Mutant ataxin1 disrupts cerebellar development in spinocerebellar ataxia type 1.
Spinocerebellar ataxia type 1 (SCA1) is an adult-onset neurodegenerative disease caused by a polyglutamine expansion in the protein ATXN1, which is involved in transcriptional regulation. Although symptoms appear relatively late in life, primarily from cerebellar dysfunction, pathogenesis begins early, with transcriptional changes detectable as early as a week after birth in SCA1-knockin mice. Given the importance of this postnatal period for cerebellar development, we asked whether this region might be developmentally altered by mutant ATXN1. We found that expanded ATXN1 stimulates the proliferation of postnatal cerebellar stem cells in SCA1 mice. These hyperproliferating stem cells tended to differentiate into GABAergic inhibitory interneurons rather than astrocytes; this significantly increased the GABAergic inhibitory interneuron synaptic connections, disrupting cerebellar Purkinje cell function in a non-cell autonomous manner. We confirmed the increased basket cell-Purkinje cell connectivity in human SCA1 patients. Mutant ATXN1 thus alters the neural circuitry of the developing cerebellum, setting the stage for the later vulnerability of Purkinje cells to SCA1. We propose that other late-onset degenerative diseases may also be rooted in subtle developmental derailments.


Game performance and intermittent hypoxic training.
Live high-train low altitude exposure simulated by hypoxic devices may improve athletic performance. In this study, intermittent normobaric hypoxia was achieved with the GO2altitude hypoxicator to determine its effects on sea level performance in rugby players. Ten players were randomly assigned to two groups. Players in each group received 14 sessions of either hypoxic (10-15% O(2)) or normoxic (21% O(2)) exposure at rest over 14 consecutive days in a single blind fashion. Various performance measures were obtained consecutively in a single testing session pre- and post-exposure. Effects of hypoxic exposure on maximum speed and sprint times were trivial (<1.0%) but unclear (90% likely range, +/-5% to +/-9%). In rugby simulation, hypoxic exposure produced impairments of peak power in two scrums (15%, +/-8%; 9%, +/-7%) and impairments of time in offensive sprints (7%, +/-8%) and tackle sprints (11%, +/-9%). Pending further research, rugby players would be unwise to use normobaric intermittent hypoxic exposure to prepare for games at sea level.


Blood safety decisions, 1982 to 1986: perceptions and misconceptions.
Although blood bankers and those who treat persons with hemophilia are supportive of most of the recommendations of the Report, the manner in which the analysis was conducted and some of the general conclusions that were reached appear flawed. The flaws may reflect the deficiencies in the process by which the Committee gathered data more than any bias on the part of its members themselves. The Report may accurately reflect the testimony heard, but it is biased by the committee's acceptance as fact the opinions of critics who claim the AIDS epidemic was mismanaged by the blood-collecting agencies, professional organizations, hemophilia organizations, and the federal government. Countervailing views on the various issues are ignored or incompletely discussed. Much testimony was taken from the victims of the transfusion-associated AIDS epidemic. Reliance seems to have been placed upon hindsight testimony (taken 10 years after the events), rather than on documentation of what was known at the time when events unfolded. The Report states that "[t]he Committee's charge did not include the development of assertions about what should have been done at the time,"l(pl:4) yet that is precisely what was done. These comments address just a few of the misconceptions we perceive in the Report. They are based on our understanding of the state of knowledge--or ignorance--at the time that decisions about the safety of the blood supply were made. If we are to avert future threats to the blood supply from emerging infectious diseases, a goal that is universally embraced, we must learn the lessons the past can teach us, as painful as they may be. However, the hazards of judging history in hindsight should be avoided. Neither allegations nor opinions should be accepted as facts without critical examination and without placement in the context of contemporary knowledge; to accept a lesser standard does a great injustice to all who were touched by this tragedy.


Airway Hygiene in COVID-19 Pneumonia: Treatment Responses of 3 Critically Ill Cruise Ship Employees.
BACKGROUND COVID-19, the disease entity caused by the novel severe acute respiratory coronavirus 2 (SARS-CoV-2), continues to pose a major therapeutic challenge for clinicians. At present, an effective treatment regimen and vaccination has not been established. Many patients develop severe symptoms requiring endotracheal intubation and a prolonged stay in the Intensive Care Unit (ICU). In early postmortem examinations of COVID-19 patients, profuse viscous secretions were observed throughout the respiratory tract. Thus, oxygen supplementation without aggressive pulmonary hygiene management may be suboptimal. In the present case series, pulmonary hygiene management encompassed mucolytics, bronchodilators, and tracheal suctioning. We report 3 severe cases of COVID-19 pneumonia in cruise ship employees who were admitted to the ICU and responded to supportive mechanical ventilation and pulmonary hygiene management. CASE REPORT Three cruise ship employees with COVID-19 underwent endotracheal intubation and were admitted to the ICU for acute hypoxemic respiratory failure. Initial chest X-rays suggested multifocal pneumonia with superimposed acute respiratory distress syndrome (ARDS). A regimen of hydroxychloroquine, azithromycin, and dexamethasone was initiated on admission in all cases. Additionally, medications used for pulmonary hygiene were administered through a metered-dose inhaler (MDI) in line with the ventilator circuit. Endotracheal suctioning was performed prior to medication administration. The duration from endotracheal intubation to extubation ranged from 9 to 24 days. All 3 patients reached 30-day survival. CONCLUSIONS The cases reported highlight the importance of the use of airway hygiene with mucolytics, bronchodilators, and tracheal suctioning for patients with COVID-19 pneumonia requiring ventilatory support.


Carbon Nanotube/Conducting Polymer Hybrid Nanofibers as Novel Organic Bioelectronic Interfaces for Efficient Removal of Protein-Bound Uremic Toxins.
Protein-bound uremic toxins (PBUTs) can cause noxious effects in patients suffering from renal failure as a result of inhibiting the transport of proteins and inducing their structural modification. They are difficult to remove through standard hemodialysis (HD) treatment. Herein, we report an organic bioelectronic HD device system for the effective removal of PBUTs through electrically triggered dissociation of protein-toxin complexes. To prepare this system, we employed electrospinning to fabricate electrically conductive quaternary composite nanofiber mats-comprising multiwalled carbon nanotubes (MWCNTs), poly(3,4-ethylenedioxythiophene):polystyrenesulfonate (PEDOT:PSS), poly(ethylene oxide) (PEO), and (3-glycidyloxypropyl)trimethoxysilane (GOPS)-on conventional polyethersulfone (PES) dialysis membranes. These composite nanofiber platforms exhibited (i) long-term water resistance (due to cross-linking among PSS, PEO, and GOPS), (ii) high adhesion strength on the PES membrane (due to GOPS functioning as an adhesion promoter), (iii) enhanced electrical properties [due to the MWCNTs and PEDOT:PSS promoting effective electrical stimulation (ES) operation in devices containing bioelectronic interfaces (BEI)], and (iv) good anticoagulant ability and negligible hemolysis of red blood cells. We employed this organic BEI electronic system as a novel single-membrane HD device to study the removal efficiency of four kinds of uremic toxins [p-cresol (PC), indoxyl sulfate, and hippuric acid as PBUTs; creatinine as a non-PBUT] as well as the effects of ES on lowering the protein binding ratio. Our organic BEI devices provided a high rate of removal of PC with low protein loss after 4 h of a simulated dialysis process. It also functioned with low complement activation, low contact activation levels, and lower amounts of platelet adsorption, suggesting great suitability for use in developing next-generation bioelectronic medicines for HD.


Small doses of droperidol do not present relevant torsadogenic actions: a double-blind, ondansetron-controlled study.
Drugs used for postoperative nausea and vomiting prophylaxis are believed to provoke torsadogenic changes in cardiac repolarization. The aim of this study was to assess the effect of small doses of droperidol on the parameters of cardiac repolarization, including the QTc interval and transmural dispersion of repolarization. A total of 75 patients were randomly allocated to receive 0.625 or 1.25 mg droperidol or 8 mg ondansetron. The QTc interval was calculated using Bazett's formula and the Framingham correction. The transmural dispersion of repolarization was determined as Tpeak -Tend time. Transient QT prolongation, corrected with both formulae, followed 1.25 mg of droperidol 10 min after administration. No change in the QTc value was observed in the other groups. When corrected with Bazett's formula, QTc was prolonged above 480 ms in two patients receiving 1.25 mg droperidol (at the 10(th) and 20(th) minute of the study) and in one receiving ondansetron. No patients developed a QTc B prolongation over 500 ms. No increase above 480 ms was observed relative to the Framingham correction method. There were no significant differences in the Tpeak -Tend time either between or within the groups. In men without cardiovascular disorders small doses (1.25 mg) of droperidol prophylaxis induced transient QTc prolongation without changes in transmural dispersion of repolarization. The apparently low risk of the drug applies only in low risk male patients with a low pro-QTc score.


Ratio of T-Helper Type 1 (Th1) to Th17 Cytokines in Whole Blood Is Associated With Human β-Defensin 3 Expression in Skin and Persistent Staphylococcus aureus Nasal Carriage.
Nasal colonization has gained attention as an effect modifier in Staphylococcus aureus vaccine trials, suggesting interference of carriage with T-cell immunity. Likewise, T-cell signals may be involved in regulating effectors of epithelial innate defense. Whole blood from 43 persistent carriers and 49 noncarriers was stimulated with viable S. aureus T-helper type 1 (Th1), Th2, and Th17 cytokine expression was measured, compared between carrier groups, and linked with data on human β-defensin 3 (hBD-3) messenger RNA (mRNA) in skin while adjusting for transcriptionally relevant promoter haplotypes. Compared with carriers, stimulated whole blood from noncarriers contained on average 60% more interferon γ mRNA (P = .031) and 19% less interleukin 17A (IL-17A) mRNA (P = .11), resulting in, on average, a 90% higher IFN-γ to IL-17A mRNA ratio (P = .003). In a multivariable model, per duplication of the mRNA template, the risk of being a carrier increased by 93% for IL-17A (odds ratio [OR], 1.93; 95% confidence interval [CI], 1.10-3.41; P = .023) and decreased by 35% for IFN-γ (OR, 0.65; 95% CI, 0.47-0.91; P = .01). Independent of carriage and DEFB promotor haplotype, a 1-unit increase in the IFN-γ to IL-17A mRNA ratio (mean ± SD, 5.93 ± 1.60) led to a 24% increase in hBD-3 transcription in experimentally wounded human skin (P = .003). A low Th1 to Th17 mRNA ratio increases the propensity for persistent S. aureus nasal colonization, with downregulated hBD-3 transcription providing a potential link.


Craniofacial profile in Southern Chinese with hypodontia.
The association between craniofacial morphology and congenitally missing teeth is at present unclear. The aims of this study were to investigate whether hypodontia is associated with changes in the sagittal skeletal profile and to identify putative relationships between the skeletal profile and the severity of hypodontia. In a cross-sectional analytical study, the craniofacial structure and profile based on two-dimensional lateral cephalograms of Southern Chinese hypodontia patients (n = 49, 24 males, 25 females, mean age 16.4 years) and a comparison group without hypodontia (n = 41, 15 males, 26 females, mean age 16.7 years) were compared. The hypodontia patients were divided into three subgroups according to the severity of hypodontia (mild: < or =5, moderate: 6-9, and severe: > or =10 congenitally missing permanent teeth). All hypodontia patients had a significantly reduced mandibular plane, ANB, and face height compared with the control group (P < 0.05). A significant increase in chin thickness was also observed in the hypodontia patients (P < 0.05). As the severity of hypodontia increased from moderate to severe, a tendency to develop a retrognathic maxilla and a Class III skeletal relationship was noted in addition to the above features, making the already thick chin even more prominent. Statistically significant correlations (Pearson's correlation coefficient) were found between the number of missing teeth and SNA, NAFH, and ANB angles, the mandibular plane, chin thickness, and face height. In Southern Chinese subjects, hypodontia was associated with a shorter face, a flatter mandibular plane, a more pronounced chin, and a Class III skeletal profile. In severe hypodontia subjects, the maxilla was more retrognathic with a greater predilection to a Class III skeletal relationship.


The neural correlates of consciousness: an analysis of cognitive skill learning.
This paper presents a functional brain-imaging strategy designed to isolate neural correlates of consciousness in humans. This strategy is based on skill learning. In the example presented (rapidly generating verbs for visually presented nouns), a cognitive skill is examined before and after practice. As shown, there are marked qualitative differences in the neural circuitry supporting performance of this task in the naive and practised state that include, importantly, both increases and decreases from the baseline activity of the brain.


Insights into ErbB signaling from the structure of the ErbB2-pertuzumab complex.
We have determined the 3.2 A X-ray crystal structure of the extracellular domain of the human epidermal growth factor receptor 2 (ErbB2 or HER2) in a complex with the antigen binding fragment of pertuzumab, an anti-ErbB2 monoclonal antibody also known as 2C4 or Omnitarg. Pertuzumab binds to ErbB2 near the center of domain II, sterically blocking a binding pocket necessary for receptor dimerization and signaling. The ErbB2-pertuzumab structure, combined with earlier mutagenesis data, defines the pertuzumab residues essential for ErbB2 interaction. To analyze the ErbB2 side of the interface, we have mutated a number of residues contacting pertuzumab and examined the effects of these mutations on pertuzumab binding and ErbB2-ErbB3 heterodimerization. We have also shown that conserved residues previously shown to be necessary for EGF receptor homodimerization may be dispensible for ErbB2-ErbB3 heterodimerization.


Preexercise static stretching effect on leaping performance in elite rhythmic gymnasts.
The aim of this study was to examine the acute effects of static stretching (SS) on technical leaps performance in rhythmic gymnastics. Thirty-eight gymnasts (age 14.13 +/- 3.2 years), competing at the international and national levels, performed vertical jumps (squat jump, countermovement jump, hopping test [HT]) and technical leaps (split leap with leg stretched [SL], split leap with ring [RG], split leap with back bend of the trunk [BBT]) assessed in 2 different conditions: after SS and after their usual typical warm-up (TWU) as control conditions. Jumps and leaps flight time (FT) and ground contact time (GCT) parameters were evaluated by OptoJump. Leap performance was simultaneously evaluated by scores awarded by judges. For each dependent variable, the effect of warm-up condition (TWU and SS) was examined by a paired-sample t-test. A multiple regression analysis determined the amount of variance in judges' scores from the FT and GCT variables. Results revealed that vertical jumps FT was not affected by SS warm-up. Ground contact time of HT significantly increased after SS warm-up (p < 0.01). Static stretching reduced significantly (p < 0.01) the technical leap FT (decrements: SL = 7.1%, RG = 7.2%, and BBT = 6.4%). The results showed no significant effects of SS on technical leaps GCT. Static stretching significantly reduced (p < 0.001) the scores awarded by the judges. The FT was the main predictor of scores of the 3 technical leaps accounting for 9-30% of variance in both warm-up conditions. This study suggests that SS before leaping performance may negatively affect rhythmic gymnastics judges' evaluation.


Patient Centered Medical Home Cooking: Community Culinary Workshops for Multidisciplinary Teams.
Ideal management of chronic disease includes team based primary care, however primary care medical staff face a lack of training when addressing nutritional counseling and lifestyle prevention. Interactive culinary medicine education has shown to improve knowledge and confidence among medical students. The aim of this study was to determine whether a culinary medicine curriculum delivered to a multidisciplinary team of primary care medical staff and medical students in a community setting would improve self-reported efficacy in nutritional counseling and whether efficacy differed between participant roles. A 4-h interactive workshop that took place within the neighborhood of a primary care medical home was delivered to medical staff and students. Participants completed a voluntary questionnaire before and after the workshop that addressed participants' attitudes and confidence in providing nutritional counseling to patients. Chi-square tests were run to determine statistically significant associations between role of participant and survey question responses. Sign Rank tests were run to determine if pre-workshop responses differed significantly from post-workshop responses. Thirteen of seventeen responses related to attitudes and efficacy demonstrated significant improvement after the workshop compared with prior to the workshop. Significant differences noted between roles prior to the workshop disappear when asking the same questions after the workshop. Delivery of culinary medicine curricula to a primary care medical home team in a community setting is an innovative opportunity to collaboratively improve nutritional education and counseling in chronic disease prevention.


RISING STARS: Mechanistic insights into maternal-fetal cross talk and islet beta-cell development.
The metabolic health trajectory of an individual is shaped as early as prepregnancy, during pregnancy, and lactation period. Both maternal nutrition and metabolic health status are critical factors in the programming of offspring toward an increased propensity to developing type 2 diabetes in adulthood. Pancreatic beta-cells, part of the endocrine islets, which are nutrient-sensitive tissues important for glucose metabolism, are primed early in life (the first 1000 days in humans) with limited plasticity later in life. This suggests the high importance of the developmental window of programming in utero and early in life. This review will focus on how changes to the maternal milieu increase offspring's susceptibility to diabetes through changes in pancreatic beta-cell mass and function and discuss potential mechanisms by which placental-driven nutrient availability, hormones, exosomes, and immune alterations that may impact beta-cell development in utero, thereby affecting susceptibility to type 2 diabetes in adulthood.


Structural insights into integrin α5β1 opening by fibronectin ligand.
Integrin α5β1 is a major fibronectin receptor critical for cell migration. Upon complex formation, fibronectin and α5β1 undergo conformational changes. While this is key for cell-tissue connections, its mechanism is unknown. Here, we report cryo-electron microscopy structures of native human α5β1 with fibronectin to 3.1-angstrom resolution, and in its resting state to 4.6-angstrom resolution. The α5β1-fibronectin complex revealed simultaneous interactions at the arginine-glycine-aspartate loop, the synergy site, and a newly identified binding site proximal to adjacent to metal ion-dependent adhesion site, inducing the translocation of helix α1 to secure integrin opening. Resting α5β1 adopts an incompletely bent conformation, challenging the model of integrin sharp bending inhibiting ligand binding. Our biochemical and structural analyses showed that affinity of α5β1 for fibronectin is increased with manganese ions (Mn2+) while adopting the half-bent conformation, indicating that ligand-binding affinity does not depend on conformation, and α5β1 opening is induced by ligand-binding.


The RCGP Quality Practice Award for primary care teams.
Accreditation is a means of improving quality through the process of externally reviewing performance against written standards. Following the introduction of clinical governance, participation in quality accreditation schemes has been encouraged. The Royal College of General Practitioners' Quality Practice Award (QPA) is an example of a quality accreditation scheme for primary care practice teams. QPA applies to the wider primary care team and is directly relevant to nursing and midwifery staff employed by or attached to practice teams. QPA supports evidence-based and reflective practice, continuing professional development and team working, all of which are integral to current nursing and midwifery practice. Nurses and midwives working in primary care teams must be aware of QPA and, where necessary, actively and collaboratively participate in this process.


In vitro viability and cytotoxicity testing and same-well multi-parametric combinations for high throughput screening.
In vitro cytotoxicity testing has become an integral aspect of drug discovery because it is a convenient, costeffective, and predictive means of characterizing the toxic potential of new chemical entities. The early and routine implementation of this testing is testament to its prognostic importance for humans. Although a plethora of assay chemistries and methods exist for 96-well formats, few are practical and sufficiently sensitive enough for application in high throughput screening (HTS). Here we briefly describe a handful of the currently most robust and validated HTS assays for accurate and efficient assessment of cytotoxic risk. We also provide guidance for successful HTS implementation and discuss unique merits and detractions inherent in each method. Lastly, we discuss the advantages of combining specific HTS compatible assays into multi-parametric, same-well formats.


Unsupervised versus Supervised Identification of Prognostic Factors in Patients with Localized Retroperitoneal Sarcoma: A Data Clustering and Mahalanobis Distance Approach.
The aim of this report is to unveil specific prognostic factors for retroperitoneal sarcoma (RPS) patients by univariate and multivariate statistical techniques. A phase I-II study on localized RPS treated with high-dose ifosfamide and radiotherapy followed by surgery (ISG-STS 0303 protocol) demonstrated that chemo/radiotherapy was safe and increased the 3-year relapse-free survival (RFS) with respect to historical controls. Of 70 patients, twenty-six developed local, 10 distant, and 5 combined relapse. Median disease-free interval (DFI) was 29.47 months. According to a discriminant function analysis, DFI, histology, relapse pattern, and the first treatment approach at relapse had a statistically significant prognostic impact. Based on scientific literature and clinical expertise, clinicopathological data were analyzed using both a supervised and an unsupervised classification method to predict the prognosis, with similar sample sizes (66 and 65, resp., in casewise approach and 70 in mean-substitution one). This is the first attempt to predict patients' prognosis by means of multivariate statistics, and in this light, it looks noticable that (i) some clinical data have a well-defined prognostic value, (ii) the unsupervised model produced comparable results with respect to the supervised one, and (iii) the appropriate combination of both models appears fruitful and easily extensible to different clinical contexts.


Targeting of lung cancer mutational hotspots by polycyclic aromatic hydrocarbons.
Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous in combustion products of organic matter, including cigarette smoke. Metabolically activated diol epoxides of these compounds, including benzo[a]pyrene diol epoxide (B[a]PDE), have been suggested as causative agents in the development of lung cancer. We previously mapped the distribution of B[a]PDE adducts within the p53 tumor suppressor gene (also known as TP53), which is mutated in 60% of human lung cancers, and found that B[a]PDE adducts preferentially form at lung cancer mutational hotspots (codons 154, 157, 158, 245, 248, and 273). Other PAHs may be important in lung cancer as well. Here we have mapped the distribution of adducts induced by diol epoxides of additional PAHs: chrysene (CDE), 5-methylchrysene (5-MCDE), 6-methylchrysene (6-MCDE), benzo[c]phenanthrene (B[c]PDE), and benzo[g]chrysene (B[g]CDE) within exons 5, 7, and 8 of the p53 gene in human bronchial epithelial cells. CDE exposure produced only low levels of adducts. Exposure of cells to the other activated PAHs resulted in DNA damage patterns similar to those previously observed with B[a]PDE but with some distinct differences. 5-MCDE, 6-MCDE, B[g]CDE, and B[c]PDE efficiently induced adducts at guanines within codons 154, 156, 157, 158, and 159 of exon 5, codons 237, 245 and 248 of exon 7, and codon 273 of exon 8, but the relative levels of adducts at each site varied for each compound. B[g]CDE, B[c]PDE, and 5-MCDE induced damage at codon 158 more selectively than 6-MCDE or B[a]PDE. The sites most strongly involved in PAH adduct formation were also the sites of highest mutation frequency (codons 157, 158, 245, 248, and 273). The data suggest that PAHs contribute to the mutational spectrum in human lung cancer.


Substrate specificity and expression of three 2,3-dihydroxybiphenyl 1,2-dioxygenases from Rhodococcus globerulus strain P6.
Rhodococcus globerulus strain P6 contains at least three genes, bphC1, bphC2, and bphC3, coding for 2,3-dihydroxybiphenyl 1,2-dioxygenases; the latter two specify enzymes of the family of one-domain extradiol dioxygenases. In order to assess the importance of these different isoenzymes for the broad catabolic activity of this organism towards the degradation of polychlorinated biphenyls (PCBs), the capacities of recombinant enzymes expressed in Escherichia coli to transform different chlorosubstituted dihydroxybiphenyls formed by the action of R. globerulus P6 biphenyl dioxygenase and biphenyl 2,3-dihydrodiol dehydrogenase were determined. Whereas both BphC2 and BphC3 showed similar activities for 2,3-dihydroxybiphenyl and all monochlorinated 2,3-dihydroxybiphenyls, BphC1 exhibited only weak activity for 2'-chloro-2,3-dihydroxybiphenyl. More highly chlorinated 2'-chlorosubstituted 2,3-dihydroxybiphenyls were also transformed at high rates by BphC2 and BphC3 but not BphC1. In R. globerulus P6, BphC2 was constitutively expressed, BphC1 expression was induced during growth on biphenyl, and BphC3 was not expressed at significant levels under the experimental conditions. Although we cannot rule out the expression of BphC3 under certain environmental conditions, it seems that the contrasting substrate specificities of BphC1 and BphC2 contribute significantly to the versatile PCB-degrading phenotype of R. globerulus P6.


Disturbances in leptin metabolism are related to energy imbalance during acute exacerbations of chronic obstructive pulmonary disease.
Previously we reported an impaired energy balance in patients with chronic obstructive pulmonary disease (COPD) during an acute disease exacerbation, but limited data are available on the underlying mechanisms. Experimental and clinical research supports the hypothesis of involvement of the hormone leptin in body weight and energy balance homeostasis. The aim of this study was to investigate the course of the energy balance in relation to leptin and the soluble tumor necrosis factor (TNF) receptors (sTNF-R) 55 and 75, plasma glucose, and serum insulin in patients with severe COPD during the first 7 d of hospitalization for an acute exacerbation (n = 17, 11 men, age mean [SD] 66 [10] yr, FEV(1) 36 [12] %pred). For reference values of the laboratory parameters, blood was collected from 23 (16 men) healthy, elderly subjects. On admission, the dietary intake/resting energy expenditure (REE) ratio was severely depressed (1.28 [0.57]), but gradually restored until Day 7 (1.65 [0. 45], p = 0.005 versus Day 1). Glucose and insulin concentrations were elevated on admission, but on Day 7 only plasma glucose was decreased. The sTNF-Rs were not different from healthy subjects and did not change. Plasma leptin, adjusted for fat mass expressed as percentage of body weight (%FM), was elevated on Day 1 compared with healthy subjects (1.82 [3.85] versus 0.32 [0.72] ng%/ml, p = 0.008), but decreased significantly until Day 7 (1.46 [3.77] ng%/ml, p = 0. 015 versus Day 1). On Day 7, sTNF-R55 was, independently of %FM, correlated with the natural logarithm (LN) of leptin (r = 0.65, p = 0.041) and with plasma glucose (r = 0.81, p = 0.015). In addition, the dietary intake/REE ratio was not only inversely related with LN leptin (-0.74, p = 0.037), but also with sTNF-R55 (r = -0.93, p = 0. 001) on day seven. In conclusion, temporary disturbances in the energy balance were seen during an acute exacerbation of COPD, related to increased leptin concentrations as well as to the systemic inflammatory response. Evidence was found that the elevated leptin concentrations were in turn under control of the systemic inflammatory response, and, presumably, the high-dose systemic glucocorticosteroid treatment.


Amino Acid Sequence Around the Catalytic Site in Glyceraldehyde-3-Phosphate Dehydrogenase from Bacillus stearothermophilus.
The tryptic peptide containing the active-site cysteine in glyceraldehyde-3-phosphate dehydrogenase from Bacillus stearothermophilus 1503 was isolated after inhibition of the enzyme with (14)C-iodoacetate. The amino acid sequence of the 20-residue peptide was determined by 19 successive cycles of dansyl-Edman degradation. The sequence shows considerable homology with its counterparts from mesophilic sources but differs by the addition of Ala-His-His at the N-terminus and by the substitution of phenylalanine for leucine in the prototype sequence.


Supported treadmill training for gait and balance in a patient with progressive supranuclear palsy.
Impaired balance, gait disturbances, and frequent falls are common problems in people with progressive supranuclear palsy (PSP). This case report describes the use of a modified body weight support treadmill training program to reduce falls and improve the balance and gait of a patient with PSP. The patient was a 62-year-old man diagnosed with PSP. His major problems were impaired balance and frequent, abrupt falls. Physical therapy included walk training, balance perturbation, and step training using body weight support with a treadmill. Training sessions lasted 11/2 hours and occurred 3 days a week for 8 weeks. Fall incidence, balance, and gait were assessed before, during, and after the program. The patient reported fewer falls during and after training. Balance and gait improved after training. This case report is the first to report fall reduction, improved gait, and improved balance following physical therapy for a person with PSP.


Combined native magnetic resonance angiography, flow-quantifying, and perfusion-imaging for impending second-stroke assessment.
This special report introduces native flow quantitative imaging for evaluating stroke risk. Moreover, the advantage of combining three imaging techniques [magnetic resonance angiography (MRA), phase-contrast (PC) flow imaging, and arterial spin-labeling imaging] is shown to be beneficial for responding to ischemia and preserving viable neurons. These quantitative imaging techniques provide authoritative information for diagnosing impending stroke and selecting appropriate treatment.


Evaluation of the protective effects of quinacrine against Bacillus anthracis Ames.
Bacillus anthracis has gained notoriety as a dangerous biological weapon because of its virulence and ability to produce highly resistant spores. In addition, the ability of this organism to produce plasmid-encoded edema toxin (EdTx) and lethal toxin (LeTx) plays a pivotal role in the pathogenesis of anthrax. In this study, the efficacy of quinacrine was evaluated against the effects of anthrax toxins in vitro and its ability to provide protection against challenge with B. anthracis Ames strain spores in an intranasal mouse and guinea pig model. Quinacrine protected murine macrophages in vitro against cytotoxicity and cAMP production induced by LeTx and EdTx, respectively, at concentrations of 40-80 microM, most likely by preventing acidification of the endosomes. However, animals dosed with human equivalent doses of quinacrine were not protected against respiratory spore challenge. The failure of quinacrine to provide protection against inhalation anthrax was attributed to our inability to attain inhibitory concentrations of the drug in the serum or tissues. After daily administration of 43.3 mg quinacrine to guinea pigs (300 g), serum levels after 96 h were only 9.9 microM, a concentration not sufficient to protect macrophages in vitro. Administration of high doses of quinacrine (86.6 mg/kg) was toxic to the animals. These results illustrate some of the difficulties in developing protective therapeutic strategies against inhalation anthrax even when antitoxic drugs appear effective in vitro.


Allogeneic stem cell transplantation using HLA-matched donors for acute myeloid leukemia with deletion 5q or monosomy 5: a study from the Acute Leukemia Working Party of the EBMT.
Deletion 5q or monosomy 5 (-5/5q-) in acute myeloid leukemia (AML) is a common high-risk feature that is referred to allogeneic stem cell transplantation. However, -5/5q- is frequently associated with other high-risk cytogenetic aberrations such as complex karyotype, monosomal karyotype, monosomy 7 (-7), or 17p abnormalities (abn (17p)), the significance of which is unknown. In order to address this question, we studied adult patients with AML harboring -5/5q- having their first allogeneic transplantation between 2000 and 2015. Five hundred and one patients with -5/5q- have been analyzed. Three hundred and thirty-eight patients (67%) were in first remission and 142 (28%) had an active disease at time of allogeneic transplantation. The 2-year probabilities of overall survival and leukemia-free survival were 27% and 20%, respectively. The 2-year probability of treatment-related mortality was 20%. We identified four different cytogenetic groups according to additional abnormalities with prognostic impact: -5/5q- without complex karyotype, monosomal karyotype or abn(17p), -5/5q- within a complex karyotype, -5/5q- within a monosomal karyotype and the combination of -5/5q- with abn(17p). In multivariate analysis, factors associated with worse overall survival and leukemia-free survival across the four groups were active disease, age, monosomal karyotype, and abn(17p). The presence of -5/5q- without monosomal karyotype or abn(17p) was associated with a significantly better survival rate while -5/5q- in conjunction with monosomal karyotype or abn(17p) translated into a worse outcome. The patients harboring the combination of -5/5q- with abn(17p) showed very limited benefit from allogeneic transplantation.


Prostate stem cell antigen polymorphisms and susceptibility to gastric cancer: a systematic review and meta-analysis.
Many studies have reported that prostate stem cell antigen (PSCA) polymorphisms (rs2294008 and/or 2976392) are significantly associated with gastric cancer (GC) risk, although the published results are inconsistent. We conducted a systematic review and meta-analysis for relevant literatures to quantitatively evaluate the relationship between PSCA polymorphisms and GC susceptibility. Extensive searches were conducted in three databases up to November 1, 2011. ORs and 95% CIs were used to assess the strength of the associations. The data were further stratified by ethnicity, histopathology, subsite, and study design. All of the associations were evaluated with dominant model and recessive model, respectively. Heterogeneity and publication bias were also assessed by Q test, I(2), and funnel plot accordingly. Nine articles including 11 case-control data sets were included, with 10,746 GC cases and 9,158 controls for rs2294008 and 6,060 cases and 4,824 controls for rs2976392. The results showed that risk allele carriers were significantly associated with GC risk compared with nonrisk allele homozygotes. In stratification analyses, these associations remained significant for majority of subgroups except for Caucasians and noncardia tumor in dominant model, and cardia tumor in both dominant and recessive model. Random model was used when heterogeneity among studies was detected. No publication bias was observed. The two loci of PSCA (rs2294008 and rs2976392) were both significantly associated with GC susceptibility and in linkage disequilibrium. More prospective studies on PSCA polymorphisms at multicenters with sufficient sample size and less heterogeneity will be needed for further validations.


Inhibitory Effects of Dietary N-Glycans From Bovine Lactoferrin on Toll-Like Receptor 8; Comparing Efficacy With Chloroquine.
Toll-like receptor 8 (TLR-8) plays a role in the pathogenesis of autoimmune disorders and associated gastrointestinal symptoms that reduce quality of life of patients. Dietary interventions are becoming more accepted as mean to manage onset, progression, and treatment of a broad spectrum of inflammatory conditions. In this study, we assessed the impact of N-glycans derived from bovine lactoferrin (bLF) on the inhibition of TLR-8 activation. We investigated the effects of N-glycans in their native form, as well as in its partially demannosylated and partially desialylated form, on HEK293 cells expressing TLR-8, and in human monocyte-derived dendritic cells (MoDCs). We found that in HEK293 cells, N-glycans strongly inhibited the ssRNA40 induced TLR-8 activation but to a lesser extent the R848 induced TLR-8 activation. The impact was compared with a pharmaceutical agent, i.e., chloroquine (CQN), that is clinically applied to antagonize endosomal TLR- activation. Inhibitory effects of the N-glycans were not influenced by the partially demannosylated or partially desialylated N-glycans. As the difference in charge of the N-glycans did not influence the inhibition capacity of TLR-8, it is possible that the inhibition mediated by the N-glycans is a result of a direct interaction with the receptor rather than a result of pH changes in the endosome. The inhibition of TLR-8 in MoDCs resulted in a significant decrease of IL-6 when cells were treated with the unmodified (0.5-fold, p < 0.0001), partially demannosylated (0.3-fold, p < 0.0001) and partially desialylated (0.4-fold, p < 0.0001) N-glycans. Furthermore, the partially demannosylated and partially desialylated N-glycans showed stronger inhibition of IL-6 production compared with the native N-glycans. This provides evidence that glycan composition plays a role in the immunomodulatory activity of the isolated N-glycans from bLF on MoDCs. Compared to CQN, the N-glycans are specific inhibitors of TLR-8 activation and of IL-6 production in MoDCs. Our findings demonstrate that isolated N-glycans from bLF have attenuating effects on TLR-8 induced immune activation in HEK293 cells and human MoDCs. The inhibitory capacity of N-glycans isolated from bLF onTLR-8 activation may become a food-based strategy to manage autoimmune, infections or other inflammatory disorders.


Nucleotide sequence of a Madagascar hepatopancreatic parvovirus (HPV) and comparison of genetic variation among geographic isolates.
A segment of Madagascar hepatopancreatic parvovirus (HPV) genomic sequence (5742 nucleotides) was determined through PCR and direct sequencing. This nucleotide sequence was compared to isolates from Australia, Thailand, Korea, and Tanzania, and the mean distance was determined to be 17%. The Madagascar HPV is closest to the Tanzania isolate (12%), followed by isolates from Korea (15%), Australia (17%) and Thailand (20%). Analysis of the genomic structure revealed that this HPV sequence is comprised of one partial Left open reading frame (ORF) (349 amino acids, aa) and complete Mid (578 aa) and Right (820 aa) ORFs. The amino acid sequences of the 3 ORFs were compared among isolates. The Right ORF was found to have the highest variation with a mean distance of 24%. This was followed by the Left and Mid ORF with distances of 13 and 7%, respectively. A phylogenetic analysis based on the amino acid sequence of the Right ORF divides 7 HPV isolates into 3 well-separated groups: Korea, Thailand, and Australia. The Madagascar HPV clustered with the Korea and Tanzania isolates. In Madagascar, HPV has been detected by histological examination since the 1990s. PCR analysis of a recent (2007) sampling showed a 100% prevalence. HPV was also detected in Mozambique with a 100% prevalence. High (95%) prevalence of HPV was found in wild Penaeus merguinesis collected from New Caledonia. These results indicate that HPV displays a high degree of genetic diversity and is distributed worldwide among populations of penaeid shrimp.


Subcellular sources of luminescence in Noctiluca.
The perivacuolar cytoplasm of Noctiluca miliaris contains approximately 10(4) microsources of luminescence, with dimensions of 0.5 to 1.5 microns, which exhibit marked fluorescence with ultraviolet excitation. Local invasion by an action potential elicits light emission (microflashes) from these sources with a coupling latency of about 2 milliseconds. Magnitudes of the microflash vary directly with the dimensions of the source. Time courses of the microflash resemble that of the macroflash emitted by the whole cell but have somewhat shorter time constants. The small discrepancy induration between micro- and macroflash can be explained by the 5- to 10-millisecond asynchrony of microsource triggering that results from the conduction time of the action potential. Reversible gradations in amplitude of the macroflash, as from potentiation or fatigue, result from parallel summation of graded changes in microflash intensity. Thus the macroflash gives a reasonably true picture of the subcellular kinetics of luminescence.


[Heavy water (D20) inhibits growth of human xenotransplanted oropharyngeal cancers. An animal experiment study in nude mice].
The large mass ratio of D to H, 2 to 1, gives rise to multiple effects in living organisms, when D is administered as heavy water (D2O). Deuterium at high concentration interferes with cell division and depresses the uptake of DNA-precursors in mammalian cells (Katz et al., 1970). However, the effects of D2O on the organism disappear after cessation of deuteration of the body water. Mammals do not survive more than 35% of D2O substitution for normal water in their body fluids for long periods of time. However, mice drinking 30% D2O have a normal life span and normal body weight (Katz et al., 1970; Hodel et al., 1982). Heavy water also exerts antineoplastic effects. We demonstrated that moderate body deuteration combined with a cytostatic drug significantly increases the survival time of mice bearing transplantable murine neoplasms (Laissue et al., 1982). Hitherto, it is not known whether heavy water affects human neoplasms in the same way. Therefore, we have studied human squamous cell carcinomas of the oropharyngeal region which were xenografted in nude mice. A moderate deuteration of body water of the mice (less than or equal to 20 atom %) drinking 30% D2O, inhibited the tumour growth markedly by a factor of 0.4 to 0.5 compared to the growth of the same tumours in control animals drinking tap water. This effect of D2O seems to be directly proportional to the grade of malignancy of the carcinomas. Histologically, no differences were detectable between the original carcinomas and the tumour grafts in animals with and without D2O.


Bottom-up Synthesis of Nitrogen-Doped Porous Graphene Nanoribbons.
Although methods for a periodic perforation and heteroatom doping of graphene sheets have been developed, patterning closely spaced holes on the nanoscale in graphene nanoribbons is still a challenging task. In this work, nitrogen-doped porous graphene nanoribbons (N-GNRs) were synthesized on Ag(111) using a silver-assisted Ullmann polymerization of brominated tetrabenzophenazine. Insights into the hierarchical reaction pathways from single molecules toward the formation of one-dimensional organometallic complexes and N-GNRs are gained by a combination of scanning tunneling microscopy (STM), atomic force microscopy (AFM) with CO-tip, scanning tunneling spectroscopy (STS), and density functional theory (DFT).


Nonlinear dynamic system identification using Chebyshev functional link artificial neural networks.
A computationally efficient artificial neural network (ANN) for the purpose of dynamic nonlinear system identification is proposed. The major drawback of feedforward neural networks, such as multilayer perceptrons (MLPs) trained with the backpropagation (BP) algorithm, is that they require a large amount of computation for learning. We propose a single-layer functional-link ANN (FLANN) in which the need for a hidden layer is eliminated by expanding the input pattern by Chebyshev polynomials. The novelty of this network is that it requires much less computation than that of a MLP. We have shown its effectiveness in the problem of nonlinear dynamic system identification. In the presence of additive Gaussian noise, the performance of the proposed network is found to be similar or superior to that of a MLP. A performance comparison in terms of computational complexity has also been carried out.


Evaluating efficiency-equality tradeoffs for mobile source control strategies in an urban area.
In environmental risk management, there are often interests in maximizing public health benefits (efficiency) and addressing inequality in the distribution of health outcomes. However, both dimensions are not generally considered within a single analytical framework. In this study, we estimate both total population health benefits and changes in quantitative indicators of health inequality for a number of alternative spatial distributions of diesel particulate filter retrofits across half of an urban bus fleet in Boston, Massachusetts. We focus on the impact of emissions controls on primary fine particulate matter (PM(2.5)) emissions, modeling the effect on PM(2.5) concentrations and premature mortality. Given spatial heterogeneity in baseline mortality rates, we apply the Atkinson index and other inequality indicators to quantify changes in the distribution of mortality risk. Across the different spatial distributions of control strategies, the public health benefits varied by more than a factor of two, related to factors such as mileage driven per day, population density near roadways, and baseline mortality rates in exposed populations. Changes in health inequality indicators varied across control strategies, with the subset of optimal strategies considering both efficiency and equality generally robust across different parametric assumptions and inequality indicators. Our analysis demonstrates the viability of formal analytical approaches to jointly address both efficiency and equality in risk assessment, providing a tool for decisionmakers who wish to consider both issues.


The intra- and inter-rater reproducibility of bone level measurements at strategic mini-implants using dental panoramic radiography.
This experimental study focused on the intra- and inter-rater reproducibility of vertical bone level (VBL) measurements at strategic mini-implants (MI) using digital panoramic radiographs (PR). VBLs of 152 MIs for removable partial denture stabilization at 50 randomly chosen PRs from a clinical trial were digitally evaluated by three ratters. Rater deviations exceeding 0.5 mm were re-examined. The intra-class correlation coefficient (ICC) was applied to estimate reliability. The smallest detectable change (SDC) was interrelated to the minimal clinically important change of 0.2 mm. The first measurement round revealed intra- and inter-rater ICCs of > 0.8. However, 28 sites (9 %) were unreadable, and 97 sites (32 %) revealed differences between observers of ≥ 0.5 mm. Following a consensus session and re-training, an additional 8 sites were excluded and all remaining VBL differences were ≤ 0.5 mm. Thus, the SDCs with 95 % credibility were improved from 0.73 to 0.31 mm in the intra-rater and from 1.52 to 0.34 mm in the inter-rater statistics. Given a 50 % credibility for this special setting, both the intra- and inter-rater SDCs were 0.11 mm. Digital PR can be reliably utilized to determine VBLs around MIs under conditions of at least two trained observers, mutual calibration sessions, and exclusion of unquantifiable radiographs. DRKS00007589, www.germanctr.de.


Evidence for self-sustaining populations of Arcuatula senhousia in the UK and a review of this species' potential impacts within Europe.
The invasive Asian date mussel (Arcuatula senhousia) inhabits diverse global coastal environments, in some circumstances posing significant ecological and economic risks. Recently recorded in the Greater North Sea ecoregion, an established population has not previously been confirmed. Combining historical and field data, we provided baseline information from the UK and recorded colonisation in a variety of habitats. Gonadal development was assessed using the gonadosomatic index (GSI) to determine if an intertidal soft-sediment population is self-sustaining. Arcuatula senhousia records from subtidal muddy/mixed-sediment within a major estuarine system from 2007 to 2016 were also analysed. First detected in 2011, spatial distribution was variable across the years within the subtidal, with individuals found at 4-9 out of 25 sites, and densities per site varying from 10 to 290 individuals per m2. The intertidal population was, in part, associated with seagrass (Zostera spp.) and attached to bivalves. In marinas, individuals were attached to concrete tiles, associated with live Mytilus edulis, and to dead Ostrea edulis. Mean GSI from the intertidal population differed across months, peaking in July before declining in September/October, but with high inter-individual variability. Arcuatula senhousia is reproducing and maintaining viable populations. Using a natural capital approach, we identify the potential impacts on Europe's functionally important habitats, fisheries and aquaculture if its spread continues.


Monte carlo simulation of macromolecular ionization by nanoelectrospray.
Electrospray ionization (ESI) is commonly used in macromolecular mass spectrometry, yet the dynamics of macromolecules in ESI droplets are not well understood. In this study, a Monte Carlo based model was developed, which can predict the efficiency of electrospray ionization for macromolecules, i.e., the number of macromolecular ions produced per macromolecules electrosprayed. The model takes into account ESI droplet evaporation, macromolecular diffusion within the droplet, droplet fissions, and the statistical nature of the ESI process. Two idealized representations of macromolecular analytes were developed, describing cluster prone, droplet surface inactive macromolecules and droplet surface active macromolecules, respectively. It was found that surface active macromolecules are preferentially ionized over surface inactive cluster prone macromolecules when the initial droplet size is large and the analyte concentration in solution is high. Simulations showed that ESI efficiency decreases with increasing initial droplet size and analyte molecular weight, and is influenced by analyte surface activity, the properties of the solvent, and the variance of the droplet size distribution. Model predictions are qualitatively supported by experimental measurements of macromolecular electrospray ionization made previously. Overall, this study demonstrates the potential capabilities of Monte Carlo based ESI models. Future developments in such models will allow for more accurate predictions of macromolecular ESI intensity.


Frequent independent duplicative transpositions activate a single VSG gene.
The expression of several surface antigen genes in Trypanosoma brucei is mediated by the duplicative transposition of a basic-copy variant surface glycoprotein (VSG) gene into an expression site. We determined that the appearance of variant 118, in a parasitemia, resulted from at least four independent duplicative transpositions of the same VSG 118 gene. Variants 117 and 118 both appeared at specific periods but resulted from multiple independent activations. Antigenic variants thus occur in an ordered manner. We show that in the duplicative transpositions of VSG genes, the ends of the transposed segments were homologous between the basic copy and the expression site. Sequences other than the previously reported 70-base-pair (bp) repeats could be involved. In one variant, 118 clone 1, the homology was between a sequence previously transposed into the expression site and a sequence located 6 kilobases upstream of the VSG 118 gene. In variant 118b the homology was presumably in 70-bp repeat arrays, while in a third 118 variant yet another sequence was involved. The possibility that the 70-bp repeats are important in the initial steps of the recombinational events was illustrated by a rearrangement involving a 70-bp repeat array. The data provide strong evidence for the notion that gene conversion mediates the duplicative transposition of VSG genes. We discuss a model that explains how the process of duplicative transposition can occur at random and still produce an ordered appearance of variants.


Accurate analysis of tumor margins using a fluorescent pH Low Insertion Peptide (pHLIP).
The recurrence of certain cancers remains quite high due to either incomplete surgical removal of the primary tumor or the presence of small metastases that are invisible to the surgeon. Near infrared (NIR) fluorescence imaging might improve surgical outcomes by providing sensitive, specific, and real-time visualization of normal and diseased tissues if agents can be found that discriminate between normal and diseased tissue and define tumor margins. We have developed a new approach for revealing tumor borders by using NIR fluorescently labeled pH Low Insertion Peptide (pHLIP) and have created a computational program for the quantitative assessment of tumor boundaries. The approach is tested in vivo by co-localization of GFP-tumors and NIR emission from the fluorescently labeled pHLIP, and it is found that boundaries are accurately reported and that sub-millimeter masses can be detected.


Update on the Management of Chronic Constipation.
Chronic constipation (CC) is a highly prevalent disorder encountered by health care providers of all specialties. The diagnosis can be confidently made by taking a careful history, evaluating for warning signs and symptoms, performing an examination, including a digital rectal exam, and using the Rome IV criteria. Treatment should begin at the first visit; most patients require few diagnostic tests to make, or confirm, the diagnosis of CC. Assuming that the patient has persistent symptoms of constipation, despite using traditional therapy (fiber, osmotic agents), then patients should be offered one of the newer treatments, rather than repeating prior treatments, which is a common practice. Lubiprostone, a chloride channel activator, has been shown to safely improve symptoms of CC. Its proven track record of success over the last decade is a common reason why many health care providers choose this as a first-line agent. Alternatively, linaclotide, which stimulates guanylate cyclase C receptors, and which has also been shown to improve symptoms of CC in large, randomized trials, is another logical choice. The decision of which agent to use first often depends upon the patient's co-payment or insurance plan. Either medication should be given a trial of at least 4-6 weeks. If a patient does not respond, then the patient should be treated with the other agent. If symptoms persist, the clinician should consider the possibility of overlapping, or predominant, pelvic floor dysfunction (PFD). The combination of high-resolution anorectal manometry and a balloon expulsion test can be used to make the diagnosis of PFD. If present, patients should be referred to a knowledgeable physical therapist for pelvic floor retraining. New treatment options are available to treat the multiple symptoms of CC. Co-existing pelvic floor dysfunction should be considered in those patients who fail medical therapy.


Hormone alterations in breast cancer: examining the hypotheses.
Many of the epidemiologic risk factors for breast cancer offer clinicians little help in anticipating who is likely to be struck with the disease or how to prevent it. There are only a handful of clinically significant risk factors for breast cancer. These include being a woman, growing older, already having breast cancer in 1 breast, and having a first-degree relative (mother, sister, or daughter) who has been diagnosed with breast cancer. Most risk factors have a weak association with breast cancer, occur too infrequently, or are physiologic events not amenable to intervention. In recent years, the search for breast cancer causes has led to the identification of genetic markers that seem to predispose some women to breast cancer. For decades, however, researchers have been assessing and analyzing hormonal changes in the hope of finding a predictable breast cancer marker or cause that can easily be manipulated to prevent or more effectively treat the disease. Nearly a dozen hormonal hypotheses of breast cancer causes have been proposed -- among them estrogen excess, low luteal-phase progestational activity, adrenal androgen deficiency, ovarian androgen excess, melatonin deficiency, prolactin excess, and thyroid insufficiency. For most, the data are equivocal and inconclusive. The androgen deficiency hypotheses, however, may have some bearing on premenopausal breast cancer, and the ovarian dysfunction hypothesis may have some bearing on postmenopausal breast cancer.


Recommendations for the standardization and interpretation of the electrocardiogram: part II: Electrocardiography diagnostic statement list: a scientific statement from the American Heart Association Electrocardiography and Arrhythmias Committee, Council on Clinical Cardiology; the American College of Cardiology Foundation; and the Heart Rhythm Society: endorsed by the International Society for Computerized Electrocardiology.
This statement provides a concise list of diagnostic terms for ECG interpretation that can be shared by students, teachers, and readers of electrocardiography. This effort was motivated by the existence of multiple automated diagnostic code sets containing imprecise and overlapping terms. An intended outcome of this statement list is greater uniformity of ECG diagnosis and a resultant improvement in patient care. The lexicon includes primary diagnostic statements, secondary diagnostic statements, modifiers, and statements for the comparison of ECGs. This diagnostic lexicon should be reviewed and updated periodically.


Novel drug targets in cell wall biosynthesis exploited by gene disruption in Pseudomonas aeruginosa.
For clinicians, Pseudomonas aeruginosa is a nightmare pathogen that is one of the top three causes of opportunistic human infections. Therapy of P. aeruginosa infections is complicated due to its natural high intrinsic resistance to antibiotics. Active efflux and decreased uptake of drugs due to cell wall/membrane permeability appear to be important issues in the acquired antibiotic tolerance mechanisms. Bacterial cell wall biosynthesis enzymes have been shown to be essential for pathogenicity of Gram-negative bacteria. However, the role of these targets in virulence has not been identified in P. aeruginosa. Here, we report knockout (k.o) mutants of six cell wall biosynthesis targets (murA, PA4450; murD, PA4414; murF, PA4416; ppiB, PA1793; rmlA, PA5163; waaA, PA4988) in P. aeruginosa PAO1, and characterized these in order to find out whether these genes and their products contribute to pathogenicity and virulence of P. aeruginosa. Except waaA k.o, deletion of cell wall biosynthesis targets significantly reduced growth rate in minimal medium compared to the parent strain. The k.o mutants showed exciting changes in cell morphology and colonial architectures. Remarkably, ΔmurF cells became grossly enlarged. Moreover, the mutants were also attenuated in vivo in a mouse infection model except ΔmurF and ΔwaaA and proved to be more sensitive to macrophage-mediated killing than the wild-type strain. Interestingly, the deletion of the murA gene resulted in loss of virulence activity in mice, and the virulence was restored in a plant model by unknown mechanism. This study demonstrates that cell wall targets contribute significantly to intracellular survival, in vivo growth, and pathogenesis of P. aeruginosa. In conclusion, these findings establish a link between cell wall targets and virulence of P. aeruginosa and thus may lead to development of novel drugs for the treatment of P. aeruginosa infection.


Variation in shape of the lingula in the adult human mandible.
The lingulae of both sides of 165 dry adult human mandibles, 131 males and 34 females of Indian origin, were classified by their shape into 4 types: 1, triangular; 2, truncated; 3, nodular; and 4, assimilated. Triangular lingulae were found in 226 (68.5%) sides, truncated in 52 (15.8%), nodular in 36 (10.9%) and assimilated in 16 (4.8%) sides. Triangular lingulae were found bilaterally in 110, truncated in 23, nodular in 17 and assimilated in 7 mandibles. Of the remaining 8 mandibles with different appearances on the 2 sides, 6 had a combination of triangular and truncated and 2 had nodular and assimilated. The incidence of triangular and assimilated types in the male and female mandibles are almost equal. In the truncated type it was double in the male mandibles while the nodular type was a little less than double in the female mandibles.


The exogenous administration of CB2 specific agonist, GW405833, inhibits inflammation by reducing cytokine production and oxidative stress.
The present study aimed to investigate the role of cannabinoid 2 (CB2) receptors in a rat model of acute inflammation. Therefore, the potential of anti-inflammatory effects of CB2 receptor agonist (GW405833), CB2 receptor antagonist (AM630), and diclofenac, were investigated in carrageenan induced paw oedema in rats: as were assessed by measuring paw oedema; myeloperoxidase (MPO) activity in paw tissue; malondialdehyde (MDA) concentration; glutathione (GSH) level in paw tissue for oxidant/antioxidant balance; cytokine (interleukin-1β, IL-1β; tumour necrosis factor-α, TNF-α) levels in serum; histopathology of paw tissue for inflammatory cell accumulations. The results showed that GW405833 or diclofenac significantly reduced carrageenan-induced paw oedema. GW405833 also inhibited the increase of MPO activity, the recruitment of total leukocytes and neutrophils, and MDA concentration during carrageenan-induced acute inflammation, along with reversed nearly to the normal levels the increased of TNF-α, and IL-1β in serum. AM630 did not affect inflammation alone however clearly reversed the effects of agonist when co-administered. The mechanism of GW405833's suppression of inflammation is supported by these results, which are achieved by the inhibition of neutrophil migration, which regulates the reduction of oxidative stress, TNF-α and IL-1β levels. Finally, the activation of CB2 receptor, by selective agonist, has a major role in peripheral inflammation, and in the near future, targeting the peripheral cannabinoid system as a promising alternative to treat inflammation diseases may be considered a novel pharmacologic approach.


A versatile platform for gas-phase molecular polaritonics.
Cavity coupling of gas-phase molecules will enable studies of benchmark chemical processes under strong light-matter interactions with a high level of experimental control and no solvent effects. We recently demonstrated the formation of gas-phase molecular polaritons by strongly coupling bright ν3, J = 3 → 4 rovibrational transitions of methane (CH4) to a Fabry-Pérot optical cavity mode inside a cryogenic buffer gas cell. Here, we further explore the flexible capabilities of this infrastructure. We show that we can greatly increase the collective coupling strength of the molecular ensemble to the cavity by increasing the intracavity CH4 number density. In doing so, we can tune from the single-mode coupling regime to a multimode coupling regime in which many nested polaritonic states arise as the Rabi splitting approaches the cavity mode spacing. We explore polariton formation for cavity geometries of varying length, finesse, and mirror radius of curvature. We also report a proof-of-principle demonstration of rovibrational gas-phase polariton formation at room temperature. This experimental flexibility affords a great degree of control over the properties of molecular polaritons and opens up a wider range of simple molecular processes to future interrogation under strong cavity-coupling. We anticipate that ongoing work in gas-phase polaritonics will facilitate convergence between experimental results and theoretical models of cavity-altered chemistry and physics.


A simple optimization can improve the performance of single feature polymorphism detection by Affymetrix expression arrays.
High-density oligonucleotide arrays are effective tools for genotyping numerous loci simultaneously. In small genome species (genome size: < approximately 300 Mb), whole-genome DNA hybridization to expression arrays has been used for various applications. In large genome species, transcript hybridization to expression arrays has been used for genotyping. Although rice is a fully sequenced model plant of medium genome size (approximately 400 Mb), there are a few examples of the use of rice oligonucleotide array as a genotyping tool. We compared the single feature polymorphism (SFP) detection performance of whole-genome and transcript hybridizations using the Affymetrix GeneChip Rice Genome Array, using the rice cultivars with full genome sequence, japonica cultivar Nipponbare and indica cultivar 93-11. Both genomes were surveyed for all probe target sequences. Only completely matched 25-mer single copy probes of the Nipponbare genome were extracted, and SFPs between them and 93-11 sequences were predicted. We investigated optimum conditions for SFP detection in both whole genome and transcript hybridization using differences between perfect match and mismatch probe intensities of non-polymorphic targets, assuming that these differences are representative of those between mismatch and perfect targets. Several statistical methods of SFP detection by whole-genome hybridization were compared under the optimized conditions. Causes of false positives and negatives in SFP detection in both types of hybridization were investigated. The optimizations allowed a more than 20% increase in true SFP detection in whole-genome hybridization and a large improvement of SFP detection performance in transcript hybridization. Significance analysis of the microarray for log-transformed raw intensities of PM probes gave the best performance in whole genome hybridization, and 22,936 true SFPs were detected with 23.58% false positives by whole genome hybridization. For transcript hybridization, stable SFP detection was achieved for highly expressed genes, and about 3,500 SFPs were detected at a high sensitivity (> 50%) in both shoot and young panicle transcripts. High SFP detection performances of both genome and transcript hybridizations indicated that microarrays of a complex genome (e.g., of Oryza sativa) can be effectively utilized for whole genome genotyping to conduct mutant mapping and analysis of quantitative traits such as gene expression levels.


Termination of responses to sympathetic nerve stimulation and to noradrenaline in a perfused arterial preparation: the role of neuronal and extraneuronal uptake.
Perfused branch vessels of bovine radial artery in vitro responded to sympathetic nerve stimulation and to noradrenaline added extralumenally and intralumenally with constrictions monitored as flow reductions. Cocaine and 17 beta-estradiol, inhibitors of neuronal and extraneuronal uptake, respectively, enhanced responses to exogenous noradrenaline given by either route. An analysis of response recovery rates, namely termination of action, revealed that the extraneuronal and neuronal uptake process each accounted for between 35 to 50% of the total inactivation capacity of the perfused neuroeffector system for noradrenaline. Responses to periarterial nerve stimulation over a 0.5 to 10 Hz frequency range, involving minimal to moderate flow reductions, were substantially magnified by cocaine and by 17 beta-estradiol. An analysis of response duration and recovery rates showed that extraneuronal processes were more significant than neuronal uptake in terminating noradrenergic transmitter action over the frequency band of 0.5 to 10 Hz.


Digital technique for in vivo assessment of internal and marginal fit of fixed dental prostheses.
Digital approaches to assess the misfit of fixed dental prostheses have been limited to in vitro evaluation. The present article describes a fully digital technique for the in vivo assessment of the fit of fixed dental prostheses by means of a chairside optical scanner and software for 3-dimensional (3D) analysis. The 3D digital capture is performed in 3 steps: an extraoral scan of the restoration, an intraoral scan of the abutment tooth, and an intraoral registration scan of the restoration positioned on the abutment tooth.


Safety and efficacy of ganaxolone in patients with CDKL5 deficiency disorder: results from the double-blind phase of a randomised, placebo-controlled, phase 3 trial.
CDKL5 deficiency disorder (CDD) is a rare, X-linked, developmental and epileptic encephalopathy characterised by severe global developmental impairment and seizures that can begin in the first few months after birth and are often treatment refractory. Ganaxolone, an investigational neuroactive steroid, reduced seizure frequency in an open-label, phase 2 trial that included patients with CDD. We aimed to further assess the efficacy and safety of ganaxolone in patients with CDD-associated refractory epilepsy. In the double-blind phase of this randomised, placebo-controlled, phase 3 trial, done at 39 outpatient clinics in eight countries (Australia, France, Israel, Italy, Poland, Russia, the UK, and the USA), patients were eligible if they were aged 2-21 years with a pathogenic or probably pathogenic CDKL5 variant and at least 16 major motor seizures (defined as bilateral tonic, generalised tonic-clonic, bilateral clonic, atonic, or focal to bilateral tonic-clonic) per 28 days in each 4-week period of an 8-week historical period. After a 6-week prospective baseline period, patients were randomly assigned (1:1) via an interactive web response system to receive either enteral adjunctive ganaxolone or matching enteral adjunctive placebo (maximum dose 63 mg/kg per day for patients weighing ≤28 kg or 1800 mg/day for patients weighing >28 kg) for 17 weeks. Patients, caregivers, investigators (including those analysing data), trial staff, and the sponsor (other than the investigational product manager) were masked to treatment allocation. The primary efficacy endpoint was percentage change in median 28-day major motor seizure frequency from the baseline period to the 17-week double-blind phase and was analysed (using a Wilcoxon-rank sum test) in all patients who received at least one dose of trial treatment and for whom baseline data were available. Safety (compared descriptively across groups) was analysed in all patients who received at least one dose of trial treatment. This study is registered with ClinicalTrials.gov, NCT03572933, and the open-label extension phase is ongoing. Between June 25, 2018, and July 2, 2020, 114 patients were screened for eligibility, of whom 101 (median age 6 years [IQR 3 to 10]) were randomly assigned to receive either ganaxolone (n=50) or placebo (n=51). All patients received at least one dose of a study drug, but seizure frequency for one patient in the ganaxolone group was not recorded at baseline and so the primary endpoint was analysed in a population of 100 patients. There was a median percentage change in 28-day major motor seizure frequency of -30·7% (IQR -49·5 to -1·9) in the ganaxolone group and of -6·9% (-24·1 to 39·7) in the placebo group (p=0·0036). The Hodges-Lehmann estimate of median difference in responses to ganaxolone versus placebo was -27·1% (95% CI -47·9 to - 9·6). Treatment-emergent adverse events occurred in 43 (86%) of 50 patients in the ganaxolone group and in 45 (88%) of 51 patients in the placebo group. Somnolence, pyrexia, and upper respiratory tract infections occurred in at least 10% of patients in the ganaxolone group and more frequently than in the placebo group. Serious adverse events occurred in six (12%) patients in the ganaxolone group and in five (10%) patients in the placebo group. Two (4%) patients in the ganaxolone group and four (8%) patients in the placebo group discontinued the trial. There were no deaths in the double-blind phase. Ganaxolone significantly reduced the frequency of CDD-associated seizures compared with placebo and was generally well tolerated. Results from what is, to our knowledge, the first controlled trial in CDD suggest a potential treatment benefit for ganaxolone. Long-term treatment is being assessed in the ongoing open-label extension phase of this trial. Marinus Pharmaceuticals.


The association between the severity of periodontitis and osteonecrosis of the jaw in patients with different cancer locations: a nationwide population-based study.
To determine the relation between the severity of periodontitis and osteonecrosis of the jaw (ONJ) occurrence among different cancer locations and estimate the effect of dental care on ONJ prevention in cancer patients. This population-based cross-sectional study was conducted through the Longitudinal Health Insurance Database, Taiwan. Patients with malignancies were collected and subdivided into groups according to their different cancer locations, the severity of periodontitis, and dental care. Multivariable logistic regression analysis was performed to assess the associations between ONJ and ONJ-related factors. A total of 8,234 ONJ patients and 32,912 control patients were investigated. Lip, oral cavity, and pharynx malignancies had the highest ONJ risk among all cancer locations (OR from 3.07 to 9.56, P < 0.01). There is a linear relationship between different severities of periodontitis and ONJ. Patients with radiotherapy and severe periodontitis had the highest ONJ risk (adjusted OR, 9.56; 95% CI, 5.34-17.1). Patients with good dental care had a lower ONJ risk. The periodontal condition and cancer location showed a significant impact on the risk of developing ONJ after adjusting for bisphosphonate use. Good dental care could decrease the risk of ONJ in cancer patients. The severity of periodontitis might be a target to predict the potency of ONJ. Dentists must be vigilant about the increased risk of ONJ in cancer patients with periodontitis, especially in the head and neck cancer population. Good dental care is advised for cancer patients with severe periodontitis.


Spliceosomal intron size expansion in domesticated grapevine (Vitis vinifera).
Spliceosomal introns are important components of eukaryotic genes as their structure, sizes and contents reflect the architecture of gene and genomes. Intron size, determined by both neutral evolution, repetitive elements activities and potential functional constraints, varies significantly in eukaryotes, suggesting unique dynamics and evolution in different lineages of eukaryotic organisms. However, the evolution of intron size, is rarely studied. To investigate intron size dynamics in flowering plants, in particular domesticated grapevines, a survey of intron size and content in wine grape (Vitis vinifera Pinot Noir) genes was conducted by assembling and mapping the transcriptome of V. vinifera genes from ESTs to characterize and analyze spliceosomal introns. Uncommonly large size of spliceosomal intron was observed in V. vinifera genome, otherwise inconsistent with overall genome size dynamics when comparing Arabidopsis, Populus and Vitis. In domesticated grapevine, intron size is generally not related to gene function. The composition of enlarged introns in grapevines indicated extensive transposable element (TE) activity within intronic regions. TEs comprise about 80% of the expanded intron space and in particular, recent LTR retrotransposon insertions are enriched in these intronic regions, suggesting an intron size expansion in the lineage leading to domesticated grapevine, instead of size contractions in Arabidopsis and Populus. Comparative analysis of selected intronic regions in V. vinifera cultivars and wild grapevine species revealed that accelerated TE activity was associated with grapevine domestication, and in some cases with the development of specific cultivars. In this study, we showed intron size expansion driven by TE activities in domesticated grapevines, likely a result of long-term vegetative propagation and intensive human care, which simultaneously promote TE proliferation and repress TE removal mechanisms such as recombination. The intron size expansion observed in domesticated grapevines provided an example of rapid plant genome evolution in response to artificial selection and propagation, and may shed light on the important genomic changes during domestication. In addition, the transcriptome approach used to gather intron size data significantly improved annotations of the V. vinifera genome.


Transforming undergraduate laboratory courses with interlinked real-world challenges.
Undergraduate laboratory course components often provide training in various techniques without connections to an interlinked real-world scenario. This article emphasizes the benefits of longitudinal integration of research techniques to enhance learning and emphasize societal relevance. An example of a biomedical engineering challenge involving a new pandemic is described.


Guided biopsy of osseous pathologies in the jaw bone using a 3D-printed, tooth-supported drilling template.
Suspicious radiological findings in the jaw bone require histopathological examination for the confirmation of a diagnosis. As pathologies in this region are difficult to reach or are in close proximity to relevant anatomical structures, e.g. tooth roots or nerves, they often represent a challenge. Such factors may adversely affect the predictability of the surgical outcome of a biopsy of the osseous tissues. This technical note introduces a novel method for performing a digitally planned, guided biopsy. For this purpose, a cone beam computed tomography scan and an intraoral scan are superimposed using specific planning software. The resulting three-dimensionally printed, tooth-supported drilling template is designed for a trephine biopsy. It allows a precise, minimally invasive approach, with an exact three-dimensional determination of the biopsy location prior to surgery. The risk of devitalization of the neighbouring teeth or possible damage to the nerve structures can be minimized. Furthermore, a small access flap can be sufficient. In summary, the method of bone biopsy presented here allows high precision and greater predictability for biopsy sampling and is minimally invasive for the patient.


Neurovascular syndromes of aortic dissection.
Dissecting aneurysm is the most common acute disease of the aorta. Men are more likely than women to develop aortic dissection. One-third of the cases show neurologic manifestations, which may be a clue to early diagnosis. Cerebral, spinal cord and peripheral nerve ischemia can occur. In general, the treatment of choice for dissection originating in the ascending aorta is prompt surgical intervention. Medical therapy to lower elevated blood pressure and to reduce the force of myocardial contractility is sufficient for dissection of the descending aorta.


Adolescent use of outpatient health services in Mexico: their health needs and associated factors.
To estimate adolescent use of outpatient services, identifying their health needs and associated socioeconomic factors. Using data from Ensanut 2018-2019, adolescents (ages 10-19) with health needs and those receiving care from health personnel (users) were identified. Needs were analyzed by sex and socioeconomic status (SES). Logistic models were used to assess the factors associated with the use of health care and choice of provider. 6% of adolescents reported health needs, of whom 64% used outpatient services. Respiratory and gastrointestinal infections were the principal health needs prompting use of services overall. However, by SES, motivations centered on pregnancy for the poor and accidental injuries for the wealthy. One in three adolescents with health needs, particularly the poorest, received no care. Living with a partner and having health insurance were the main predictors of use. Greater schooling among household heads and higher SES correlated with the use of private services. Despite being aware of their health needs, adolescents are the group that uses health services the least in Mexico. Promoting preventative and timely treatment for this population would encourage youths to seek care more often.


The Charcot Marie Tooth disease protein LITAF is a zinc-binding monotopic membrane protein.
LITAF (LPS-induced TNF-activating factor) is an endosome-associated integral membrane protein important for multivesicular body sorting. Several mutations in LITAF cause autosomal-dominant Charcot Marie Tooth disease type 1C. These mutations map to a highly conserved C-terminal region, termed the LITAF domain, which includes a 22 residue hydrophobic sequence and flanking cysteine-rich regions that contain peptide motifs found in zinc fingers. Although the LITAF domain is thought to be responsible for membrane integration, the membrane topology of LITAF has not been established. Here, we have investigated whether LITAF is a tail-anchored (TA) membrane-spanning protein or monotopic membrane protein. When translated in vitro, LITAF integrates poorly into ER-derived microsomes compared with Sec61β, a bona fide TA protein. Furthermore, introduction of N-linked glycosylation reporters shows that neither the N-terminal nor C-terminal domains of LITAF translocate into the ER lumen. Expression in cells of an LITAF construct containing C-terminal glycosylation sites confirms that LITAF is not a TA protein in cells. Finally, an immunofluorescence-based latency assay showed that both the N- and C-termini of LITAF are exposed to the cytoplasm. Recombinant LITAF contains 1 mol/mol zinc, while mutation of predicted zinc-binding residues disrupts LITAF membrane association. Hence, we conclude that LITAF is a monotopic membrane protein whose membrane integration is stabilised by a zinc finger. The related human protein, CDIP1 (cell death involved p53 target 1), displays identical membrane topology, suggesting that this mode of membrane integration is conserved in LITAF family proteins.


Robust Regression with Density Power Divergence: Theory, Comparisons, and Data Analysis.
Minimum density power divergence estimation provides a general framework for robust statistics, depending on a parameter α , which determines the robustness properties of the method. The usual estimation method is numerical minimization of the power divergence. The paper considers the special case of linear regression. We developed an alternative estimation procedure using the methods of S-estimation. The rho function so obtained is proportional to one minus a suitably scaled normal density raised to the power α . We used the theory of S-estimation to determine the asymptotic efficiency and breakdown point for this new form of S-estimation. Two sets of comparisons were made. In one, S power divergence is compared with other S-estimators using four distinct rho functions. Plots of efficiency against breakdown point show that the properties of S power divergence are close to those of Tukey's biweight. The second set of comparisons is between S power divergence estimation and numerical minimization. Monitoring these two procedures in terms of breakdown point shows that the numerical minimization yields a procedure with larger robust residuals and a lower empirical breakdown point, thus providing an estimate of α leading to more efficient parameter estimates.


On the frequency and amplitude spectrum and the fluctuations at the output of a communication receiver.
A mixer cascaded with a low-pass filter is the basic piece of any communication receiver. The frequency and amplitude fluctuations of beat signals recorded at the output of the mixer plus filter set-up are investigated experimentally and explained on the basis of number theory in relation to the Riemann problem concerning the distribution of prime numbers. The frequency of the beat signal is obtained from a diophantine approximation of the frequency ratio of the input oscillators. The amplitude is defined globally from the position of resolved fractions with respect to an equally spaced graduation. Time series analysis methods show that frequency fluctuations present a transition from white frequency noise to 1/f frequency noise close to resonance; the latter is compatible with an underlying fractal attractor. The same transition is observed in the case of time series computed from continued fraction expansions.


GALK inhibitors for classic galactosemia.
Classic galactosemia is an inherited metabolic disease for which, at present, no therapy is available apart from galactose-restricted diet. However, the efficacy of the diet is questionable, since it is not able to prevent the insurgence of chronic complications later in life. In addition, it is possible that dietary restriction itself could induce negative side effects. Therefore, there is a need for an alternative therapeutic approach that can avert the manifestation of chronic complications in the patients. In this review, the authors describe the development of a novel class of pharmaceutical agents that target the production of a toxic metabolite, galactose-1-phosphate, considered as the main culprit for the cause of the complications, in the patients.


Nicotine is a competitive antagonist for H1-receptors in smooth muscles.
The effects of nicotine on histamine-induced contractions in the aortic strip from rabbit and in the intestinal and airway smooth muscles from guinea-pig were examined. Nicotine inhibited the contractile effects of histamine in a concentration-dependent manner in all investigated smooth muscle preparations having identical pA2 values. Data obtained in this study indicate that nicotine has a histamine H1-receptor antagonistic property with a lower activity than mepyramine for H1-receptors as verified by the calculated pA2 values.


The 1996 Helene Hudson Memorial Lecture. Work redesign and re-engineering: a challenge for professional nursing practice.
Work redesign and re-engineering have become the buzzwords of the 1990s as all sectors of the health care arena struggle to meet the demands of patient care while coping with increasing fiscal constraint. Redesign and re-engineering are terms that describe a wide range of strategies in health care and radically different models of care delivery. These new approaches to care are shifting the way we view care delivery and how it is structured. This paper describes the principles of redesign and re-engineering, common applications in health care organizations, outcomes and evaluation. Multiskilling and use of genetic health care workers are addressed. The potential impact on the practice of oncology nurses is explored as well as strategies to meet the challenges of today's health care environment.


Optical properties of Nannochloropsis sp and their application to remote estimation of cell mass.
The absorption and scattering coefficients and reflectance spectra of ultra-high density Nannochloropsis occulata cultures were investigated in detail to identify the optical properties of the cultures and devise algorithms for remote estimation of dry cell mass in ultra-high cell density cultures. High-spectral resolution measurements of apparent absorption and attenuation as well as reflectance from 400 to 900 nm were carried out in relation to the dry weight, cell count, and pigment concentration in outdoor cultures. Indices calculated as (R(NIR) - R(red))/(R(NIR) + R(red)) and R(NIR)/R(red), in which R(NIR) is reflectance in the range from 750 to 800 nm and R(red) is reflectance in the range 670-680 nm, were used for remote assessment of dry cell mass. Remote estimation in the range 1 to 8 g/L was accomplished with an error of less than 0.66 g/L. A different index, i.e., (R(NIR) - R(red)) was employed for estimation of cell-chlorophyll concentration. This is the first report of in vivo specific absorption coefficient of chlorophyll-a and specific scattering coefficient per dry algal weight of Nannochloropsis sp., providing a basis for remote monitoring of dense phytoplankton masses.


Intron-exon swapping of transglutaminase mRNA and neuronal Tau aggregation in Alzheimer's disease.
In order to understand the mechanism for insoluble neurotoxic protein polymerization in Alzheimer's disease (AD) brain neurons, we examined protein and gene expression for transglutaminase (TGase 2; tissue transglutaminase (tTG)) in hippocampus and isocortex. We found co-localization of tTG protein and activity with tau-positive neurofibrillary tangles, whereas mRNA and sequence analysis indicated an absolute increase in tTG synthesized. Although apoptosis in AD hippocampus is now an established mode of neuronal cell death, no definite underlying mechanism(s) is known. Since TGase-mediated protein aggregation is implicated in polyglutamine ((CAG)(n)/Q(n) expansion) disorder apoptosis, and expanded Q(n) repeats are excellent TGase substrates, a role for TGase in AD is possible. However, despite such suggestions almost 20 years ago, the molecular mechanism remained elusive. We now present one possible molecular mechanism for tTG-mediated, neurotoxic protein polymerization leading to neuronal apoptosis in AD that involves not its substrates (like Q(n) repeats) but rather the unique presence of alternative transcripts of tTG mRNA. In addition to a full-length (L) isoform in aged non-demented brains, we found a short isoform (S) lacking a binding domain in all AD brains. Our current results identify intron-exon "switching" between L and S isoforms, implicating G-protein-coupled signaling pathways associated with tTG that may help to determine the dual roles of this enzyme in neuronal life and death processes.


The parent-of-origin effect of 10q22 in pre-eclamptic females coincides with two regions clustered for genes with down-regulated expression in androgenetic placentas.
By affected sib-pair linkage analysis of 24 families with pre-eclampsia, we confirm a susceptibility locus on chromosome 10q22.1 in Dutch females: a multipoint non-parametric linkage score of 3.6 near marker D10S1432 was obtained. Haplotype analysis showed a parent-of-origin effect: maximal allele sharing in the affected sibs was found for maternally derived alleles in all families, but not for the paternally derived alleles. As matrilineal inheritance suggests the presence of maternally expressed imprinted genes, while imprinting operates predominantly in (extra)embryonic tissues, all genes (n=132) known on 10q22 between GATA121A08 and D10S580 were screened for seven sequence-related features associated with imprinting and subsequently tested for expression in first trimester placenta. Placental expression of genes selected in this way (n=55) was compared with expression in androgenetic placentas of identical gestational age. Two regions on 10q22 were identified with developmentally co-repressed genes with non-random chromosomal distribution. Interestingly, these two clusters, near CTNNA3 and KCNMA1 and each containing five genes with down-regulated expression in androgenetic placentas, coincided with the regions with maximal maternal allele sharing seen in the pre-eclamptic sisters. Our linkage and expression data are compatible with the concept that pre-eclampsia involves maternally expressed imprinted genes that operate in the first trimester placenta.


Problems related to the use of pesticides.
Potentiation and additive effects of pesticides are considered major problems in the determination of safe daily intakes of these chemicals. Examples of potentiation between two organophosphates, such as ethyl-p-nitrophenyl benzenethionophosphonate (EPN) and malathion, delnav and malathion, and guthion and dipterex are considered. Although this type of potentiation could give rise to acute accidents, the most important point in evaluation centres on the possible effects of small amounts of potentiating compounds in the daily diet. Likewise, certain drugs of the phenothiazine type potentiate the toxic effects of organophosphates. A review of the data for both types of potentiation indicates that the consumption of raw agricultural products containing pesticides at tolerance levels constitutes a minimal hazard. A two-year rat-feeding investigation using a combination of eight flavouring agents and six pesticides, at use levels and higher, did not cause a significant alteration of toxicity in comparison with the toxicity of the individual substances.


Bowel dysfunction and colon transit time in brain-injured patients.
To report the defecation patterns of brain-injured patients and evaluate the relationship between functional ability and colon transit time (CTT) in stroke patients. A total of 55 brain-injured patients were recruited. Patient interviews and medical records review of pattern of brain injury, anatomical site of lesion, bowel habits, constipation score, and Bristol scale were conducted. We divided the patients into constipation (n=29) and non-constipation (n=26) groups according to Rome II criteria for constipation. The CTTs of total and segmental colon were assessed using radio-opaque markers Kolomark® and functional ability was evaluated using the functional independence measure (FIM). Constipation scores in constipation and non-constipation groups were 7.32±3.63 and 5.04±2.46, respectively, and the difference was statistically significant. The CTTs of the total colon in both groups were 46.6±18.7 and 32.3±23.5 h, respectively. The CTTs of total, right, and left colon were significantly delayed in the constipation group (p<0.05). No significant correlation was found between anatomical location of brain injury and constipation score or total CTT. Only the CTT of the left colon was delayed in the patient group with pontine lesions (p<0.05). The constipation group had significantly elevated constipation scores and lower Bristol stool form scale, with prolonged CTTs of total, right, and left colon. In classification by site of brain injury, we did not find significantly different constipation scores, Bristol stool form scale, or CTTs between the groups with pontine and suprapontine injury.