Exploring the Fatty Acid Binding Pocket in the SARS-CoV-2 Spike Protein - Confirmed and Potential Ligands.
Severe Acute Respiratory syndrome 2 (SARS-CoV-2) is a respiratory virus responsible for coronavirus disease 19 (COVID-19) and the still ongoing and unprecedented global pandemic. The key viral protein for cell infection is the spike glycoprotein, a surface-exposed fusion protein that both recognizes and mediates entry into host cells. Within the spike glycoprotein, a fatty acid binding pocket (FABP) was confirmed, with the crystallization of linoleic acid (LA) occupying a well-defined site. Importantly, when the pocket is occupied by a fatty acid, an inactive conformation is stabilized, and cell recognition is hindered. In this review, we discuss ligands reported so far for this site, correlating their activity predicted through in silico studies with antispike experimental activity, assessed by either binding assays or cell-infection assays. LA was the first confirmed ligand, cocrystallized in a cryo-EM structure of the spike protein, resulting in increased stability of the inactive conformation of the spike protein. The next identified ligand, lifitegrast, was also experimentally confirmed as a ligand with antiviral activity, suggesting the potential for diverse chemical scaffolds to bind this site. Finally, SPC-14 was also confirmed as a ligand, although no inhibition assays were performed. In this review, we identified 20 studies describing small-molecule compounds predicted to bind the pocket in in silico studies and with confirmed binding or in vitro activity, either inhibitory activity against the spike-ACE2 interaction or antiviral activity in cell-based assays. When considering all ligands confirmed with in vitro assays, a good overall occupation of the pocket should be complemented with the ability to make direct interactions, both hydrophilic and hydrophobic, with key amino acid residues defining the pocket surface. Among the active compounds, long flexible carbon chains are recurrent, with retinoids capable of binding the FABP, although bulkier systems are also capable of affecting viral fitness. Compounds able to bind this site with high affinity have the potential to stabilize the inactive conformation of the SARS-CoV-2 spike protein and therefore reduce the virus's ability to infect new cells. Since this pocket is conserved in highly pathogenic human coronaviruses, including MERS-CoV and SARS-CoV, this effect could be exploited for the development of new antiviral agents, with broad-spectrum anticoronavirus activity.


Factors important to patients' quality of life at the end of life.
When curative treatments are no longer options for patients dying of cancer, the focus of care often turns from prolonging life to promoting quality of life (QOL). Few data exist on what predicts better QOL at the end of life (EOL) for advanced cancer patients. The purpose of this study was to determine the factors that most influence QOL at the EOL, thereby identifying promising targets for interventions to promote QOL at the EOL. Coping With Cancer is a US multisite, prospective, longitudinal cohort study of 396 advanced cancer patients and their informal caregivers who were enrolled from September 1, 2002, through February 28, 2008. Patients were followed up from enrollment to death a median of 4.1 months later. Patient QOL in the last week of life was a primary outcome of Coping With Cancer and the present report. The following set of 9 factors, preceded by a sign indicating the direction of the effect and presented in rank order of importance, explained the most variance in patients' QOL at the EOL: 1 = (-) intensive care unit stays in the final week (explained 4.4% of the variance in QOL at the EOL), 2 = (-) hospital deaths (2.7%), 3 = (-) patient worry at baseline (2.7%), 4 = (+) religious prayer or meditation at baseline (2.5%), 5 = site of cancer care (1.8%), 6 = (-) feeding-tube use in the final week (1.1%), 7 = (+) pastoral care within the hospital or clinic (1.0%), 8 = (-) chemotherapy in the final week (0.8%), and 9 = (+) patient-physician therapeutic alliance at baseline (0.7%). The vast majority of the variance in QOL at the EOL, however, remained unexplained. Advanced cancer patients who avoid hospitalizations and the intensive care unit, who are less worried, who pray or meditate, who are visited by a pastor in the hospital/clinic, and who feel a therapeutic alliance with their physicians have the highest QOL at the EOL.


Visually improved image compression by combining a conventional wavelet-codec with texture modeling.
Human observers are very sensitive to a loss of image texture in photo-realistic images. For example a portrait image without the fine skin texture appears unnatural. Once the image is decomposed by a wavelet transformation, this texture is represented by many wavelet coefficients of low- and medium-amplitude. The conventional encoding of all these coefficients is very bitrate expensive. Instead, such an unstructured or stochastic texture can be modeled by a noise process and be characterized with very few parameters. Thus, a hybrid scheme can be designed that encodes the structural image information by a conventional wavelet codec and the stochastic texture in a model-based manner. Such a scheme, called WITCH (Wavelet-based Image/Texture Coding Hybrid), is proposed. It implements such an hybrid coding approach, while nevertheless preserving the features of progressive and lossless coding. Its low computational complexity and the parameter coding costs of only 0.01 bpp make it a valuable extension of conventional codecs. A comparison with the JPEG2000 image compression standard showed that the WITCH-scheme achieves the same subjective quality while increasing the compression ratio by more than a factor of two.


Solvent- and pH-Stable Eu(III)-Based Metal-Organic Framework with Phosphate-Ratio Fluorescence Sensing and Significant Proton Conduction.
Lanthanide-based metal-organic frameworks show good potential for applications due to their unique structures and functional properties. A highly thermally and acid-base stable Eu-MOF was synthesized by a solvothermal method with the molecular formula {[(CH3)2NH2]2[Eu2(NDDP)2(H2O)2]·H2O}n (Eu-MOF, H4NDDP = 5,5'-(naphthalene-2,6-diyl)diisophthalic acid). Eu-MOF takes a three-dimensional (4,4,8)-connected topology. The water molecules involved in the coordination, free water molecules, and [(CH3)2NH2]+ cations in the pore can be used as proton carriers. The proton conductivity attains 1.25 × 10-4 S cm-1 at room temperature and 2.42 × 10-3 S cm-1 at 70 °C and 98% relative humidity. Combined with the dual-emission properties from the ligands and Eu(III) ions enables Eu-MOF to be used as a ratiometric fluorescent sensor for phosphate efficiently and rapidly, with a limit of detection of 0.12 μM in the Tris-HCl buffer solution. These results provide a new approach for the construction of MOFs with high proton conductivity and a ratiometric fluorescence response.


[Economic growth and health inequities].
This essay reviews the relation between health inequities and economic growth. The general meaning of these and ancillary concepts (economic development, health inequalities) is briefly reviewed. Some studies illustrating different hypotheses on the long-run historical evolution of health inequalities are presented, and three case studies -the United States in 1920-1940 and in recent years, Finland during the expansion of the 1980s and the recession of the 1990s- are reviewed to demonstrate the evolution of health inequalities during the periods of expansion and recession in markets economies that conform to the so-called business cycle. Health inequities between ethnic groups and social classes are often found in modern societies, and some of these disparities seem to be widening. Periods of economic expansion do not seem favorable for the lessening of health inequalities. Contrarily, and counter-intuitively, evidence rather suggests that it is during periods of recession that gaps in health between privileged and disadvantaged groups tend to narrow.


The appropriateness of drug use in an older nondemented and demented population.
To assess the extent of inappropriateness of drug use in an older nondemented and demented population. Descriptive analysis based on data from a sample of older subjects age 81 years and older. Data were collected from the second follow-up conducted in 1994-1996. A population-based study of the Kungsholmen project in Stockholm, Sweden. Drug information was obtained from 681 subjects with a mean age of 86.9 years. The subjects were predominantly women (78%). Thirteen percent resided in institutions and 27.6% were diagnosed with dementia. Dementia diagnosis based on DSM III-R. Criteria for inappropriateness of drug use: use of drugs with potent anticholinergic properties, drug duplication, potential drug-drug and drug-disease interactions, and inappropriate drug dosage. The mean number of drugs used was 4.6: 4.5 drugs for nondemented and 4.8 for demented subjects. Nondemented subjects more commonly used cardiovascular-system drugs and demented subjects used nervous-system drugs. Demented subjects were more commonly exposed to drug duplication and to drugs with potent anticholinergic properties, both involving the use of psychotropic drugs. Nondemented subjects were more commonly exposed to potential drug-disease interactions, mostly with the use of cardiovascular drugs. The most common drug combination leading to a potential interaction was the use of digoxin with furosemide, occurring more frequently among nondemented subjects. The most common drug-disease interaction was the use of beta-blockers and calcium antagonists in subjects with congestive heart failure. The doses of drugs taken by both nondemented and demented subjects were mostly lower than the defined daily dose. There was substantial exposure to presumptive inappropriateness of drug use in this very old nondemented and demented population. The exposure of demented subjects to psychotropic drugs and nondemented subjects to cardiovascular drugs reflect the high frequency of prescribing these drugs in this population.


Traumatic stress in Japanese broadcast journalists.
Job-related traumatic stress experienced by broadcast journalists in Japan was investigated. A questionnaire inquiring about the most traumatic event they faced when covering the news and the Impact of Event Scale-Revised (IES-R) were administered to 270 journalist participants working for Japanese news companies. Of these, 6% met the IES-R criterion for potential posttraumatic stress disorder (PTSD). Moreover, posttraumatic stress reactions at the time of the survey were strongly related to symptoms of stress experienced during the stressful assignment. The early assessment of stress symptoms in journalists that experience traumatic news coverage is important for preventing the development of PTSD symptoms.


Full 2D displacement vector and strain tensor estimation for superficial tissue using beam-steered ultrasound imaging.
Ultrasound strain imaging is used to measure local tissue deformations. Usually, only strains along the ultrasound beam are estimated, because those estimates are most precise, due to the availability of phase information. For estimating strain in other directions we propose to steer the ultrasound beam at an angle, which allows estimating different projections of the 2D strain tensor, while phase information remains available. This study investigates beam steering at maximally three different angles to determine the full 2D strain tensor. The method was tested on simulated and experimental data of an inclusion phantom and a vessel phantom. The combination of data from a non-steered acquisition and acquisitions at a large positive and an equally large but negative steering angle enabled the most precise estimation of the strain components. The method outperforms conventional methods that do not use beam steering.


Cytoreductive surgery and hyperthermic intrapleural chemotherapy for malignant pleural diseases: preliminary experience.
Cytoreductive surgery and hyperthermic-intraoperative-intrapleural-chemotherapy (HITHOC) is a known approach for malignant pleural diseases (MPD). This study was started to clarify the role of cytoreductive surgery and HITHOC in MPD. Criteria of inclusion were early-stage disease in malignant pleural mesothelioma (MPM), young age, good condition and selected stage-M1a lung cancer. Six patients with MPM and two patients with lung cancer were enrolled. After surgical debulking, intrapleural cisplatin was administered for 60 min at 42.5°C. Wedge, rib resection and repaired diaphragm were added in three, one and one patient, respectively. Morbidity, toxicity and mortality was nil. Hospital stay was 8 days. Mean survival is 13.6 months. This experience confirms that cytoreductive surgery and HITHOC is a good option in the treatment of MPD. A randomized controlled trial is necessary.


High-dose ursodeoxycholic acid therapy for nonalcoholic steatohepatitis: a double-blind, randomized, placebo-controlled trial.
In uncontrolled clinical studies, ursodeoxycholic acid (UDCA) had a beneficial effect on nonalcoholic steatohepatitis (NASH). However, a large controlled trial using UDCA (13-15 mg/kg/day) was unable to confirm these results. Accordingly, a randomized, placebo-controlled study was initiated with a high dose of UDCA (23-28 mg/kg/day). The allocation of patients and the evaluation of liver histology were performed according to a modified Brunt score and the nonalcoholic fatty liver disease activity score (NAS). With the modified Brunt score, 185 patients with histologically proven NASH were randomized [intention to treat (ITT)], and 147 were treated per protocol (PP). With the NAS, 137 patients were confirmed to have NASH, 48 had borderline NASH, and 1 did not have NASH. The treatment time was 18 months. At entry, the treatment groups were comparable. A second biopsy sample was obtained from 139 of 185 patients (NAS: 107/137). The primary criterion for evaluation was a change in the liver histology; the secondary criteria were single histological variables and liver biochemistry. Significant differences in the overall histology could not be detected between the two treatment groups with the modified Brunt score (P = 0.881) or NAS (P = 0.355). Only lobular inflammation improved significantly (P for the modified Brunt score = 0.011, P for NAS = 0.005). In subgroup analyses, significant improvements in lobular inflammation were also observed in males, younger patients up to 50 years of age, slightly overweight patients, and patients with hypertension and an increased histology score. The fibrosis score did not change (P for ITT = 0.133, P for PP = 0.140). With the exception of gamma-glutamyl transferase, UDCA did not improve laboratory data. High-dose UDCA failed to improve the overall histology in patients with NASH in comparison with placebo.


Permanent prostate seed implant brachytherapy: report of the American Association of Physicists in Medicine Task Group No. 64.
There is now considerable evidence to suggest that technical innovations, 3D image-based planning, template guidance, computerized dosimetry analysis and improved quality assurance practice have converged in synergy in modern prostate brachytherapy, which promise to lead to increased tumor control and decreased toxicity. A substantial part of the medical physicist's contribution to this multi-disciplinary modality has a direct impact on the factors that may singly or jointly determine the treatment outcome. It is therefore of paramount importance for the medical physics community to establish a uniform standard of practice for prostate brachytherapy physics, so that the therapeutic potential of the modality can be maximally and consistently realized in the wider healthcare community. A recent survey in the U.S. for prostate brachytherapy revealed alarming variance in the pattern of practice in physics and dosimetry, particularly in regard to dose calculation, seed assay and time/method of postimplant imaging. Because of the large number of start-up programs at this time, it is essential that the roles and responsibilities of the medical physicist be clearly defined, consistent with the pivotal nature of the clinical physics component in assuring the ultimate success of prostate brachytherapy. It was against this background that the Radiation Therapy Committee of the American Association of Physicists in Medicine formed Task Group No. 64, which was charged (1) to review the current techniques in prostate seed implant brachytherapy, (2) to summarize the present knowledge in treatment planning, dose specification and reporting, (3) to recommend practical guidelines for the clinical medical physicist, and (4) to identify issues for future investigation.


Multipurpose modular experimental station for the DiProI beamline of Fermi@Elettra free electron laser.
We present a compact modular apparatus with a flexible design that will be operated at the DiProI beamline of the Fermi@Elettra free electron laser (FEL) for performing static and time-resolved coherent diffraction imaging experiments, taking advantage of the full coherence and variable polarization of the short seeded FEL pulses. The apparatus has been assembled and the potential of the experimental setup is demonstrated by commissioning tests with coherent synchrotron radiation. This multipurpose experimental station will be open to general users after installation at the Fermi@Elettra free electron laser in 2011.


A cytochrome c methyltransferase from Crithidia oncopelti.
The mitochondrial cytochrome c-557 of Crithidia oncopelti contains two lysine residues and an N-terminal proline residue that are methylated in vivo by the methyl group of methionine. The purified cytochrome can act as a methyl acceptor for a methyltransferase activity in the cell extract that uses S-adenosylmethionine as methyl donor. Crithidia cytochrome c-557 is by far the best substrate for this methyltransferase of those tested, in spite of the fact that methylation sites are already almost fully occupied. The radioactive uptake of [14C]methyl groups from S-adenosylmethionine occurred only at a lysine residue (-8) and the N-terminal proline residue. This methyltransferase appears to differ from that of Neurospora and yeast [Durban, Nochumson, Kim, Paik & Chan (1978) J. Biol. Chem. 253, 1427-1435; DiMaria, Polastro, DeLange, Kim & Paik (1979) J. Biol. Chem. 254, 4645-4652] in that lysine-72 of horse cytochrome c is a poor acceptor. Also, the Crithidia methyltransferase appears to be stable to carry lysine methylation much further to completion than do the enzymes from yeast and Neurospora, which produce very low degrees of methylation in native cytochromes c.


Maintenance costs of serotiny in a variably serotinous pine: The role of water supply.
Serotiny is an important adaptation for plants in fire-prone environments. However, different mechanisms also induce the opening of serotinous cones in the absence of fire in variably serotinous species. Xeriscence -cone opening driven by dry and hot conditions- is considered to be mediated only by the external environment, but endogenous factors could also play a significant role. Using the variably serotinous Pinus halepensis as our model species, we determined the effects of cone age and scales density in cone opening, and using in-situ and ex-situ manipulative experiments we investigated the role of water availability in the opening of serotinous cones. We hypothesized that loss of connection between the cones and the branch through the peduncles or the absence of water supply could induce a faster cone opening. Results showed that older cones lost more water and opened at lower temperatures, with no influence of scales density. Both field and chamber manipulative experiments (using paired cones of the same whorl) confirmed that water intake through the peduncles affected significantly the pace of cone opening, such that lack of water supply speeded up cone dehiscence. However, this was true for weakly serotinous provenances-more common in this species-, while highly serotinous provenances were indifferent to this effect in the field test. All our results support that cone serotiny in P. halepensis involves the allocation of water to the cones, which is highly consistent with the previously observed environmental effects. Importantly, the existence of maintenance costs of serotinous cones has strong implications on the effects of climate change in the resilience of natural populations, via modifications of the canopy seed banks and recruitment after stand-replacing fires. Moreover, evolutionary models for serotiny in P. halepensis must take into account the significant contribution of maintenance costs to the complex interaction between genotype and the environment.


A high affinity interaction of plasminogen with fibrin is not essential for efficient activation by tissue-type plasminogen activator.
Fibrin (Fn) enhances plasminogen (Pg) activation by tissue-type plasminogen activator (tPA) by serving as a template onto which Pg and tPA assemble. To explore the contribution of the Pg/Fn interaction to Fn cofactor activity, Pg variants were generated and their affinities for Fn were determined using surface plasmon resonance (SPR). Glu-Pg, Lys-Pg (des(1-77)), and Mini-Pg (lacking kringles 1-4) bound Fn with K(d) values of 3.1, 0.21, and 24.5 μm, respectively, whereas Micro-Pg (lacking all kringles) did not bind. The kinetics of activation of the Pg variants by tPA were then examined in the absence or presence of Fn. Whereas Fn had no effect on Micro-Pg activation, the catalytic efficiencies of Glu-Pg, Lys-Pg, and Mini-Pg activation in the presence of Fn were 300- to 600-fold higher than in its absence. The retention of Fn cofactor activity with Mini-Pg, which has low affinity for Fn, suggests that Mini-Pg binds the tPA-Fn complex more tightly than tPA alone. To explore this possibility, SPR was used to examine the interaction of Mini-Pg with Fn in the absence or presence of tPA. There was 50% more Mini-Pg binding to Fn in the presence of tPA than in its absence, suggesting that formation of the tPA-Fn complex exposes a cryptic site that binds Mini-Pg. Thus, our data (a) indicate that high affinity binding of Pg to Fn is not essential for Fn cofactor activity, and (b) suggest that kringle 5 localizes and stabilizes Pg within the tPA-Fn complex and contributes to its efficient activation.


Is medical abortion acceptable to all American women: the impact of sociodemographic characteristics on the acceptability of mifepristone-misoprostol abortion.
To determine whether mifepristone-misoprostol medical abortion is more acceptable to some American women than to others. Using previously reported acceptability data from a large US multicenter study (n = 2121), we conducted a more detailed analysis to test whether characteristics such as race/ethnicity, education, age, gestational age, and geographic location affect acceptability. In the United States, Asian women were more than twice as likely as other women to choose this method because they believed it was safer, while white women were twice as likely to select it because they considered it more natural. More educated women were likely to select this method to show support for greater choice and were more pleased about being able to avoid surgery. Particular features of the protocol also appealed to different women. White and African-American women as well as women with more education would feel significantly more comfortable taking one or both of the drugs at home than they would in the clinic. Nearly all groups of women were equally satisfied with this method and found it highly acceptable. The data showed surprisingly few differences among women's overall satisfaction level, their willingness to choose the method again, or to recommend it to others. Mifepristone-misoprostol medical abortion has widespread appeal to a broad range of women, but different women choose and prefer this method for different reasons.


Esophageal injury associated with anterior cervical spine surgery.
Anterior cervical spinal surgery has been used to treat a variety of conditions including spondylosis, fracture, tumor, infection, trauma, and instability. Esophageal perforation, a rare and unusual complication of anterior cervical procedures, has been largely relegated to only incidental case reports with few large retrospective studies performed to determine true incidence, treatment, etiology, and outcome. More than 3000 anterior cervical spine surgeries conducted over a 30-year period by 5 active practicing surgeons were reviewed. There were 3 cases of esophageal injury identified with subsequent critical evaluation to determine presentation, diagnosis, risk factors, management, and outcomes. In addition, incidence rates were calculated based on overall occurrence and antecedent risk factors. Two of the patients with esophageal injury had predisposing risk factors, including diverticula or cervical spine trauma. The third patient had no antecedent risk factors. Symptoms included axial spine pain, odynophagia, dysphagia, purulent spondylitis, and sepsis. Treatment consisted of one or more of the following: reoperation with exploration and repair, esophageal diversion, esophageal rest, antibiotic administration, and wound drainage. Functional outcomes were achieved in all cases with no deaths. Esophageal injury incidence based on overall occurrence in this study was 0.1%. Patients with no antecedent risk factors had an incidence of 0.03%. Our results compare favorably with those of the Cervical Spine Research Society survey from 1989, which predicted an incidence of 0.25% based on questionnaires filed by surgeons, representing 1 of only 2 reports that included more than 1000 patients.


Effect of attenuation mismatches in time of flight PET reconstruction.
While the pursuit of better time resolution in positron emission tomography (PET) is rapidly evolving, little work has been performed on time of flight (TOF) image quality at high time resolution in the presence of modelling inconsistencies. This works focuses on the effect of using the wrong attenuation map in the system model, causing perturbations in the reconstructed radioactivity image. Previous work has usually considered the effects to be local to the area where there is attenuation mismatch, and has shown that the quantification errors in this area tend to reduce with improved time resolution. This publication shows however that errors in the PET image at a distance from the mismatch increase with time resolution. The errors depend on the reconstruction algorithm used. We quantify the errors in the hypothetical case of perfect time resolution for maximum likelihood reconstructions. In addition, we perform reconstructions on simulated and patient data. In particular, for respiratory-gated reconstructions from a wrong attenuation map, increased errors are observed with improved time resolutions in areas close to the lungs (e.g. from 13.3% in non-TOF to up to 20.9% at 200 ps in the left ventricle).


Appropriateness of soldiers' referrals to the emergency department.
Emergency departments (ED) in Israel deal with a large number of referrals from the Israel Defense Force. In a recent study, we observed that only 3% of soldiers referred to the ED in Soroka Medical Center were hospitalized. The purpose of this study was to determine the appropriateness of soldier referrals to the ED, using a proposed set of criteria for ED appropriateness. A retrospective assessment of 707 discharge letters of soldiers from the ED was performed. A set of criteria to assess the appropriateness of soldier's referrals to the ED was developed. The criteria were as follows: (1) resuscitation, (2) hospitalization, (3) laboratory examination, (4) X-ray examination, (5) medical treatment, and (6) specialist consultation. A referral was considered appropriate if at least one criterion was positive. According to the proposed criteria of ED military appropriateness, 586 of 707 referrals (83%) were considered as appropriate. Appropriateness of the referrals according to sections of the ED was as follows: orthopedics (97%), internal medicine (63%), ophthalmology (100%), and neurosurgery (100%). A discrepancy exists between the high proportion of appropriate soldiers' referrals to the ED and the low hospitalization proportion. Organizational steps should be undertaken in the Israel Defense Force to decrease excess military use of civilian ED services.


Foliar-applied ethephon enhances the content of anthocyanin of black carrot roots (Daucus carota ssp. sativus var. atrorubens Alef.).
Black carrots (Daucus carota ssp. sativus var. atrorubens Alef.) constitute a valuable source of anthocyanins, which are used as natural red, blue and purple food colourants. Anthocyanins and phenolic compounds are specialised metabolites, accumulation of which often requires elicitors, which act as molecular signals in plant stress responses. In the present study, ethephon, an ethylene-generating compound was explored as enhancer of anthocyanin and phenolic contents during growth of 'Deep Purple' black carrots. The effects of ethephon on several parameters were investigated, and the expression of biosynthetic anthocyanin genes was studied during growth and anthocyanin accumulation. Roots of ethephon-treated carrot plants exhibited an increase in anthocyanin content of approximately 25%, with values ranging from 2.25 to 3.10 mg g-1 fresh weight, compared with values ranging from 1.50 to 1.90 mg g-1 fresh weight in untreated roots. The most rapid accumulation rate for anthocyanins, phenolic compounds, soluble solids and dry matter was observed between 10 and 13 weeks after sowing in both untreated and ethephon-treated carrots. The differences in anthocyanin contents between untreated and treated carrots increased for several weeks after the ethephon treatment was terminated. Five cyanidin-based anthocyanin forms were identified, with variable relative abundance values detected during root growth. Overall, the expression of the anthocyanin biosynthetic genes analysed (PAL1, PAL3, F3H1, DFR1, LDOX2) increased in response to ethephon treatment, as did the expression of the MYB1 transcription factor, which is associated with activation of the phenylpropanoid pathway under stress conditions. In addition, a correlation was proposed between ethylene and sugar contents and the induction of anthocyanin synthesis. This study presents a novel method for enhancing anthocyanin content in black carrots. This finding is of economic importance as increased pigment concentration per unit of biomass implies improved profitability parameters in food colour production. We provide new insight into the accumulation patterns of the different cyanidin-based anthocyanins and phenolic compounds during root growth. Moreover, we show that enhanced anthocyanin content in ethephon-treated carrots is accompanied by increased expression of anthocyanin biosynthetic genes.


Speeding disease gene discovery by sequence based candidate prioritization.
Regions of interest identified through genetic linkage studies regularly exceed 30 centimorgans in size and can contain hundreds of genes. Traditionally this number is reduced by matching functional annotation to knowledge of the disease or phenotype in question. However, here we show that disease genes share patterns of sequence-based features that can provide a good basis for automatic prioritization of candidates by machine learning. We examined a variety of sequence-based features and found that for many of them there are significant differences between the sets of genes known to be involved in human hereditary disease and those not known to be involved in disease. We have created an automatic classifier called PROSPECTR based on those features using the alternating decision tree algorithm which ranks genes in the order of likelihood of involvement in disease. On average, PROSPECTR enriches lists for disease genes two-fold 77% of the time, five-fold 37% of the time and twenty-fold 11% of the time. PROSPECTR is a simple and effective way to identify genes involved in Mendelian and oligogenic disorders. It performs markedly better than the single existing sequence-based classifier on novel data. PROSPECTR could save investigators looking at large regions of interest time and effort by prioritizing positional candidate genes for mutation detection and case-control association studies.


Distinction between medical and non-medical usages of short forms in clinical narratives.
The short forms of medical concepts or expressions (i.e., acronyms/abbreviations) are prevalent in clinical documentation. Given the limited number of potential short forms, they are also highly ambiguous. Resolving the ambiguity of short forms is essential in clinical natural language processing (NLP). However, one prerequisite for resolving ambiguity of short forms is to have a sense inventory. This paper outlines our process of identifying 141 potential short forms with randomly sampled phrases from a large clinical corpus. We assessed various features in their ability to disambiguate medical and non-medical usages. We identified 68% of our short forms as primarily serving medical usages, whereas 12% had non-medical usages. The remaining 19% showed alternating usage based upon case form. Our short forms had an average of 3.58 senses. Usages could be distinguished using basic trigram/bigram/line information. Our initial findings will be applicable for automatic usage/sense resolution.


Masked priming with orthographic neighbors: a test of the lexical competition assumption.
In models of visual word identification that incorporate inhibitory competition among activated lexical units, a word's higher frequency neighbors will be the word's strongest competitors. Preactivation of these neighbors by a prime is predicted to delay the word's identification. Using the masked priming paradigm (K. I. Forster & C. Davis, 1984, J. Segui and J. Grainger (1990) reported that, consistent with this prediction, a higher frequency neighbor prime delayed the responses to a lower frequency target, whereas a lower frequency neighbor prime did not delay the responses to a higher frequency target. In the present experiments, using English stimuli, it was found that this pattern held only when the primes and targets had few neighbors; when the primes and targets had many neighbors, lower frequency primes delayed responses to higher frequency targets essentially as much as higher frequency primes delayed responses to lower frequency targets. Several possible explanations for these findings are discussed along with their theoretical implications. Considered together, the results are most consistent with activation-based accounts of the masked priming effect.


Modulation of tooth regeneration through opposing responses to Wnt and BMP signals in teleosts.
Most vertebrate species undergo tooth replacement throughout adult life. This process is marked by the shedding of existing teeth and the regeneration of tooth organs. However, little is known about the genetic circuitry regulating tooth replacement. Here, we tested whether fish orthologs of genes known to regulate mammalian hair regeneration have effects on tooth replacement. Using two fish species that demonstrate distinct modes of tooth regeneration, threespine stickleback (Gasterosteus aculeatus) and zebrafish (Danio rerio), we found that transgenic overexpression of four different genes changed tooth replacement rates in the direction predicted by a hair regeneration model: Wnt10a and Grem2a increased tooth replacement rate, whereas Bmp6 and Dkk2 strongly inhibited tooth formation. Thus, similar to known roles in hair regeneration, Wnt and BMP signals promote and inhibit regeneration, respectively. Regulation of total tooth number was separable from regulation of replacement rates. RNA sequencing of stickleback dental tissue showed that Bmp6 overexpression resulted in an upregulation of Wnt inhibitors. Together, these data support a model in which different epithelial organs, such as teeth and hair, share genetic circuitry driving organ regeneration.


The nephrotic syndrome: from the simple to the complex.
The nephrotic syndrome represents a series of physiological results ensuing from the occurrence of significant urinary loss of protein. Although the common initiating event of proteinuria has long been established, the list of problems that result has continued to grow as our understanding of the pathophysiology of nephrosis has increased. This article discusses this pathophysiology, emphasizing the consequences of nephrotic-range proteinuria. These consequences include not only acute and subacute manifestations of disrupted homeostasis, but also ways in which nephrosis itself may amplify mechanisms by which progressive renal nephron loss occurs. New insights into the factors that initiate and maintain glomerulosclerosis and interstitial fibrosis may offer potential approaches to preventing or ameliorating chronic renal insufficiency.


Interprofessional grant writing seminar for early career faculty in a small, isolated teaching center.
Small, isolated teaching centers have difficulty mentoring interprofessional junior faculty in research methods and grant writing. Peer mentoring programs for grant writing at larger institutions have been successful. In this short report, we describe our program that leveraged mentor experience using four framing seminars followed by project refinement in three-person peer groups and monthly mentored works in progress meetings. In its first year, ten faculty from medicine, psychology, and pharmacy completed the program and successfully obtained six funded grants. Five of the projects transitioned from single profession applications to interprofessional applications as participants connected and profession-specific expertise was identified. Refinements for future cohorts are discussed.


Increasing Ketamine Use for Refractory Status Epilepticus in US Pediatric Hospitals.
Ketamine is an emerging therapy for pediatric refractory status epilepticus. The circumstances of its use, however, are understudied. The authors described pediatric refractory status epilepticus treated with ketamine from 2010 to 2014 at 45 centers using the Pediatric Hospital Inpatient System database. For comparison, they described children treated with pentobarbital. The authors estimated that 48 children received ketamine and pentobarbital for refractory status epilepticus, and 630 pentobarbital without ketamine. Those receiving only pentobarbital were median age 3 [interquartile range 0-10], and spent 30 [18-52] days in-hospital, including 17 [9-28] intensive care unit (ICU) days; 17% died. Median cost was $148 000 [81 000-241 000]. The pentobarbital-ketamine group was older (7 [2-11]) with longer hospital stays (51 [30-93]) and more ICU days (29 [20-56]); 29% died. Median cost was $298 000 [176 000-607 000]. For 71%, ketamine was given ≥1 day after pentobarbital. Ketamine cases per half-year increased from 2 to 9 ( P < .05). Ketamine is increasingly used for severe pediatric refractory status epilepticus, typically after pentobarbital. Research on its effectiveness is indicated.


The role of the CN vibration in the activated dynamics of LiNC<−>LiCN isomerization in an argon solvent at high temperatures.
The isomerization between CN-Li and Li-CN in an argon bath provides a paradigmatic example of a reaction in a solvent with tunable coupling. In previous work, we found that the rates exhibited a turnover with the density of the argon bath in the limit that the CN bond was held fixed [P. L. Garcia-Muller, R. Hernandez, R. M. Benito, and F. Borondo, J. Chem. Phys. 137, 204301 (2012)]. Here, we report the effect of the CN bond vibration on the dynamics and the persistence of the turnover. As hypothesized earlier, the CN bond is indeed weakly coupled with the reaction path despite the presence of the argon cage.


Complex Coronary Hemodynamics - Simple Analog Modelling as an Educational Tool.
Invasive coronary angiography remains the cornerstone for evaluation of coronary stenoses despite there being a poor correlation between luminal loss assessment by coronary luminography and myocardial ischemia. This is especially true for coronary lesions deemed moderate by visual assessment. Coronary pressure-derived fractional flow reserve (FFR) has emerged as the gold standard for the evaluation of hemodynamic significance of coronary artery stenosis, which is cost effective and leads to improved patient outcomes. There are, however, several limitations to the use of FFR including the evaluation of serial stenoses. In this article, we discuss the electronic-hydraulic analogy and the utility of simple electrical modelling to mimic the coronary circulation and coronary stenoses. We exemplify the effect of tandem coronary lesions on the FFR by modelling of a patient with sequential disease segments and complex anatomy. We believe that such computational modelling can serve as a powerful educational tool to help clinicians better understand the complexity of coronary hemodynamics and improve patient care.


Central nervous system mechanisms in blood pressure control.
Much of our knowledge about the CNS control of blood pressure is derived from animal studies using techniques such as intracerebroventricular administration of drugs, stereotactic ablation of specific brain nuclei, and biochemical analysis of these nuclei. These methods have identified numerous specific brain nuclei in the brain stem and a meshwork of interconnecting neurones involved in cardiovascular control. The main neurotransmitter involved is noradrenaline but recent interest has focused on several laterally situated nuclei which are capable of synthesizing adrenaline. Centrally acting antihypertensive drugs are thought to act by stimulating central alpha 2-adrenoceptors either by the parent drug itself (clonidine) or via the formation of an active metabolite (alpha-methyldopa). This leads to decreased peripheral sympathetic activity and a hypotensive response but the latter is often attained at the expense of central side-effects such as drowsiness or dry mouth. The mechanism of the antihypertensive effect of beta-blockers remains uncertain although the balance of evidence is against a central effect. The central administration of propranolol causes decreased peripheral sympathetic activity in animals, but plasma catecholamine levels are little altered by beta-blockers in man. In equipotent antihypertensive doses, central alpha-agonists cause a much greater reduction in plasma noradrenaline than beta-blockers.


Strategies to prevent HIV transmission among heterosexual African-American women.
BACKGROUND: African-American women are disproportionately affected by HIV, accounting for 60% of all cases among women in the United States. Although their race is not a precursor for HIV, the socioeconomic and cultural disparities associated with being African American may increase their risk of infection. Prior research has shown that interventions designed to reduce HIV infection among African-American women must address the life demands and social problems they encounter. The present study used a qualitative exploratory design to elicit information about strategies to prevent HIV transmission among young, low-income African-American women. METHODS: Twenty five low income African American women, ages 18-29, participated in five focus groups of five women each conducted at a housing project in Houston, Texas, a large demographically diverse metropolitan area that is regarded as one of the HIV/AIDS epicenters in the United States. Each group was audiotaped, transcribed, and analyzed using theme and domain analysis. RESULTS: The participants revealed that they had most frequently placed themselves at risk for HIV infection through drugs and drinking and they also reported drug and alcohol use as important barriers to practicing safer sex. The women also reported that the need for money and having sex for money to buy food or drugs had placed them at risk for HIV transmission. About one-third of the participants stated that a barrier to their practicing safe sex was their belief that there was no risk based on their being in a monogamous relationship and feeling no need to use protection, but later learning that their mate was unfaithful. Other reasons given were lack of concern, being unprepared, partner's refusal to use a condom, and lack of money to buy condoms. Finally, the women stated that they were motivated to practice safe sex because of fear of contracting sexually transmitted diseases and HIV, desire not to become pregnant, and personal experience with someone who had contracted HIV. CONCLUSION: This study offers a foundation for further research that may be used to create culturally relevant HIV prevention programs for African-American women.


Immunoassay techniques for detection of the herbicide simazine based on use of oppositely charged water-soluble polyelectrolytes.
Linear water-soluble polyelectrolytes, i.e., poly(methacrylate) polyanion and poly(N-ethyl-4-vinylpyridinium) polycation, were used as carriers for the reactants in immunoassay. The strength of ionic forces through distance and the cooperative binding of oppositely charged chains, the carriers interact with each other at an extremely high rate and affinity. These properties of the polyelectrolytes made it possible to carry out the immunochemical steps of the assay in true solution and then to quickly separate the resulting products from the reaction mixtures. The above approach was applied to an assay for the herbicide simazine. Both enzyme-linked immunosorbent assay (ELISA) and dot blot formats of the immunoassay were evaluated. In the ELISA format, the polycation was adsorbed on the surface of a microtiter plate. A tracer antigen (simazine) was allowed to interact in solution with components of the reaction mixture containing simazine-peroxidase conjugate, specific antibodies, and staphylococcal protein A conjugated with the polyanion, and then the mixture was added to the immobilized polycation. Quick separation of the immunoreactants was achieved due to formation of interpolyelectrolyte complexes between polycation and polyanion molecules. After washing, the microplate wells were filled with a solution of substrate, and the optical density of the reaction products was measured. In the second format, a solution of the same reaction mixture (after incubation) was filtered through a porous membrane, with the polycation adsorbed. The subsequent addition of substrate led to the development of colored spots. Sensitivity of the dot blot format was close to that of the traditional ELISA format using the same reactants, i.e., 0.5 ng/mL. However, the assay was much faster (assay time decreased from 100-120 to 45 min). Sensitivities of the dot immunoassay were 1 ng/mL for densitometric detection and 10 ng/mL for visual detection with a duration of 20 min. The techniques developed here were used for simazine determination in water, milk, and juices.


What is appropriate follow-up for the patient with colorectal cancer?
Overall recurrence rates after curative resection for colorectal cancer average 30%. The value of intensive follow-up protocols is assessed from a review of the literature, which reveals that recurrent colorectal cancer can be detected at an earlier stage (average 3 months) and that repeat resection rates for cure can be increased (twofold or more) by intensive follow-up. Despite this, no improvement in survival has been documented. As nonsurgical modalities of therapy for recurrence become further refined, earlier detection of recurrent tumour will become increasingly important. An intensive follow-up regimen is therefore proposed, based on a knowledge of the patterns and timing of recurrence, the relative merits of the investigational modalities available and the assumption that at some point earliest detection and subsequent therapy will improve survival.


Incubators: Building community networks and developing open educational resources to integrate bioinformatics into life science education.
While it is essential for life science students to be trained in modern techniques and approaches, rapidly developing, interdisciplinary fields such as bioinformatics present distinct challenges to undergraduate educators. In particular, many educators lack training in new fields, and high-quality teaching and learning materials may be sparse. To address this challenge with respect to bioinformatics, the Network for the Integration of Bioinformatics into Life Science Education (NIBLSE), in partnership with Quantitative Undergraduate Biology Education and Synthesis (QUBES), developed incubators, a novel collaborative process for the development of open educational resources (OER). Incubators are short-term, online communities that refine unpublished teaching lessons into more polished and widely usable learning resources. The resulting products are published and made freely available in the NIBLSE Resource Collection, providing recognition of scholarly work by incubator participants. In addition to producing accessible, high-quality resources, incubators also provide opportunities for faculty development. Because participants are intentionally chosen to represent a range of expertise in bioinformatics and pedagogy, incubators also build professional connections among educators with diverse backgrounds and perspectives and promote the discussion of practical issues involved in deploying a resource in the classroom. Here we describe the incubator process and provide examples of beneficial outcomes. Our experience indicates that incubators are a low cost, short-term, flexible method for the development of OERs and professional community that could be adapted to a variety of disciplinary and pedagogical contexts.


Estrogen-related receptor β deletion modulates whole-body energy balance via estrogen-related receptor γ and attenuates neuropeptide Y gene expression.
Estrogen-related receptors (ERRs) α, β and γ are orphan nuclear hormone receptors with no known ligands. Little is known concerning the role of ERRβ in energy homeostasis, as complete ERRβ-null mice die mid-gestation. We generated two viable conditional ERRβ-null mouse models to address its metabolic function. Whole-body deletion of ERRβ in Sox2-Cre:ERRβ(lox/lox) mice resulted in major alterations in body composition, metabolic rate, meal patterns and voluntary physical activity levels. Nestin-Cre:ERRβ(lox/lox) mice exhibited decreased expression of ERRβ in hindbrain neurons, the predominant site of expression, decreased neuropeptide Y (NPY) gene expression in the hindbrain, increased lean body mass, insulin sensitivity, increased energy expenditure, decreased satiety and decreased time between meals. In the absence of ERRβ, increased ERRγ signaling decreased satiety and the duration of time between meals, similar to meal patterns observed for both the Sox2-Cre:ERRβ(lox/lox) and Nestin-Cre:ERRβ(lox/lox) strains of mice. Central and/or peripheral ERRγ signaling may modulate these phenotypes by decreasing NPY gene expression. Overall, the relative expression ratio between ERRβ and ERRγ may be important in modulating ingestive behavior, specifically satiety, gene expression, as well as whole-body energy balance.


Whole blood assays for interferon-gamma: practicalities and potential for use as diagnostic tests in the field.
Recent years have seen the introduction of a number of whole-blood assays, in which unseparated heparinized blood is stimulated with antigen either overnight or for as long as 6 days, and cytokine production is measured in the plasma or supernatant. These assays have potential for use in the field as immunodiagnostic assays, as they require only a small blood sample and basic laboratory facilities. Use of these assays in a large study of the immunological effects of BCG vaccination in Malawi has shown that the diluted blood, 6-day whole-blood assays is robust, and can be used to assess T-cell responses to both crude and recombinant antigens. If used with antigens specific to Mycobacterium leprae, these assays could be used to measure exposure of M. leprae within communities or populations, or to aid the early diagnosis of leprosy.


Determination of trans-arachidonic acid isomers in human blood plasma.
Endogenous trans fatty acids originate from diet, but recent studies also suggest that cis-trans isomerization of fatty acids is possible by nitrogen dioxide radical, a product of NO and nitrite oxidation. We developed a method for quantitative analysis of four trans-arachidonic acids (TAA) in human plasma using isotopic dilution gas chromatography/mass spectrometry (GC/MS) with deuterium-labeled internal standard. Esterification of the plasma fatty acid extract with pentafluorobenzyl (PFB) bromide followed by high-performance liquid chromatography purification yielded a fairly pure fraction containing TAA-PFB esters that was analyzed by GC/MS. Partial separation of the TAA isomers was obtained on various GC columns. Comparison of the retention time with the synthetic standards revealed that all four TAA isomers are present in human plasma. The mean concentration of TAA in human plasma was 20.2ng/ml. The levels of isomers were 12.48+/-1.28, 2.75+/-0.39, and 4.99+/-0.74ng/ml for 5E-AA + 11E-AA, 8E-AA, and 14E-AA, respectively. The identification of TAA in plasma suggests that isomerization of arachidonic acid occurs in vivo. Our method allows distinguishing between the dietary and the NO(2)-dependent mechanisms of trans fatty acid formation and will be useful in defining the role of TAA as an in vivo marker of nitrooxidative stress in clinical and experimental settings.


Codifying knowledge to improve patient safety: a qualitative study of practice-based interventions.
Although it is well established that health care professionals use tacit and codified knowledge to provide front-line care, less is known about how these two forms of knowledge can be combined to support improvement related to patient safety. Patient safety interventions involving the codification of knowledge were co-designed by university and hospital-based staff in two English National Health Service (NHS) hospitals to support the governance of medication safety and mortality and morbidity (M&M) meetings. At hospital A, a structured mortality review process was introduced into three clinical specialities from January to December 2010. A qualitative approach of observing M&M meetings (n = 30) and conducting interviews (n = 40) was used to examine the impact on meetings and on front-line clinicians and hospital managers. At hospital B, a medication safety 'scorecard' was administered on a general medicine and elderly care ward from September to November 2011. Weekly feedback meetings were observed (n = 18) and interviews with front-line staff conducted (n = 10) to examine how knowledge codification influenced behaviour. Codification was shown to support learning related to patient safety at the micro (front-line service) level by structuring the sharing of tacit knowledge, but the presence of professional and managerial boundaries at the organisational level affected the codification initiatives' implementation. The findings suggest that codifying knowledge to support improvement presents distinct challenges at the group and organisational level; translating knowledge across these levels is contingent on the presence of enabling organisational factors, including the alignment of learning from clinical practice with its governance.


Molecular virology of hepatitis B virus for clinicians.
This article reviews the molecular biology of the hepatitis B virus in an effort to explain its natural history from a molecular perspective. The life cycle of the virus, with special attention to virus replication, polypeptide production, and morphogenesis, is described. The way in which these steps may influence the natural history of viral pathogenesis, as well as the effectiveness of interventions, receives special consideration.


Fabrication and Characterization of a Flexible Fluxgate Sensor with Pad-Printed Solenoid Coils.
This paper presents the fabrication and characterization of a flexible, flat, miniaturized fluxgate sensor with a thin amorphous rectangular magnetic core fabricated by the pad/printing technique. Both the design and the various printing steps of the sensor are presented. The fluxgate sensor comprises of solenoid coils, and to the best of our knowledge, is the first to be printed with a conventional micro-printing technique. The magnetic core is a non-printed component, placed between the printed layers. The sensor's linear measuring range is ±40 µT with 2% full-scale linearity error, at 100 kHz excitation frequency. The highest measured sensitivity reaches 14,620 V/T at 200 kHz, while the noise of the sensor was found to be 10 nT/ Hz at 1 Hz.


Pulse amplification of a single-frequency cr:forsterite laser.
Stable single-longitudinal-mode operation of a tunable gain-switched Cr:forsterite oscillator-amplifier system is reported. A novel coupled-cavity oscillator configuration provides a low-threshold fluence of 0.3 J cm(-2), making the system attractive for laser media with low gain or high parasitic loss. Nearly transform-limited pulses with instrument-limited bandwidths of 150 MHz have been obtained across a 100-nm wavelength range, limited by the cavity optics. A range of forsterite crystals with Cr(4+) concentration in the 3-21 x 10(18)-cm(-3) range has been used for investigation of amplifier performance in single- and double-pass configurations.


Incidence of Myelotoxicity and Other Adverse Effects Related to Thiopurine Starting in Patients with Inflammatory Bowel Disease: Retrospective Observational Study in a Third-Level Hospital.
Thiopurines are an effective treatment for the maintenance of remission in inflammatory bowel disease (IBD). They can present adverse effects (AEs), with myelotoxicity being the most relevant. This study aims to determine the incidence of AEs related to the starting of thiopurines in our centre. Retrospective study. The AEs in patients that were started on thiopurines between January 2016 and June 2020 were registered, with a two-year follow-up. The mean and standard deviation were used to describe the quantitative variables, and percentages and confidence intervals were used for the qualitative variables. The statistical significance was set at a p-value < 0.05. 98 patients were included, with 64 AEs detected in 48 patients (49%). Most of the AEs appeared in the first 6 months. The most relevant were: 21 neutropenia (21.4%), 19 hypertransaminasemia (19.4%), 13 digestive intolerances (13.2%), 6 acute pancreatitis (6.12%), 3 phototoxicity (3%), and 2 unknown origin fevers (2%). In 29 patients (29.4%) the treatment had to be suspended due to AEs. In 11 cases (11.2%), azathioprine (AZA) was switched to 6-mercaptopurine (6 MP) as 5 showed tolerance and 6 patients needed suspension due to AEs. Eight patients required hospital admission, but none of them needed intensive care unit admission. There were no fatal adverse effects. Thiopurines are a safe drug with few AEs, especially after the first months of treatment. These results suggest that periodic analytic follow-up may not be necessary after the initial period of treatment.


A comprehensive review of the novel therapeutic targets for the treatment of diabetic cardiomyopathy.
Diabetic cardiomyopathy (DCM) is characterized by structural and functional abnormalities in the myocardium affecting people with diabetes. Treatment of DCM focuses on glucose control, blood pressure management, lipid-lowering, and lifestyle changes. Due to limited therapeutic options, DCM remains a significant cause of morbidity and mortality in patients with diabetes, thus emphasizing the need to develop new therapeutic strategies. Ongoing research is aimed at understanding the underlying molecular mechanism(s) involved in the development and progression of DCM, including oxidative stress, inflammation, and metabolic dysregulation. The goal is to develope innovative pharmaceutical therapeutics, offering significant improvements in the clinical management of DCM. Some of these approaches include the effective targeting of impaired insulin signaling, cardiac stiffness, glucotoxicity, lipotoxicity, inflammation, oxidative stress, cardiac hypertrophy, and fibrosis. This review focuses on the latest developments in understanding the underlying causes of DCM and the therapeutic landscape of DCM treatment.


Dendritic cytoskeletal protein expression in mental retardation: an immunohistochemical study of the neocortex in Rett syndrome.
Many syndromes associated with mental retardation (MR) are characterized by cortical dendritic anomalies. Despite their morphological similarity, these changes appear to involve different stages of dendritic development. The neuronal cytoskeleton, which includes microfilaments, neurofilaments and microtubules, is essential for these developmental processes. Levels and phosphorylation of microtubule-associated proteins (MAPs), which stabilize microtubules, seem to determine different stages of dendritic formation with certain MAPs (e.g. MAP-2) appearing to mediate the effects of external modulators upon these processes. Early studies on neuronal cytoskeleton in MR, which have shown a selective reduction in MAP-2 expression, have focused on Rett syndrome (RS). Here, by a semiquantitative immunohistochemical analysis of the pericentral cortex, we examine the contribution of specific neuronal populations to these changes in cytoskeletal proteins. Decreased MAP-2 staining in RS was more marked in layers V-VI, while increased nonphosphorylated neurofilament immunoreactivity was found in layers II-III in RS. Age-related increases in dendritic MAP-2 immunoreactivity in layers V-VI were also absent in RS. The specificity of these cytoskeletal protein changes, their significance for RS pathogenesis and plasticity, as well as their implications for other MR-associated disorders, are also discussed.


Surgical results of corrosive injuries involving esophagus to jejunum.
Severe corrosive injury involving esophagus to jejunum remains an unique surgical problem which is associated with high mortality and morbidity. Herein we report the outcomes of 28 caustic patients who underwent resections of the stomach, duodenum, a segment of jejunum, and adjacent involving organs. In all of these patients except one, esophagectomy was also performed. The concomitant procedures included pancreaticojejunostomy (n = 24), choledochojejunosotmy (n = 4), cholecystostomy (n = 4), common bile duct or pancreatic duct drainage, feeding and drainage jejunostomies, and cervical esophagostomy. Major complications consisted of bile leakage (n = 10), bile-bronchial fistula (n = 2), internal bleeding due to vessel necrosis (n = 5), peritonitis (n = 4), acute renal failure (n = 4), and septicemia (n = 4). There were 13 hospital deaths (46.4%) and three late deaths. Eight out of 12 survivors underwent subsequent reconstruction of esophagus. The remaining four survivors depended on jejunostomy feeding. Early approaches and appropriate procedures can save a number of patients with corrosive injury involving esophagus to jejunum.


The N Terminus of the PduB Protein Binds the Protein Shell of the Pdu Microcompartment to Its Enzymatic Core.
Bacterial microcompartments (MCPs) are extremely large proteinaceous organelles that consist of an enzymatic core encapsulated within a complex protein shell. A key question in MCP biology is the nature of the interactions that guide the assembly of thousands of protein subunits into a well-ordered metabolic compartment. In this report, we show that the N-terminal 37 amino acids of the PduB protein have a critical role in binding the shell of the 1,2-propanediol utilization (Pdu) microcompartment to its enzymatic core. Several mutations were constructed that deleted short regions of the N terminus of PduB. Growth tests indicated that three of these deletions were impaired MCP assembly. Attempts to purify MCPs from these mutants, followed by gel electrophoresis and enzyme assays, indicated that the protein complexes isolated consisted of MCP shells depleted of core enzymes. Electron microscopy substantiated these findings by identifying apparently empty MCP shells but not intact MCPs. Analyses of 13 site-directed mutants indicated that the key region of the N terminus of PduB required for MCP assembly is a putative helix spanning residues 6 to 18. Considering the findings presented here together with prior work, we propose a new model for MCP assembly.IMPORTANCE Bacterial microcompartments consist of metabolic enzymes encapsulated within a protein shell and are widely used to optimize metabolic process. Here, we show that the N-terminal 37 amino acids of the PduB shell protein are essential for assembly of the 1,2-propanediol utilization microcompartment. The results indicate that it plays a key role in binding the outer shell to the enzymatic core. We propose that this interaction might be used to define the relative orientation of the shell with respect to the core. This finding is of fundamental importance to our understanding of microcompartment assembly and may have application to engineering microcompartments as nanobioreactors for chemical production.


Regulation of autolysis-dependent extracellular DNA release by Enterococcus faecalis extracellular proteases influences biofilm development.
Enterococci are major contributors of hospital-acquired infections and have emerged as important reservoirs for the dissemination of antibiotic resistance traits. The ability to form biofilms on medical devices is an important aspect of pathogenesis in the hospital environment. The Enterococcus faecalis Fsr quorum system has been shown to regulate biofilm formation through the production of gelatinase, but the mechanism has been hitherto unknown. Here we show that both gelatinase (GelE) and serine protease (SprE) contribute to biofilm formation by E. faecalis and provide clues to how the activity of these proteases governs this developmental process. Confocal imaging of biofilms suggested that GelE(-) mutants were significantly reduced in biofilm biomass compared to the parental strain, whereas the absence of SprE appeared to accelerate the progression of biofilm development. The phenotype observed in a SprE(-) mutant was linked to an observed increase in autolytic rate compared to the parental strain. Culture supernatant analysis and confocal microscopy confirmed the inability of mutants deficient in GelE to release extracellular DNA (eDNA) in planktonic and biofilm cultures, whereas cells deficient in SprE produced significantly more eDNA as a component of the biofilm matrix. DNase I treatment of E. faecalis biofilms reduced the accumulation of biofilm, implying a critical role for eDNA in biofilm development. In conclusion, our data suggest that the interplay of two secreted and coregulated proteases--GelE and SprE--is responsible for regulating autolysis and the release of high-molecular-weight eDNA, a critical component for the development of E. faecalis biofilms.


Cleavage of p65/RelA of the NF-kappaB pathway by Chlamydia.
Chlamydia trachomatis is a bacterial pathogen that infects the eyes and urogenital tract. Ocular infection by this organism is the leading cause of preventable blindness worldwide. The infection is also a leading cause of sexually transmitted disease in the United States. As obligate intracellular pathogens, chlamydiae have evolved sophisticated, yet undefined, mechanisms to maintain a favorable habitat for intracellular growth while avoiding harm to the host. We show here that chlamydiae have the ability to interfere with the NF-kappaB pathway of host inflammatory response. We found that Chlamydia infection did not promote IkappaBalpha degradation, a prerequisite for NF-kappaB nuclear translocation/activation, nor induce p65/RelA nuclear redistribution. Instead, it caused p65 cleavage into an N terminus-derived p40 fragment and a p22 of the C terminus. The activity was specific because no protein cleavage or degradation of NF-kappaB pathway components was detected. Moreover, murine p65 protein was resistant to cleavage by both human and mouse biovars. The chlamydial protein that selectively cleaved p65 was identified as a tail-specific protease (CT441). Importantly, expression of either this protease or the p40 cleavage product could block NF-kappaB activation. A hallmark of chlamydial STD is its asymptomatic nature, although inflammatory cellular response and chronic inflammation are among the underlying mechanisms. The data presented here demonstrate that chlamydiae have the ability to convert a regulatory molecule of host inflammatory response to a dominant negative inhibitor of the same pathway potentially to minimize inflammation.


Crystal structure of a nicotinate phosphoribosyltransferase from Thermoplasma acidophilum.
We have determined the crystal structure of nicotinate phosphoribosyltransferase from Themoplasma acidophilum (TaNAPRTase). The TaNAPRTase has three domains, an N-terminal domain, a central functional domain, and a unique C-terminal domain. The crystal structure revealed that the functional domain has a type II phosphoribosyltransferase fold that may be a common architecture for both nicotinic acid and quinolinic acid (QA) phosphoribosyltransferases (PRTase) despite low sequence similarity between them. Unlike QAPRTase, TaNAPRTase has a unique extra C-terminal domain containing a zinc knuckle-like motif containing 4 cysteines. The TaNAPRTase forms a trimer of dimers in the crystal. The active site pocket is formed at dimer interfaces. The complex structures with phosphoribosylpyrophosphate (PRPP) and nicotinate mononucleotide (NAMN) showed, surprisingly, that functional residues lining on the active site of TaNAPRTase are quite different from those of QAPRTase, although their substrates are quite similar to each other. The phosphate moiety of PRPP and NAMN is anchored to the phosphate-binding loops formed by backbone amides, as found in many alpha/beta barrel enzymes. The pyrophosphate moiety of PRPP is located at the entrance of the active site pocket, whereas the nicotinate moiety of NAMN is located deep inside. Interestingly, the nicotinate moiety of NAMN is intercalated between highly conserved aromatic residues Tyr(21) and Phe(138). Careful structural analyses combined with other NAPRTase sequence subfamilies reveal that TaNAPRTase represents a unique sequence subfamily of NAPRTase. The structures of TaNAPRTase also provide valuable insight for other sequence subfamilies such as pre-B cell colony-enhancing factor, known to have nicotinamide phosphoribosyltransferase activity.


The molecular basis of frictional loads in the in vitro motility assay with applications to the study of the loaded mechanochemistry of molecular motors.
Molecular motors convert chemical energy into mechanical movement, generating forces necessary to accomplish an array of cellular functions. Since molecular motors generate force, they typically work under loaded conditions where the motor mechanochemistry is altered by the presence of a load. Several biophysical techniques have been developed to study the loaded behavior and force generating capabilities of molecular motors yet most of these techniques require specialized equipment. The frictional loading assay is a modification to the in vitro motility assay that can be performed on a standard epifluorescence microscope, permitting the high-throughput measurement of the loaded mechanochemistry of molecular motors. Here, we describe a model for the molecular basis of the frictional loading assay by modeling the load as a series of either elastic or viscoelastic elements. The model, which calculates the frictional loads imposed by different binding proteins, permits the measurement of isotonic kinetics, force-velocity relationships, and power curves in the motility assay. We show computationally and experimentally that the frictional load imposed by alpha-actinin, the most widely employed actin binding protein in frictional loading experiments, behaves as a viscoelastic rather than purely elastic load. As a test of the model, we examined the frictional loading behavior of rabbit skeletal muscle myosin under normal and fatigue-like conditions using alpha-actinin as a load. We found that, consistent with fiber studies, fatigue-like conditions cause reductions in myosin isometric force, unloaded sliding velocity, maximal power output, and shift the load at which peak power output occurs.


Deciphering source contributions of trace metal contamination in urban soil, road dust, and foliar dust of Guangzhou, southern China.
Trace metal contamination prevails in various compartments of the urban environment. Understanding the roles of various anthropogenic sources in urban trace metal contamination is critical for pollution control and city development. In this study, the source contribution from various contamination sources to trace metal contamination (e.g., Cu, Pb, Zn, Co, Cr and Ni) in different environmental compartments in a typical megacity, Guangzhou, southern China, was investigated using the receptor model (Absolute Principal Component Scores-Multiple Linear Regression, APCS-MLR) coupled with the Kriging technique. Lead isotopic data and APCS-MLR analysis identified industrial and traffic emissions as the major sources of trace metals in surface soil, road dust, and foliar dust in Guangzhou. Lead isotopic compositions of road dust and foliar dust exhibited similar ranges, implying their similar sources and potential metal exchange between them. Re-suspended soil contributed to 0-38% and 25-58% of the trace metals in the road dust and foliar dust, respectively, indicating the transport of the different terrestrial dust. Spatial distribution patterns implied that Cu in the road dust was a good indicator of traffic contamination, particularly with traffic volume and vehicle speed. Lead and Zn in foliar dust indicated mainly industrial contamination, which decreased from the emission source (e.g., a power plant and steel factory) to the surrounding environment. The spatial influence of industry and traffic on the contamination status of road dust/foliar dust was successfully separated from that of other anthropogenic sources. This study demonstrated that anthropogenic inputs of trace metals in various environmental compartments (e.g., urban soil, road dust, and foliar dust) can be evaluated using a combined APCS-MLR receptor model and geostatistical analysis at a megacity scale. The coupled use of APCS-MLR analysis, geostatistics, and Pb isotopes successfully deciphered the spatial influence of the contamination sources in the urban environment matrix, providing some important information for further land remediation and health risk assessment.


Exercise and osteoarthritis.
Exercise remains an extremely popular leisure time activity in many countries throughout the western world. It is widely promoted in the lay press as having salutory benefits for weight control, disease management advantages for cardiovascular disease and diabetes, in addition to improving psychological well-being amongst an array of other benefits. In contrast, however, the lay press and community perception is also that exercise is potentially deleterious to one's joints. The purpose of this review is to consider what osteoarthritis (OA) is and provide an overview of the epidemiology of OA focusing on validated risk factors for its development. In particular the role of both exercise and occupational activity in OA will be described as well as the role of exercise to the joints' tissues (particularly cartilage) and the role of exercise in disease management. Despite the common misconception that exercise is deleterious to one's joints, in the absence of joint injury there is no evidence to support this notion. Rather it would appear that exercise has positive salutory benefits for joint tissues in addition to its other health benefits.


Mercury accumulation efficiency of different biomonitors in indoor environments: the case study of the Central Italian Herbarium (Florence, Italy).
Biomonitoring studies are often employed to track airborne pollutants both in outdoor and indoor environments. In this study, the mercury (Hg) sorption by three biomonitors, i.e., Pinus nigra bark, Pseudovernia furfuracea lichen, and Hypnum cupressiforme moss, was investigated in controlled (indoor) conditions. In comparison to outdoor environments, controlled conditions offer the opportunity to investigate more in detail the variables (humidity, temperature, pollutants speciation, etc.) that control Hg uptake. The biomonitors were exposed in two distinct periods of the year for 2 and 12 months respectively, in the halls of the Central Italian Herbarium (Natural History Museum of the University of Florence, Italy), which are polluted by Hg, due to past plant sample treatments. The Hg sorption trend was monitored every 3 weeks by recording: (i) the Hg content in the substrata, (ii) gaseous elemental mercury (GEM) concentrations in the exposition halls, (iii) temperature, (iv) humidity, and (v) particulate matter (PM) concentrations. At the end of the experiment, Hg concentrations in the biomonitors range from 1130 ± 201 to 293 ± 45 μg kg-1 (max-min) in barks, from 3470 ± 571 to 648 ± 40 μg kg-1 in lichens, and from 3052 ± 483 to 750 ± 127 μg kg-1 in mosses. All the biomonitors showed the highest Hg accumulation after the first 3 weeks of exposure. Mercury concentrations increased over time showing a continuous accumulation during the experiments. The biomonitors demonstrated different Hg accumulation trends in response to GEM concentrations and to the different climatic conditions (temperature and humidity) of the Herbarium halls. Barks strictly reflected the gaseous Hg pollution, while lichen and moss accumulation was also influenced by the climatic conditions of the indoor environment. Mercury bound to PM seemed to provide a negligible contribution to the biomonitors final uptake.


Prospective study of adult onset diabetes mellitus (type 2) and risk of colorectal cancer in women.
The remarkable similarity of lifestyle and environmental risk factors for type 2 (non-insulin-dependent) diabetes mellitus and colon cancer has led to the hypothesis that diabetes may increase the risk of this cancer. We prospectively examined the relationship between diabetes and risk of colorectal cancer in a cohort of 118403 women aged 30 through 55 years who were without previously diagnosed cancer at baseline in 1976. The women, who were enrolled in the Nurses' Health Study, were assessed for history of diabetes at baseline and during follow-up by use of biennial questionnaires. Self-reported diabetes was validated by information obtained from a supplemental questionnaire on symptoms and treatment and was confirmed by medical record review in a sample of the participants. Incident cases of colorectal cancer were ascertained through medical record review. All reported P values are two-sided. During 18 years of follow-up (201061 person-years), we documented 892 new cases of colorectal cancer. After adjustment for age, body mass index (weight in kg/height in m2), physical activity, and other covariates, relative risks (RRs) were 1.43 (95% confidence interval [CI] = 1.10-1.87; P = .009) for colorectal cancer, 1.49 (95% CI = 1.09-2.06; P = .01) for colon cancer, 1.11 (95% CI = 0.56-2.21; P = .76) for rectal cancer, 1.56 (95% CI = 1.07-2.28; P = .02) for advanced colorectal cancer, and 2.39 (95% CI = 1.46-3.92; P = .0005) for fatal colorectal cancer. Our data provide support for the hypothesis that diabetes is associated with an increased risk of colorectal cancer in women.


Preserving Continuity of Behavioral Health Clinical Care to Patients Using Mobile Devices.
The current model for treating behavioral health patients requires that providers and patients be in the same location for appointments. However, deploying warfighters present a challenge to this current model. Recent advances in technology make telehealth or virtual visits a viable option to replace the current model. This project leveraged mobile technology to see if performing tele-behavioral health visits presented a viable option to the current in-person model for future deployed warfighters. At the time of this publication, the authors note the current pandemic lends all the more urgency to the need for enhancing our video communication platforms for remote monitoring with the Military Health System. The research team assessed existing Internet protocol-based desktop teleconferencing solutions, generically known as a Web Real-Time Communications (WebRTC) system, for establishing a secure connection to a Service Members personal mobile device outside of the Department of Defense (DoD) network. Of the five existing WebRTC systems evaluated, only the backbone component to the existing Defense Information Systems Agency Global Video Services (DISA GVS) known as Vidyo, was suitable to meet DoD security requirements and still connect with both major operating systems (OS) on mobile devices. An existing DoD program of record mobile application, mCare, was integrated with Vidyo desktop technologies to form what the research team called "Mobile Connect." Deployment of the Mobile Connect product yielded distinct differences and high levels or variability between the .osd.mil and the army.mil network connections over time. These network differences impacted quality of service solution where Mobile Connect could not be used to provide care between the .mil and patient's personal mobile devices from a osd.mil domain connection. The current DoD WebRTC systems offer potential solutions but presently cannot connect with personal mobile devices in their current configurations. Additionally, any WebRTC system used by the DoD for future connections to personal mobile device must leverage commercial Single Socket Layer certificates (e.g., not DoD issued), or the communications with the mobile device will fail as a result of an authentication error. It is technically feasible to provide desktop Video Tele-Conference capabilities from a .mil computer to a personal mobile device without compromising DoD security and information assurance requirements using future WebRTC systems. Approved ports, protocols, and system settings must be configured to accept both inbound and outbound, encrypted traffic to/from personal mobile devices to maintain consistent quality of service with all DoD networks. Of the current DoD WebRTC options, working with the DISA GVS Program Manager to expand services to support commercial mobile devices has the highest probability of future success.


The Effectiveness of Palate Re-Repair for Treating Velopharyngeal Insufficiency: A Systematic Review and Meta-Analysis.
Palate re-repair has been proposed as an effective treatment for velopharyngeal insufficiency (VPI) with a low risk of obstructive sleep apnea (OSA). The authors conducted a systematic review and meta-analysis to determine the proportion of patients achieving normal speech resonance following palate re-repair for VPI, the proportion developing OSA, and the criteria for patient selection that are associated with increased effectiveness. PubMed, Embase, and Scopus were searched from inception through April 2018 for English language articles evaluating palate re-repair for the treatment of VPI in patients with a repaired cleft palate. Inclusion criteria included reporting of hypernasality, nasal air emission, nasometry, additional VPI surgery, and/or OSA outcomes. Meta-analysis was conducted using random effects models. Risk of bias was assessed regarding criteria for patient selection, blinding of outcome assessors, and validity of speech assessment scale. Eighteen studies met inclusion criteria. The incidence of achieving no consistent hypernasality follow palate re-repair was 61% (95% confidence interval [CI]: 44%-75%). The incidence of additional surgery for persistent VPI symptoms was 21% (95% CI: 12%-33%). The incidence of OSA was 28% (95% CI: 13%-49%). Criteria for selecting patients to undergo re-repair varied, with anterior/sagittal position of palatal muscles (33%) and small velopharyngeal gap (22%) being the most common. No specific patient selection criteria led to superior speech outcomes (P = .6572). Palate re-repair achieves normal speech resonance in many but not all patients with VPI. Further research is needed to identify the specific examination and imaging findings that predict successful correction of VPI with re-repair.


Production of volatiles by the red seaweed Gelidium arbuscula (Rhodophyta): emission of ethylene and dimethyl sulfide.
The effects of different light conditions and exogenous ethylene on the emission of volatile compounds from the alga Gelidium arbuscula Bory de Saint-Vincent were studied. Special emphasis was placed on the possibility that the emission of ethylene and dimethyl sulfide (DMS) are related through the action of dimethylsulfoniopropionate (DMSP) lyase. The conversion of DMSP to DMS and acrylate, which is catalyzed by DMSP lyase, can indirectly support the synthesis of ethylene through the transformation of acrylate to ethylene. After mimicking the desiccation of G. arbuscula thalli experienced during low tides, the volatile compounds emitted were trapped in the headspace of 2 mL glass vials for 1 h. Two methods based on gas chromatography/mass spectrometry revealed that the range of organic volatile compounds released was affected by abiotic factors, such as the availability and spectral quality of light, salinity, and exogenous ethylene. Amines and methyl alkyl compounds were produced after exposure to white light and darkness but not after exposure to exogenous ethylene or red light. Volatiles potentially associated with the oxidation of fatty acids, such as alkenes and low-molecular-weight oxygenated compounds, accumu-lated after exposure to exogenous ethylene and red light. Ethylene was produced in all treatments, especially after exposure to exogenous ethylene. Levels of DMS, the most abundant sulfur-compound that was emitted in all of the conditions tested, did not increase after incubation with ethylene. Thus, although DMSP lyase is active in G. arbuscula, it is unlikely to contribute to ethylene synthesis. The generation of ethylene and DMS do not appear to be coordinated in G. arbuscula.


Role differentiation in copulating cicada killer wasps.
Copulating male and female cicada killer wasps have distinct behavioral roles independent of reproduction. Males terminate copulation, and females initiate the copulatory flight in which the pair in copulation escape potential danger. Separation and escape behavior are mutually exclusive. Separation occurs because the female clings to the substrate and fails to join the male in his frequent attempts at flight; thus he eventually pulls free. Escape occurs when the female begins flight, which the male readily joins. Differences in thresholds for flight probably largely determine both roles. There appears to be an evolutionary balance in escape and separation behavior determined by the behavior of the female, and illustrative of behavioral homeostasis. The female remains still in the presence of mild stimuli, such as attempted male flights, and thereby aids in separation; she initiates escape in the presence of strong stimuli such as potential enemies.


Spinal Epidural Abscess Complicated by Meningitis, Sepsis and Thrombocytopenia in a Patient Lacking Traditional Risk Factors.
Spinal epidural abscess is a rare diagnosis with a classic triad of fever, spinal pain and neurologic deficits. Only a small proportion of patients have all three findings, making the diagnosis challenging. Here we present a case of cervical and thoracic spinal epidural abscess complicated by meningitis, sepsis and thrombocytopenia in a patient lacking traditional risk factors. The patient was initially treated non-operatively secondary to thrombocytopenia but subsequently required transfer to a tertiary care facility for surgical drainage after clinical deterioration. This case report highlights the need for a high index of suspicion and low threshold for imaging when considering this rare but potentially deadly condition.


Efficacy and Safety of Ramucirumab/nab-paclitaxel for Previously Treated Advanced Gastric Cancer in Community Hospitals.
As the nanoparticle albumin-bound paclitaxel (nab-PTX) is free of ethanol and premedication, the duration of administration is shorter and patients can drive themselves to and from the hospital. In the 2018 Japanese gastric cancer treatment guidelines, ramucirumab (RAM) plus weekly nab-PTX is conditionally recommended for previously treated patients with advanced gastric cancer. Here, we retrospectively analysed the efficacy and safety of RAM+nab-PTX for such patients in community hospitals. From January 2018 to December 2019, 43 patients with metastatic and recurrent gastric cancer received RAM+nab-PTX treatment. Six patients (13.9%) were older than 80 years and 9 patients (20.9%) showed ECOG-PS 2. Progression-free survival (PFS), overall survival (OS), overall response rate (ORR), disease control rate (DCR), and adverse events (AEs) were reviewed retrospectively. Median PFS was 114 days (95% confidence interval [CI]: 84-190) and median OS was 297 days (95% CI: 180-398). ORR and DCR were 32.4% and 72.2%, respectively. The incidence rates of ≥grade 3 neutropenia and febrile neutropenia were 53.5% and 2.3%, respectively. No treatment-related deaths occurred. RAM plus nab-PTX combination therapy demonstrated manageable toxicity even patients who were elderly or had an ECOG-PS 2. This treatment is useful in community hospital settings.


Frequent attenders' consulting patterns with general practitioners.
Despite the growing literature on frequent attendance, little is known about the consulting patterns of frequent attenders with different doctors. To develop appropriate intervention strategies and to improve the clinical care of frequent attenders, a full understanding of these consulting patterns is essential. This paper has three aims: to determine whether frequent attenders consult more with some doctors than others; to determine how many different doctors frequent attenders consult with; and to determine whether frequent attenders exhibit greater continuity of care than non-frequent attenders. Analysis of a validated dataset of 592,028 consultations made by 61,055 patients from four practices over 41 months. Comparisons between the consulting patterns of the frequent attenders, defined as the most frequently consulting 3% of the population by practice, with non-frequent attenders and the overall practice populations. There was considerable variation in the numbers and proportions of consultations with frequent attenders between individual doctors. Most of the frequent attenders consulted with most or all of the doctors within practices over the timeframe. Frequent attenders exhibited more continuity of care than non-frequent attenders. The reasons why some doctors have more consultations with frequent attenders is unclear. Some doctors may actively encourage frequent attendance. While many frequent attenders have clear allegiances to one doctor, many also consult widely with a large number of doctors. The consequences of such behaviour are unknown. These findings have important implications in the development of appropriate interventions for reducing problematic frequent attendance.


[Pain relief by low-intensity frequency-modulated millimeter waves acting on the acupuncture points].
Analgetic effect of low-intensive frequency-modulated millimetric waves (MW) was studied in mice with formalin induced nociceptive behavior reaction (licking of defeat hindpaw). MW were applied to the acupoint E 36 of the defeat hindpaw. The following MW were used: 60 GHz (1) and 118 GHz (2) which were modulated by 4 Hz; noise MW within the range of 42-95 GHz (3) and 90-140 GHz (4) which were modulated in accidental order by frequencies 1-60 Hz; combinations of fixed frequencies with noise - 60 GHz + noise 42-95 GHz (5) and 118 GHz + noise 90-140 GHz (6). All used MW combinations suppressed licking of the defeat hindpaw and increased duration of sleep and eating. The strongest analgesia was achieved in series 1-3 (42.4-69.7%), the weakest in series 6 and 4 of the experiment (12.2-19.7%).


[The action of pilocarpine on the accommodation properties of isolated phasic neurons of the rat hippocampus].
Using of voltage and current clamp methods in conditions of perforated membrane fragment, we investigated the accommodation properties of isolated neurones from different zones of hippocampus. The investigated neurones could be separated into two main groups concerning their responses to prolonged pulses of suprathreshold depolarization--neurones which after generation of several action potentials (AP) stopped their generation (phasic, F-neurones), and (tonic, T-neurones), which did not accommodate and continued AP generation during entire depolarizing pulse. Comparison of ionic current composition in neurones of both groups has shown, that T-neurones demonstrate more expressed inactivation of outward currents that can be due to the presence of a fast inactivating A-type potassium current, or decreased inactivation of inward currents. Pilocarpine, well-known epileptogenic compound increased accommodation and induced significant slow-down of outward current inactivation in F- neurones. It can be suggested that the presence of A-type potassium current and/or a slowly inactivating component of inward current can play main role in determining the accommodation effect in hippocampal neurones.


Does the Severity of the Curve in Lenke 1 and 2 Adolescent Idiopathic Scoliosis Patients Affect the Distance and Position of the Aorta From Vertebra?
Retrospective study of a prospectively-collected data. To determine whether the severity of the curve magnitude in Lenke 1 and 2 Adolescent Idiopathic Scoliosis (AIS) patients affects the distance and position of the aorta from the vertebra. There were studies that looked into the position of the aorta in scoliotic patients but none of them documented the change in distance of the aorta to the vertebra in relation to the magnitude of the scoliosis. Patients with Lenke 1 and 2 AIS who underwent posterior spinal fusion using pedicle screw construct and had a preoperative computed tomography (CT) scan performed were recruited. The radiological parameters measured on preoperative CT scan were: Aortic-Vertebral Distance (AVD), Entry-Aortic Distance (EAD), Aortic-Vertebral angle (AVA), Pedicle Aorta angle/Aortic Alpha angle (α angle), and Aortic Beta angle (β angle). Thirty-nine patients were recruited. Significant moderate to strong positive correlation was found between AVD and Cobb angle from T8 to T12 vertebrae (r = 0.360 to 0.666). The EAD was generally small in the thoracic region (T4-T10) with mean EAD of less than 30 mm. Among all apical vertebrae, the mean AVD was 5.9 ± 2.2 mm with significant moderate-strong positive correlation to Cobb angle (r = 0.580). The mean α angle was 37.7 ± 8.7° with significant weak positive correlation with Cobb angle (r = 0.325). The larger the scoliotic curve, the aorta was located further away from the apical vertebral wall. The aorta has less risk of injury from the left lateral pedicle screw breach in larger scoliotic curve at the apical region. The distance from the pedicle screw entry point to the wall of the aorta was generally small (less than 30 mm) in the thoracic region (T4-T10). 4.


Role of Ca2+-independent phospholipase A2gamma in Ca2+-induced mitochondrial permeability transition.
Our laboratory previously demonstrated Ca2+-independent phospholipase A2gamma (iPLA2gamma) is localized to mitochondria and that iPLA2 inhibition blocks cisplatin-induced caspase-mediated apoptosis. Whereas the mitochondrial permeability transition (MPT) is a key control point for apoptosis, the role of mitochondrial iPLA2gamma in MPT has not been established. In the present study, we addressed this issue. Ca2+-induced renal cortex mitochondrial (RCM) swelling was blocked by the MPT inhibitor cyclosporine A. The R-isomer of bromoenol lactone (R-BEL), which enantiospecifically inhibits iPLA2gamma, inhibited Ca2+-induced RCM MPT, whereas S-BEL (negative control) had no effect. Ca2+ treatment resulted in a significant increase in free arachidonic acid (AA) (>50 microM) in the RCM suspension that was blocked by pretreatment with BEL. No increases in free myristic, palmitic, stearic, oleic, linoleic, or docosahexaenoic acid were detected after Ca2+ treatment. The addition of AA (18 microM) to Ca2+-treated RCM with inhibited iPLA2gamma activity restored MPT. We also determined that RCM iPLA2gamma displays higher activity against plasmenylcholine with AA in the sn-2 position than oleic acid. Ca2+ exposure significantly increased RCM iPLA2gamma activity; however, the Ca2+-induced activation of iPLA2gamma was not the result of mitochondrial membrane potential dissipation, opening of the MPT pore, or mitochondrial swelling. Taken together these findings provide strong evidence that Ca2+-induced RCM MPT is mediated by iPLA2gamma-catalyzed AA liberation.