New frontiers in headache therapy.
There are numerous headache therapies available for our patients, more for migraine than for any of the other primary headache disorders. Only four medications have been approved for migraine prevention in the last few decades in the US and onabotulinumtoxinA was recently approved in the UK and the US for chronic migraine. We have been more fortunate in the acute care arena where in the US we have had seven triptans and one nonsteroidal anti-inflammatory medication approved by the FDA and currently available. There are several other acute care medications in various stages of development and there are two new methods of administering a triptan and others under investigation. We are always looking for faster, easier and more efficient administration of medications with fewer adverse events, as optimal migraine therapy requires these characteristics. What follows is a brief review of the progress in development for four of the many new acute care medications being investigated: the CGRP antagonist tablet telcagepant, the sumatriptan iontophoretic patch, sumatriptan powder for use in the OptiNose apparatus and the dihydroergotamine oral inhaler. I will not include transcranial magnetic stimulation, a 5-HT(1F) agonist, large conductance calcium-activated potassium channel openers, glial modulators or other medications and devices in early stages of development [1].


Supercritical extraction of volatile and fixed oils from Petroselinum crispum L. seeds: chemical composition and biological activity.
The supercritical fluid extraction (SFE) of volatile and fixed oil from milled parsley (Petroselinum crispum L.) seeds, using CO2 as solvent, is presented in this study. Extraction experiments were carried out in two steps: at pressures of (90 or 300) bar and temperature of 40 °C. The first extraction step, performed at 90 bar, produced a volatile fraction mainly formed by apiole (82.1%) and myristicin (11.4%). The volatile oil yield was 2.6% by weight of the charge. The second extraction step, carried out at 300 bar produced a fixed oil at a yield of 0.4% by weight. The most represented fatty acids in P. crispum fixed oil were 18:1 n-12 (49.9%), 18:2 n-6 (18.2%), 18:1 n-9 (11.8%), and 16:0 (7.4%). In particular, the unsaturated fatty acids 18:1 n-12 and 18:1 n-9 averaged 182.2 mg/g and 92.1 mg/g of oil extract, respectively. The quality of the oils extracted by SFE, in terms of its chemical composition, was compared to the oils obtained by hydrodistillation (HD) in a Clevenger apparatus and by solvent extraction (SE) using n-hexane in a Soxhlet apparatus. The antioxidant properties were determined by means of the ABTS assay. The results indicated that the fixed oil possessed low antioxidant activity (EC50 = 0.4 mg/mL) and the volatile oil had no antioxidant activity. The total phenolic content, expressed as concentration of gallic acid (gallic acid equivalent, GAE), of the fixed oil was 1.5 mg/g. The fixed oil found to have inhibitory effects against α-glucosidase, the volatile oil is active on acetylcholinesterase (AChE), tyrosinase, and α-glucosidase. Both samples have weak inhibitory activity on α-amylase and no activity on butyrylcholinesterase (BChE).


Building the Neurology Pipeline With Undergraduate Students in Research and Clinical Practice.
There is currently a shortage of neurologists in the United States. Multiple efforts are underway to increase the strength of the neurology workforce. One potential approach is early exposure to neurology research and clinical care to pique interest and promote the specialty as a career choice. This study details the rewarding experience of working with undergraduate students, both in clinical research and clinical care. The logistics, benefits to students, and positive aspects for neurologists are outlined. Examples provided by undergraduate students who have participated in neurology research and clinical care are presented. The ultimate goals of this work are to encourage and inspire academic neurologists to involve undergraduate students in research and clinical care, to facilitate this process by outlining the steps needed to make this pairing successful, and to ultimately promote a pathway to build the neurology pipeline.


[Effect of streptozotocin on the glycemic and lipid profiles and oxidative stress in hamsters].
This study evaluated the effects of streptozotocin on glycemic and lipid profiles and oxidative stress status in hamsters. Male Golden Syrian hamsters were divided in diabetic group (D) which received a streptozotocin single injection (STZ - 50 mg/kg), and control group (C) which received a single injection of the vehicle citrate buffer. Animals were euthanized after 10 days of experiment and blood, liver and kidneys were collected. The diabetic group had higher levels of glucose, triacylglycerols and cholesterol in serum and thiobarbituric acid reactive substances (TBARS) concentration increased in the liver and kidneys. Diabetes induced a significant increase in glutathione concentration in the liver and decreased paraoxonase and superoxide dismutase activities. Hamsters provide a novel animal model for diabetes mellitus and oxidative stress, similar to the human syndrome, which may be suitable for the testing of antidiabetic compounds.


Long-term results of a postoperative pneumonia prevention program for the inpatient surgical ward.
Pneumonia is the third most common complication in postoperative patients and is associated with significant morbidity and high cost of care. Prevention has focused primarily on mechanically ventilated patients. This study outlines the results of the longest-running postoperative pneumonia prevention program for nonmechanically ventilated patients, to our knowledge. To present long-term results (2008-2012) of a standardized postoperative ward-acquired pneumonia prevention program introduced in 2007 on the surgical ward of our hospital and compare our postintervention pneumonia rates with those captured in the American College of Surgeons National Surgical Quality Improvement Program (ACS-NSQIP). We also estimate the cost savings attributable to the pneumonia prevention program. Retrospective cohort study at a university-affiliated Veterans Affairs hospital of all noncardiac surgical patients with ward-acquired postoperative pneumonia. A previously described standardized postoperative pneumonia prevention program for patients on the surgical ward. Ward-acquired postoperative pneumonia. Between 2008 and 2012, there were 18 cases of postoperative pneumonia among 4099 at-risk patients hospitalized on the surgical ward, yielding a case rate of 0.44%. This represents a 43.6% decrease from our preintervention rate (0.78%) (P = .01). The pneumonia rates in all years were lower than the preintervention rate (0.25%, 0.50%, 0.58%, 0.68%, and 0.13% in 2008-2012, respectively). The overall pneumonia rate in ACS-NSQIP was 2.56% (14,033 cases of pneumonia among 547,571 at-risk patients), which is 582% higher than the postintervention rate at our ward. Using a national average of $46,400 in attributable health care cost of postoperative pneumonia and a benchmark of a 43.6% decrease in pneumonia rate achieved at our facility over the 5-year study period, a similar percentage of decrease in pneumonia occurrence at ACS-NSQIP hospitals would represent approximately 6118 prevented pneumonia cases and a cost savings of more than $280 million. The standardized pneumonia prevention program achieved substantial and sustained reduction in postoperative pneumonia incidence on our surgical ward; its wider adoption could improve postoperative outcomes and reduce overall health care costs.


Antinociceptive activity of a hydroalcoholic extract obtained from aerial parts of Sebastiania schottiana (Euphorbiaceae).
This study analyzed the antinociceptive effects of a hydroalcoholic extract obtained from the aerial parts of Sebastiania schottiana, a Brazilian medicinal plant used to treat various painful diseases. For this purpose, the writhing test, capsaicin and formalin induced-pain in mice were used. The results showed that the hydroalcoholic extract exhibited considerable antinociception in all the models studied, being more potent than aspirin.


Cell-autonomous expression of barley Mla1 confers race-specific resistance to the powdery mildew fungus via a Rar1-independent signaling pathway.
The barley Mla locus encodes 28 characterized resistance specificities to the biotrophic fungal pathogen barley powdery mildew. We describe a single-cell transient expression assay using entire cosmid DNAs to pinpoint Mla1 within the complex 240-kb Mla locus. The MLA1 cDNA encodes a 108-kD protein containing an N-terminal coiled-coil structure, a central nucleotide binding domain, and a C-terminal leucine-rich repeat region; it also contains a second short open reading frame at the 5' end that has a possible regulatory function. Although most Mla-encoded resistance specificities require Rar1 for their function, we used the single-cell expression system to demonstrate that Mla1 triggers full resistance in the presence of the severely defective rar1-2 mutant allele. Wheat contains an ortholog of barley Mla, designated TaMla, that is tightly linked to (0.7 centimorgan) but distinct from a tested resistance specificity at the complex Pm3 locus to wheat powdery mildew. Thus, the most polymorphic powdery mildew resistance loci in barley and wheat may have evolved in parallel at two closely linked homeoloci. Barley Mla1 expressed in wheat using the single-cell transformation system failed to trigger a response to any of the wheat powdery mildew Avr genes tested, indicating that AvrMla1 is not genetically fixed in wheat mildew strains.


Effect of pulmonary vein isolation on the distribution of complex fractionated electrograms in humans.
Targeting of complex fractionated electrograms (CFEs) has been used as an adjunctive strategy to pulmonary vein isolation (PVI) in patients with persistent atrial fibrillation (AF). However, it is unclear whether CFEs should be targeted before or after PVI. The purpose of this study was to examine the effect of PVI on CFE distribution in humans. We compared left atrial (LA) CFE maps acquired using the NavX system before and after PVI in patients with persistent AF. CFE maps were constructed from bipolar electrograms acquired from a circular mapping catheter. At each point, the mean AF cycle length (CL) was calculated automatically by averaging the intervals between deflections over a 4-second window. Sites with mean CL < or =120 ms were considered CFE+. A total of 22 consecutive patients (82% male, age 58 +/- 9 years) were studied. At baseline, 47% of the LA was encompassed by electrograms with CL <120 ms. PVI had a significant effect on CFE characteristics, with an increase in mean LA AF CL (144 ms pre-PVI vs. 214 ms post-PVI; P <.01) and a decrease in CFE+ LA surface area (47% vs 23%; P <.01). There was significant reduction in CFE burden after PVI in both PV (50% vs. 6%; P <.01) and non-PV (61% vs. 39%; P <.01) regions. In patients with persistent AF, PVI results in a significant decrease in both PV and non-PV areas of CFE. To limit extensive LA ablation, PVI should be performed before targeting CFE when a combined approach is pursued.


Smoking in Pregnancy and Child ADHD.
There is a well-documented association between maternal smoking during pregnancy and offspring attention-deficit/hyperactivity disorder (ADHD). The degree to which this reflects causal intrauterine effects or is due to unmeasured confounding is not clear. We sought to compare the association between maternal smoking during pregnancy and offspring ADHD with the associations with paternal smoking, grandmother's smoking when pregnant with mother, and maternal smoking in previous pregnancies. Each of these exposures is expected to be influenced by much of the same confounding factors as maternal smoking during pregnancy, but cannot have direct intrauterine effects. A sibling control design was also used. The current study used data from the Norwegian Mother and Child Cohort Study (n > 100 000 children). Mothers and fathers reported on smoking during pregnancy, and mothers reported on smoking in previous pregnancies and their mother's smoking when pregnant with them. Mothers reported on child ADHD symptoms at 5 years of age. Information about child ADHD diagnosis was obtained from the Norwegian Patient Registry. Maternal smoking during pregnancy was not more strongly associated with offspring ADHD diagnosis than was paternal smoking, grandmother's smoking when pregnant with mother, or maternal smoking in previous pregnancies. Sibling control analyses showed no association between maternal smoking in pregnancy and child ADHD symptoms among siblings discordant for maternal smoking. These results suggest that the association between maternal smoking during pregnancy and offspring ADHD is not due to causal intrauterine effects, but reflects unmeasured confounding.


The use of indocyanine green and near-infrared imaging in laparoscopic completion cholecystectomy for the management of stump cholecystitis: A case series.
Stump cholecystitis is managed by performing a completion cholecystectomy, which can be done either laparoscopically or by an open method. The use of indocyanine green (ICG) is known to improve the identification of the biliary tree anatomy, facilitating Calot's triangle dissection and shortening surgery, thereby reducing the risk of bile duct injuries and making laparoscopic cholecystectomy safer. A retrospective analysis was performed of prospectively collected data from 15 patients at our institution from March 2016 to March 2021. Magnetic resonance cholangiopancreatography was performed in all 15 cases, showing remnant gall bladder in all cases with calculi within. Four cases had a dilated common bile duct (CBD) with CBD calculi. Endoscopic retrograde cholangiopancreatography (ERCP) and stone removal followed by CBD stenting were performed in the four patients with CBD calculi. These four cases were scheduled for surgery 4 weeks post-ERCP. All 15 patients underwent laparoscopic completion cholecystectomy. The mean operating time was 80 min. The post-operative period of all cases was uneventful, and the patients were discharged on post-operative day 2 or day 3. All patients remained asymptomatic during 1-5 years of follow-up. Laparoscopic completion cholecystectomy was performed safely in cases of stump cholecystitis and resulted in symptom relief during short-term follow-up. The use of ICG and near-infrared imaging in such cases helps identify the biliary anatomy, may contribute to the safety of laparoscopic completion cholecystectomy and might reduce the duration of surgery.


Cell-mediated immunity to Plasmodium falciparum infection: evidence against the involvement of cytotoxic lymphocytes.
Blood mononuclear cells (PBMC) recognizing soluble malaria antigens (SPag) are present in the peripheral blood of individuals clinically immune to malaria, and they proliferate after exposure to such antigens. To test whether these cells have effector activity against Plasmodium falciparum, we stimulated PBMC from malaria-immune donors by SPag and purified protein derivative (PPD) in culture for 7 days. The PBMC were then co-incubated with P. falciparum for 48 h, and parasitaemia was determined by microscopy. Parasite growth was only significantly impaired after incubation with PBMC stimulated by either SPag or PPD in the presence of immune serum. Studies on subpopulations of PBMC indicated that the inhibitory cells resided among the adherent cell fraction. Furthermore we tested PBMC for cytotoxic activity against P. falciparum-infected autologous or heterologous erythrocytes. Experiments were done both in the absence and the presence of immune serum. Neither fresh PBMC nor PBMC activated by SPag or PPD for 7 days prior to assay were cytotoxic, indicating that cytotoxic T cells, natural killer (NK) cells, and K cells did not possess cytotoxic activity directed against parasitized erythrocytes. These data support the hypothesis that activated monocytes are the most important effector cells in the peripheral blood of malaria immune individuals.


Regulation of PPAR-alpha pathway by Dicer revealed through proteomic analysis.
Dicer is a crucial RNase III enzyme in miRNA biogenesis pathway. Although numerous studies have been carried out to investigate the role of miRNAs and Dicer in the regulation of biological processes, few studies have examined proteomic alterations upon knockout of Dicer. We employed a Cre-loxP-based inducible knockout mouse system to investigate the proteome regulated by Dicer-dependent miRNAs. We utilized spiked liver lysates from metabolically labeled mice to quantify the subtle changes in the liver proteome upon deletion of Dicer. We identified 2137 proteins using high resolution tandem mass spectrometry analysis. The upregulated proteins included several enzymes involved in peroxisomal β-oxidation of fatty acids and a large majority of the upregulated proteins involved in lipid metabolism were known PPARα targets. MRM-based assays were carried out to confirm the upregulation of enzymes including peroxisomal bifunctional enzyme, phosphoenolpyruvate carboxykinase 1, cytochrome P450 3A13, cytochrome P450 3A41 and myristoylated alanine-rich protein kinase C substrate. Further, miRNA-124 which is predicted to regulate expression of peroxisomal bifunctional enzyme was confirmed to be downregulated in the Dicer knockout mice. Our study demonstrates the strength of coupling knockout mouse models and quantitative proteomic strategies to investigate functions of individual proteins in vivo. Dicer dependent miRNA biogenesis is the major pathway for generation of mature miRNAs. We developed SILAC mouse-based proteomics screen to identify protein targets of Dicer-dependent miRNAs in liver of Dicer knockout mice. We spiked liver lysates of induced and uninduced Dicer knockout mice with liver lysate of SILAC labeled mice for identification of dysregulated proteome. We quantitated 1217 proteins of which 257 were upregulated in induced Dicer knockout mice. We observed enrichment of PPAR-α targets and proteins involved in lipid metabolism among upregulated proteins. We further carried out MRM-based validation of peroxisomal bifunctional enzyme, phosphoenolpyruvate carboxykinase 1, Cyp3A13, Cyp3A41 and myristoylated alanine-rich protein kinase C substrate. We further validated upregulation of peroxisomal bifunctional enzyme using Western blot analysis and downregulation of its predicted upstream miRNA, miR-124 using qRT-PCR. Our study demonstrates that upon ablation of Dicer, certain Dicer-dependent miRNAs are dysregulated which result in dysregulation of their target proteins such as proteins associated with lipid metabolism. Our study illustrates the use of SILAC strategy for quantitative proteomic investigations of animal model systems.


Acute clinical care and care coordination for traumatic brain injury within Department of Defense.
The nature of current combat situations that U.S. military forces encounter and the use of unconventional weaponry have dramatically increased service personnel's risks of sustaining a traumatic brain injury (TBI). Although the true incidence and prevalence of combat-related TBI are unknown, service personnel returning from deployment have reported rates of concussion between 10% and 20%. The Department of Defense has recently released statistics on TBI dating back to before the wars in Iraq and Afghanistan to better elucidate the impact and burden of TBI on America's warriors and veterans. Patients with severe TBI move through a well-established trauma system of care, beginning with triage of initial injury by first-responders in the war zone to acute care to rehabilitation and then returning home and to the community. Mild and moderate TBIs may pose different clinical challenges, especially when initially undetected or if treatment is delayed because more serious injuries are present. To ensure identification and prompt treatment of mild and moderate TBI, the U.S. Congress has mandated that military and Department of Veterans Affairs hospitals screen all service personnel returning from combat. Military health professionals must evaluate them for concussion and then treat the physical, emotional, and cognitive problems that may surface. A new approach to health management and care coordination is needed that will allow medical transitions between networks of care to become more centralized and allow for optimal recovery at all severity levels. This article summarizes the care systems available for the acute management of TBI from point of injury to stateside military treatment facilities. We describe TBI assessment, treatment, and overall coordination of care, including innovative clinical initiatives now used.


Facilitating Perinatal Access to Resources and Support (PeARS): a feasibility study with external pilot of a novel intervention.
Up to 50% of women in areas of high socio-economic deprivation are at risk of developing depressive symptoms in pregnancy. Feeling well supported, can facilitate good mental health perinatally. A brief, innovative intervention to facilitate access to support and resources was developed and tested. This included one antenatal and one postnatal session, each with three evidence-based components: i) support from a non-professional peer to enable a woman to identify her needs; ii) information about local community services and signposting; and iii) development of a personalised If-Then plan to access that support. The aims were to evaluate the intervention and research methods for feasibility and acceptability for perinatal women, maternity care providers and peers, and provide preliminary effectiveness indications. Pregnant women living in an area of high deprivation were recruited from community-based antenatal clinics and randomised to intervention or control condition (a booklet about local resources). Outcome measures included women's use of community services by 34 + weeks gestation and 6 months postnatally; mental health and wellbeing measures, and plan implementation. Interviews and focus groups were conducted with women participants, providers, and peers. Data were analysed using framework analysis. Recruitment and retention of peers and participants, intervention fidelity, and acceptability of outcome measures were recorded. Peer facilitators could be recruited, trained, retained and provide the intervention with fidelity. One hundred twenty six women were recruited and randomised, 85% lived in the 1% most deprived UK areas. Recruitment constituted 39% of those eligible, improving to 54% after midwifery liaison. Sixty five percent were retained at 6 months postnatally. Women welcomed the intervention, and found it helpful to plan access to community services. Providers strongly supported the intervention philosophy and integrated this easily into services. The study was not powered to detect significant group differences but there were positive trends in community service use, particularly postnatally. No differences were evident in mental health and wellbeing. This intervention was well received and easily integrated into existing services. Women living in highly deprived areas could be recruited, randomised and retained. Measures were acceptable. Peer facilitators were successfully trained and retained. Full effectiveness studies are warranted.


Quantification of circulating Mycobacterium tuberculosis antigen peptides allows rapid diagnosis of active disease and treatment monitoring.
Tuberculosis (TB) is a major global health threat, resulting in an urgent unmet need for a rapid, non-sputum-based quantitative test to detect active Mycobacterium tuberculosis (Mtb) infections in clinically diverse populations and quickly assess Mtb treatment responses for emerging drug-resistant strains. We have identified Mtb-specific peptide fragments and developed a method to rapidly quantify their serum concentrations, using antibody-labeled and energy-focusing porous discoidal silicon nanoparticles (nanodisks) and high-throughput mass spectrometry (MS) to enhance sensitivity and specificity. NanoDisk-MS diagnosed active Mtb cases with high sensitivity and specificity in a case-control study with cohorts reflecting the complexity of clinical practice. Similar robust sensitivities were obtained for cases of culture-positive pulmonary TB (PTB; 91.3%) and extrapulmonary TB (EPTB; 92.3%), and the sensitivities obtained for culture-negative PTB (82.4%) and EPTB (75.0%) in HIV-positive patients significantly outperformed those reported for other available assays. NanoDisk-MS also exhibited high specificity (87.1-100%) in both healthy and high-risk groups. Absolute quantification of serum Mtb antigen concentration was informative in assessing responses to antimycobacterial treatment. Thus, a NanoDisk-MS assay approach could significantly improve the diagnosis and management of active TB cases, and perhaps other infectious diseases as well.


From Mediterranean diet to Mediterranean lifestyle: a narrative review.
To summarize the recent scientific evidence regarding the wellness-promoting capacity of the Mediterranean lifestyle (ML), with a special focus on physical, social and environmental wellness. Narrative review of English-language publications in PubMed, Scopus and Embase, from 1 January 2010 to 31 October 2018. Prospective cohort studies, interventional studies, meta-analyses and reviews of those investigating the effect of at least one component of the ML on wellness parameters. General population. Although an explicit definition of ML is missing, compliance with various combinations of its components improves metabolic health and protects against or ameliorates disease state. However, there is heterogeneity in the healthy behaviours that the ML-focused studies include in their design and the way these are assessed. Also, despite that features of the ML could contribute to other wellness dimensions, there are no studies exploring the effect this healthy lifestyle could confer to them. Chronic lifestyle diseases are of multifactorial aetiology and they warrant multifaceted approaches targeting the general way of living. ML, if thoroughly evaluated, can provide a valuable tool to holistically promote health and wellness.


Determination of N-nitrosodimethylamine levels in some Canadian 2,4-D amine formulations.
Levels of N-nitrosodimethylamine (NDMA) were determined in 112 samples of 2,4-dichlorophenoxyacetic acid, (2,4-D), formulated as the dimethylamine salt, collected over a 2 year period from products on the Canadian market. A sample aliquot is partitioned with dichloromethane, and the co-extracted dimethylamine is removed by cleanup on a silica gel column. The eluates containing NDMA are concentrated, an internal standard of N-nitrosodipropylamine is added, and nitrosamine levels are determined using a gas chromatograph interfaced with a thermal energy analyzer. Recoveries of NDMA and N-nitrosodiethylamine spiked into samples were 103 +/- 16 and 96.3 +/- 9.8%, respectively. Of the 112 samples analyzed, 92 were below 1 part per million (ppm) relative to the amount of 2,4-D in the samples, 16 were between 1 and 5 ppm, and 4 were greater than 5 ppm. The gas chromatographic column used is compared to a conventional packing material for volatile nitrosamine analysis. Formation of NDMA during cleanup and analysis was shown not to occur.


Validation of a Postoperative Nomogram Predicting Recurrence in Patients with Conventional Clear Cell Renal Cell Carcinoma.
Clear cell renal cell carcinoma (RCC) continues to be the most commonly diagnosed subtype and is associated with more aggressive behavior than papillary and chromophobe RCC. Predicting disease recurrence after surgical extirpation is important for counseling and targeting those at high risk for adjuvant therapy clinical trials. To validate a postoperative nomogram predicting 5-yr recurrence-free probability (RFP) for clinically localized clear cell RCC. We identified all patients who underwent nephrectomy for clinically localized clear cell RCC from 1990 to 2009 at Memorial Sloan Kettering Cancer Center. After excluding patients with bilateral renal masses, familial RCC syndromes, and T3c or T4 tumors due to the limited number, 1642 participants were available for analysis. Partial or radical nephrectomy. Disease recurrence was defined as any new tumor after nephrectomy or kidney cancer-specific mortality, whichever occurred first. A postoperative nomogram was used to calculate the predicted 5-yr RFP, and these values were compared with the actual 5-yr RFP. Nomogram performance was evaluated by concordance index and calibration plot. Median follow-up was 39 mo (interquartile range: 14-79 mo), and disease recurrence was observed in 50 patients. The nomogram concordance index was 0.81. The calibration curve showed that the nomogram underestimated the actual 5-yr RFP. We updated the nomogram by including the entire patient population, which maintained performance and significantly improved calibration. The updated clear cell RCC postoperative nomogram performed well in the combined cohort. Underestimation of actual 5-yr RFP by the original nomogram may be due to increased surgeon experience and other unknown variables. We updated a valuable prediction tool used for assessing the disease recurrence probability after nephrectomy for clear cell renal cell carcinoma.


Using functional magnetic resonance imaging to determine if cerebral hemodynamic responses to pain change following thoracic spine thrust manipulation in healthy individuals.
Case series. To use blood oxygenation level-dependent functional magnetic resonance imaging (fMRI) to determine if supraspinal activation in response to noxious mechanical stimuli varies pre- and post-thrust manipulation to the thoracic spine. Recent studies have demonstrated the effectiveness of thoracic thrust manipulation in reducing pain and improving function in some individuals with neck and shoulder pain. However, the mechanisms by which manipulation exerts such effects remain largely unexplained. The use of fMRI in the animal model has revealed a decrease in cortical activity in response to noxious stimuli following manual joint mobilization. Supraspinal mediation contributing to hypoalgesia in humans may be triggered following spinal manipulation. Ten healthy volunteers (5 women, 5 men) between the ages of 23 and 48 years (mean, 31.2 years) were recruited. Subjects underwent fMRI scanning while receiving noxious stimuli applied to the cuticle of the index finger at a rate of 1 Hz for periods of 15 seconds, alternating with periods of 15 seconds without stimuli, for a total duration of 5 minutes. Subjects then received a supine thrust manipulation directed to the midthoracic spine and were immediately returned to the scanner for reimaging with a second delivery of noxious stimuli. An 11-point numeric pain rating scale was administered immediately after the application of noxious stimuli, premanipulation and postmanipulation. Blood oxygenation level-dependent fMRI recorded the cerebral hemodynamic response to the painful stimuli premanipulation and postmanipulation. The data indicated a significant reduction in subjects' perception of pain (P<.01), as well as a reduction in cerebral blood flow as measured by the blood oxygenation level-dependent response following manipulation to areas associated with the pain matrix (P<.05). There was a significant relationship between reduced activation in the insular cortex and decreased subjective pain ratings on the numeric pain rating scale (r = 0.59, P<.05). This study provides preliminary evidence that suggests that supraspinal mechanisms may be associated with thoracic thrust manipulation and hypoalgesia. However, because the study lacked a control group, the results do not allow for the discernment of the causative effects of manipulation, which may also be related to changes in levels of subjects' fear, anxiety, or expectation of successful outcomes with manipulation. Future investigations should strive to elicit more conclusive findings in the form of randomized clinical trials.


BRCC36A is epistatic to BRCA1 in DNA crosslink repair and homologous recombination in Arabidopsis thaliana.
BRCA1 is a well-known tumor suppressor protein in mammals, involved in multiple cellular processes such as DNA repair, chromosome segregation and chromatin remodeling. Interestingly, homologs of BRCA1 and several of its complex partners are also found in plants. As the respective mutants are viable, in contrast to mammalian mutants, detailed analyses of their biological role is possible. Here we demonstrate that the model plant Arabidopsis thaliana harbors two homologs of the mammalian BRCA1 interaction partner BRCC36, AtBRCC36A and AtBRCC36B. Mutants of both genes as well as the double mutants are fully fertile and show no defects in development. We were able to show that mutation of one of the homologs, AtBRCC36A, leads to a severe defect in intra- and interchromosomal homologous recombination (HR). A HR defect is also apparent in Atbrca1 mutants. As the Atbrcc36a/Atbrca1 double mutant behaves like the single mutants of AtBRCA1 and AtBRCC36A both proteins seem to be involved in a common pathway in the regulation of HR. AtBRCC36 is also epistatic to AtBRCA1 in DNA crosslink repair. Upon genotoxic stress, AtBRCC36A is transferred into the nucleus.


Discrimination between 8-oxo-2'-deoxyguanosine and 2'-deoxyguanosine in DNA by the single nucleotide primer extension reaction with adap triphosphate.
The adenosine derivative of 2-oxo-1,3-diazaphenoxazine (Adap) exhibits a superb ability to recognize and form base pairs with 8-oxo-2'-deoxyguanosine (8-oxo-dG) in duplex DNA. In this study, the triphosphate of Adap (dAdapTP) was synthesized and tested for single nucleotide incorporation into primer strands using the Klenow Fragment. The efficiency of dAdapTP incorporation into 8-oxo-dG-containing templates was more than 36-fold higher than with dG-containing templates, and provides better discrimination than does the incorporation of natural 2'-deoxyadenosine triphosphate (dATP). The selective incorporation of dAdapTP into 8-oxo-dG templates was therefore applied to the detection of 8-oxo-dG in human telomeric DNA sequences extracted from H2 O2 -treated HeLa cells. The enzymatic incorporation of dAdapTP into 8-oxo-dG-containing templates may provide a novel basis for sequencing oxidative DNA damage in the genome.


Two p90 ribosomal S6 kinase isoforms are involved in the regulation of mitotic and meiotic arrest in Artemia.
There are multiple isoforms of p90 ribosomal S6 kinase (RSK), which regulate diverse cellular functions such as cell growth, proliferation, maturation, and motility. However, the relationship between the structures and functions of RSK isoforms remains undetermined. Artemia is a useful model in which to study cell cycle arrest because these animals undergo prolonged diapauses, a state of obligate dormancy. A novel RSK isoform was identified in Artemia, which was termed Ar-Rsk2. This isoform was compared with an RSK isoform that we previously identified in Artemia, termed Ar-Rsk1. Ar-Rsk2 has an ERK-docking motif, whereas Ar-Rsk1 does not. Western blot analysis revealed that Ar-Rsk1 was activated by phosphorylation, which blocked meiosis in oocytes. Knockdown of Ar-Rsk1 reduced the level of phosphorylated cdc2 and thereby suppressed cytostatic factor activity. This indicates that Ar-Rsk1 regulates the cytostatic factor in meiosis. Expression of Ar-Rsk2 was down-regulated in Artemia cysts in which mitosis was arrested. Knockdown of Ar-Rsk2 resulted in decreased levels of cyclin D3 and phosphorylated histone H3, and the production of pseudo-diapause cysts. This indicates that Ar-Rsk2 regulates mitotic arrest. PLK and ERK RNAi showed that Ar-Rsk2, but not Ar-Rsk1, could be activated by PLK-ERK in Artemia. This is the first study to report that RSK isoforms with and without an ERK-docking motif regulate mitosis and meiosis, respectively. This study provides insight into the relationship between the structures and functions of RSK isoforms.


Evaluation of ricin A chain-containing immunotoxins directed against different epitopes on the delta-chain of cell surface-associated IgD on murine B cells.
Over the past decade, immunotoxins (IT) composed of mAb covalently coupled to toxins or their subunits have been developed for the treatment of malignancies and autoimmune diseases. Despite specific binding to target cells, not every mAb makes a therapeutically potent ricin A chain-containing IT (IT-A). A number of variables influence the potency of a mAb as an IT-A, including the affinity of the mAb, the nature and density of the cell surface Ag, and the type of target cell used. The present report investigates the influence of the epitope specificity of a mAb on the effectiveness of that mAb as an IT-A. Seven mAb directed against different regions of the mouse delta H chain of surface IgD, were conjugated to deglycosylated ricin A chain, and tested for their ability to kill murine B cells. The panel of IT-A had similar A chain activities and similar binding avidities. However, the mAb directed against epitopes in the Fc portion of surface IgD made more effective IT-A than those directed against epitopes in the Fd region. Overall, the anti-Fc-A were approximately 60- to 150-fold more toxic than the anti-Fd-A. Taken together with previous studies, these findings suggest that the epitope on a target Ag recognized by a given mAb is an important variable in determining the potency of a mAb as an IT-A.


Induction of leukemia cell apoptosis by cheliensisin A involves down-regulation of Bcl-2 expression.
To investigate the apoptosis-inducing effect of cheliensisin A (GC-51), a novel styryl-lactone isolated from Goniothalamus cheliensis, on human promyelocytic leukemia HL-60 cells and the mechanism of action involved. Apoptotic cell death was determined by morphological examination and DNA agarose gel electrophoresis. The activity of caspase-3 was assessed using Western blotting and the expression of Bcl-2 and Bax genes was analyzed using the reverse transcription-polymerase chain reaction (RT-PCR) method. GC-51 significantly inhibited the proliferation of HL-60 cells with an IC50 of 2.4+/-0.2 micromol/L and effectively induced apoptosis in HL-60 cells. Exposure of HL-60 cells to 10 micromol/L GC-51 for 8 h resulted in approximately 53% of the cells undergoing apoptosis. Caspase-3 was activated in GC-51-treated cells, which was manifested by the appearance of the 17 kDa active form of caspase-3 and the cleavage of poly(ADP-ribose) polymerase (PARP). Meanwhile, GC-51 markedly reduced the expression of the anti-apoptotic gene Bcl-2 and increased the expression of the pro-apoptotic gene Bax. The apoptosis-inducing effect of GC-51 was cAMP-dependent protein kinase (PKA) dependent because PKA, but not the protein kinase C, specific inhibitor H-89, blocked the induction of apoptosis by GC-51 in HL-60 cells. The results demonstrate that GC-51 effectively induces apoptosis in HL-60 cells and that this effect is PKA-dependent and involves the downregulation of Bcl-2 expression and the activation of caspase-3.


Stimulation of acetylcholine release from myenteric neurons of guinea pig small intestine by forskolin and cyclic AMP.
Forskolin, an activator of adenylate cyclase, was used to examine the regulation of [3H]acetylcholine (ACh) release by cyclic AMP (cAMP)-related mechanisms in myenteric plexus-longitudinal muscle preparations of guinea pig small intestine. Forskolin evoked a dose-related increase in [3H]ACh release. Both dibutyryl-cAMP and 8-Br-cAMP significantly elevated [3H]ACh secretion. In the presence of phosphodiesterase inhibitors (theophylline and 3-isobutyl-1-methylxanthine), the basal [3H]ACh output was increased. There was a significantly greater stimulation when forskolin was used to incite endogenous cAMP synthesis and phosphodiesterase inhibitors were simultaneously applied to prevent cAMP breakdown. The enhancement of forskolin-stimulated release by theophylline or 3-isobutyl-1-methylxanthine strongly implicates a synergistic interaction between the two. These findings suggest that forskolin acts to increase ACh release by a modulation of endogenous cAMP and further support a cAMP-mediated mechanism in the secretion of ACh from myenteric cholinergic neurons.


Serological and molecular investigation of Ehrlichia spp. and Anaplasma spp. in ticks and blood of dogs, in the Thrace Region of Turkey.
In recent years, tick-borne diseases like ehrlichiosis and anaplasmosis became widespread worldwide threatening the health of both human and companion animals. Therefore, the aim of this study was to determine the presence of Anaplasma spp., and Ehrlichia spp. in dogs and ticks in the Thrace Region of Turkey. A total of 400 blood samples and 912 ticks were collected from dogs living in shelters that are located in four cities (Istanbul, Edirne, Tekirdag and Kirklareli) of the Thrace Region. Blood and buffy coat smears were prepared for microscopic examination. Hematologic and serologic analyses were performed using cell counter and commercial Snap3Dx test kit, respectively. Eight hundred fifty of collected ticks were classified as Rhipicephalus sanguineus, 33 as Rhipicephalus turanicus and 29 as Ixodes ricinus. After DNA extraction from blood samples and pooled ticks (127 tick pools, in total), nested PCR was performed to detect the DNA of Anaplasma spp., and Ehrlichia spp. The seroprevalence of Ehrlichia canis was 27.25% (109) by Snap3Dx test and the total molecular positivity was 11.75% (47) in dog blood samples and 21.25% (27) in tick pools by nested PCR. The frequencies of the infected blood samples with E. canis, Anaplasma phagocytophilum and Anaplasma platys were detected as 6%, 4% and 6%, respectively. E. canis and A. platys were detected in R. sanguineus pools with a ratio of 15.75% and 0.7%, respectively. In addition, A. platys was also detected in R. turanicus pools (0.7%). A. phagocytophilum was found only in I. ricinus pools (3.93%). Morulae of three species were detected in buffy coat and blood smears. While anemia was observed in dogs infected with E. canis and co-infected (with one or more species), thrombocytopenia was observed only in co-infected dogs. This is the first study providing evidence for the presence of Anaplasma spp. and Ehrlichia spp. in dogs and ticks in the Thrace Region of Turkey. Based on the results of the tests used in this study, we recommend the combined use of serologic, molecular, cytologic, hematologic analyses and physical examination of tick exposure for an accurate diagnosis of ehrlichiosis and anaplasmosis.


Lactobacillus paracasei endocarditis of bioprosthetic aortic valve presenting with recurrent embolic strokes.
Lactobacillus prosthetic valve endocarditis is a rare infection caused by Lactobacillus bacteria. This bacterium is found in the normal flora of the human mouth, gastrointestinal tract and female genital tract. While there have been isolated cases of Lactobacillus bacteraemia and endocarditis, the infections are associated with comorbidities, immune deficiency, dental manipulation procedures and other medical history. This case of bioprosthetic valve endocarditis caused by Lactobacillus paracasei is unusual, as the patient was immune-competent and treated with pre-procedural antibiotics. We present a case of a 65-year-old male who underwent a dental extraction. He presented after 3 months of fever, chills and fatigue. On initial presentation, blood cultures were positive for alpha-haemolytic streptococcus bacteraemia. He was treated with IV penicillin and underwent aortic valve replacement with a bioprosthetic valve and excision of the mitral vegetation with repair of the mitral valve. Two years later, he had a tooth extraction after being treated properly with antibiotics. Three months later he presented with difficulty speaking, left leg weakness and increased drooling. All testing was normal. Three months later he presented with left side lower extremity weakness and expressive aphasia. He was diagnosed with bioprosthetic aortic valve endocarditis and was treated with IV penicillin and gentamicin for 6 weeks and then switched to oral penicillin. He remained stable. L. paracasei can potentially be a cause of complicated endocarditis in patients with prosthetic heart valves undergoing dental procedures. Timely culture-guided antibiotic therapy is critical and may obviate the need for valve surgery.


Reliability and responsiveness of the Danish version of The Chelsea Critical Care Physical Assessment tool (CPAx).
Measurement instruments are important in clinical practice and research for assessing physical function in critically ill patients in the intensive care unit (ICU). To investigate inter-rater reliability and responsiveness of the Danish version of the CPAx (CPAx-D). Critically ill patients from three Danish ICUs were included. Patients were assessed with CPAx-D by two blinded testers during a regular physiotherapy session. Follow-up tests were performed in patients who stayed in the ICU for more than 24 hours, were not transferred to another hospital or received palliative care. Floor and ceiling effects were examined in all assessments.Results For the reliability analysis 66 patients were included. Results Showed no significant difference between raters. For the total score, intra class correlation coefficient (ICC) was 0.996 (95% CI: 0.993; 0.997), standard error of measurement was 0.72 point and minimal detectable change 2.0 points. Bland-Altman plot revealed no heteroscedacity. The responsiveness results of 24 patients showed that the effect size was 1.2 and the standardized response mean 1.1, which was in accordance with the hypothesis. No ceiling or floor effect was revealed. The CPAx-D showed excellent inter-rater reliability and responsiveness.


The optimal use of steroids in paediatric acute lymphoblastic leukaemia: no easy answers.
Glucocorticoids are an integral component of therapy for acute lymphoblastic leukaemia (ALL), but usage differs between cooperative group protocols. All groups use glucocorticoids during induction but vary on whether to use dexamethasone or prednisone. Issues to consider in the choice of induction steroid include impact on event-free and overall survival, acute morbidity such as infection risk, diabetes, and behavioural disturbances and long-term complications such as avascular necrosis. It is generally agreed that dexamethasone is the steroid of choice for groups using a delayed intensification phase, but dosing schedules (intermittent versus continuous) vary. There is no consensus on the potential benefit of steroid administration during maintenance therapy. This review will summarize the current available data on steroid use in paediatric ALL, highlighting outcomes as well as major toxicities.


Adolescent Reproductive Health Care: Views and Practices of Pediatric Hospitalists.
Many hospitalized adolescents are at increased risk for pregnancy complications due to an underlying medical condition, however sexual risk assessment is not consistently performed in this setting. While adolescents and their parents are supportive of sexual health discussion in the inpatient setting, a thorough understanding of factors that influence provision of this care among pediatric hospital physicians is lacking. This formative information is needed to facilitate efforts to improve and standardize clinical care provision. Our objective is to assess the frequency and factors that influence the provision of adolescent sexual and reproductive care by pediatric hospitalists. We performed a cross-sectional computerized survey of hospitalists at 5 pediatric hospitals who cared for ≥1 adolescent (14-21 years old) in the past year. Sexual and reproductive care practices were assessed by using a 76-item novel survey informed by the theory of planned behavior. We used descriptive statistics to summarize the data. Sixty-eight pediatric hospitalists participated (49% response rate): 78% were women and 65% were aged <40 years. Most (69%) reported treating >46 adolescents annually, including many who are at an increased risk for pregnancy complications due to teratogenic medication use or a comorbid condition. A majority felt that sexual and reproductive services are appropriate, although many endorsed barriers, including concern about follow-up after emergency contraception (63%) and time constraints (53%). Most reported insufficient knowledge regarding contraception (59%), desired contraception education (57%), and were likely to increase contraceptive provision if provided education (63%). Hospitalists rarely provided condoms or referral for an intrauterine device. Pediatric hospitalists frequently care for adolescents who are at risk for pregnancy complications and generally agree that reproductive care is appropriate in the inpatient setting. With these findings, we highlight the critical need for effective comprehensive reproductive health service interventions that are tailored to address the numerous actionable barriers identified in this study.


Studies in involuntary civil commitment and involuntary electroconvulsive therapy.
On January 2, 1979, the state of Missouri enacted new mental health laws. In this tandem study the impact of the change in law at an acute urban mental health center in St. Louis, Missouri, was examined in two specific areas: involuntary civil commitment and involuntary electroconvulsive therapy (ECT). The first part of this examination was conducted by comparing all patients (N = 86) who were referred to court to determine their need for involuntary civil commitment from July 1, 1980, to June 30, 1982, to a nonadjudicated control group (N = 87). The second part of this examination was conducted by comparing all patients (N = 19) who were referred to court to determine their need for involuntary ECT from July 1, 1979, to June 30, 1983, to a voluntary ECT recipient control group (N = 24). A large series of statistical comparisons were made. The major finding to emerge from these data was that clinicians were selectively referring to court a group of patients with histories of recurrently becoming both dangerous and noncompliant with hospitalization. In addition, it appears that voluntary ECT is used conservatively and involuntary ECT is used very conservatively.


Predictors and long-term prognostic significance of angiographically visible distal embolization during primary percutaneous coronary intervention.
We aimed to identify the predictors of angiographically visible distal embolization (AVDE) during primary percutaneous coronary intervention (p-PCI) as well as to assess its impact on short- and long-term clinical outcomes in patients with acute ST-segment elevation myocardial infarction (STEMI). We retrospectively enrolled 2007 patients with STEMI who underwent p-PCI. We assessed the clinical and angiographic characteristics of patients in order to identify the predictors of AVDE and compared the outcomes of patients with and without AVDE during p-PCI. Distal embolization developed in 135 (6.7%) patients. Age (for each 10- year increase, Odds Ratio (OR) 1.34, 95% Confidence Interval (CI) 1.16-1.52, p<0.001), treatment of right coronary artery (OR 2.52, 95% CI 1.30-4.87, p=0.034), repeated balloon dilatation (OR 1.84, 95% CI 1.16-2.94, p=0.009), cut-off occlusion pattern (OR 2.17, 95% CI 1.38-3.42, p=0.001), lesion length >15 mm (OR 1.67, 95% CI 1.09-2.58, p=0.019), and reference vessel diameter >3.5 mm (OR 5.08, 95% CI 3.32-7.65, p<0.001) were independent predictors of AVDE. In-hospital (8.1% vs. 3.8%, p=0.014) and one-month (10.8% vs. 4.9%, p=0.004) all-cause mortality rates were higher in patients with AVDE. At the long-term follow-up (median: 42 months), both all-cause (21.5% vs. 10.4%, p<0.001) and cardiac mortality rates (18.4% vs. 8.0%, p<0.001) were higher in patients with AVDE. AVDE is associated with worse clinical outcome at both the short- and long-term follow-up of STEMI patients treated early with p-PCI.


Autonomic dysreflexia-induced reversible posterior leukoencephalopathy syndrome in patients with spinal cord injury: two case reports.
To report two cases of reversible posterior leukoencephalopathy syndrome (RPLS) in patients with traumatic cervical spinal cord injury. Case report. Rehabilitation Inpatient Unit, Pusan National University Hospital, Yangsan-si, Korea. Two men with spastic tetraplegia developed autonomic dysreflexia. Use of antihypertensive medication and Foley catheter insertion prevented further episodes of acute arterial hypertension and development of new symptoms. RPLS can occur in the setting of autonomic dysreflexia in patients with traumatic cervical cord injury. The prompt recognition of this syndrome is of importance to prevent further morbidity and mortality in patients with spinal cord injury.


New insights into the molecular mechanism of intestinal fatty acid absorption.
Dietary fat is one of the most important energy sources of all the nutrients. Fatty acids, stored as triacylglycerols (also called triglycerides) in the body, are an important reservoir of stored energy and derived primarily from animal fats and vegetable oils. Although the molecular mechanisms for the transport of water-insoluble amphipathic fatty acids across cell membranes have been debated for many years, it is now believed that the dominant means for intestinal fatty acid uptake is via membrane-associated fatty acid-binding proteins, that is, fatty acid transporters on the apical membrane of enterocytes. These findings indicate that intestinal fatty acid absorption is a multistep process that is regulated by multiple genes at the enterocyte level, and intestinal fatty acid absorption efficiency could be determined by factors influencing intraluminal fatty acid molecules across the brush border membrane of enterocytes. To facilitate research on intestinal, hepatic and plasma triacylglycerol metabolism, it is imperative to establish standard protocols for precisely and accurately measuring the efficiency of intestinal fatty acid absorption in humans and animal models. In this review, we will discuss the chemical structure and nomenclature of fatty acids and summarize recent progress in investigating the molecular mechanisms underlying the intestinal absorption of fatty acids, with a particular emphasis on the physical chemistry of intestinal lipids and the molecular physiology of intestinal fatty acid transporters. A better understanding of the molecular mechanism of intestinal fatty acid absorption should lead to novel approaches to the treatment and the prevention of fatty acid-related metabolic diseases that are prevalent worldwide.


D-mannose induces regulatory T cells and suppresses immunopathology.
D-mannose, a C-2 epimer of glucose, exists naturally in many plants and fruits, and is found in human blood at concentrations less than one-fiftieth of that of glucose. However, although the roles of glucose in T cell metabolism, diabetes and obesity are well characterized, the function of D-mannose in T cell immune responses remains unknown. Here we show that supraphysiological levels of D-mannose safely achievable by drinking-water supplementation suppressed immunopathology in mouse models of autoimmune diabetes and airway inflammation, and increased the proportion of Foxp3+ regulatory T cells (Treg cells) in mice. In vitro, D-mannose stimulated Treg cell differentiation in human and mouse cells by promoting TGF-β activation, which in turn was mediated by upregulation of integrin αvβ8 and reactive oxygen species generated by increased fatty acid oxidation. This previously unrecognized immunoregulatory function of D-mannose may have clinical applications for immunopathology.


Complete sequence analysis of the genome of the bacterium Mycoplasma pneumoniae.
The entire genome of the bacterium Mycoplasma pneumoniae M129 has been sequenced. It has a size of 816,394 base pairs with an average G+C content of 40.0 mol%. We predict 677 open reading frames (ORFs) and 39 genes coding for various RNA species. Of the predicted ORFs, 75.9% showed significant similarity to genes/proteins of other organisms while only 9.9% did not reveal any significant similarity to gene sequences in databases. This permitted us tentatively to assign a functional classification to a large number of ORFs and to deduce the biochemical and physiological properties of this bacterium. The reduction of the genome size of M. pneumoniae during its reductive evolution from ancestral bacteria can be explained by the loss of complete anabolic (e.g. no amino acid synthesis) and metabolic pathways. Therefore, M. pneumoniae depends in nature on an obligate parasitic lifestyle which requires the provision of exogenous essential metabolites. All the major classes of cellular processes and metabolic pathways are briefly described. For a number of activities/functions present in M. pneumoniae according to experimental evidence, the corresponding genes could not be identified by similarity search. For instance we failed to identify genes/proteins involved in motility, chemotaxis and management of oxidative stress.


Disability and chronic pain after open mesh and laparoscopic inguinal hernia repair.
Proponents of laparoscopic inguinal hernia repair maintain that the associated costs and risks are offset by faster recovery and less postoperative pain. It was our hypothesis that the incidence of chronic pain in both groups of our patients was not as high as reported in the literature. Patients for the study were identified from a community hospital medical record database. A total of 229 patients were available and agreed to participate in the study. Data collected included the patient's current pain level at the hernia site, pain medication currently used, narcotics currently used, return to normal work, and return to normal activity. Overall, 19.7 per cent of patients complained of mild pain, but only 2.2 per cent classified this as moderate or severe. Mild pain was noted more often in the open repair patients compared with the laparoscopic group. However, there was no difference in the frequency of moderate or severe pain. The time to return to work was longer in the open repair group than the laparoscopic repair group, but there were large ranges in both groups. The inability to return to full preoperative activity was infrequent and equivalent in both open and laparoscopic hernia repair groups. In our study of 229 patients undergoing elective open or laparoscopic inguinal hernia repair at a community hospital, we have found a low incidence of moderate or severe chronic pain. In addition, we found that this procedure did not interfere with return to work at 6 months or return to daily activities in either the laparoscopic or open repair group.


Fate and behavior of extended-spectrum β-lactamase-producing genes in municipal sewage treatment plants.
Extended-spectrum β-lactamases (ESBLs) have the capability of hydrolyzing a variety of the newer β-lactam antibiotics, including the third-generation cephalosporins and monobactams known as a rapidly evolving group of ESBLs. The purpose of this study was to investigate the occurrence and fate of β-lactamase producing genes (CTX-M type 1, type2, CTX-M probe for all groups except CTX-M-1, and TEM, SHV, OXA) through wastewater treatment utilities. β-lactamase producing genes in influent, digested sludge, activated sludge, and disinfected effluent were monitored. The results showed that influent contained high level of all target genes, and all CTX-M types, SHV, and OXA gene decreased significantly in biological treatment process such as activated sludge process and anaerobic digestion, however, TEM type was not effectively eliminated. Possibly, host microbes of TEM could be most resistant in target genes or to some extent gene transfer occurred in wastewater treatment processes. All target genes were significantly reduced during disinfection. Consequently, wastewater treatment process apparently reduced host microbes carrying β-lactamase producing genes effectively, although they are selectively removed in biological processes. In addition, the significant reduction during disinfection was shown, although slightly differences of removal efficiency were observed in resistance.


Neonatal hydrocephalus: an atypical presentation of malignant infantile osteopetrosis.
Autosomal recessive osteopetrosis has a variable presentation, most commonly including failure to thrive, hypocalcemia, seizures, hepatosplenomegaly, hydrocephalus, vision or hearing loss, and cytopenias. Multiple symptoms are usually seen at presentation. The variability of presentation often delays diagnosis and subsequent treatment. Here, we present a case of an infant with this condition who initially presented with triventricular hydrocephalus with Chiari I malformation. This alone is not a common presentation of this disease, and we present this case to highlight autosomal recessive osteopetrosis as a potential diagnosis in infants presenting with hydrocephalus and discuss the other associated symptoms, management, and prognosis of this condition. The patient was a full-term infant with a routine newborn period. At 6 months, the infant had macrocephaly and frontal bossing with a bulging fontanelle. She was found to have hydrocephalus with moderate ventriculomegaly involving the third and lateral ventricles with an associated Chiari 1 malformation. The infant was asymptomatic at the time. The infant was promptly referred to neurosurgery and underwent an uncomplicated ventriculoperitoneal shunt placement. Post-operative X-rays showed increased density of the skull with other bone changes suggestive of autosomal recessive osteopetrosis. Subsequent lab work and imaging studies were consistent with this condition. The diagnosis was confirmed by genetic testing, and the patient has undergone treatment with hematopoietic stem cell transplant. Hydrocephalus is a common feature of this condition, typically seen in conjunction with other systemic symptoms and laboratory findings. Our patient had a limited initial presentation of triventricular hydrocephalus with Chiari I malformation and was otherwise clinically asymptomatic. There is limited literature of such a presentation, and we highlight this case to increase awareness, as timely diagnosis of these patients is critical for treatment and future outcomes.


Understanding humerus fractures in young children: Abuse or not abuse?
Fractures are the second most common abusive injury occurring in young children, particularly those under 2 years of age. The humerus is often affected. To better identify factors discriminating between abusive and non-abusive humerus fractures, this retrospective study examined the characteristics and mechanisms of injuries causing humerus fractures in children less than 18 months of age. Electronic medical records were reviewed for eligible patients evaluated between September 1, 2007 and January 1, 2012 at two children's hospitals in Chicago, IL. The main outcome measures were the type of fracture and the etiology of the fracture (abuse vs not abuse). The 97 eligible patients had 100 humerus fractures. The most common fracture location was the distal humerus (65%) and the most common fracture type was supracondylar (48%). Child Protection Teams evaluated 44 patients (45%) and determined that 24 of those had 25 fractures caused by abuse (25% of the total study population).Among children with fractures determined to have been caused by abuse, the most common location was the distal humerus (50%) and the most common types were transverse and oblique (25% each); however, transverse and oblique fractures were also seen in patients whose injuries were determined to have been non-abusive. A younger age, non-ambulatory developmental stage, and the presence of additional injuries were significantly associated with abusive fractures. Caregivers did not provide a mechanism of injury for half of children with abusive fractures, whereas caregivers provided some explanation for all children with non-abusive fractures.


Preimplantation development of giant triploid zygotes in the mouse.
Giant oocytes or two-cell embryos have been reported in various mammalian species. They may arise during multiplication of oogonia, after fusion of two oogonia or, more probably, when nuclear division is not accompanied by cytoplasmic division. The ultimate fate of these giant embryos is not well known. In our laboratory, giant two-cell mouse embryos have been occasionally observed. Recently, we observed two giant one-cell zygotes in the same species. Both showed two female pronuclei and one male pronucleus, as well as two second polar bodies localized at opposite poles of the embryo. These two giant zygotes showed normal viability and developmental capacity. Their triploid nature was confirmed by cytogenetic analysis. In order to study this interesting phenomenon in more detail, we produced giant oocytes containing two germinal vesicles by cell fusion and cultured them in vitro. About one third of them extruded two first polar bodies; in the second group only one polar body was observed, whilst the last group was without polar bodies. When parthenogenetically activated, the consistent answer analogical to that observed in "in vivo" oocytes was only observed when oocytes with two polar bodies were activated. The implication for IVF technologies is discussed.


Brain development and scholastic achievement in the Education Quality Measurement System tests in Chilean school-aged children.
Head circumference (HC), the anthropometric index of both brain development and nutritional background, has been described to be significantly associated with scholastic achievement (SA). The aim of this study was to determine the impact of nutritional background and current nutritional status parameters on SA in the Education Quality Measurement System (SIMCE) tests. A representative sample of 33 schools was randomly chosen in the Metropolitan Region of Chile. The sample consisted of 1,353 school-aged children of both sexes, from the fifth grade of elementary school and from the first grade of high school who in 2009 took the SIMCE tests. Nutritional status was assessed through anthropometric parameters. Brain development was measured through the HC expressed as HC-for-age Z-score (Z-HC). Students with Z-HC < -2 SD and >2 SD obtained low and high SA, respectively, both in the language and the mathematics tests (P < 0.001). In general, in both grades, those students with Z-HC ≥ 0 SD increase more than double the probability to obtain language and mathematics SA scores ≥ the median (P < 0.0001). We confirm the hypothesis that HC is the most relevant physical index associated with SA; therefore, children with the lowest scores in the SIMCE tests probably have lower brain development.


The physiology of leukocyte recruitment: an in vivo perspective.
The mechanisms of leukocyte recruitment have been studied extensively in vitro and have shed light on the basic molecular structure-function relationship of adhesion and signaling molecules involved in this essential immune response. This review will summarize how these in vitro observations extend to leukocyte behavior in inflamed blood vessels in the microcirculation. We highlight physiological results that might not have been predicted from in vitro systems. Special attention is placed on the physiology of rolling, adhesion, and intralumenal crawling in blood vessels. The importance of the glycocalyx, secondary tethers, shear, and the microenvironment are discussed. Docking structures forming rings of adhesion molecules together with a novel endothelial dome-like structure in vivo during transmigration are highlighted. Transcellular and paracellular emigration out of inflamed blood vessels is also discussed. The last section highlights leukocyte recruitment in some organs that do not always follow the accepted paradigm of leukocyte recruitment.


HER2 status in hormone receptor positive premenopausal primary breast cancer adds prognostic, but not tamoxifen treatment predictive, information.
Overexpression of human epidermal growth factor receptor 2 (HER2) or amplification of its gene is a prognostic factor in primary breast cancer and a predictor for tamoxifen treatment efficacy in oestrogen receptor (ER) positive disease. In the present study we explored a defined cohort of breast cancer patients included in a randomised trial in order to assess prognostic and tamoxifen treatment information yielded by HER2 status. Premenopausal breast cancer patients with stage II tumours (n = 564) were included and allocated to 2 years of adjuvant tamoxifen treatment versus no adjuvant treatment. ER, progesterone receptor (PR) status and HER2 status was determined by immunohistochemistry using a tissue microarray. HER2 amplification was analysed by fluorescent in situ hybridisation and tumours being amplified and/or HER2 3+ were considered HER2+. HER2 status was evaluable in 83% of the patients and 12.6% were HER2+. In untreated patients, HER2 was a negative prognostic factor in ER+ patients, HR 2.95; 95% CI: 1.61-5.38, p < 0.001, but not in ER- patients, HR 0.67; 95% CI: 0.28-1.61, p = 0.4, and a significant interaction between the two markers was found, p < 0.01. HER2 status was not related to tamoxifen treatment efficacy in ER+ patients (term of interaction p = 0.95). When stratifying for PR status, similar results were achieved. HER2+ and ER+ breast cancer constituted a subgroup of tumours with poor prognosis in premenopausal breast cancer, whereas no treatment interaction was found between HER2 status and tamoxifen in ER+ tumours. The poor prognosis in HER2+ and ER+ patients may interfere with the interpretation of HER2 data in non-randomised trials of adjuvant tamoxifen.


Slipping Anchor? Testing the Vignettes Approach to Identification and Correction of Reporting Heterogeneity.
We propose tests of the two assumptions under which anchoring vignettes identify heterogeneity in reporting of categorical evaluations. Systematic variation in the perceived difference between any two vignette states is sufficient to reject vignette equivalence. Response consistency - the respondent uses the same response scale to evaluate the vignette and herself - is testable given sufficiently comprehensive objective indicators that independently identify response scales. Both assumptions are rejected for reporting of cognitive and physical functioning in a sample of older English individuals, although a weaker test resting on less stringent assumptions does not reject response consistency for cognition.


Twenty years of physical mapping of major ribosomal RNA genes across the teleosts: A review of research.
Molecular cytogenetic data on the number and position of 45S ribosomal RNA genes (rDNA; located in nucleolus organizing regions, NORs) detected by FISH in 330 species of 77 families and 22 orders of bony fishes (Teleostei) and, additionally, 11 species of basal ray-finned fishes are compiled and analyzed. The portion of species with single rDNA sites in the sample amounts to 72%. The percentage of species with multiple NORs decreases with increasing numbers of rDNA loci per genome, i.e. scarcely 3% of species carry 4 or more rDNA-bearing chromosome pairs. 43% of all rDNA sites analyzed occur terminally on the short arms of chromosomes or constitute them. In general, terminal rDNA sites account for 87% of all examined cases. Interspecific variation in the location of single rDNA sites among related taxa, polymorphisms of multiple NORs in some groups of teleosts and analytical outcomes on the subject are reviewed.


Diphacinone and zinc phosphide toxicity in a flock of Peafowl.
Toxicity probably due to a combination of diphacinone and zinc phosphide was diagnosed in a flock of peafowl, in which 35 birds in a flock of 80 died over a span of 10 days without any apparent clinical signs. Chickens and guinea fowl, 30 each on the same premises, were not affected. Plastic tubes containing diphacinone and zinc phosphide were used on the premises to control ground squirrels. Most of the six dead peafowl, which ranged in age from 6 months to 4 years, had an accumulation of serosanguinous fluid in the abdominal cavity, semi-clotted blood over the liver lobes, increased pericardial fluid, and enlarged and pale kidneys. Pellets of diphacinone and zinc phosphide were found in the crop and gizzard contents from most of the birds. Microscopically, most of the birds had mild to moderate centrolobular degeneration of hepatocytes and multifocal degeneration of myofibres in the heart with infiltration by a few mononuclear cells. Acute nephrosis and mucosal oedema in the oesophagus and crop were also observed. Toxicological analysis of the crop and gizzard contents revealed the presence of diphacinone and phosphine gas, and analysis of the crop contents from two birds for heavy metals revealed zinc levels of up to 6600 parts/10 6 . It was suspected that only the peafowl and not the chickens and guinea fowl were affected because peafowl, with their longer necks, were able to reach into the plastic tubes and eat the pellets.


[Induction by kopsinine of hepatic mixed-function oxidase in mice].
Kopsinine is an indole alkaloid. Oral administration of kopsinine 200 mg/kg once daily for 3 d significantly increased liver microsomal protein, cytochrome P-450, cytochrome b5, NADPH-cytochrome C reductase, aminopyrine demethylase and benzo(a)pyrine hydroxylase activities in mice. kopsinine only induced cytochrome P-450 in rough endoplasmic reticulum of liver. SDS-polyamine gel electrophoresis analysis showed that the protein bands of microsomes from kopsinine treated mice were similar to that induced by phenobarbital in mice. Metyrapon, a specific inhibitor of cytochrome P-450, partially antagonized aminopyrine demethylase activity of microsomes from mice treated with kopsinine. The results suggest that kopsinine yields a pentobarbital-like induction on liver mixed function oxidase in mice. In addition, kopsinine was found to shorten the barbital-induced sleeping time in mice.


Pubertal maturation and timing effects on resting state electroencephalography in autistic and comparison youth.
Autistic and comparison individuals differ in resting-state electroencephalography (EEG), such that sex and age explain variability within and between groups. Pubertal maturation and timing may further explain variation, as previous work has suggested alterations in pubertal timing in autistic youth. In a sample from two studies of 181 autistic and 94 comparison youth (8 years to 17 years and 11 months), mixed-effects linear regressions were conducted to assess differences in EEG (midline power for theta, alpha, and beta frequency bands). Alpha power was analyzed as a mediator in the relation between pubertal maturation and timing with autistic traits in the autistic groups to understand the role of puberty in brain-based changes that contribute to functional outcomes. Individuals advanced in puberty exhibited decreased power in all bands. Those who experienced puberty relatively early showed decreased power in theta and beta bands, controlling for age, sex, and diagnosis. Autistic individuals further along in pubertal development exhibited lower social skills. Alpha mediated the relation between puberty and repetitive behaviors. Pubertal maturation and timing appear to play unique roles in the development of cognitive processes for autistic and comparison youth and should be considered in research on developmental variation in resting-state EEG.


Dietary fat, telomere length and cognitive function: unravelling the complex relations.
The review aims to explore the recent evidence on the associations between different dietary fat intake and cognitive function, and to understand the role of telomere length in this relationship. Clinical and preclinical studies included in this review suggest that dietary fat intake is associated with cognitive function and telomere length. High intake of saturated fats and trans fats, commonly found in ultra-processed foods, appears to have negative effects on cognitive function and telomere length, while other dietary fats, such as omega-3 polyunsaturated fatty acids and monounsaturated fatty acids are associated with improved cognitive performance and reduced telomere attrition. Controversial results related to omega-6 polyunsaturated fatty acids intake and its impact on cognitive function were found. Dietary fats may affect telomere length and cognition through oxidative stress, inflammation, and insulin resistance. The current review illustrated the relationship between dietary fat and cognitive function by focusing on the role of telomere length as a potential intermediator. More future studies are required, however, in order to develop targeted interventions aimed at preserving cognitive well-being throughout life.


Synthesis and properties of human beta-endorphin-(1-9) and its analogs.
Four analogs of human beta-endorphin (beta h-EP) were synthesized by the solid-phase method: beta h-EP-(1-9) (I), [D-Ala2]-beta h-EP-(1-9) (II), [Gln8]-beta h-EP-(1-9) (III), and [D-Ala2, Gln8]-beta h-EP-(1-9) (IV). Measurement in a radioreceptor binding assay with use of tritiated beta h-EP as primary ligand gave relative potencies as follows: Met-enkephalin, 100; I, 76; II, 100; III, 200; IV, 200. Two new amino acid derivatives were prepared and used for synthesis of the analogs: N alpha-t-butyloxycarbonyl-O-(cyclopentyl)-tyrosine and N alpha-t-butyloxycarbonyl-gamma-(cyclopentyl)-glutamic acid.


Effect of preoperative administration of celecoxib on postoperative pain after laparoscopic cholecystectomy.
For laparoscopic cholecystectomy, pain is most frequent complaint and the most common cause of delayed discharge. The aim of this study was to determine the effect of preoperative administration of celecoxib on the level of postoperative pain in patient undergoing laparoscopic cholecystectomy. We enrolled 60 patients ASA class I and II, scheduled for elective laparoscopic cholecystectomy. The patients were randomized to receive celecoxib 200 mg, celecoxib 400 mg or placebo two hour before the induction of anesthesia. The patients received the same anesthetics. The intensities of abdominal pain were assessed using VAS (visual analog scale) at 1, 2, 4, 12, 24 hours after surgery. In celecoxib 200 mg group, VAS score of somatic pain compared to control group decreased at 1, 2, and 4 hours after surgery. In celecoxib 400 mg group, VAS score of somatic pain compared to control group decreased at 1, 2, and 4 hours after surgery. There was no difference between celecoxib 200 mg and celecoxib 400 mg in pain scores of somatic pain. Dosage of meperidine in two celecoxib groups after surgery were each 31 mg and 26 mg and that of control group was 72 mg. There was no difference between celecoxib groups and placebo group in pain scores of visceral pain. The preoperative administration of celecoxib reduces the level of postoperative pain after laparoscopic cholecystectomy without adverse effects.


Data-driven prediction of thresholded time series of rainfall and self-organized criticality models.
We study the occurrence of events, subject to threshold, in a representative self-organized criticality (SOC) sandpile model and in high-resolution rainfall data. The predictability in both systems is analyzed by means of a decision variable sensitive to event clustering, and the quality of the predictions is evaluated by the receiver operating characteristic (ROC) method. In the case of the SOC sandpile model, the scaling of quiet-time distributions with increasing threshold leads to increased predictability of extreme events. A scaling theory allows us to understand all the details of the prediction procedure and to extrapolate the shape of the ROC curves for the most extreme events. For rainfall data, the quiet-time distributions do not scale for high thresholds, which means that the corresponding ROC curves cannot be straightforwardly related to those for lower thresholds. In this way, ROC curves are useful for highlighting differences in predictability of extreme events between toy models and real-world phenomena.


Activity-based probes for rhomboid proteases discovered in a mass spectrometry-based assay.
Rhomboid proteases are evolutionary conserved intramembrane serine proteases. Because of their emerging role in many important biological pathways, rhomboids are potential drug targets. Unfortunately, few chemical tools are available for their study. Here, we describe a mass spectrometry-based assay to measure rhomboid substrate cleavage and inhibition. We have identified isocoumarin inhibitors and developed activity-based probes for rhomboid proteases. The probes can distinguish between active and inactive rhomboids due to covalent, reversible binding of the active-site serine and stable modification of a histidine residue. Finally, the structure of an isocoumarin-based inhibitor with Escherichia coli rhomboid GlpG uncovers an unusual mode of binding at the active site and suggests that the interactions between the 3-substituent on the isocoumarin inhibitor and hydrophobic residues on the protease reflect S' subsite binding. Overall, these probes represent valuable tools for rhomboid study, and the structural insights may facilitate future inhibitor design.


Impact of sterile leukocyturia on outcome of critically ill patients with severe acute kidney injury.
The role of immunological mechanisms on renal regeneration and functional recovery after an episode of Acute Kidney Injury (AKI) is still understudied. We aim to evaluate the impact of sterile leukocyturia on outcomes of critically-ill AKI patients. A retrospective analysis of critically-ill patients with stage ≥2 AKI by KDIGO was performed. Patients with urinary tract infection, previous renal replacement therapy, chronic kidney disease stage >3 and kidney, urinary tract or prostatic cancer were excluded. Sterile leukocyturia was defined as a positive leukocyte esterase value. 108 patients with stage ≥2 AKI were included, 39.8% of which had sterile leukocyturia. AKI patients with sterile leukocyturia were older, had more cardiovascular disease and a lower baseline renal function (p < 0.05). They had a higher serum creatinine and leukocytosis at admission, were more frequently septic (p < 0.05) and had more persistent AKI by both KDIGO criteria at multivariable analysis (OR 6.130, 95% CI 2.007-18.747). Sterile leukocyturia was associated with different patient baseline and AKI characteristics and more persistent AKI by both KDIGO criteria. Sterile leukocyturia may represent a surrogate marker of renal inflammation during AKI.


A microfluidic platform integrated with field-effect transistors for enumeration of circulating tumor cells.
Circulating tumor cells (CTCs) are one of the promising cancer biomarkers whose concentrations are measured not only in the initial diagnostic stages, but also as treatment progresses. However, the existing methods for CTC detection are relatively time-consuming and labor-intensive. In this study, a new microfluidic platform integrated with field-effect transistors (FETs) and chambers for the trapping of CTCs was developed. This novel design could not only trap CTCs from whole blood samples, but also enumerate them via FET sensing of CTC-specific aptamer-CTC complexes. The FET output signal was experimentally found to increase with the increasing number of captured CTCs. More importantly, the enumeration of spiked CTCs in blood samples could be achieved in accordance with the signals measured on the FET devices. We therefore believe that this automated system could be a useful tool for enumeration of CTCs.


Agents in early development for treatment of bladder dysfunction - promise of drugs acting at TRP channels?
Introduction: In the lower urinary tract (LUT) several members of the TRP superfamily are involved in nociception and mechanosensory transduction. Animal studies have suggested a therapeutic potential of some of these channels, including TRPV1, TRPV4, TRPM8, TRPA1, and TRPM4, for treatment of bladder over- and underactivity and bladder pain disorders, but translation of this information to clinical application has been slow. Areas covered: An update on and discussion of current information on the potential clinical use of TRP channel agonists/antagonists in the treatment of different types of bladder dysfunction. The electronic databases PubMed and Scopus were used to identify relevant clinical and animal studies. Expert opinion: The therapeutic effect of TRPV1 channel desensitizing agonists (capsaicin, resiniferatoxin, given intravesically) has been convincingly demonstrated in some forms of bladder overactivity. However, so far, the potential of any of the small-molecule TRP channel blockers developed for non-bladder indications and tested in early human trials for safety has not been explored clinically in LUT dysfunction. The adverse effects of hyperthermia and reduction of noxious heat sensation of the first generation TRPV1 blockers have delayed development. Despite lack of translational information, TRP channels remain interesting targets for future LUT drugs.


Asking terminally ill patients about their preferences concerning place of care and death.
Asking patients with palliative care needs about their end-of-life (EoL) preferences is widely acknowledged as an important aspect of EoL care. However, the issue of how to ask patients these questions has not been fully explored. Most prior studies in this area do not differentiate between patients' pragmatic preferences and ideal preferences, and between preferences concerning place of care (PoC) and place of death (PoD). The aim of this study was to examine possible differences between pragmatic and ideal preferences of terminally ill patients, as well as differences between asking patients about preferences concerning PoC and PoD. Structured interviews were performed with terminally ill cancer patients at inclusion and a follow-up questionnaire was completed 1 month later. Answers were compared using kappa (k) statistics and Pearson's c2-test. Among 96 cancer patients, agreement between pragmatic and ideal preferences was statistically significantly different (p=<0.001). Agreement between preferences for PoC and PoD was high (k:0.76-0.85). Differences exist between pragmatic and ideal EoL preferences, whereas preferences for PoC and PoD were found to be similar. These findings highlight the importance of the phrasing of questions when uncovering patients' preferences for EoL care.


Intravenous carotid arteriography using line-scanned digital radiography.
Digital subtraction and an experimental system for line-scanned radiography were used to image the bifurcations of the carotid arteries. The subtracted images were obtained before and after injections of contrast media. The anatomy of the extracranial carotid arteries was demonstrated in most patients, and verified by selective catheter arteriograms. Unsuccessful studies were attributed to improper timing while obtaining the images, and to venous occlusion. Artifacts produced by motion limit the temporal subtraction method.


Rebuilding immunity in cancer patients.
Rebuilding and maintaining immunity are paramount to the success of cancer immunotherapy and hematopoietic stem cell transplantation. If immune surveillance indeed can protect from cancer, the very manifestation of malignancy means that the disease has prevailed over immunity. Yet, often, tumor-specific T cells can be found in cancer patients irrespective of vaccination. Interestingly, patients suffering from malignancy often harbor unexpectedly high levels of immature CD14(+)HLA-DR(-) monocytes, although the abundance of CD4(+) cells, CD8(+) cells and CD4(+)CD25(high) cells may be normal. It is plausible that in cancer such cells suppress T cell function, analogous to CD14(+)HLA-DR(-) cells in sepsis and major trauma, in addition to their likely failure to re-present tumor-associated antigens once dendritic cells have initiated the T cell response. Recent evidence indicates that tumor-borne adenosine, lactate and hypoxia in the tumor environment may modulate tumor-specific immunity to a significant extent, but their effects on myeloid cell function are unclear. Thus, understanding and controlling these factors may appreciably impact the success of rebuilding and maintaining immunity in cancer patients.


Hydroxytyrosol protects against myocardial ischemia reperfusion injury by inhibiting mitochondrial permeability transition pore opening.
Hydroxytyrosol (HT), a phenolic compound extracted from olive oil, is reported to protect against myocardial ischemia reperfusion injury (MIRI), but its mechanism has not been fully elucidated. The mitochondria permeability transition pore (MPTP) is an important therapeutic target for MIRI. The present study aimed to investigate the role of MPTP in the cardioprotection of HT. Isolated rat hearts were mounted on a Langendorff apparatus and subjected to 30 min of ischemia followed by 120 min of reperfusion to mimic a MIRI model. Isolated hearts were pretreated with different doses of HT (10, 100 and 1,000 µM) for 10 min prior to ischemia. Myocardial infarct size was detected using TTC staining. Changes in myocardial cell structure were observed using hematoxylin and eosin staining. MPTP opening was detected spectrophotometrically. Myocardial cell apoptosis was observed with terminal deoxynucleotidyl-transferase-mediated dUTP nick end labeling assays. The expression of apoptosis-associated proteins was measured by western blot analysis. The data revealed that HT (100 and 1,000 µM) treatment significantly alleviated pathological damage in ischemic myocardium and reduced myocardial infarct size compared with the untreated control. However, no significant difference was observed in the 10 µM HT treatment group compared with the untreated control. It was further revealed that HT decreased the B cell lymphoma-2 (Bcl-2)-like protein 4 (Bax)/Bcl-2 ratio, suppressed MPTP opening and subsequently decreased the expression of cytochrome c, cleaved caspase-9 and -3, thereby inhibiting apoptosis. Additionally, the beneficial effects of HT on MIRI were reversed by atractyloside, which induces MPTP opening. In conclusion, the present study demonstrated that HT inhibited MPTP opening, partially via modulation of Bax and Bcl-2, thereby protecting against MIRI and thereby providing a pharmacological basis for future research and treatment of MIRI.


Survival Outcomes in Cancer Patients Predicted by a Partial EMT Gene Expression Scoring Metric.
Metastasis is a significant contributor to morbidity and mortality for many cancer patients and remains a major obstacle for effective treatment. In many tissue types, metastasis is fueled by the epithelial-to-mesenchymal transition (EMT)-a dynamic process characterized by phenotypic and morphologic changes concomitant with increased migratory and invasive potential. Recent experimental and theoretical evidence suggests that cells can be stably halted en route to EMT in a hybrid E/M phenotype. Cells in this phenotype tend to move collectively, forming clusters of circulating tumor cells that are key tumor-initiating agents. Here, we developed an inferential model built on the gene expression of multiple cancer subtypes to devise an EMT metric that characterizes the degree to which a given cell line exhibits hybrid E/M features. Our model identified drivers and fine-tuners of epithelial-mesenchymal plasticity and recapitulated the behavior observed in multiple in vitro experiments across cancer types. We also predicted and experimentally validated the hybrid E/M status of certain cancer cell lines, including DU145 and A549. Finally, we demonstrated the relevance of predicted EMT scores to patient survival and observed that the role of the hybrid E/M phenotype in characterizing tumor aggressiveness is tissue and subtype specific. Our algorithm is a promising tool to quantify the EMT spectrum, to investigate the correlation of EMT score with cancer treatment response and survival, and to provide an important metric for systematic clinical risk stratification and treatment. Cancer Res; 77(22); 6415-28. ©2017 AACR.


Microwave measurements of carbon monoxide on titan.
The ratio of the flux density of Titan was measured in two 200-megahertz bands, one centered on the (1-0) rotation line of carbon monoxide at 115.3 gigahertz and the other 2600 megahertz lower. The measurements were made with a complex-correlation technique on the new millimeter-wavelength interferometer at the Owens Valley Radio Observatory, Big Pine, California. The excess flux in the carbon monoxide band is interpreted as a strong detection of carbon monoxide and a mixing ratio, assumed constant, of 6 x 10(-5). The brightness temperature of Titan at 112.6 gigahertz is 69 +/- 10 kelvins, consistent with atmospheric emission from just below the tropopause.


Elemental composition and metal pollution in Egyptian Red Sea mangrove sediments: Characterization and origin.
The present work was conducted to characterize the mangrove sediments along the Egyptian Red Sea in terms of elemental composition and to assess the extent of pollution and its sources. A total of 26 samples of mangrove sediments were collected from three different areas: Sharm El Madfea, Sowmaa Mangrove and Abu Fasi. The samples were analyzed using the inductively coupled plasma - mass spectrometry ICP-MS and atomic-emission spectrometry ICP-AES. Mass fractions of a total of 58 major and trace elements were determined in the mangrove samples. Univariate and multivariate statistical analyses were performed to determine the origin of trace and major elements in the mangrove sediments. The normalized values show that the elements above the background can be indicated in descending order as follows: P > Cd > Sr > Ca > U > Se > As > Sn > Cu > Sb > Pb > Mo > Ag. Several pollution indices were also calculated. Principal component analysis revealed three clusters of the studied sediment samples. The analysis of the ratio indicators shows that the origin of the sediments mostly falls near continental island arcs (CIA). The pollution indices show remarkable pollution levels and enriched elements. The data obtained can serve as baseline data for the sediments of the mangrove environment and can be used to study possible changes in the future.


Identifying Behavioural Traits and Underlying Personality Dimensions in Domestic Ferrets (Mustela putorius furo).
The aim of this study was to examine the personality structure of domestic ferrets (Mustela putorius furo) by using owner-based reporting of personality traits. A total of 743 ferret owners participated in an online questionnaire, with a total of 1029 ferrets being assessed. Respondents rated 67 adjectives based on their ferret(s) behavioural traits and personality. Principal component analysis (PCA) of these trait ratings identified four underlying personality dimensions, which accounted for 47.1% of the total variance. These were labelled according to the traits that they encompass: Extraversion, Sociability, Attentiveness, and Neuroticism. Details about ferret sex, de-sexing status, age, and coat colour were also sought, and General Linear Mixed Models were used to test the main effects of these characteristics on the personality dimensions. It was found that sex (p < 0.01) and age (p < 0.001) significantly influenced certain personality components, whereas de-sexing did not. Sociability, Attentiveness, and Neuroticism were found to differ based on sex, whereby males were rated as more sociable than females, but females were rated higher on the Attentiveness and Neuroticism subscales. Finally, Extraversion was found to generally decrease with age. We can use the findings of this study to make cross-species comparisons and further inform the discussion regarding the adaptive relevance of animal personality. Identifying differences in personality types can improve welfare by using this information to satisfy the different needs of individuals.


Triple metallic stenting in hilar malignant biliary obstructions.
To research the effects of triple stenting on primary patency rates and on clinical and biochemical findings in patients with high-level malignant biliary obstruction. We analyzed eight patients who had undergone triple stenting for hilar malignant biliary obstruction, mainly with the percutaneous approach, between January 2009, and September 2009. Pre-interventional bilirubin levels and the existence of pruritus or cholangitis were recorded. Patients were examined 15 days and three months post-intervention. Procedure-related mortality and 30-day mortality rates were recorded. Changes in the serum bilirubin levels, pruritis and cholangitis were examined. Primary patency rates were calculated with the Kaplan-Meier method. Pruritis and cholangitis improved within 15 days. There was a significant decrease in serum bilirubin levels, which were very near to normal limits. Two patients died: one in the fourth month and the other in the eighth month. The mean patency rate was 179±18.81 days. There were no procedure-related or 30-day mortalities in the study group. Triple metallic stenting did not significantly improve primary patency rates in hilar malignant biliary obstructions. However, the beneficial effects of triple stenting included the rapid improvement in clinical and biochemical signs in select patients. Triple stenting will be beneficial in preventing isolation that might cause cholangitis. Malignant biliary obstruction in patients with a trifurcation anomaly in the hilar region may necessitate triple stenting.