of total hospital charges for SCAIF were 32% lower than that of FTT (P = .0001). This is among the first studies to compare SCAIF with FTT in a large cohort analysis. We find decreased operating room times for SCAIF vs FTT, with similar length of stay and wound healing. Other outcomes between SCAIF and FTT were also comparable.


Microform cleft lip associated with a complete cleft palate.
Clefting of the lip with or without an associated cleft palate may be present in varying degrees of severity. The so-called microform cleft lip or forme fruste has been characteristically described as having the appearance of a repaired cleft lip. The following case describes a patient with microform cleft lip and a complete cleft of the hard and soft palates. To the best of our knowledge, this is the first report of such an association.


Aging effect on the ocular vestibular-evoked myogenic potentials.
This study applied ocular vestibular-evoked myogenic potential (oVEMP) tests in healthy subjects of various ages to measure the effect of aging on oVEMPs quantitatively. Prospective study. University hospital. Seventy healthy subjects with the ages ranging from 24 to 76 years were divided into 6 groups by decade. Each group consisted of 10 subjects in 1 decade except 20 subjects in the group of 60 to 69 years. All subjects underwent oVEMP tests using bone-conducted vibration stimuli. All subjects in groups of 20-59 years showed 100% response rates of clear oVEMPs, while 22 (55%) of 40 eyes in group of 60-69 years and 8 (40%) of 20 eyes in group of >or= 70 years had clear oVEMPs, exhibiting significant differences between those below and over 60 years of age. The mean nI and pI latencies also showed significantly prolonged in those over 60 years, whereas the mean nI-pI amplitude reduced significantly in subjects over 40 years. However, the asymmetry ratios of oVEMPs did not differ significantly among the age groups. Analyses between the age factor and characteristic parameters revealed that significant correlations existed between the age factor and the nI latency, pI latency and nI-pI amplitude, but not the nI-pI interval. Aging effect (> 60 years old) on the oVEMPs includes reduced response rate, prolonged nI and pI latencies, and decreased nI-pI amplitude. Thus, age factor should be taken into consideration when interpreting the oVEMP results.


Restricting indications for sinonasal computed tomography in children with cystic fibrosis.
OBJECTIVES To evaluate whether a low rate of exposure to sinonasal computed tomographic (CT) scans can be achieved when strict criteria are applied for their use in children with cystic fibrosis (CF) and to emphasize the importance of limiting radiation exposure in the context of the current longer life expectancy in this group of patients. DESIGN Retrospective chart review. SETTING Tertiary care children's hospital. PATIENTS The study included 277 children who were regularly followed up in the CF clinic in the last 11 years (mean duration of follow up, 7.87 years), 33 of whom underwent sinonasal CT. MAIN OUTCOME MEASURES Indications used for scanning, health professional (ear, nose, and throat specialist or pulmonologist) ordering the test, eventual modifications of ongoing treatment according to CT results, and time lapse between CT scanning and surgery. RESULTS Of 277 children with CF, 33 (12%) underwent a total of 39 sinonasal CT scans during the follow-up period (0.018 scans per patient per year of follow-up). Twenty-nine of the CT scans (74% of all cases, 90% of CT scans ordered by the ear, nose, and throat surgeon) were performed in the preoperative context and demonstrated the extent of the polypoid disease whenever present (26 cases [90%]) and the cause of nasal obstruction (20 cases [69%]). The mean period between the scanning and the surgery was 57 days (range, 0.10-173 days). Computed tomographic scans that were not meant for preoperative planning were performed in 10 cases (26%). The indications were disease evaluation (10%), ruling out a mucocele (5%); pre-lung transplantation status (5%), ruling out an intraorbital complication (3%); and headache investigation (3%). The results of the scans did not modify the management of the disease in those patients. CONCLUSIONS With the use of stringent criteria, it is possible to achieve a low rate of exposure to sinonasal CT scans in the population of children with CF. The main indication should be the preoperative planning regarding anatomy, extent of disease, and sites of nasal obstruction. The use of CT scans for disease evaluation does not seem to appreciably modify the treatment course and could be avoided.


Plasma free and protein-bound testosterone in hirsutism.
Blood samples were obtained from 145 preovulatory and postovulatory hirsute women and the plasma was analyzed for total testosterone (T), T index, and free T concentration as well as the fraction of free T percentage and that bound by testosterone-estradiol binding globulin. Fifty-six patients were subjected to a 2-week dexamethasone (DXM) suppression test to delineate the source of androgen excess. Presuppression and postsuppression samples were also analyzed for plasma T concentrations and its free and bound fractions. The total T, T index, and free T were elevated in 58, 66, and 55% of hirsute women, respectively. The percentage of bound T fraction was decreased in only 39% of hirsute women, whereas the percentage of free T fraction was increased in 30% and decreased in 11% of hirsute women. The postovulatory hirsute women had a normal percentage of free and bound T fractions, although their total plasma concentrations were elevated. They also had a lower frequency of abnormal levels in all T measurements than preovulatory hirsute women. The DXM seems to deplete the plasma binding globulin; thus the bound T fractions in the post-DXM samples were lower than the pre-DXM values. The total T may be a better measure than the free T values in evaluation of the DXM suppression test for hirsute women.


From cells to muropeptide structures in 24 h: peptidoglycan mapping by UPLC-MS.
Peptidoglycan (PGN) is ubiquitous in nearly all bacterial species. The PGN sacculus protects the cells against their own internal turgor making PGN one of the most important targets for antibacterial treatment. Within the last sixty years PGN composition has been intensively studied by various methods. The breakthrough was the application of HPLC technology on the analysis of muropeptides. However, preparation of pure PGN relied on a very time consuming method of about one week. We established a purification protocol for both Gram-positive and Gram-negative bacteria which can be completely performed in plastic reaction tubes yielding pure muropeptides within 24 hours. The muropeptides can be analyzed by UPLC-MS, allowing their immediate determination. This new rapid method provides the feasibility to screen PGN composition even in high throughput, making it a highly useful tool for basic research as well as for the pharmaceutical industry.


A truncating mutation of CEP135 causes primary microcephaly and disturbed centrosomal function.
Autosomal-recessive primary microcephaly (MCPH) is a rare congenital disorder characterized by intellectual disability, reduced brain and head size, but usually without defects in cerebral cortical architecture, and other syndromic abnormalities. MCPH is heterogeneous. The underlying genes of the seven known loci code for centrosomal proteins. We studied a family from northern Pakistan with two microcephalic children using homozygosity mapping and found suggestive linkage for regions on chromosomes 2, 4, and 9. We sequenced two positional candidate genes and identified a homozygous frameshift mutation in the gene encoding the 135 kDa centrosomal protein (CEP135), located in the linkage interval on chromosome 4, in both affected children. Post hoc whole-exome sequencing corroborated this mutation's identification as the causal variant. Fibroblasts obtained from one of the patients showed multiple and fragmented centrosomes, disorganized microtubules, and reduced growth rate. Similar effects were reported after knockdown of CEP135 through RNA interference; we could provoke them also by ectopic overexpression of the mutant protein. Our findings suggest an additional locus for MCPH at HSA 4q12 (MCPH8), further strengthen the role of centrosomes in the development of MCPH, and place CEP135 among the essential components of this important organelle in particular for a normal neurogenesis.


Analysis of quaternary ammonium compounds in urban stormwater samples.
A method for benzalkonium analysis has been developed to measure benzalkonium concentration in dissolved and particulate fractions from urban runoff samples. The analysis was performed by liquid chromatography coupled with mass spectrometry (LC-MS/MS). The dissolved matrix was extracted by Solid Phase Extraction (SPE), with cationic exchange and the particles by microwave extraction with acidified methanol. Recovery percentages were closed to 100% for benzalkonium C12 and C14. The protocol was applied to roof runoff samples collected after a roof demossing treatment, and to separative stormwater samples from a 200 ha catchment. The results illustrate an important contamination of the roof runoff, with a maximum concentration close to 27 mg/L during the first rain. The benzalkonium concentration (sum of C12 and C14) stayed high (up to 1 mg/L) even 5 months after the treatment. Benzalkonium concentration measured in stormwaters was low (0.2 μg/L) but with contaminated suspended solids (up to 80 μg/g).


Validation of abdominal aortic aneurysm repair codes in Ontario administrative data.
To determine the positive predictive values (PPV) of Ontario administrative data codes for the identification of open (OSR) and endovascular (EVAR) repairs of elective (eAAA) and ruptured (rAAA) abdominal aortic aneurysms. We randomly identified 319 eAAA and rAAA repairs at two Toronto hospitals between April 2003 and March 2015, using administrative health data in Ontario, Canada. International Statistical Classification of Diseases and Related Health Problems 10th Revision (ICD-10) codes I71.3 and I71.4, were used to identify rAAA and eAAA patients, respectively. A blinded retrospective chart review was conducted and served as the gold standard comparator. Re-abstracted records were compared to Canadian Classification of Health Interventions (CCI) and Ontario Health Insurance Plan (OHIP) codes in the Canadian Institute for Health Information Discharge Abstract Database (CIHI-DAD) and OHIP databases. We calculated the PPV and 95% confidence intervals (95% CI) of individual and combined procedure and billing codes for elective and ruptured OSR and EVAR (eOSR, eEVAR, rOSR, and rEVAR). Permutation of codes allowed identification of eOSR with 95% PPV (95% CI 88, 98), eEVAR with 96% PPV (95% CI 90, 99), rOSR with 87% PPV (95% CI 79, 93) and rEVAR with 91% PPV (95% CI 59, 100). Diagnostic, procedure and billing code combinations allow identification of eOSR, eEVAR, rOSR and rEVAR patients in Ontario administrative data with a high degree of certainty.


Therapeutic implications of HPA axis abnormalities in Alzheimer's disease: review and update.
The adaptive and maladaptive roles of the hypothalamic-pituitary-adrenal (HPA) axis in stressful conditions and in disorders such as major depression, posttraumatic stress disorder, and Cushing's syndrome, have been the subject of substantial, ongoing study. In particular, HPA disturbances have been associated with memory impairments, and hypercortisolemic conditions with atrophy of the hippocampus, a limbic structure closely associated with declarative memory. Recent discoveries support a more complicated picture of HPA axis function and pathology in acquiring, retrieving, and consolidating new memories. These findings include: the existence of an 'inverted U-shaped relationship" between stimulation of brain glucocorticoid receptors and memory performance; that distinct areas of the hippocampus have been found to respond differently to cortisol stimulation; and that hippocampal atrophy has been found to be potentially reversible in some conditions, although whether such atrophy is a cause or effect of these pathological conditions is currently unclear. More longitudinal studies of HPA axis function in aging normal individuals, those with mild cognitive impairment,and individuals with Alzheimer' disease, examining pertinent variables such as APOEe-4 status, are needed to help clarify these new findings. Antiglucocorticoid agents appear to have therapeutic value in particular conditions. These results are relevant for understanding and treating memory dysfunction in individuals with Alzheimer's disease, a disorder prominently and invariably characterized by early hippocampal lesions and memory impairment. Given the burden of this disease, we feel it timely to encourage controlled trials of antiglucocorticoid agents in the treatment of mild cognitive impairment and Alzheimers disease.


Expression and thermal stability of simian virus 40 tumor-specific transplantation antigen and tumor antigen in wild type- and tsA mutant-transformed cells.
We have explored aspects of a suggested relationship between the expression of simian virus 40 tumor-specific transplantation antigen (TSTA) and tumor antigen (TA). A unique rat embryo cell line transformed by a temperature-sensitive A mutant that loses TA during exposure to the nonpermissive temperature (A28-RE) was found to lose TSTA. On the other hand, a typical control tsA-transformed cell line (A239-MB) expressed both TA and TSTA at the non-permissive temperature. TA in lysates obtained from A239-MB cells was found to be three to four times more thermolabile by covwt-mb) when incubated at either 33 or 40 degrees C. These data complement previous reports using TA from lytic infection and are consistent with the suggestion that TA is virus encoded. In contrast to TA, which even in wild-type-transformed cells was completely destroyed in less than 10 min at 50 degrees C, TSTA, assayed in vivo by tumor rejection, and tumor-specific surface antigen(s) TSSA) defined by an in vitro cytolytic assay, were thermostabile. Even after 24 h of incubation of extracts of 50 degrees C, high levels of TSTA and TSSA activity were present. Since these surface antigens when obtained from cells transformed by tsA mutants were also thermostabile, they could not be distinguished from the wild-type antigens. These results (i) indicate a coordinate expression of TA and TSTA in transformed cells; (ii) confirm that TA is virus encoded; and (iii) confirm that tha antigenic and immunogenic determinants that characterize TA and TSTA activities are distinct. However, the possibility that TSTA, like TA, is of viral rather than cellular origin is not excluded.


"You cannot perform music without taking care of your body": a qualitative study on musicians' representation of body and health.
To identify professional musicians' representation of health and illness and to identify its perceived impact on musical performance. A total of 11 professional musicians participated in this phenomenological study. Five of the musicians were healthy, and the others suffered debilitating physical health problems caused by playing their instruments. Semi-structured interviews were conducted, transcribed verbatim and analysed. Thematic analysis, including a six-step coding process, was performed (ATLAS-ti 6). Three major themes emerged from the data: music as art, the health of musicians, and learning through experience. The first theme, music as art, was discussed by both groups; they talked about such things as passion, joy, sense of identity, sensitivity, and a musician's hard life. Discussions of the second theme, the health of musicians, revealed a complex link between health and performance, including the dramatic impact of potential or actual health problems on musical careers. Not surprisingly, musicians with health problems were more concerned with dysfunctional body parts (mostly the hand), whereas healthy musicians focused on maintaining the health of the entire person. The third theme, learning through experience, focused on the dynamic nature of health and included the life-long learning approach, not only in terms of using the body in musical performance but also in daily life. The centre of a musician's life is making music in which the body plays an important part. Participants in this study evidenced a complex link between health and musical performance, and maintaining health was perceived by these musicians as a dynamic balance. Our results suggest that learning through experience might help musicians adapt to changes related to their bodies.


Simultaneous quantification of peroxidase and ascorbic acid in biosamples with an automatic system based on a Fe(iii)/methylthymol blue-carbon dot simulative enzyme.
Peroxidase (POD) and ascorbic acid (AsA) usually coexist in organisms to synergistically protect them from reactive oxygen damage, and their contents undergo dynamic changes under different physiological conditions. What's more, the response of POD-catalytic activity in spectrophotometry has to be corrected using the content of concomitant AsA because we found that there is an extinction reaction between AsA and chromogenic products obtained from POD catalysis. With these implications, by skilfully using the chromogenic and the extinction phenomena in the guaiacol/POD/H2O2 reaction, an automatic analysis system for simultaneous quantification of POD (73-440 U L-1) and AsA (4-60 mg L-1) was successfully established based on flow injection analysis (FIA). Furthermore, under acidic conditions (0.5 mol L-1 of HCl), hydrothermal synthesis (250 °C for 1 h) was used for synthesizing new carbon dots (sPOD-CDs) of methylthymol blue (0.08 g L-1)/FeCl3 (0.8 g L-1), which is a simulative enzyme for POD, and it was first used for catalyzing the guaiacol/H2O2 reaction within the FIA system to replace natural HRP in the extinction reaction. This sPOD-CD solution has no background absorption and its concentration shows excellent correlation with simulative POD-activity. Finally, after optimization, this FIA system was utilized to testify that the reducibility of AsA is due to ascorbate ions and to determine POD and AsA in some plant samples. The standard addition recovery experiment showed that there was no interference from the matrix in real samples (recoveries: 95%-105%), and the obtained POD and AsA results were also consistent with the reference experiments (relative deviation ≤ 2.80%, t-test ≥ 0.07). The proposed FIA system is characterized by high sample-throughput (40 samples per h), better repeatability (relative standard deviation ≤ 1.4%), etc.


Hub GABA neurons mediate gamma-frequency oscillations at ictal-like event onset in the immature hippocampus.
Gamma-frequency oscillations (GFOs, >40 Hz) are a general network signature at seizure onset at all stages of development, with possible deleterious consequences in the immature brain. At early developmental stages, the simultaneous occurrence of GFOs in different brain regions suggests the existence of a long-ranging synchronizing mechanism at seizure onset. Here, we show that hippocamposeptal (HS) neurons, which are GABA long-range projection neurons, are mandatory to drive the firing of hippocampal interneurons in a high-frequency regime at the onset of epileptiform discharges in the intact, immature septohippocampal formation. The synchronized firing of interneurons in turn produces GFOs, which are abolished after the elimination of a small number of HS neurons. Because they provide the necessary fast conduit for pacing large neuronal populations and display intra- and extrahippocampal long-range projections, HS neurons appear to belong to the class of hub cells that play a crucial role in the synchronization of developing networks.


Acid ceramidase as a chemotherapeutic target to overcome resistance to the antitumoral effect of choline kinase α inhibition.
We have analyzed the response of primary cultures derived from tumor specimens of non small cell lung cancer (NSCLC) patients to choline kinase α (ChoKα) inhibitors. ChoKα inhibitors have been demonstrated to increase ceramides levels specifically in tumor cells, and this increase has been suggested as the mechanism that explain its proapoptotic effect in cancer cells. Here, we have investigated the molecular mechanism associated to the intrinsic resistance, and found that other enzyme involved in lipid metabolism, acid ceramidase (ASAH1), is specifically upregulated in resistant tumors. NSCLC cells with acquired resistance to ChoKα inhibitors also display increased levels of ASAH1. Accordingly, ASAH1 inhibition synergistically sensitizes lung cancer cells to the antiproliferative effect of ChoKα inhibitors. Thus, the determination of the levels of ASAH1 predicts sensitivity to targeted therapy based on ChoKα specific inhibition and represents a model for combinatorial treatments of ChoKα inhibitors and ASAH1 inhibitors. Considering that ChoKα inhibitors have been recently approved to enter Phase I clinical trials by the Food and Drug Administration (FDA), these findings are anticipating critical information to improve the clinical outcome of this family of novel anticancer drugs under development.


Domain evolution with electric field and delineation of extrinsic contributions in (K, Na, Li)(Nb, Ta, Sb)O3 single crystal.
Extrinsic contributions play an important role in the functionalities of ferroelectric materials, while domain structure evolution is crucial for understanding the extrinsic dielectric and piezoelectric responses. In this work, domain configuration changes with an electric field applied along [001] C in the tetragonal (K, Na, Li)(Nb, Sb, Ta)O3 single crystal were studied by means of polarizing light microscopy. Results show that parts of the spontaneous polarizations in the (001) C plane are switched to [001] C direction, while others still stay in the (001) C plane due to high induced internal stresses. Single domain state cannot be achieved even under a high electric field. After being poled along [001] C , the volume fraction of domains with polarzations in the (001) C plane is still about 25.2%. The extrinsic contributions to the dielectric constant are 15.7% and 27.2% under the E field of 1 kV/cm and under 2 kV/cm, respectively, estimated by the Rayleigh analysis.


A multitude of kinases--which are the best targets in treating rheumatoid arthritis?
Small-molecule kinase inhibitors are increasingly taking center stage in the quest for new drugs for the treatment of rheumatoid arthritis (RA). By targeting kinases, small-molecule inhibitors can exert potent anti-inflammatory and immunomodulatory effects; the success of small-molecule kinase inhibitors in the treatment of cancer has spurred efforts to identify kinases that could be targeted for the treatment of chronic inflammatory disorders, such as RA. Although many kinase inhibitors have proved efficacious in the treatment of inflammatory arthritis in animals few have been tested in RA clinical trials. This article discusses the challenges and progress in the pursuit of small-molecule kinase inhibitors for RA, including lessons learned from the failure of erstwhile frontrunner inhibitors and the promise of inhibitors making their debut on the RA stage.


Pediatric stroke initiatives and preliminary studies: What is known and what is needed?
Cerebrovascular disorders are increasingly recognized as important causes of mortality and morbidity in the pediatric population. However, there have been no clinical trials performed to assess the safety and tolerability of acute interventions or secondary preventative treatments. In 2002, the International Pediatric Stroke Study was launched to create a network of investigators, with an interest in developing standards of practice, as well as design and implement the first-ever clinical trials in pediatric stroke. This article reviews existing studies in pediatric stroke epidemiology, risk factors, outcomes, as well as experience with antithrombotic trials in children. From there, current and future initiatives in the development of clinical trials in pediatric stroke are evaluated.


Improving evolutionary models for mitochondrial protein data with site-class specific amino acid exchangeability matrices.
Adequate modeling of mitochondrial sequence evolution is an essential component of mitochondrial phylogenomics (comparative mitogenomics). There is wide recognition within the field that lineage-specific aspects of mitochondrial evolution should be accommodated through lineage-specific amino-acid exchangeability matrices (e.g., mtMam for mammalian data). However, such a matrix must be applied to all sites and this implies that all sites are subject to the same, or largely similar, evolutionary constraints. This assumption is unjustified. Indeed, substantial differences are expected to arise from three-dimensional structures that impose different physiochemical environments on individual amino acid residues. The objectives of this paper are (1) to investigate the extent to which amino acid evolution varies among sites of mitochondrial proteins, and (2) to assess the potential benefits of explicitly modeling such variability. To achieve this, we developed a novel method for partitioning sites based on amino acid physiochemical properties. We apply this method to two datasets derived from complete mitochondrial genomes of mammals and fish, and use maximum likelihood to estimate amino acid exchangeabilities for the different groups of sites. Using this approach we identified large groups of sites evolving under unique physiochemical constraints. Estimates of amino acid exchangeabilities differed significantly among such groups. Moreover, we found that joint estimates of amino acid exchangeabilities do not adequately represent the natural variability in evolutionary processes among sites of mitochondrial proteins. Significant improvements in likelihood are obtained when the new matrices are employed. We also find that maximum likelihood estimates of branch lengths can be strongly impacted. We provide sets of matrices suitable for groups of sites subject to similar physiochemical constraints, and discuss how they might be used to analyze real data. We also discuss how the general approach might be employed to improve a variety of mitogenomic-based research activities.


Coronavirus Disease 2019 Return to Work Guidance and Recommendations for Vestibular Clinicians.
As states begin issuing progressive deconfinement guidelines, hospitals and institutions are starting to reopen for elective procedures and consultations. Vestibular clinicians are opening their practices to evaluate, test, or treat patients with dizziness and balance problems. The following document, requested by the American Balance Society, collates the current information about the virus, including transmission from asymptomatic carriers, decontamination, and other safety protocols, and provides a return to work guidance for clinicians caring for this population of patients, promoting provider, patient, and staff safety.


Gut immunity: its development and reasons and opportunities for modulation in monogastric production animals.
The intestine performs the critical roles of nutrient acquisition, tolerance of innocuous and beneficial microorganisms, while retaining the ability to respond appropriately to undesirable microbes or microbial products and preventing their translocation to more sterile body compartments. Various components contribute to antimicrobial defenses in the intestine. The mucus layer(s), antimicrobial peptides and IgA provide the first line of defense, and seek to trap and facilitate the removal of invading microbes. If breached, invading microbes next encounter a single layer of epithelial cells and, below this, the lamina propria with its associated immune cells. The gut immune system has developmental stages, and studies from different species demonstrate that innate capability develops earlier than acquired. In addition, various factors may influence the developmental process; for example, the composition and activity of the gut microbiota, antimicrobials, maternally derived antibodies, host genetics, and various stressors (e.g. feed deprivation). Therefore, it is clear that particularly younger (meat-producing) animals are reliant on innate immune responses (as well as passive immunity) for a considerable period of their productive life, and thus focusing on modulating appropriate innate responses should be an intervention priority. The gut microbiota is probably the most influential factor for immune development and capability. Interventions (e.g. probiotics, prebiotics, antibodies, etc.) that appropriately modulate the composition or activity of the intestinal microbiota can play an important role in shaping the desired functionality of the innate (and acquired) response. In addition, innate immune mediators, such as toll-like receptor agonists, cytokines, etc., may provide more specific ways to suitably modulate the response. A better understanding of mucosal immunology, signaling pathways, and processes, etc., will provide even more precise methods in the future to boost innate immune capability and minimize any associated (e.g. nutrient) costs. This will provide the livestock industry with more effective options to promote robust and efficient productivity.


Homosexual and heterosexual sex-role orientation on six sex-role scales.
A comparison was made of the sex roles of homosexual and heterosexual men and women on the Bem Sex Role Inventory, Personality Attributes Questionnaire, Personality Research Form Androgyny Scale, Adjective Checklist Masculinity and Femininity Scales, Extended Personality Attributes Questionnaire and Undesirable Characteristics Scale. The results indicated that homosexuals and heterosexuals differ in their response to different aspects of sex roles. The most consistent difference was the greater femininity of male homosexuals in respect to male heterosexuals. Other differences were scale-specific and the low interscale comparability indicated such scales should not be used interchangeably. Differences between results of studies comparing sex roles of the homosexuals and heterosexuals appear attributable to sample heterogeneity and distinctions between sex-role scales.


Hypothermia for neuroprotection in adults after cardiac arrest.
Good neurological outcome after cardiac arrest is difficult to achieve. Interventions during the resuscitation phase and treatment within the first hours after the event are critical for a favourable prognosis. Experimental evidence suggests that therapeutic hypothermia is beneficial, and several clinical studies on this topic have been published. This review was originally published in 2009; updated versions were published in 2012 and 2016. To evaluate the benefits and harms of therapeutic hypothermia after cardiac arrest in adults compared to standard treatment. We used standard, extensive Cochrane search methods. The latest search date was 30 September 2022. We included randomised controlled trials (RCTs) and quasi-RCTs in adults comparing therapeutic hypothermia after cardiac arrest with standard treatment (control). We included studies with adults cooled by any method, applied within six hours of cardiac arrest, to target body temperatures of 32 °C to 34 °C. Good neurological outcome was defined as no or only minor brain damage allowing people to live an independent life. We used standard Cochrane methods. Our primary outcome was 1. neurological recovery. Our secondary outcomes were 2. survival to hospital discharge, 3. quality of life, 4. cost-effectiveness and 5. We used GRADE to assess certainty. We found 12 studies with 3956 participants reporting the effects of therapeutic hypothermia on neurological outcome or survival. There were some concerns about the quality of all the studies, and two studies had high risk of bias overall. When we compared conventional cooling methods versus any type of standard treatment (including a body temperature of 36 °C), we found that participants in the therapeutic hypothermia group were more likely to reach a favourable neurological outcome (risk ratio (RR) 1.41, 95% confidence interval (CI) 1.12 to 1.76; 11 studies, 3914 participants). The certainty of the evidence was low. When we compared therapeutic hypothermia with fever prevention or no cooling, we found that participants in the therapeutic hypothermia group were more likely to reach a favourable neurological outcome (RR 1.60, 95% CI 1.15 to 2.23; 8 studies, 2870 participants). The certainty of the evidence was low. When we compared therapeutic hypothermia methods with temperature management at 36 °C, there was no evidence of a difference between groups (RR 1.78, 95% CI 0.70 to 4.53; 3 studies; 1044 participants). The certainty of the evidence was low. Across all studies, the incidence of pneumonia, hypokalaemia and severe arrhythmia was increased amongst participants receiving therapeutic hypothermia (pneumonia: RR 1.09, 95% CI 1.00 to 1.18; 4 trials, 3634 participants; hypokalaemia: RR 1.38, 95% CI 1.03 to 1.84; 2 trials, 975 participants; severe arrhythmia: RR 1.40, 95% CI 1.19 to 1.64; 3 trials, 2163 participants). The certainty of the evidence was low (pneumonia, severe arrhythmia) to very low (hypokalaemia). There were no differences in other reported adverse events between groups. Current evidence suggests that conventional cooling methods to induce therapeutic hypothermia may improve neurological outcomes after cardiac arrest. We obtained available evidence from studies in which the target temperature was 32 °C to 34 °C.


Machine Learning-Based Fragment Selection Improves the Performance of Qualitative PRM Assays.
Targeted mass spectrometry-based platforms have become a valuable tool for the sensitive and specific detection of protein biomarkers in clinical and research settings. Traditionally, developing a targeted assay for peptide quantification has involved manually preselecting several fragment ions and establishing a limit of detection (LOD) and a lower limit of quantitation (LLOQ) for confident detection of the target. Established thresholds such as LOD and LLOQ, however, inherently sacrifice sensitivity to afford specificity. Here, we demonstrate that machine learning can be applied to qualitative PRM assays to discriminate positive from negative samples more effectively than a traditional approach utilizing conventional methods. To demonstrate the utility of this method, we trained an ensemble machine learning model using 282 SARS-CoV-2 positive and 994 SARS-CoV-2 negative nasopharyngeal swabs (NP swab) analyzed using a targeted PRM method. This model was then validated using an independent set of 200 positive and 150 negative samples and achieved a sensitivity of 92% relative to results obtained by RT-PCR, which was superior to a traditional approach that resulted in 86.5% sensitivity when analyzing the same data. These results demonstrate that machine learning can be applied to qualitative PRM assays and results in superior performance relative to traditional methods.


Development of Probiotic Formulation for the Treatment of Iron Deficiency Anemia.
Probiotics are increasingly more present both as functional foods, and in pharmaceutical preparations with multiple levels of action that contribute to human health. Probiotics realize their positive effects with a proper dose, and by maintaining a declared number of probiotics cells by the expiration date. Important precondition for developing a probiotic product is the right choice of clinically proven probiotic strain, the choice of other active components, as well as, the optimization of the quantity of active component of probiotic per product dose. This scientific paper describes the optimization of the number of probiotics cells in the formulation of dietary supplement that contains probiotic culture Lactobacillus plantarum 299v, iron and vitamin C. Variations of the quantity of active component were analyzed in development batches of the encapsulated probiotic product categorized as dietary supplement with the following ingredients: probiotic culture, sucrosomal form of iron and vitamin C. Optimal quantity of active component L. plantarum of 50 mg, was selected. The purpose of this scientific paper is to select the optimal formulation of probiotic culture in a dietary supplement that contains iron and vitamin C, and to also determine its expiration date by the analysis of the number of viable probiotic cells.


Reengineering biocatalysts: Computational redesign of chondroitinase ABC improves efficacy and stability.
Maintaining biocatalyst stability and activity is a critical challenge. Chondroitinase ABC (ChABC) has shown promise in central nervous system (CNS) regeneration, yet its therapeutic utility is severely limited by instability. We computationally reengineered ChABC by introducing 37, 55, and 92 amino acid changes using consensus design and forcefield-based optimization. All mutants were more stable than wild-type ChABC with increased aggregation temperatures between 4° and 8°C. Only ChABC with 37 mutations (ChABC-37) was more active and had a 6.5 times greater half-life than wild-type ChABC, increasing to 106 hours (4.4 days) from only 16.8 hours. ChABC-37, expressed as a fusion protein with Src homology 3 (ChABC-37-SH3), was active for 7 days when released from a hydrogel modified with SH3-binding peptides. This study demonstrates the broad opportunity to improve biocatalysts through computational engineering and sets the stage for future testing of this substantially improved protein in the treatment of debilitating CNS injuries.


Optical brain imaging in vivo: techniques and applications from animal to man.
Optical brain imaging has seen 30 years of intense development, and has grown into a rich and diverse field. In-vivo imaging using light provides unprecedented sensitivity to functional changes through intrinsic contrast, and is rapidly exploiting the growing availability of exogenous optical contrast agents. Light can be used to image microscopic structure and function in vivo in exposed animal brain, while also allowing noninvasive imaging of hemodynamics and metabolism in a clinical setting. This work presents an overview of the wide range of approaches currently being applied to in-vivo optical brain imaging, from animal to man. Techniques include multispectral optical imaging, voltage sensitive dye imaging and speckle-flow imaging of exposed cortex, in-vivo two-photon microscopy of the living brain, and the broad range of noninvasive topography and tomography approaches to near-infrared imaging of the human brain. The basic principles of each technique are described, followed by examples of current applications to cutting-edge neuroscience research. In summary, it is shown that optical brain imaging continues to grow and evolve, embracing new technologies and advancing to address ever more complex and important neuroscience questions.