Pediatric Critical Care Neurologists in the United States and Canada: A Survey of Clinical Practice Experience.
To describe the characteristics of pediatric intensive care neurologists and their practice in the United States and Canada. We performed a survey-based study of child neurologists who self-identify as 'intensive care neurologists'. The survey included questions about demographics, training, pediatric neurocritical care service and job structure, teaching, academics, challenges, and views on the future of pediatric neurocritical care. We analyzed 55 surveys. Most respondents were 31-50 years of age with ≤10 years of practice experience. Fifty-four percent identified as female. Most completed subspecialty training after child neurology residency. The majority practice at highly resourced centers with >45 intensive care unit beds. Respondents cover a variety of inpatient (critical and noncritical care) services, at times simultaneously, for a median of 19.5 weeks/y and work >70 hours/wk when on service for pediatric neurocritical care. The top 3 challenges reported were competing demands for time, excess volume, and communication with critical care medicine. Top priorities for the "ideal pediatric neurocritical care service" were attendings with training in pediatric neurocritical care or a related field and joint rounding with critical care medicine. We report a survey-based analysis of the demographics and scope of practice of pediatric critical care neurologists. We highlight challenges faced and provide a framework for the further development of this rapidly growing field.


Mithramycin encapsulated in polymeric micelles by microfluidic technology as novel therapeutic protocol for beta-thalassemia.
This report shows that the DNA-binding drug, mithramycin, can be efficiently encapsulated in polymeric micelles (PM-MTH), based on Pluronic(®) block copolymers, by a new microfluidic approach. The effect of different production parameters has been investigated for their effect on PM-MTH characteristics. The compared analysis of PM-MTH produced by microfluidic and conventional bulk mixing procedures revealed that microfluidics provides a useful platform for the production of PM-MTH with improved controllability, reproducibility, smaller size, and polydispersity. Finally, an investigation of the effects of PM-MTH, produced by microfluidic and conventional bulk mixing procedures, on the erythroid differentiation of both human erythroleukemia and human erythroid precursor cells is reported. It is demonstrated that PM-MTH exhibited a slightly lower toxicity and more pronounced differentiative activity when compared to the free drug. In addition, PM-MTH were able to upregulate preferentially γ-globin messenger ribonucleic acid production and to increase fetal hemoglobin (HbF) accumulation, the percentage of HbF-containing cells, and their HbF content without stimulating α-globin gene expression, which is responsible for the clinical symptoms of β-thalassemia. These results represent an important first step toward a potential clinical application, since an increase in HbF could alleviate the symptoms underlying β-thalassemia and sickle cell anemia. In conclusion, this report suggests that PM-MTH produced by microfluidic approach warrants further evaluation as a potential therapeutic protocol for β-thalassemia.


Cognitive Dysfunction in Insomnia Phenotypes: Further Evidence for Different Disorders.
Study Objectives: To determine cognitive profiles in individuals with short sleep duration insomnia (SSDI) and normal sleep duration insomnia (NSDI; also, paradoxical insomnia), compared to healthy sleepers. Method: Polysomnographic (PSG) and neuropsychological data were analysed from 902 community-based Raine Study participants aged 22 ± 0.6 years of whom 124 met criteria for insomnia (53 with NSDI and 71 with or SSDI) and 246 were classified as healthy with normal sleep (i.e., without insomnia or other sleep disorders). Measurements of self- report (attention and memory) and laboratory-assessed (attention, episodic memory, working memory, learning, and psychomotor function) cognition and mood, and PSG-based sleep stages (% total sleep time; %TST) were compared between these 3 groups. Results: In comparison to the healthy sleeper group, both insomnia groups had poorer self-reported attention, memory, mood, and sleep, and poorer laboratory-assessed attention (inconsistency). The NSDI group had less consistent working memory reaction time than healthy-sleepers or those with SSDI. The SSDI group had more inconsistency in executive function (shifting), and showed greater %TST in stage N1 and N3, and less REM sleep than either healthy-sleepers or those with NSDI. Conclusions: Individuals with NSDI demonstrated greater working memory inconsistency, despite no laboratory assessed sleep problems, implicating early signs of pathophysiology other than disturbed sleep. Those with SSDI demonstrated different sleep architecture, poorer attention (inconsistency), and greater executive function (inconsistency) compared to healthy-sleepers and those with NSDI, implicating sleep disturbance in the disease process of this phenotype.


[Interrupted alcohol treatment and liver: free radical homeostasis, nitric oxide, adaptive mechanisms].
Alcohol administration can result in liver damage. Reactive oxygen species (ROS), nitric oxide (NO) and their interaction are crucial factors in this process. The aim of work was to investigate, free radical state and mechanisms of adaptation of the antioxidant system (AOS) to stress, caused by interrupted alcohol intake. Repeated cycles of alcoholization caused an imbalance between production and utilization of various ROS. This imbalance was due to impairments in the system superoxide dismutase/catalase. Nevertheless, in most experimental groups there was clear reduction of lipid peroxidation (LPO) products evaluated by thiobarbituric acid reactive substances. This might be attributed to the antioxidant effect of NO. However, there was an increased level of transaminases in blood plasma. After 28 days of this experimental scheme all the parameters studied normalized.


Loss of Endothelial Cytochrome P450 Reductase Induces Vascular Dysfunction in Mice.
POR (cytochrome P450 reductase) provides electrons for the catalytic activity of the CYP (cytochrome P450) monooxygenases. CYPs are dual-function enzymes as they generate protective vasoactive mediators derived from polyunsaturated fatty acids but also reactive oxygen species. It is not known in which conditions the endothelial POR/CYP system is beneficial versus deleterious. Here, the activity of all CYP enzymes was eliminated in the vascular endothelium to examine its impact on vascular function. An endothelial-specific, tamoxifen-inducible POR knockout mouse (ecPOR-/-) was generated. Vascular function was studied by organ chamber experiments. eNOS (endothelial nitric oxide synthase) activity was accessed by heavy arginine/citrulline LC-MS/MS detection and phosphorylation of serine1177 in aortic rings. CYP-derived epoxyeicosatrienoic acids and prostanoids were measured by LC-MS/MS. Gene expression of aorta and endothelial cells was profiled by RNA sequencing. Blood pressure was measured by telemetry. Acetylcholine-induced endothelium-dependent relaxation was attenuated in isolated vessels of ecPOR-/- as compared with control mice. Additionally, ecPOR-/- mice had attenuated eNOS activity and eNOS/AKT phosphorylation. POR deletion reduced endothelial stores of CYP-derived epoxyeicosatrienoic acids but increased vascular prostanoids. This phenomenon was paralleled by the induction of genes implicated in eicosanoid generation. In response to Ang II (angiotensin II) infusion, blood pressure increased significantly more in ecPOR-/- mice. Importantly, the cyclooxygenase inhibitor Naproxen selectively lowered the Ang II-induced hypertension in ecPOR-/- mice. POR expression in endothelial cells maintains eNOS activity and its loss results in an overactivation of the vasoconstrictor prostanoid system. Through these mechanisms, loss of endothelial POR induces vascular dysfunction and hypertension.


Catalytic and framework mutations in the neuraminidase active site of influenza viruses that are resistant to 4-guanidino-Neu5Ac2en.
Here we report the isolation of influenza virus A/turkey/Minnesota/833/80 (H4N2) with a mutation at the catalytic residue of the neuraminidase (NA) active site, rendering it resistant to the novel NA inhibitor 4-guanidino-Neu5Ac2en (GG167). The resistance of the mutant stems from replacement of one of three invariant arginines (Arg 292-->Lys) that are conserved among all viral and bacterial NAs and participate in the conformational change of sialic acid moiety necessary for substrate catalysis. The Lys292 mutant was selected in vitro after 15 passages at increasing concentrations of GG167 (from 0.1 to 1,000 microM), conditions that earlier gave rise to GG167-resistant mutants with a substitution at the framework residue Glu119. Both types of mutants showed similar degrees of resistance in plaque reduction assays, but the Lys292 mutant was more sensitive to the inhibitor in NA inhibition tests than were mutants bearing a substitution at framework residue 119 (Asp, Ala, or Gly). Cross-resistance to other NA inhibitors (4-amino-Neu5Ac2en and Neu5Ac2en) varied among mutants resistant to GG167, being lowest for Lys292 and highest for Asp119. All GG167-resistant mutants demonstrated markedly reduced NA activity, only 3 to 50% of the parental level, depending on the particular amino acid substitution. The catalytic mutant (Lys292) showed a significant change in pH optimum of NA activity, from 5.9 to 5.3. All of the mutant NAs were less stable than the parental enzyme at low pH. Despite their impaired NA activity, the GG167-resistant mutants grew as well as parental virus in Madin-Darby canine kidney cells or in embryonated chicken eggs. However, the infectivity in mice was 500-fold lower for Lys292 than for the parental virus. These findings demonstrate that amino acid substitution in the NA active site at the catalytic or framework residues, followed by multiple passages in vitro, in the presence of increasing concentrations of the NA inhibitor GG167, generates GG167-resistant viruses with reduced NA activity and decreased infectivity in animals.


Why and How Schools Make Nutrition Education Programs "Work".
There are many potential health benefits to having nutrition education programs offered by expert outside sources in schools. However, little is known about why and how schools initiate, implement, and institutionalize them. Gaining this understanding may allow the impact and reach of nutrition and other health education programs in schools to be extended. A total of 22 school community members from 21 purposefully selected New York City public elementary schools were interviewed using a semistructured interview protocol about their schools' experiences initiating, implementing, and institutionalizing nutrition education programs. Interviews were audiotaped and transcribed. Chronological narratives were written detailing each school's experience and passages highlighting key aspects of each school's experience were identified. These passages (N = 266) were sorted into domains and themes which were regrouped, resorted, and adjusted until all researchers agreed the domains and themes represented the collective experiences of the schools. The interviews elicited 4 broad domains of action: building motivation, choosing programs, developing capacity, and legitimizing nutrition education. Within each domain, themes reflecting specific actions and thoughts emerged. The identified domains of action and their themes highlight specific, practical actions that school health advocates can use to initiate, implement, and institutionalize nutrition education programs in schools.


Metabolism and covalent binding of vic-dihaloalkanes, vinyl halides and acrylonitrile.
The roles of various metabolic pathways in DNA and protein alkylation are discussed here. Simple vinyl halides are oxidized to 2-haloethylene oxides and 2-haloacetaldehydes, which alkylate DNA and proteins, respectively. Polysubstituted vinyl halides are oxidized with group transfer to yield halocarbonyl compounds which alkylate proteins. Oxidation of vic-dihaloalkanes results in protein alkylation while glutathione conjugates alkylate DNA. Acrylonitrile, without previous activation, alkylates proteins and glutathione. Oxidation of acrylonitrile yields a relatively stable epoxide which can react with DNA in vitro, but alkylation by this epoxide does not occur readily in vivo. Hard-soft acid-base theory is of some use in understanding why some adducts are formed in preference to others.


Production of humic acids from oil palm empty fruit bunch by submerged fermentation with Trichoderma viride: cellulosic substrates and nitrogen sources.
The novelty of this study was to produce humic acids by submerged fermentation of empty fruit bunch (EFB) with Trichoderma viride and to investigate the effects of the cellulosic substrates and the organic sources of nitrogen on the biotechnological production of these acids. The results obtained indicate the potential application of EFB, a waste of oil palm processing, for humic acids production. Because EFB contains cellulose, hemicellulose and lignin, fermentations were also performed using these polymers as carbon sources, separately or in combination. After 120 h of fermentation, significant production of humic acids was observed only in cultures containing either EFB or a mixture of the three polymers. Use of either potato peptone or yeast extract as a nitrogen source yielded nearly identical patterns of fungal growth and production of humic acids. The data obtained from microscopic imaging of T. viride growth and sporulation in EFB, coupled with the determined rates of production of humic acids indicated that the production of these acids is related to T. viride sporulation.


The 1994 G. Brohée Lecture (Los Angeles). The scientification of gastroenterology during the 20th century.
At the occasion of the Georges Brohée Lecture presented at the 10th World Congress of Gastroenterology in Los Angeles (1994), the author makes a brief history of Georges Brohée (1887-1957), Belgian Surgeon and Radiologist who promoted modern Gastroenterology, in particular by founding the Belgian Society of Gastroenterology in 1928 and by organizing the first International Congress of Gastroenterology in Brussels in 1935. His continuing efforts culminated in the constitution of the "Organisation Mondiale de Gastro-entérologie" (OMG) which prepared the first World Congress of Gastroenterology for 1958. In the second part, the author continues his description of the scientific development of Gastroenterology during the 20th century. The following topics are treated: gastrointestinal endoscopy, enteric microflora with emphasis on the discovery of Helicobacter pylori, of Tropheryma Whippleii and of antibacterial agents, gastrointestinal immunology.


Insulin-like growth factor and growth hormone receptor in nephrotic rats.
In an attempt to elucidate the mechanism of growth retardation in the nephrotic syndrome, specific serum and hepatic growth factors were measured in Sprague-Dawley rats in which nephrotic syndrome was produced by administration of puromycin (1.5 mg.100 g body wt-1.day-1) for 12 days. On the 13th day, the results of these nephrotic animals were compared with those of an equal number of pair-fed and control animals: the mean dietary intake of the nephrotic group was 71% that of the control group (P < 0.001). Serum insulin-like growth factor (IGF) binding protein-3 was significantly reduced (P < 0.005) in the nephrotic rats, compared with the pair-fed and the control groups. Hepatic IGF-I mRNA in the nephrotic rats averaged 36% that of control (P < 0.001) and 46% that of the pair-fed animals (P < 0.001). Hepatic growth hormone receptor (GHr) mRNA in the nephrotic rats averaged 19% of that of the control (P < 0.001) and 27% of that of the pair-fed rats (P < 0.001). These results indicate that the growth retardation of the nephrotic rats may be associated with the significant decrease in IGF-I mRNA and reduction in GHr mRNA.


Molecular engineering of DNA: molecular beacons.
Molecular beacons (MBs) are specifically designed DNA hairpin structures that are widely used as fluorescent probes. Applications of MBs range from genetic screening, biosensor development, biochip construction, and the detection of single-nucleotide polymorphisms to mRNA monitoring in living cells. The inherent signal-transduction mechanism of MBs enables the analysis of target oligonucleotides without the separation of unbound probes. The MB stem-loop structure holds the fluorescence-donor and fluorescence-acceptor moieties in close proximity to one another, which results in resonant energy transfer. A spontaneous conformation change occurs upon hybridization to separate the two moieties and restore the fluorescence of the donor. Recent research has focused on the improvement of probe composition, intracellular gene quantitation, protein-DNA interaction studies, and protein recognition.


Bilateral locked posterior shoulder dislocation overlooked for 15 months treated with the modified McLaughlin procedure: A case report.
Neglected bilateral posterior shoulder dislocation is a rare injury caused primarily by an epileptic seizure. The injury is usually associated with a reverse Hill-Sachs lesion in the anteromedial aspect of the humeral head. The modified McLaughlin technique may avoid instability and osteoarthritis when 20 to 40% of the articular surface is affected by reverse Hill-Sachs. In this article, we present the clinical results of a case overlooked in the literature for the longest time, i.e., for 15 months. A 46-year-old male patient was receiving treatment for epilepsy for five years. There was no fall or trauma in the four seizures he had during this time. The last seizure was 15 months ago. When the patient presented to our clinic, both shoulders were symmetrically deformed, the anterior shoulder contour disappeared (empty socket sign), and there was a loss of upper contour. The computed tomography (CT) scan revealed a posteriorly locked dislocation with a reverse Hill-Sachs lesion in 32% of the left shoulder and 36% of the right shoulder. We applied the modified McLaughlin procedure to the dominant right shoulder and, two months later, we used it to the left shoulder (with a graft taken from the anterior superior iliac spine). At one-year of follow-up, both shoulders were moderately functional: forward elevation left 70° and right 50°, abduction left 40° and right 60°, and internal rotation: the back of the hand could touch the fifth lumbar vertebra. Meanwhile, the patient did not suffer from recurrent dislocation. The pre- and postoperative Constant-Murley Scores for the right and left shoulder were 30/52 and 11/48, respectively. Although the operational outcomes using the modified McLaughlin technique were not ideal, with no recurrence, the patient seemed to be satisfied with this outcome. In conclusion, in neglected locked shoulder fracture-dislocations, the modified McLaughlin technique is a method that can respond to the pathophysiology by eliminating reverse Hill-Sachs lesion.


Monitoring the hepatitis A virus in urban wastewater from Rio de Janeiro, Brazil.
Hepatitis A is a viral disease with a significant public health impact, especially in developing countries. Improvements in sewage services could prevent hepatitis A virus (HAV) dissemination into the environment and minimize the risk of infection. The aim of this study was to monitor HAV for one year in urban sewage samples from the largest wastewater treatment plant in Rio de Janeiro, Brazil, to assess environmental contamination with HAV and its dissemination after treatment by an activated sludge process. For this purpose, 48 samples (24 raw sewage samples and 24 treated effluent samples) were collected from August 2009 to July 2010 for HAV detection. Using quantitative real-time PCR 14 (58%) raw sewage samples were positive for HAV, and the highest viral genome loads were detected in the spring and summer. HAV was not detected in treated effluent samples, which suggests that the viral loads observed could be easily removed by the activated sludge process, thus preventing the dissemination of HAV into the environment. All of the HAV strains sequenced belonged to subgenotype IA, which clustered closely with Brazilian and Argentine HAV strains. These data demonstrate that environmental monitoring can be a useful tool in epidemiological studies.


Six3 repression of Wnt signaling in the anterior neuroectoderm is essential for vertebrate forebrain development.
In vertebrate embryos, formation of anterior neural structures requires suppression of Wnt signals emanating from the paraxial mesoderm and midbrain territory. In Six3(-/-) mice, the prosencephalon was severely truncated, and the expression of Wnt1 was rostrally expanded, a finding that indicates that the mutant head was posteriorized. Ectopic expression of Six3 in chick and fish embryos, together with the use of in vivo and in vitro DNA-binding assays, allowed us to determine that Six3 is a direct negative regulator of Wnt1 expression. These results, together with those of phenotypic rescue of headless/tcf3 zebrafish mutants by mouse Six3, demonstrate that regionalization of the vertebrate forebrain involves repression of Wnt1 expression by Six3 within the anterior neuroectoderm. Furthermore, these results support the hypothesis that a Wnt signal gradient specifies posterior fates in the anterior neural plate.


Antimicrobial and Phytotoxic Activity of Origanum heracleoticum and O. majorana Essential Oils Growing in Cilento (Southern Italy).
There is a growing interest in a potential use of essential oils (EOs) as a replacement for traditional pesticides and herbicides. The aims of this study were to: (i) Identify the chemical composition of the two EOs derived from Origanum heracleoticum L. and O. majorana L., (ii) evaluate the in vitro antifungal activity of the EOs against some postharvest phytopathogens (Botrytis cinerea, Penicillium expansum, Aspergillus niger and Monilinia fructicola), (iii) evaluate the in vitro antibacterial activity against Bacillus megaterium, Clavibacter michiganensis, Xanthomonas campestris, Pseudomonas fluorescens and P. syringae pv. phaseolicola, (iv) evaluate the effect of both studied EOs on the spore germination percentage and their minimum inhibitory concentration (MIC) against M. fructicola, and (v) study the possible phytotoxicity of the two EOs and their major constituents, carvacrol for O. heracleoticum and terpinen-4-ol for O. majorana, against tha germination and initial radicle growth of radish, lettuce, garden cress and tomato. The two EOs demonstrated promising in vitro antimicrobial and antifungal activities against all tested microorganisms. EOs showed high inhibition of spore germination percentage at the minimal inhibitory concentration of 500 and 2000 µg/mL, respectively. Moreover, both germination and radical elongation of selected plant species were sensitive to the oils.


Sodium alginate microencapsulation improves the short-term oral bioavailability of cannabidiol when administered with deoxycholic acid.
Cannabidiol (CBD) confers therapeutic effects in some neurological disorders via modulation of inflammatory, oxidative and cell-signalling pathways. However, CBD is lipophilic and highly photooxidative with low oral bioavailability in plasma and brain. In this study, we aimed to design and test a CBD microencapsulation method as a drug delivery strategy to improve the absorption of CBD. Additionally, we evaluated the brain uptake of CBD capsules when administered alongside capsules containing a permeation-modifying bile acid, deoxycholic acid (DCA). Microcapsules containing either CBD or DCA were formed using the ionic gelation method with 1.5% sodium alginate formulations and 100 mM calcium chloride. C57BL/6J wild type mice randomly assigned to three treatment groups (3-4 mice per group) were administered CBD in the following preparations: 1) CBD capsules, 2) CBD capsules + DCA capsules and 3) naked CBD oil (control). To assess the short-term bioavailability of CBD, plasma and brain samples were collected at 0.3, 1 and 3 hours post administration and CBD levels were analysed with liquid chromatography mass spectrometer. We produced spherical capsules at 400 ± 50 μm in size. The CBD capsules were calculated to have a drug loading of 2% and an encapsulation efficiency of 23%. Mice that received CBD capsules + DCA capsules showed a 40% and 47% increase in CBD plasma concentration compared to mice on CBD capsules and naked CBD oil, respectively. Furthermore, the CBD capsules + DCA capsules group showed a 48% and 25% increase in CBD brain concentration compared to mice on CBD capsules and naked CBD oil, respectively. In mice treated with CBD capsules + DCA capsules, the brain CBD concentration peaked at 0.3 hours with a 300% increased availability compared to CBD capsules and naked CBD oil groups, which peaked at 1 hour after administration. The microencapsulation method combined with a permeation enhancer, DCA increased the short-term bioavailability of CBD in plasma and brain.


Covariate-based constrained randomization of group-randomized trials.
Group-randomized study designs are useful when individually randomized designs are either not possible, or will not be able to estimate the parameters of interest. Blocked and/or stratified (for example, pair-matched) designs have been used, and their properties statistically evaluated by many researchers. Group-randomized trials often have small numbers of experimental units, and strong, geographically induced between-unit correlation, which increase the chance of obtaining a "bad" randomization outcome. This article describes a procedure--random selection from a list of acceptable allocations--to allocate treatment conditions in a way that ensures balance on relevant covariates. Numerous individual- and group-level covariates can be balanced using exact or caliper criteria. Simulation results indicate that this method has good frequency properties, but some care may be needed not to overly constrain the randomization. There is a trade-off between achieving good balance through a highly constrained design, and jeopardizing the appearance of impartiality of the investigator and potentially departing from the nominal Type I error.


Hematopoietic stem cell transplantation without in vivo T-cell depletion for pediatric aplastic anemia: A single-center experience.
For young patients, HLA-MRD HSCT is the first-line treatment of SAA. However, due to China's birth control policy, few patients could find suitable sibling donors and HLA-MUD. More and more transplantation centers have used Haplo-D as the donor source for young adult and pediatric patients. However, studies with larger amount of pediatric patients are rare. We retrospectively analyzed the data of children with AA who were treated with allogeneic HSCT and compared the therapeutic efficacy of Haplo-HSCT and MRD/MUD group. A total of 62 patients were enrolled. Implantation was successfully performed in 58 patients. There was no significant difference in the time for reconstruction of hematopoietic function between patients in the two groups. Thirty-two had grade I-IV aGVHD with incidence of 51.61%. The incidence of aGVHD was 79.41% for patients in the Haplo-HSCT, significantly higher than that of 17.86% for patients in the MRD/MUD group (P < .01). However, the incidence of cGVHD was not significantly different between patients in the two groups (26.47% vs 10.71%, P = .09), the incidence of CMV infection was 28.57% and 52.94% for patients in the MRD/MUD and Haplo group, respectively, showing no significant difference (P = .053). The incidence of EBV infection was 47.06% for patients in the Haplo group and 28.57% for patients in the MRD/MUD group, showing no significant difference (P = .11). However, the 3- and 5-year cumulative OS and FFS rates showed statistically significant difference in the two groups, P = .012 and .045, respectively. Compared to Haplo-HSCT, MRD/MUD is more economic. In this study, we achieved good Haplo transplantation results. The incidences of cGVHD and CMV/EBV were not significantly different between Haplo group and MRD/MUD group. Although OS and FFS of the Haplo group were not as good as those of the MRD/MUD group, it is still acceptable as an alternative treatment under emergency.


The association between reasons for first using cannabis, later pattern of use, and risk of first-episode psychosis: the EU-GEI case-control study.
While cannabis use is a well-established risk factor for psychosis, little is known about any association between reasons for first using cannabis (RFUC) and later patterns of use and risk of psychosis. We used data from 11 sites of the multicentre European Gene-Environment Interaction (EU-GEI) case-control study. 558 first-episode psychosis patients (FEPp) and 567 population controls who had used cannabis and reported their RFUC.We ran logistic regressions to examine whether RFUC were associated with first-episode psychosis (FEP) case-control status. Path analysis then examined the relationship between RFUC, subsequent patterns of cannabis use, and case-control status. Controls (86.1%) and FEPp (75.63%) were most likely to report 'because of friends' as their most common RFUC. However, 20.1% of FEPp compared to 5.8% of controls reported: 'to feel better' as their RFUC (χ2 = 50.97; p < 0.001). RFUC 'to feel better' was associated with being a FEPp (OR 1.74; 95% CI 1.03-2.95) while RFUC 'with friends' was associated with being a control (OR 0.56; 95% CI 0.37-0.83). The path model indicated an association between RFUC 'to feel better' with heavy cannabis use and with FEPp-control status. Both FEPp and controls usually started using cannabis with their friends, but more patients than controls had begun to use 'to feel better'. People who reported their reason for first using cannabis to 'feel better' were more likely to progress to heavy use and develop a psychotic disorder than those reporting 'because of friends'.


Effects of Cancer Genetic Panel Testing on at-Risk Individuals.
To evaluate the role of screening patients at increased risk for hereditary cancer syndromes with an extended panel of cancer predisposition genes to identify actionable genetic mutations. A retrospective chart review was conducted of all patients presenting to a multidisciplinary cancer program for genetic counseling and testing from January 2015 to December 2016. Individuals presenting to the program were identified as at-risk by a personal or family history of cancer, by their health care provider, or by self-referral. All participants met current National Comprehensive Cancer Network criteria for genetic risk evaluation for hereditary cancer. The results of testing and its implications for management, based on National Comprehensive Cancer Network guidelines, were recorded. Of 670 at-risk patients who underwent genetic testing, 66 (9.9%) had BRCA-limited testing; of these, 26 of 670 (3.9%) had a deleterious or likely pathogenic mutation. Expanded panel testing was done for 560 of the 670 patients (83.4%), and abnormal results were found in 65 of 670 (9.7%); non-BRCA mutations (predominantly CHEK2) were found in 49 of the 65 (75%). Abnormal genetic testing was associated with increased surveillance in 96% of those with deleterious mutations, whereas negative testing for a known familial mutation in 45 patients was associated with a downgrade of their risk and reduction of subsequent surveillance and management. Guideline-based management is frequently altered by genetic testing, including panel testing, in patients at risk for cancer. We recommend that obstetrics and gynecology providers routinely refer at-risk patients for genetic counseling and testing when clinically appropriate.


Transcriptional activation of cathepsin D gene expression by growth factors.
Insulin-like growth factor-I (IGF-I), transforming growth factor alpha (TGFalpha) and epidermal growth factor (EGF) induced cathepsin D gene expression and reporter gene activity in MCF-7 human breast cancer cells transiently transfected with a construct (pCD1) containing a -2576 to -124 cathepsin D gene promoter insert. In contrast, IGF-I, but not TGFalpha or EGF, induced reporter gene activity in cells cotransfected with wild-type estrogen receptor (ER) expression plasmid and a construct (pCD2) containing estrogen-responsive downstream elements from -208 to -101. Promoter deletion and mutational analysis experiments identified four GC-rich sites and an imperfect palindromic estrogen responsive element required for IGF-I activation of the ER (ligand-independent). Subsequent studies with the mitogen-activated protein kinase (MAPK) inhibitor, PD98059, and a serine(118(-ER mutant confirmed the role of the MAPK pathway for IGF-I activation of the ER in MCF-7 cells. Thus, growth factor activation of ER can mediate transactivation vs ER/Sp1 binding to GC-rich sites and represents a novel pathway for ligand-independent ER action. The divergent pathways for IGF-I and TGFalpha/EGF activation of the ER observed in MCF-7 cells contrast with previous data indicating that pathways for growth factor activation of the ER are dependent on the gene and/or gene promoter and on cell context.


Surgery for lung adenocarcinoma with smokers' polycythemia: a case report.
Smoking is a cause of cancer and polycythemia. Therefore, surgeons who treat patients with cancer may also encounter patients with polycythemia. However, few cases of surgical patients with polycythemia have been reported; in particular, a surgical case involving smokers' polycythemia has never been reported. We herein report a patient with lung cancer and smokers' polycythemia who successfully underwent lobectomy with control of hematocrit based on a modified formula in the perioperative period. A 67-year-old man underwent abdominoperineal resection for rectal carcinoma in June 2008. A ground glass opacity had been identified in the upper lobe of the right lung and was gradually enlarging. In March 2012, bronchoscopic cytology for investigation of the mass revealed non-small cell lung cancer, suggesting primary lung non-small cell carcinoma (T1bN0M0, Stage IA). When he was referred to our hospital for surgery, his complete blood count showed a red blood cell level of 6.50×106/μL, hemoglobin of 21.0 g/dL, and hematocrit of 60.1%. The hematologists' diagnosis was secondary polycythemia due to heavy smoking (smokers' polycythemia) because the white blood cell and platelet counts were within normal limits and the erythropoietin was not increased. We calculated the appropriate phlebotomy and infusion volumes based on a formula that we modified. After 550 g of blood was phlebotomized to reduce the hematocrit to approximately 55%, video-assisted right lung upper lobectomy with lymph node dissection was performed in April 2012. The hematocrit was maintained at <50% postoperatively, and the patient was uneventfully discharged on postoperative day 7. The predictive hematocrit and measured hematocrit were very closely approximated in this case. We experienced a patient with smokers' polycythemia who underwent right upper lobectomy for adenocarcinoma. The findings in this case report are meaningful for surgeons treating cancer patients because there are few reports discussing the perioperative care of surgical patients with polycythemia.


Skin decontamination of mustards and organophosphates: comparative efficiency of RSDL and Fuller's earth in domestic swine.
Research in skin decontamination and therapy of chemical warfare agents has been a difficult problem due to the simultaneous requirement of rapid action and non-aggressive behaviour. The aim of this study was to compare the performance of two decontaminating systems: the Canadian Reactive Skin Decontaminant Lotion (RSDL) and the Fuller's Earth (FE). The experiment was conducted with domestic swine, as a good model for extrapolation to human skin. RSDL and FE were tested against sulphur mustard (SM), a powerful vesicant, and VX, a potent and persistent cholinesterase inhibitor. When used 5 min after contamination, the results clearly showed that both systems were active against SM (10.1 mg/cm(2)) and VX (0.06 mg/cm(2)). The potency of the RSDL/sponge was statistically better than FE against skin injury induced by SM, observed 3 days post-exposure. RSDL was rather more efficient than FE in reducing the formation of perinuclear vacuoles and inflammation processes in the epidermis and dermis. Against a severe inhibition (67%) of plasmatic cholinesterases induced by VX poisoning, the potencies of the RSDL/sponge and FE were similar. Both systems completely prevented cholinesterase inhibition, which indirectly indicates a prevention of toxic absorption through the skin.


The Role of Navigated Transcranial Magnetic Stimulation Motor Mapping in Adjuvant Radiotherapy Planning in Patients With Supratentorial Brain Metastases.
Purpose: In radiotherapy (RT) of brain tumors, the primary motor cortex is not regularly considered in target volume delineation, although decline in motor function is possible due to radiation. Non-invasive identification of motor-eloquent brain areas is currently mostly restricted to functional magnetic resonance imaging (fMRI), which has shown to lack precision for this purpose. Navigated transcranial magnetic stimulation (nTMS) is a novel tool to identify motor-eloquent brain areas. This study aims to integrate nTMS motor maps in RT planning and evaluates the influence on dosage modulations in patients harboring brain metastases. Materials and Methods: Preoperative nTMS motor maps of 30 patients diagnosed with motor-eloquent brain metastases were fused with conventional planning imaging and transferred to the RT planning software. RT plans of eleven patients were optimized by contouring nTMS motor maps as organs at risk (OARs). Dose modulation analyses were performed using dose-volume histogram (DVH) parameters. Results: By constraining the dose applied to the nTMS motor maps outside the planning target volume (PTV) to 15 Gy, the mean dose (Dmean) to the nTMS motor maps was significantly reduced by 18.1% from 23.0 Gy (16.9-30.4 Gy) to 18.9 Gy (13.5-28.8 Gy, p < 0.05). The Dmean of the PTV increased by 0.6 ± 0.3 Gy (1.7%). Conclusion: Implementing nTMS motor maps in standard RT planning is feasible in patients suffering from intracranial metastases. A significant reduction of the dose applied to the nTMS motor maps can be achieved without impairing treatment doses to the PTV. Thus, nTMS might provide a valuable tool for safer application of RT in patients harboring motor-eloquent brain metastases.


Electrophysiological effects of multiple instillation of Haemophilus influenzae type b endotoxin on the inner ear.
An analysis of auditory brainstem response (ABR) thresholds and ABR-based frequency tuning curves was performed in 15 Sprague-Dawley rats exposed to Haemophilus influenzae type b endotoxin; 5 microg/50 microl toxin was instilled every second day, altogether five times, into the middle ear cavity through a small perforation in the tympanic membrane. ABR was measured 48 h after the second application and 24 h, 48 h, 5 days and 10 days after the fifth instillation. Five applications of toxin had no statistical effect on ABR thresholds and no changes in TC configuration were observed. It is concluded that Haemophilus influenzae type b endotoxin, instilled repeatedly through the tympanic membrane into the middle ear, does not affect cochlear electrophysiology.


Tuberculosis in five basset hounds.
Five Basset Hounds (2 females and 3 males) under the age of 5 years, acquired systemic tuberculosis. We suspected tuberculosis in one dog, because it had histologic lesions similar to those in 4 dogs in which bacteria were identified as Mycobacterium avium complex. A review of canine tuberculosis revealed a similar diagnosis in a Basset Hound. The association of this infection in Basset Hounds suggests an inherited immunologic defect. Results of our survey suggest that the defect might exist in cell-mediated immunity.


Direct androgen receptor regulation of sexually dimorphic gene expression in the mammalian kidney.
Mammalian organs exhibit distinct physiology, disease susceptibility and injury responses between the sexes. In the mouse kidney, sexually dimorphic gene activity maps predominantly to proximal tubule (PT) segments. Bulk RNA-seq data demonstrated sex differences were established from 4 and 8 weeks after birth under gonadal control. Hormone injection studies and genetic removal of androgen and estrogen receptors demonstrated androgen receptor (AR) mediated regulation of gene activity in PT cells as the regulatory mechanism. Interestingly, caloric restriction feminizes the male kidney. Single-nuclear multiomic analysis identified putative cis-regulatory regions and cooperating factors mediating PT responses to AR activity in the mouse kidney. In the human kidney, a limited set of genes showed conserved sex-linked regulation while analysis of the mouse liver underscored organ-specific differences in the regulation of sexually dimorphic gene expression. These findings raise interesting questions on the evolution, physiological significance, and disease and metabolic linkage, of sexually dimorphic gene activity.


A 9-year follow-up study of participants and nonparticipants in sigmoidoscopy screening: importance of self-selection.
Self-selection may compromise cost-effectiveness of screening programs. We hypothesized that nonparticipants have generally higher morbidity and mortality than participants. A Swedish population-based random sample of 1,986 subjects ages 59 to 61 years was invited to sigmoidoscopy screening and followed up for 9 years by means of multiple record linkages to health and population registers. Gender-adjusted cancer incidence rate ratio (IRR) and overall and disease group-specific and mortality rate ratio (MRR) with 95% confidence intervals (95% CI) were estimated for nonparticipants relative to participants. Cancer and mortality rates were also estimated relative to the age-matched, gender-matched, and calendar period-matched Swedish population using standardized incidence ratios and standardized mortality ratios. Thirty-nine percent participated. The incidence of colorectal cancer (IRR, 2.2; 95% CI, 0.8-5.9), other gastrointestinal cancer (IRR, 2.7; 95% CI, 0.6-12.8), lung cancer (IRR, 2.2; 95% CI, 0.8-5.9), and smoking-related cancer overall (IRR, 1.4; 95% CI, 0.7-2.5) tended to be increased among nonparticipants relative to participants. Standardized incidence ratios for most of the studied cancers tended to be >1.0 among nonparticipants and <1.0 among participants. Mortality from all causes (MRR, 2.4; 95% CI, 1.7-3.4), neoplastic diseases (MRR, 1.9; 95% CI, 1.1-3.5), gastrointestinal cancer (MRR, 4.7; 95% CI, 1.1-20.7), and circulatory diseases (MRR, 2.3; 95% CI, 1.2-4.2) was significantly higher among nonparticipants than among participants. Standardized mortality ratio for the studied outcomes tended to be increased among nonparticipants and was generally decreased among participants. Individuals who might benefit most from screening are overrepresented among nonparticipants. This self-selection may attenuate the cost-effectiveness of screening programs on a population level.


The contribution of evoked potentials to the diagnosis of demyelinating diseases.
Examination of evoked potentials can be a useful diagnostic tool in the diagnosis and evaluation of patients with Multiple Sclerosis (MS). A description of the most frequently used EPs is given: BAEP, checkerboard VEP and short latency median nerve SSEP, together with an impression of their sensitivity. It is obvious that more clinical trials are necessary to evaluate also the other EP modalities before a balanced conclusion about the usefulness of EPs in MS can be formulated.


Selective polarization imager for contrast enhancements in remote scattering media.
Conventional intensity imaging through turbid media suffers from rapid loss of image contrast due to light scattering from particles or random variations of refractive index. This paper features the development of an active imaging, snapshot, system design and postprocessing algorithms that differentiate between radiation that scatters or reflects from remote, obscured objects and the radiation from the scattering media itself through a combination of polarization difference imaging, channel blurring, and Fourier spatial filtering. The produced sensor acquires and processes image data in real time, yielding improved image contrasts by factors of 10 or greater for dense water vapor obscurants.


Rivaroxaban for Thromboprophylaxis After Fracture-Related Orthopedic Surgery in Routine Clinical Practice.
The risk of venous thromboembolism is high in patients undergoing fracture-related major orthopedic surgery, but data on pharmacological thromboprophylaxis are limited. This analysis evaluated the effectiveness and safety of rivaroxaban after fracture-related orthopedic surgery in routine care compared with other pharmacological thromboprophylaxis (standard of care [SOC]). The study population comprised a subset of patients with lower-limb fracture from XArelto in the prophylaxis of post-surgical venous thromboembolism after elective Major Orthopaedic Surgery of hip or knee (XAMOS; a phase IV noninterventional study) and a XAMOS extension study (XAMOS-Extra). The study participants included patients who underwent surgery for hip/femur or lower-leg fractures (below-knee lower-leg fractures, eg, the tibia or foot). All adverse events were recorded, including symptomatic thromboembolic events and bleeding events. Data from 790 patients were available for analysis (n = 350 for rivaroxaban and n = 440 for SOC). The incidence of symptomatic thromboembolic events 3 months postsurgery was 0.57% (2 of the 350) in the rivaroxaban group and 1.14% (5 of the 440) in the SOC group (odds ratio [OR]: 0.50; 95% confidence interval [CI]: 0.10-2.59). Treatment-emergent major bleeding events occurred in 0.29% (1 of the 350) of patients receiving rivaroxaban and 0.45% (2 of the 440) of patients receiving SOC (OR: 0.63; 95% CI: 0.06-6.95). There were no cases of fatal or critical bleeding in either treatment group. The incidences of wound complications and any other adverse events were numerically lower with rivaroxaban compared with SOC. These data from routine practice demonstrate that rivaroxaban can provide effective thromboprophylaxis after fracture-related orthopedic surgery of the lower limb with a favorable safety profile.


Song practice promotes acute vocal variability at a key stage of sensorimotor learning.
Trial by trial variability during motor learning is a feature encoded by the basal ganglia of both humans and songbirds, and is important for reinforcement of optimal motor patterns, including those that produce speech and birdsong. Given the many parallels between these behaviors, songbirds provide a useful model to investigate neural mechanisms underlying vocal learning. In juvenile and adult male zebra finches, endogenous levels of FoxP2, a molecule critical for language, decrease two hours after morning song onset within area X, part of the basal ganglia-forebrain pathway dedicated to song. In juveniles, experimental 'knockdown' of area X FoxP2 results in abnormally variable song in adulthood. These findings motivated our hypothesis that low FoxP2 levels increase vocal variability, enabling vocal motor exploration in normal birds. After two hours in either singing or non-singing conditions (previously shown to produce differential area X FoxP2 levels), phonological and sequential features of the subsequent songs were compared across conditions in the same bird. In line with our prediction, analysis of songs sung by 75 day (75d) birds revealed that syllable structure was more variable and sequence stereotypy was reduced following two hours of continuous practice compared to these features following two hours of non-singing. Similar trends in song were observed in these birds at 65d, despite higher overall within-condition variability at this age. Together with previous work, these findings point to the importance of behaviorally-driven acute periods during song learning that allow for both refinement and reinforcement of motor patterns. Future work is aimed at testing the observation that not only does vocal practice influence expression of molecular networks, but that these networks then influence subsequent variability in these skills.


Serum ferritin and stool occult blood and colon cancer screening.
A group of 531 participants age 50 years or older were evaluated for colonic polyps and malignancy with stool occult blood testing, hemoglobin, hematocrit, serum ferritin, flexible fiberoptic sigmoidoscopy, and colonoscopy. Screening revealed 1 colorectal cancer, 2 malignant polyps, 30 hyperplastic polyps, 69 adenomatous polyps, 4 villous adenomas, and 2 villous components. Low serum ferritin, an indicator of iron deficiency without anemia, improved the detection of colonic malignancies and polyps when used in combination with stool occult blood testing. The program was acceptable to participants and attrition was low; the attrition rate for the screening program at 1 year was 14.5%.


Young offenders' perspectives on their literacy and communication skills.
Research has revealed that the youth offending population has low language ability when assessed on standardized language measures. However, little is known about the perceptions young offenders (YOs) have of their own literacy ability and their communicative interactions with others. Such knowledge might further our understanding of the possible association between language, literacy and offending behaviour. This study investigates the perceptions and experiences YOs have of using literacy and communicating with others. It addresses the following questions. How satisfied are YOs with their own literacy and communication skills and how important do YOs perceive these to be? How much do YOs believe they understand others in their communicative interactions? How satisfied are YOs with their communicative interactions with others and how does this influence conflict at home, school, and in the youth justice system? An opportunity sample of 31 YOs on court orders were recruited from a local youth offending service, excluding any who did not have English as a first language or were in receipt of current speech and language therapy provision. Twenty-six qualitative individual semi-structured interviews and two focus group interviews were carried out and analysed using a framework analysis method. Themes revealed participants were dissatisfied with their communication and literacy ability. Other themes identified were difficulty in understanding others, a perceived lack of support and respect gained from others, and a negative impact of communication on self-esteem. The findings suggest that YOs often found themselves in disputes with authority figures, but that they avoided using positive communication to solve such conflicts and also avoided confiding in others. The findings support the results found from quantitative research on the language abilities of YOs. This emphasizes the value in adopting qualitative methodology to understand the relationship between literacy, communication skills and offending behaviour in YOs. The findings highlight a need for increased language, literacy and communication training, and support for YOs, and for the staff who work alongside them.


THE DETERMINATION OF THE EQUIVALENT WEIGHT OF PROTEINS.
The magnitude of the correction in the fifth column of Table III may be open to some doubt, as are all corrections of such a character, and the significance of the above experiment in the author's mind lies not so much in the actual magnitude of the values given in the last column of this table as in their comparative magnitudes. For this reason the entire experiment reported was performed in a single session using the same gelatin solution, so that, whatever the magnitude of the correction, it would be the same in all cases. Actually the results in the case of the acid titrations are in fair agreement with those of Hitchcock (8). In the present experiment it is seen that, within the limits of experimental error, one gets the same value for the number of cc. of tenth normal acid bound by 1 gm. of gelatin whether one titrates with the acid or with the gelatin. In the case of the base there is a small difference, due probably to carbon dioxide, but this effect is in a direction opposite to that which one would expect on the assumption that it is due to appreciable adsorption. From this it is concluded that the binding due to adsorption in the case of gelatin is not significant compared to that due to chemical neutralization. The author realizes that gelatin is a poor choice for a basis of generalizations, and similar work is at present in progress on various other proteins. He does feel, however, that the conclusions of Hoffman and Gortner from their work on the prolamines may also be too widely generalized, and that, on the whole, the acid or alkali bound by adsorption in the case of proteins will not constitute the large majority of the total amounts bound, though certainly one will expect a certain amount of such binding in all cases. It also seems that before placing undue emphasis on the conclusions of these workers the possibilities of equivocal results due to specific technique should be considered. This technique consisted in introducing weighed amounts of dry protein into a definite volume of standard acid or base at the equilibrium temperature, in general, and, "after about 15 minutes, during which time the flask was shaken several times," determining the pH of the equilibrium solution. Is it possible that the actual speed of solution of the protean is such that, even though reproducible results are obtained using identical technique, actual equilibrium conditions are approached only when comparatively high concentrations of acid or alkali are employed, in which cases the solution velocity of the protein may he expected to be greater, other factors remaining constant?


The typing of Morganella morgani by bacteriocin production and sensitivity.
A typing scheme for Morganella morgani based on bacteriocin (morganocin) production and sensitivity is described. These characteristics were determined by testing 160 strains in all combinations and permitted their differentiation into 90 types. Morganocin production was induced with mitomycin C and morganocin sensitivity determined with a diluted inoculum on Lab-lemco agar at 30 degrees C. Most strains (82.5%) produced morganocins and 49 different types were defined. Most strains (97.5%) were sensitive to morganocins and usually to several different types. The scheme is more discriminating than other reported methods. The finding in an epidemiological survey of the carriage of certain strains in the bowel for several weeks suggests that in practice the method is stable and reproducible.


Use of monoclonal antibody probes against rat hepatic cytochromes P-450c and P-450d to detect immunochemically related isozymes in liver microsomes from different species.
Nine distinct monoclonal antibodies raised against purified rat liver cytochrome P-450c react with six different epitopes on the antigen, and one of these epitopes is shared by cytochrome P-450d. None of these monoclonal antibodies recognize seven other purified rat liver isozymes (cytochromes P-450a, b, and e-i) or other proteins in the cytochrome P-450 region of "Western blots" of liver microsomes. Each of the monoclonal antibodies was used to probe "Western blots" of liver microsomes from untreated, or 3-methylcholanthrene-, or isosafrole-treated animals to determine if laboratory animals other than rats possess isozymes immunochemically related to cytochromes P-450c and P-450d. Two protein-staining bands immunorelated to cytochromes P-450c and P-450d were observed in all animals treated with 3-methylcholanthrene (rabbit, hamster, guinea pig, and C57BL/6J mouse) except the DBA/2J mouse, where no polypeptide immunorelated to cytochrome P-450c was detected. The conservation of the number of rat cytochrome P-450c epitopes among these species varied from as few as two (guinea pig) to as many as five epitopes (C57BL/6J mouse and rabbit). The relative mobility in sodium dodecyl sulfate-gels of polypeptides immunorelated to cytochromes P-450c and P-450d was similar in all species examined except the guinea pig, where the polypeptide related to cytochrome P-450c had a smaller Mr than cytochrome P-450d. With the use of both monoclonal and polyclonal antibodies, we were able to establish that purified rabbit cytochromes P-450 LM4 and P-450 LM6 are immunorelated to rat cytochromes P-450d and P-450c, respectively.


Signals for local and systemic responses of plants to pathogen attack.
Activation of plant defences following recognition of pathogen attack involves complex reiterative signal networks with extensive signal amplification and cross-talk. The results of two approaches that have been taken to analyse signalling in plant-microbe interactions are discussed here. Activation tagging with T-DNA harbouring multiple 35S enhancer elements was employed as a gain-of-function approach to dissect signalling related to bacterial pathogen resistance in Arabidopsis thaliana. From a screen of approximately 5000 activation tagged lines, one line was identified as harbouring a T-DNA leading to over-expression of an apoplastic aspartic protease (CDR-1), that resulted in resistance to normally virulent Pseudomonas syringae. The second approach was to screen for loss-of-function mutants in T-DNA tagged populations. From a screen of 11 000 lines, one line, defective in induced resistance-1 (dir-1) lost resistance to normally avirulent P. syringae. Models for action of the products of the CDR-1 and DIR-1 genes suggest involvement of peptide and lipid signals in systemic disease resistance responses in A. thaliana.


Cell-free synthesis of the precursor polypeptide for avian myeloblastosis virus DNA polymerase.
High molecular weight RNA (35S) isolated from avian myeloblastosis virus directs the cell-free synthesis of two prominent polypeptides of 180,000 and 76,000 molecular weight. The latter polypeptide has previously been identified as the precursor to the group-specific antigens of the virus ("gag" proteins) [Vogt, V. M., Eisenman, R. & Diggelmann, H. (1975) J. Mol. Biol. 96, 471-493]. Two-dimensional tryptic peptide analyses of the [35S]methionine-labeled peptides demonstrate that the 180,000-dalton product is a polyprotein that can account for all the peptides of the avian myeloblastosis virus DNA polymerase (DNA nucleotidyltransferase, EC 184.108.40.206) and those of the gag viral proteins. This is direct confirmation of the genomic order of the viral structural genes, placing the polymerase gene adjacent to the 5'-proximal gag gene of the virus. Furthermore, our findings suggest that the primary polymerase gene product is the beta subunit of the enzyme. These results are discussed in relation to the proposed structural gene map for the avian retraviruses and suggest a model for the in vivo processing of the viral polymerase.


Deciphering the Crosstalk Between Myeloid-Derived Suppressor Cells and Regulatory T Cells in Pancreatic Ductal Adenocarcinoma.
Pancreatic ductal adenocarcinoma (PDAC) is a fatal disease with rising incidence and a remarkable resistance to current therapies. The reasons for this therapeutic failure include the tumor's extensive infiltration by immunosuppressive cells such as myeloid-derived suppressor cells (MDSCs) and regulatory T cells (Tregs). By using light sheet fluorescent microscopy, we identified here direct interactions between these major immunoregulatory cells in PDAC. The in vivo depletion of MDSCs led to a significant reduction in Tregs in the pancreatic tumors. Through videomicroscopy and ex vivo functional assays we have shown that (i) MDSCs are able to induce Treg cells in a cell-cell dependent manner; (ii) Treg cells affect the survival and/or the proliferation of MDSCs. Furthermore, we have observed contacts between MDSCs and Treg cells at different stages of human cancer. Overall our findings suggest that interactions between MDSCs and Treg cells contribute to PDAC immunosuppressive environment.


Expression of vascular cell adhesion molecule-1 in human renal allografts.
The expression of vascular cell adhesion molecule-1 (VCAM-1) in 11 human renal allograft biopsies and 3 normal kidney specimens was investigated by immunocytochemistry. VCAM-1 expression was correlated with the degree of CD3+ T cell infiltration and the clinicopathologic diagnosis of acute rejection. CD3+ infiltrates were seen in all biopsies with rejection, but not in normal biopsies or one with acute tubular necrosis, and were accompanied by CD68+ monocyte/macrophage infiltrates. In normal biopsies, VCAM-1 was present on occasional tubules, where its expression was patchy and restricted to the basolateral surface of cells with slight cytoplasmic staining. The total number of tubules expressing VCAM-1 significantly increased in specimens infiltrated with CD3+ T cells. Moreover, in these infiltrated biopsy specimens, VCAM-1 was present throughout the cytoplasm of tubular cells concentrated on the basolateral surface. VCAM-1 was also observed on vascular endothelial cells where its expression correlated with the degree of CD3+ infiltrate. Mean scores (0 to 3+) for endothelial VCAM-1 expression increased from 0 (CD3+ score, 0) to a mean score of 2.25 in association with CD3+ T cell infiltrates (CD3+ score, 3). Endothelial VCAM-1 was predominantly on vessels in areas of infiltrate, including peritubular capillaries, venules, and arterioles, but was notably absent on glomerular endothelium. VCAM-1 also stained mesangial cells in an occasional CD3+ infiltrated specimen. It was concluded that the expression of VCAM-1 is increased on renal tubules and renovascular endothelium in rejecting renal allografts in association with CD3+ infiltrates.(ABSTRACT TRUNCATED AT 250 WORDS)


[A clinical study of aortic dissecting].
The clinical features, etiologies, pathogenesis, treatment and prognosis of 70 patients with aortic dissecting were studied. Patients with age ranging from 31 to 70 years consisted 93% of the total. The proportion of proximal to distal types is 0.8: 1. Hypertension was present in 67% and Marfan's syndrome 18% of the patients. The pathologic finding was scattered rupture with myxomatous and cystic necrosis of the mecial elastic fibers. 80% of the patients suffered from the disease without any inducing cause. Of the 76% patients had chest pain, it was very severe in 69%. Chest X-ray, ultrasound, computed tomography and magnetic resonance imaging together with clinical manifestations may accurately diagnose the disease. The sensitivity and specificity of the ultrasound are 97% and 94% respectively. The mortality in hospital with surgical and medical therapy of the proximal type was 14% and 22% respectively, while that of the distal type was 63% and 27% respectively. Rupture of the aneurysm was the cause of death in 68% of the patients.


Spaceflight results in reduced mRNA levels for tissue-specific proteins in the musculoskeletal system.
The purpose of the present study in growing rats was to investigate the effects of short-term spaceflight on gene expression in bone and muscle and on cortical bone histomorphometry. Two experiments were carried out; Physiological Systems Experiments 1 and 2 were 4- and 10-day flights, respectively. Radial bone growth in the humerus was unchanged during the 4-day flight and decreased during the 10-day flight. Expression of mRNA for glyceraldehyde-3-phosphate dehydrogenase was unchanged in biceps, calvarial periosteum, and long-bone periosteum after spaceflight. Similarly, no changes in ribosomal RNA levels were observed in long-bone or calvarial periosteum after spaceflight. In contrast, spaceflight decreased steady-state mRNA levels for actin in muscle (4-day flight). Osteocalcin (both spaceflights) and the prepro-alpha 2[I] chain of type I precollagen (10-day flight) mRNA levels were decreased in long-bone and calvarial periosteum after spaceflight. These results indicate that the effects of spaceflight on the musculoskeletal system include decreased expression of some muscle- and bone-specific genes as well as decreased bone formation. Interestingly, detectable reductions in gene expression for bone matrix proteins preceded histological evidence for decreased bone formation.


Ubiquitin-proteasome system in diabetic retinopathy.
Diabetic retinopathy (DR) is the most common complication of diabetes, being the most prevalent reason for blindness among the working-age population in the developed world. Despite constant improvement of understanding of the pathogenesis of DR, identification of novel biomarkers of DR is needed for improvement of patient risk stratification and development of novel prevention and therapeutic approaches. The ubiquitin-proteasome system (UPS) is the primary protein quality control system responsible for recognizing and degrading of damaged proteins. This review aims to summarize literature data on modifications of UPS in diabetes and DR. First, we briefly review the structure and functions of UPS in physiological conditions. We then describe how UPS is involved in the development and progression of diabetes and touch upon the association of UPS genetic factors with diabetes and its complications. Further, we focused on the effect of diabetes-induced hyperglycemia, oxidative stress and hypoxia on UPS functioning, with examples of studies on DR. In other sections, we discussed the association of several other mechanisms of DR (endoplasmic reticulum stress, neurodegeneration etc) with UPS modifications. Finally, UPS-affecting drugs and remedies are reviewed. This review highlights UPS as a promising target for the development of therapies for DR prevention and treatment and identifies gaps in existing knowledge and possible future study directions.


The path of a click stimulus from ear canal to umbo.
The tympanic membrane (TM) has a key role in transmitting sounds to the inner ear, but a concise description of how the TM performs this function remains elusive. This paper probes TM operation by applying a free field click stimulus to the gerbil ear and exploring the consequent motions of the TM and umbo. Motions of the TM were measured both on radial tracks starting close to the umbo and on a grid distal and adjacent to the umbo. The experimental results confirmed the high fidelity of sound transmission from the ear canal to the umbo. A delay of 5-15 μs was seen in the onset of TM motion between points just adjacent to the umbo and mid-radial points. The TM responded with a ringing motion, with different locations possessing different primary ringing frequencies. A simple analytic model from the literature, treating the TM as a string, was used to explore the experimental results. The click-based experiments and analysis led to the following description of TM operation: A transient sound pressure on the TM causes a transient initial TM motion that is maximal ∼ at the TM's radial midpoints. Mechanical forces generated by this initial prominent TM distortion then pull the umbo inward, leading to a delayed umbo response. The initial TM deformation also gives rise to prolonged mechanical ringing on the TM that does not result in significant umbo motion, likely due to destructive interference from the range of ringing frequencies. Thus, the umbo's response is a high-fidelity representation of the transient stimulus. Because any sound can be considered as a consecutive series of clicks, this description is applicable to any sound stimulus.


Incidence and complications of perioperative atrial fibrillation after non-cardiac surgery for malignancy.
Perioperative atrial fibrillation (POAF) is one of the common arrhythmias in the setting of non-cardiac surgeries for malignancy. As POAF may cause subsequent adverse events, it is important to confirm its characteristics and risk factors. The prospective cohort study of surveillance for perioperative atrial fibrillation recurrence (PREDICT AF RECURRENCE) is an ongoing prospective, single-center, observational study that aims to illustrate the clinical impact of POAF in major non-cardiac surgery for malignancy. Patients who planned to undergo non-cardiac surgery for definitive/suspected malignancy were registered. Those with a history of AF and atrial flutter were excluded. Any 30-day complications included acute myocardial infarction, congestive heart failure, bleeding, thrombosis, any infection, and acute kidney injury. The primary endpoint was an incidence of POAF. The present study included 799 patients (age, 68 ± 11; male, 62%). Of these, 80 patients (10.0%) developed POAF. Notably, 66 patients (83%) had no symptoms. Any 30-day complications occurred in 180 patients (23%) (with POAF: 34 (43%); without POAF: 146 (20%); p < 0.001). POAF in 17 patients (50%) was preceded by the development of complications. No patient developed cardiogenic shock and/or acute heart failure. The association between 30-day complications and POAF development were analyzed using the multivariate adjusted model (odds ratio: 2.84; 95% confidence interval: 1.74-4.62; p < 0.001). Ten percent of patients who underwent non-cardiac surgery for malignancy developed POAF, which was strongly associated with perioperative complications. As a majority were asymptomatic, careful observation using electrocardiography monitoring is important to avoid oversights. UMIN ID: UMIN000016146.


Impact of Renal Dysfunction on Results of Transcatheter Aortic Valve Replacement Outcomes in a Large Multicenter Cohort.
Patients with advanced chronic renal dysfunction were excluded from randomized trials of transcatheter aortic valve replacement (TAVR). The potential impact of chronic renal disease on TAVR prognosis is not fully understood. We aim to evaluate outcomes within a large cohort of patients who underwent TAVR distinguished by renal function. Baseline characteristics, procedural data, and clinical follow-up findings were collected from 10 high-volume TAVR centers in Europe, Israel, and Japan. Data were analyzed according to renal function. Patients (n = 1,204) were divided into 4 groups according to pre-TAVR-estimated glomerular filtration rate (eGFR): group I (eGFR >60), n = 288 (female 45%), group II (eGFR 31 to 60), n = 452 (female 61%), group III (eGFR ≤30), n = 398 (female 61%), and group IV (dialysis), n = 66 (female 31%). Mean Society of Thoracic Surgeons score was higher in patients with lower preprocedural eGFR. All-cause mortality at 1 year was higher in patients with lower eGFR (9.0%, 12.1%, 24.3%, and 24.2% for group I, II, III, and IV, respectively, p <0.001). Multivariate analysis demonstrated that eGFR ≤30, but not eGFR 31 to 60, was associated with increased risk of death (odds ratio 3), bleeding (odds ratio 5.2), and device implantation failure (hazard ratio 2.28). For each 10 ml/min decrease in eGFR, there was an associated relative increase in the risk of death (35%; p <0.001), cardiovascular death (14%; p = 0.018), major bleeding 35% (p <0.001), and transcatheter valve failure (16%; p = 0.007). Renal dysfunction was not associated with stroke or need for pacemaker implantation. In conclusion, among patients who underwent TAVR, baseline renal dysfunction is an important independent predictor of morbidity and mortality.


Extracellular hippocampal acetylcholine level controls amygdala function and promotes adaptive conditioned emotional response.
Ample data indicate that tone and contextual fear conditioning differentially require the amygdala and the hippocampus. However, mechanisms subserving the adaptive selection among environmental stimuli (discrete tone vs context) of those that best predict an aversive event are still elusive. Because the hippocampal cholinergic neurotransmission is thought to play a critical role in the coordination between different memory systems leading to the selection of appropriate behavioral strategies, we hypothesized that this cholinergic signal may control the competing acquisition of amygdala-mediated tone and contextual conditioning. Using pavlovian fear conditioning in mice, we first show a higher level of hippocampal acetylcholine release and a specific pattern of extracellular signal-regulated kinase 1/2 (ERK1/2) activation within the lateral (LA) and basolateral (BLA) amygdala under conditions in which the context is a better predictor than a discrete tone stimulus. Second, we demonstrate that levels of hippocampal cholinergic neurotransmission are causally related to the patterns of ERK1/2 activation in amygdala nuclei and actually determine the selection among the context or the simple tone the stimulus that best predicts the aversive event. Specifically, decreasing the hippocampal cholinergic signal not only impaired contextual conditioning but also mimicked conditioning to the discrete tone, both in terms of the behavioral outcome and the LA/BLA ERK1/2 activation pattern. Conversely, increasing this cholinergic signal not only disrupted tone conditioning but also promoted contextual fear conditioning. Hence, these findings highlight that hippocampal cholinergic neurotransmission controls amygdala function, thereby leading to the selection of relevant emotional information.


Conotoxins: review and docking studies to determine potentials of conotoxin as an anticancer drug molecule.
It is known that potassium channels are important for cell proliferation. HERG, a potassium channel protein, is a transmembrane protein, which increases in concentration on the cell surface of cancer cells. Apart from cancer cells, this protein is found only in the brain & heart tissue, in very low number. The proliferation of cells in cancer is dependent on activation of this protein, and it has been noted that blocking of this protein with drug molecule, helps inhibit the proliferation of the cells further. The current work aims to study the binding potentials of κ-PVIIA, conotoxin isolated from Conus purpurascens venom with HERG K+ channel of tumor cells, where HERG mutation has been noted. The toxin under consideration i.e. κ-conotoxins-PVIIA (κ-PVIIA) is a 27 residue peptide. The docking studies suggest that the conotoxin binds stably to the HERG protein. The study shows that the peptide interacts with the charged extracellular unit of the HERG protein, i.e. the extracellular portion of the S5 domain named S5-P extracellular linker. Study of binding of toxins of similar origin, with normal potassium channels has been studied in silico. Further, wet laboratory work needs to be conducted for development of a drug molecule from this toxin, to treat some number of cancers.


Winogradskyella psychrotolerans sp. nov., a marine bacterium of the family Flavobacteriaceae isolated from Arctic sediment.
A novel Gram-negative, rod-coccus shaped, non-motile, strain, RS-3(T), was isolated from a sediment sample collected from the marine transect of Kongsfjorden, Ny-Ålesund, Svalbard, Arctic. Colonies and broth cultures were yellowish in colour due to the presence of carotenoids. Strain RS-3(T) was positive for oxidase, aesculinase, caseinase, gelatinase and urease activities and negative for amylase, catalase, lipase, lysine decarboxylase, ornithine decarboxylase, DNase and β-galactosidase activities. The predominant fatty acids were iso-C15 : 0 (18.0), anteiso-C15 : 0 (16.8), iso-C15 : 1 G (14.2), anteiso-C15 : 1 A (6.0) and iso-C15 : 0 3-OH (6.8). Strain RS-3(T) contained MK-6 (72.42 %) and MK-7 (27.58 %) as the major respiratory quinones and phosphatidylethanolamine, two unidentified aminolipids and two unidentified lipids make up the polar lipid composition. The DNA G+C content of strain RS-3(T) was 34.7±1.2 mol%. The 16S rRNA gene sequence analysis indicated that Winogradskyella pacifica and Winogradskyella thalassocola are the most closely related species with sequence similarities to the type strains of these species of 98.5 and 97.7 %, respectively. However, DNA-DNA hybridization with Winogradskyella pacifica KCTC 22997(T) and Winogradskyella thalassocola DSM 15363(T) showed a relatedness of 22 and 42.5 % with respect to strain RS-3(T). Based on the DNA-DNA hybridization values, phenotypic and chemotaxonomic characteristics and phylogenetic inference, strain RS-3(T) is proposed as a novel species of the genus Winogradskyella, for which the name Winogradskyella psychrotolerans sp. nov. is proposed. The type strain of Winogradskyella psychrotolerans sp. nov. is RS-3(T) ( = CIP 110154(T) = NBRC 106169(T)). An emended description of the genus Winogradskyella is provided.


Adiponectin receptor agonist AdipoRon improves skeletal muscle function in aged mice.
The loss of skeletal muscle function with age, known as sarcopenia, significantly reduces independence and quality of life and can have significant metabolic consequences. Although exercise is effective in treating sarcopenia it is not always a viable option clinically, and currently, there are no pharmacological therapeutic interventions for sarcopenia. Here, we show that chronic treatment with pan-adiponectin receptor agonist AdipoRon improved muscle function in male mice by a mechanism linked to skeletal muscle metabolism and tissue remodeling. In aged mice, 6 weeks of AdipoRon treatment improved skeletal muscle functional measures in vivo and ex vivo. Improvements were linked to changes in fiber type, including an enrichment of oxidative fibers, and an increase in mitochondrial activity. In young mice, 6 weeks of AdipoRon treatment improved contractile force and activated the energy-sensing kinase AMPK and the mitochondrial regulator PGC-1a (peroxisome proliferator-activated receptor gamma coactivator one alpha). In cultured cells, the AdipoRon induced stimulation of AMPK and PGC-1a was associated with increased mitochondrial membrane potential, reorganization of mitochondrial architecture, increased respiration, and increased ATP production. Furthermore, the ability of AdipoRon to stimulate AMPK and PGC1a was conserved in nonhuman primate cultured cells. These data show that AdipoRon is an effective agent for the prevention of sarcopenia in mice and indicate that its effects translate to primates, suggesting it may also be a suitable therapeutic for sarcopenia in clinical application.


Ponicidin suppresses HT29 cell growth via the induction of G1 cell cycle arrest and apoptosis.
Ponicidin is a diterpenoid extracted from the Chinese herb Isodon adenolomus, which has been reported as a therapeutic cytotoxic drug that may be used to treat various types of human cancer. The present study aimed to determine the antitumor effects of ponicidin, and to investigate its underlying mechanisms in colorectal cancer. The HT29 colorectal cancer cell line was used to detect the cytotoxicity of various doses of ponicidin. Cell proliferation was measured using a Cell Counting kit‑8 assay. Cell cycle and apoptosis analyses were performed using flow cytometry and fluorescent microscopy. Western blot analysis was used to measure the expression levels of apoptosis‑associated proteins following treatment with ponicidin. Treatment with ponicidin significantly suppressed HT29 cell growth by inducing G1 cell cycle arrest and apoptosis. The AKT and MEK signaling pathways were also suppressed by ponicidin; however, the p38 signaling pathway was significantly activated. The expression levels of caspase 3 and Bax protein were markedly upregulated following treatment with ponicidin. These results suggest that ponicidin exerts significant antitumor effects via the induction of cell cycle arrest and apoptosis in colorectal cells. In conclusion, ponicidin acted as an inducer of apoptosis, and may be used as a therapeutic cytotoxic drug to treat human cancer, including colorectal cancer.


The Ataxin-2 protein is required for microRNA function and synapse-specific long-term olfactory habituation.
Local control of mRNA translation has been proposed as a mechanism for regulating synapse-specific plasticity associated with long-term memory. We show here that glomerulus-selective plasticity of Drosophila multiglomerular local interneurons observed during long-term olfactory habituation (LTH) requires the Ataxin-2 protein (Atx2) to function in uniglomerular projection neurons (PNs) postsynaptic to local interneurons (LNs). PN-selective knockdown of Atx2 selectively blocks LTH to odorants to which the PN responds and in addition selectively blocks LTH-associated structural and functional plasticity in odorant-responsive glomeruli. Atx2 has been shown previously to bind DEAD box helicases of the Me31B family, proteins associated with Argonaute (Ago) and microRNA (miRNA) function. Robust transdominant interactions of atx2 with me31B and ago1 indicate that Atx2 functions with miRNA-pathway components for LTH and associated synaptic plasticity. Further direct experiments show that Atx2 is required for miRNA-mediated repression of several translational reporters in vivo. Together, these observations (i) show that Atx2 and miRNA components regulate synapse-specific long-term plasticity in vivo; (ii) identify Atx2 as a component of the miRNA pathway; and (iii) provide insight into the biological function of Atx2 that is of potential relevance to spinocerebellar ataxia and neurodegenerative disease.


A case with concurrent duplication, triplication, and uniparental isodisomy at 1q42.12-qter supporting microhomology-mediated break-induced replication model for replicative rearrangements.
Complex genomic rearrangements (CGRs) consisting of interstitial triplications in conjunction with uniparental isodisomy (isoUPD) have rarely been reported in patients with multiple congenital anomalies (MCA)/intellectual disability (ID). One-ended DNA break repair coupled with microhomology-mediated break-induced replication (MMBIR) has been recently proposed as a possible mechanism giving rise to interstitial copy number gains and distal isoUPD, although only a few cases providing supportive evidence in human congenital diseases with MCA have been documented. Here, we report on the chromosomal microarray (CMA)-based identification of the first known case with concurrent interstitial duplication at 1q42.12-q42.2 and triplication at 1q42.2-q43 followed by isoUPD for the remainder of chromosome 1q (at 1q43-qter). In distal 1q duplication/triplication overlapping with 1q42.12-q43, variable clinical features have been reported, and our 25-year-old patient with MCA/ID presented with some of these frequently described features. Further analyses including the precise mapping of breakpoint junctions within the CGR in a sequence level suggested that the CGR found in association with isoUPD in our case is a triplication with flanking duplications, characterized as a triplication with a particularly long duplication-inverted triplication-duplication (DUP-TRP/INV-DUP) structure. Because microhomology was observed in both junctions between the triplicated region and the flanking duplicated regions, our case provides supportive evidence for recently proposed replication-based mechanisms, such as MMBIR, underlying the formation of CGRs + isoUPD implicated in chromosomal disorders. To the best of our knowledge, this is the first case of CGRs + isoUPD observed in 1q and having DUP-TRP/INV-DUP structure with a long proximal duplication, which supports MMBIR-based model for genomic rearrangements. Molecular cytogenetic analyses using CMA containing single-nucleotide polymorphism probes with further analyses of the breakpoint junctions are recommended in cases suspected of having complex chromosomal abnormalities based on discrepancies between clinical and conventional cytogenetic findings.


Workplace impact on employees: A Lifelines Corona Research Initiative on the return to work.
A large proportion of the global workforce migrated home during the COVID-19 pandemic and subsequent lockdowns. It remains unclear what the exact differences between home workers and non-home workers were, especially during the pandemic when a return to work was imminent. How were building, workplace, and related facilities associated with workers' perceptions and health? What are the lessons to be learned? Lifelines Corona Research Initiative was used to compare employees' workplaces and related concerns, facilities, work quality, and health in a complete case analysis (N = 12,776) when return to work was imminent. Mann-Whitney U, logistic regression, and Wilcoxon matched-pairs were used for analyses. Notwithstanding small differences, the results show that home workers had less favourable scores for concerns about and facilities of on-site buildings and workplaces upon return to work, but better scores for work quality and health than non-home workers. However, additional analyses also suggest that building, workplace, and related facilities may have had the capacity to positively influence employees' affective responses and work quality, but not always their health.


Serum antibodies to native and denatured type I and III collagen in patients with periodontal disease.
Serum IgG antibodies to collagen were investigated by using enzyme linked immunosorbent assay (ELISA) in patients with chronic periodontal disease. Patients with varying forms of periodontal disease including gingivitis, juvenile periodontitis, and adult periodontitis were compared with the normal subjects. The mean serum IgG levels of ELISA antibodies to native type I or III collagen in patients with juvenile periodontitis were significantly higher than those of the normal subjects, but no difference was found between the patients with either gingivitis or adult periodontitis and the normal subjects. In addition, the mean serum IgG levels of ELISA antibodies to denatured type I or III collagen in patients with juvenile or adult periodontitis were significantly higher than those of the normal subjects. These findings suggest that antibodies to native and denatured type I or III collagen may be associated with different forms or severities of periodontal disease, especially advanced periodontal destruction.


Increased resting energy expenditure in hemodialysis patients with severe hyperparathyroidism.
Several metabolic derangements, including enhanced protein catabolism, have been suggested to be associated with increased circulating parathyroid hormone (PTH) in patients with secondary hyperparathyroidism (HPT). Such conditions, therefore, might lead to an increase in energy expenditure. The present study examined by indirect calorimetry the resting energy expenditure (REE) of 15 hemodialysis patients who have severe HPT (PTH = 1457 +/- 676 pg/ml) and were pair-matched for age and gender to 15 hemodialysis patients with mild to moderate HPT (PTH = 247 +/- 196 pg/ml). Both groups were also pair-matched for age and gender to a group of 15 healthy adult subjects (control). In six patients from the severe HPT group submitted to total parathyroidectomy, REE was determined 6 mo after the surgery. The groups were not different regarding lean body mass (LBM) measured by bioelectric impedance, serum C-reactive protein, and bicarbonate. Thyroid-stimulating hormone was within the normal range in all groups. Nonadjusted REE was significantly higher in the severe HPT group (1674 +/- 337 kcal/d) compared with patients with mild to moderate HPT (1388 +/- 229 kcal/d; P < 0.05). Both groups did not differ from the control group (1468 +/- 323 kcal/d). When adjustment of REE for LBM was performed using the multiple regression analysis, patients with mild to moderate HPT and control subjects had significantly lower REE (-231 and -262 kcal, respectively) than that of the severe HPT group. Considering all patients together, nonadjusted REE correlated directly with LBM (r = 0.61; P < 0.01). PTH correlated strongly with LBM in the severe HPT group (r = -0.82; P < 0.01). In the multiple linear regression analysis, only LBM and PTH were independent determinants of REE (n = 30; R(2) = 0.47). REE decreased significantly in the six patients who were evaluated 6 mo after parathyroidectomy (from 1617 +/- 339 to 1226 +/- 253; P = 0.02). These results demonstrate that hemodialysis patients with severe HPT have increased REE that might be reduced after parathyroidectomy.


Effects of essential amino acid deficiency: down-regulation of KCC2 and the GABAA receptor; disinhibition in the anterior piriform cortex.
The anterior piriform cortex (APC) is activated by, and is the brain area most sensitive to, essential (indispensable) amino acid (IAA) deficiency. The APC is required for the rapid (20 min) behavioral rejection of IAA deficient diets and increased foraging, both crucial adaptive functions supporting IAA homeostasis in omnivores. The biochemical mechanisms signaling IAA deficiency in the APC block initiation of translation in protein synthesis via uncharged tRNA and the general amino acid control kinase, general control nonderepressing kinase 2. Yet, how inhibition of protein synthesis activates the APC is unknown. The neuronal K(+) Cl(-) cotransporter, neural potassium chloride co-transporter (KCC2), and GABAA receptors are essential inhibitory elements in the APC with short plasmalemmal half-lives that maintain control in this highly excitable circuitry. After a single IAA deficient meal both proteins were reduced (vs. basal diet controls) in western blots of APC (but not neocortex or cerebellum) and in immunohistochemistry of APC. Furthermore, electrophysiological analyses support loss of inhibitory elements such as the GABAA receptor in this model. As the crucial inhibitory function of the GABAA receptor depends on KCC2 and the Cl(-) transmembrane gradient it establishes, these results suggest that loss of such inhibitory elements contributes to disinhibition of the APC in IAA deficiency. The circuitry of the anterior piriform cortex (APC) is finely balanced between excitatory (glutamate, +) and inhibitory (GABA, -) transmission. GABAA receptors use Cl(-), requiring the neural potassium chloride co-transporter (KCC2). Both are rapidly turning-over proteins, dependent on protein synthesis for repletion. In IAA (indispensable amino acid) deficiency, within 20 min, blockade of protein synthesis prevents restoration of these inhibitors; they are diminished; disinhibition ensues. GCN2 = general control non-derepressing kinase 2, eIF2α = α-subunit of the eukaryotic initiation factor 2.


Effect of garlic on lead contents in chicken tissues.
Lead has been indicted to be involved in the aetiology of human and animal diseases. In view of earlier literature indicating that garlic antagonized lead toxicity, we have investigated the possible use of garlic feeding to clean up lead contents from chickens which had been exposed to natural or experimental lead pollution and consequently eliminate one of the sources of lead pollution to human consumers. Groups of chickens (10 birds each) were given lead alone (lead acetate equivalent to 5 mg lead/kg B.W.) or both lead and garlic simultaneously or lead followed by garlic post-treatment or garlic alone or distilled water. Lead concentrations were reduced in muscle and liver tissues of chickens given both lead and garlic simultaneously or as a post-treatment. Reduction in tissue-lead concentrations were greater in birds given garlic as a post-treatment than those given garlic simultaneously with lead. The results indicate that garlic contain chelating compounds capable of enhancing elimination of lead. Garlic feeding can be exploited to safeguard human consumers by minimizing lead concentrations in meat of food animals which had been grown in a lead polluted environment.


Precise label-free leukocyte subpopulation separation using hybrid acoustic-optical chip.
Leukocyte subpopulations contain crucial physiological information; hence, precise and specific leukocyte separation is very important for leukemia diagnosis and analysis. However, conventional centrifugation and immunofluorescence-based separation methods are inaccurate and inconvenient due to the overlapping cell size and density or complex marking processes. Herein, we report a new label-free technology for precise leukocyte subpopulation separation by synergy of acoustic and optical technologies. Standing surface acoustic wave (SSAW) solved the problem of gentle and precise focusing of cells in optical systems. In addition, SSAW was used for the separation of granulocytes, which have evident size distinction from other components. In case of lymphocytes and monocytes, which have overlap in size/density, optical force could distinguish them accurately based on the RI difference, with the convenience of acoustic pre-focusing. In this experiment, separation of three types of leukocyte subtypes with considerable throughput and purity was conducted, through which we obtained 99% pure lymphocytes, 98% pure monocytes, and 95% pure granulocytes. Experimental results prove that the device has robust ability in separating leukocyte phenotypes and have the advantages of being non-invasive, label-free and precise. In the future, this convenient hybrid method will be a potential powerful tool for auxiliary clinical diagnosis and analysis.


[Cow's milk protein allergy: a challenging diagnosis].
Cow's Milk Protein Allergy (CMPA) is the most common food allergy in infants. It can manifest itself through a wide variety of symptoms depending on the type of immune response is IgE or non-IgE mediated. In most cases acquisition of tolerance to cow's milk protein (CMP) occurs until the second year of life. To evaluate the cases of CMPA followed in our Outpatient Clinic of Pediatric Gastroenterology and compare IgE and non-IgE mediated CMPA. CMPA patients observed between December 1992 and December 2009 were included in this prospective study and followed-up according to our CMPA protocol. 199 children were included in the protocol, but only 130 cases were confirmed the diagnoses of CMPA, 77 were IgE mediated (group A) and 53 non-IgE mediated (group B). Comparing the two groups, 45% vs 36% had positive family history of atopy. The median age of onset was 10 and six weeks respectively, and the interval between introduction of PLV into the diet and the onset of symptoms was four and five days. Infant formula was, in both groups, the main dairy product that triggered symptoms (71% vs 81%). In group A over half the children did the first dose of PLV in first 24 hours of life. The value of specific IgE to the major PLV was identified in 76 children and only 15.6% were positive. The most frequent symptoms in both groups were cutaneous (85.7% vs 50.6%), especially urticaria and angioedema, gastrointestinal symptoms (40% vs 38%), including vomiting and diarrhea.


Multiple subsets of HIV-specific cytotoxic T lymphocytes in humans and in mice.
The human immunodeficiency virus type 1 (HIV-1) induces a strong cytotoxic T lymphocyte (CTL) response in humans following infection. HIV-specific CTL can be detected directly in the blood and lungs of infected patients, and can be expanded in vitro by stimulation with autologous HIV-infected lymphoblasts. Furthermore, CTL specific for HIV envelope glycoprotein gp160 have been obtained in mice by immunization with recombinant vaccinia virus (VV) that carry the HIV env gene. In this study, we show that mice also produce strong CTL responses to gag and nef proteins following immunization with VV recombinants, thus providing a convenient model system to study T lymphocyte immunity to defined HIV antigens. To determine the specificity of circulating HIV-immune CTL in humans, a panel of doubly transfected mouse P815 tumor cells was produced which express the human HLA-A2 or HLA-A3 transplantation antigen gene and one HIV-1 gene (env, gag or nef). Using these cells as targets to CTL, we show that HIV-infected humans carry co-existing CTL sub-populations of different specificities. Each subpopulation appears to vary in intensity among different individuals. Surprisingly, CTL specific for regulatory, non-structural nef protein appear to be a major constituent of the human immune response to HIV.


A Meta-Regression Analysis of Atrial Fibrillation Ablation in Patients with Systolic Heart Failure.
Meta-analyses of randomized controlled trials comparing atrial fibrillation (AF) ablation to medical therapy in patients with heart failure (HF) reported improvement in left ventricular ejection fraction (LVEF), quality of life using the Minnesota Living with HF Questionnaire (MLWHFQ), and 6-minute walk test (6MWT). Nonetheless, there was significant heterogeneity not accounted for suggesting that not all HF patients derive the same effect from AF ablation. To evaluate if baseline LVEF or the etiology of the cardiomyopathy would moderate the efficacy of AF ablation. We performed random effects meta-regression using the mean baseline LVEF and total percentage of patients with non-ischemic cardiomyopathy (NICMP) in the placebo arms as moderator variables. Six trials with a total of 687 patients were included. The baseline LVEF in the control arm of trials ranged from 25% - 42.9%, and the percentage of patients with NICMP within each trial varied from 35% to 100%. When baseline LVEF was used as the moderator variable, no significant change in heterogeneity was observed for any of the outcomes of interest (R2 0.00 - 0.02). However, when controlling for NICMP, heterogeneity dropped substantially for the outcomes of LVEF (I2 44.7%, R2 0.91), and MLWHFQ (I2 0.00%, R2 1.00) but not 6MWT (I2 67.4%, R2 0.00). This indicates that improvement in LVEF and MLWHFQ was greater in the AF ablation group when more patients with NICMP were included in the trials. In patients with systolic HF, AF ablation may be more beneficial in patients with NICMP.