Gradualness facilitates knowledge refinement.
To facilitate knowledge refinement, a system should be designed so that small changes in the knowledge correspond to small changes in the function or performance of the system. Two sets of experiments show the value of small, heuristically guided changes in a weighted rule base. In the first set, the ordering among numbers (reflecting certainties) makes their manipulation more straightforward than the manipulation of relationships. A simple credit assignment and weight adjustment strategy for improving numbers in a weighted, rule-based expert system is presented. In the second set, the rearrangement of predicates benefits from additional knowledge about the ``ordering'' among predicates. A third set of experiments indicates the importance of the proper level of granularity when augmenting a knowledge base. Augmentation of one knowledge base by analogical reasoning from another knowledge base did not work with only binary relationships, but did succeed with ternary relationships. To obtain a small improvement in the knowledge base, a substantial amount of structure had to be treated as a unit.


Swarming of Pseudomonas aeruginosa is a complex adaptation leading to increased production of virulence factors and antibiotic resistance.
In addition to exhibiting swimming and twitching motility, Pseudomonas aeruginosa is able to swarm on semisolid (viscous) surfaces. Recent studies have indicated that swarming is a more complex type of motility influenced by a large number of different genes. To investigate the adaptation process involved in swarming motility, gene expression profiles were analyzed by performing microarrays on bacteria from the leading edge of a swarm zone compared to bacteria growing in identical medium under swimming conditions. Major shifts in gene expression patterns were observed under swarming conditions, including, among others, the overexpression of a large number of virulence-related genes such as those encoding the type III secretion system and its effectors, those encoding extracellular proteases, and those associated with iron transport. In addition, swarming cells exhibited adaptive antibiotic resistance against polymyxin B, gentamicin, and ciprofloxacin compared to what was seen for their planktonic (swimming) counterparts. By analyzing a large subset of up-regulated genes, we were able to show that two virulence genes, lasB and pvdQ, were required for swarming motility. These results clearly favored the conclusion that swarming of P. aeruginosa is a complex adaptation process in response to a viscous environment resulting in a substantial change in virulence gene expression and antibiotic resistance.


Antiarrhythmic effects of Org 6001 in rats: correlation with plasma and tissue drug concentrations.
The antiarrhythmic effects of Org 6001 following oral administration in the rat have been assessed and correlated with plasma, myocardial and skeletal muscle drug concentrations. Arrythmias were induced by coronary artery ligation in anaesthetized rats. Org 6001 (10, 20, 50 and 100 mg/kg given 1 h before ligation) significantly reduced mortality and the incidence of ventricular fibrillation in the 0-30 min post ligation period. Only the highest dose of drug also significantly reduced the number of ventricular ectopic beats following ligation. A linear relationship was observed between the oral dose and the Org 6001 concentrations in plasma and skeletal muscle determined 90 min after drug administration. The Org 6001 concentration in the myocardium was not linearly related to the administered dose and Org 6001 appeared to be concentrated to a higher extent in cardiac than in skeletal muscle at that time. No statistically significant difference in drug levels in the ischaemic left ventricle and normal right ventricle plus septum was observed. The antiarrythmic effect of Org 6001, as measured by changes in the incidence of ventricular fibrillation, correlated with myocardial concentrations of the drug.


Survival of Young Versus Old Patients With Oral Cavity Squamous Cell Carcinoma: A Meta-Analysis.
To assess whether young patients with oral cavity squamous cell carcinoma (OCSCC) demonstrate worse oncologic outcomes than older patients after definitive therapy. Systematic review and meta-analysis. A medical librarian composed a search strategy to identify relevant studies in Medline, Embase, Scopus, and other major databases (Prospero registration number CRD42019127974). Inclusion criteria were adults with histologically diagnosed OCSCC that underwent treatment, comparator groups with an age cutoff of 40 years old, and reported survival outcomes. Articles were excluded if they contained patients with oropharyngeal squamous cell carcinoma or patients treated for palliative intent. Overall survival hazard ratios were analyzed with a meta-analysis. There were 23,382 patients with OCSCC that were treated with definitive therapy from 22 included studies. The pooled cohort contained 2,238 (10%) patients ≤40 years of age. Oral tongue was the most common subsite in both the younger (n = 1,961, 91%) and older (n = 18,047, 88%) cohorts. The majority of OCSCCs were either T1 or T2, representing 859 (80%) malignancies in younger patients and 8,126 (77%) malignancies in older patients. A meta-analysis of nine studies demonstrated that younger patients did not experience worse survival outcomes than older patients (hazard ratio = 0.97, 95% confidence interval = 0.66-1.41). Young adults with OCSCC experienced similar oncologic outcomes as older patients with OCSCC after definitive treatment. Until compelling evidence demonstrates clinically relevant differences between these two cohorts, their approach to management should be similar. Future studies should consider comorbidities and using age 40 as a standard age cutoff to provide more uniform data moving forward. Laryngoscope, 131:1310-1319, 2021.


Sympathoadrenal stress reactivity is a predictor of future blood pressure: an 18-year follow-up study.
In the present study we hypothesized that arterial catecholamine concentrations during rest and 2 laboratory stress tests were independent predictors of blood pressure at an 18-year follow-up. At entry, blood pressure, heart rate, and arterial plasma epinephrine and norepinephrine concentrations were measured in 99 healthy men (age: 19.3+/-0.4 years, mean+/-SD) at rest, during a mental arithmetic test, and during a cold pressor test. After 18.0+/-0.9 years of follow-up, resting blood pressure was measured. The norepinephrine and epinephrine concentrations during the mental arithmetic explained 12.7% of the variation of future systolic blood pressure after adjusting for initial resting blood pressure, family history, body mass index, and systolic blood pressure during the stress test in a multiple regression analysis (adjusted R(2)=0.651; P<0.001). To conclude, the present study shows that sympathetic nervous activity during mental arithmetic predicts future blood pressure, indicating a possible causal factor in the development of essential hypertension independent of the initial blood pressure.


[Magnetic resonance tomography of the orbit using surface coils. Study technic, anatomy and initial clinical experience].
MRI of the orbit is strongly improved by the use of surface coils due to a higher signal-to-noise ratio. Oblique views without moving the patient present the optic nerve in full length on one slice. First experience with a small number of cases demonstrates normal anatomy and lesions in detail only at T1-weighted pulse sequences. Losses in contrast variation and detail accuracy are caused by movements of the eyeballs. Edge artifacts due to chemical shifting impair the image quality. So far there are no pointers towards tissue-specific signal intensity behaviour. Procedure and most favourable parameters at 1 tesla are given.


Identification of a stimulant for Lactobacillus casei produced by Streptococcus lactis.
A compound stimulatory to the growth of Lactobacillus casei was isolated from cell extracts of Streptococcus lactis, purified, and characterized. The stimulant was identified as a small peptide with a molecular weight of approximately 4,500 daltons. The purified peptide gave negative tests for nucleic acids, phosphorus, glucosamine, and carbohydrates. Sixteen amino acids were detected in acid hydrolysates of this peptide. Serine, proline, glycine, alanine, leucine, and glutamic acid were present in hydrolysates in greatest abundance.


The effect of red clover isoflavone supplementation over vasomotor and menopausal symptoms in postmenopausal women.
To evaluate the effect of red clover isoflavone supplementation over vasomotor and overall menopausal symptoms in postmenopausal women. One hundred and nine postmenopausal women aged 40 or more were assigned to randomly receive either two daily capsules of the active compound (80 mg red clover isoflavones, Group A) or placebo of equal appearance (Group B) for a 90-day period. After a washout period of 7 days, medication was crossed over and taken for 90 days more. Daily hot flush and night sweat frequency and overall menopausal symptom intensity (Kupperman Index) were measured at baseline, 90, 97 and 187 days. Daily hot flush/night sweat frequency and Kupperman Index values were similar in both studied groups at baseline. All indices significantly decreased after red clover phase in Group A, corresponding, respectively to a 73.5%, 72.2% and 75.4% average decrement. These decrements were significantly higher than those observed for Group B after placebo phase (8.2%, 0.9% and 6.7% respectively). In Group A, after washout and placebo phases all values significantly increased. In Group B, all indices remained similar after placebo and washout phases, however significantly dropping after red clover treatment. These values were also significantly lower than those observed in Group A after placebo phase. No side effects were encountered after treatment with the active compound or placebo. Red clover isoflavone supplementation was more effective than placebo in reducing daily vasomotor frequency and overall menopausal intensity in postmenopausal women.


[Analysis of the surveillance system of larval traps in the municipality of Mariel (1984-1987)].
An analysis is done of the species of culicides captured by the surveillance system of Aedes aegypti, with larvitraps, at the Mariel Municipality, during 1984-1987 period. The study shows an increased presence of Aedes mediovittatus in the system of larvitraps during the working period, demonstrating it the usefulness of sing larvitraps in the surveillance programmes of dengue in order to detect other species besides Aedes aegypti.


The Th17 Lineage: From Barrier Surfaces Homeostasis to Autoimmunity, Cancer, and HIV-1 Pathogenesis.
The T helper 17 (Th17) cells represent a subset of CD4+ T-cells with unique effector functions, developmental plasticity, and stem-cell features. Th17 cells bridge innate and adaptive immunity against fungal and bacterial infections at skin and mucosal barrier surfaces. Although Th17 cells have been extensively studied in the context of autoimmunity, their role in various other pathologies is underexplored and remains an area of open investigation. This review summarizes the history of Th17 cell discovery and the current knowledge relative to the beneficial role of Th17 cells in maintaining mucosal immunity homeostasis. We further discuss the concept of Th17 pathogenicity in the context of autoimmunity, cancer, and HIV infection, and we review the most recent discoveries on molecular mechanisms regulating HIV replication/persistence in pathogenic Th17 cells. Finally, we stress the need for novel fundamental research discovery-based Th17-specific therapeutic interventions to treat pathogenic conditions associated with Th17 abnormalities, including HIV infection.


Retrospective cohort study comparing activated partial thromboplastin time versus anti-factor Xa activity nomograms for therapeutic unfractionated heparin monitoring in pediatrics.
Unfractionated heparin (UFH) is widely used to treat thromboembolic disease, but monitoring in children is challenging. Both activated partial thromboplastin time (aPTT) and anti-factor Xa activity (anti-Xa) are utilized, but a comparison of dosing nomograms has not been reported in pediatrics. To compare the performance of aPTT and anti-Xa for UFH monitoring in pediatric patients. A retrospective cohort study was conducted in patients ≤ 21 years old treated with UFH at Johns Hopkins Hospital from January 2009 to May 2011. For monitoring, an aPTT nomogram was used for the initial 15 months, and an anti-Xa nomogram was used for the subsequent 12 months. Clinical characteristics, laboratory data and outcomes were analyzed. Thirty-four patients were monitored with aPTT and 26 patients with anti-Xa. There was no significant difference in median time to therapeutic range (11.6 h aPTT, 95%CI = 6.0-17.0; 9.9 h anti-Xa, 95%CI = 7.3-20.7) or per cent of patients achieving therapeutic measurements at 24 (79% aPTT, 95%CI = 62-91; 73% anti-Xa, 95%CI = 52-88) and 48 h (88% aPTT, 95%CI = 73-97; 96% anti-Xa, 95%CI = 80-100). However, anti-Xa measurements were more frequently therapeutic than aPTT (74% [95%CI = 69-78] vs. 54% [95%CI = 50-59]). Variance between anti-Xa and aPTT measurements was high (R(2) = 0.236). No significant difference was seen in bleeding incidence (9% aPTT, 95%CI = 2-24; 15% anti-Xa, 95%CI = 4-35). The time to achieve therapeutic measures and bleeding outcomes were not significantly different between anti-Xa and aPTT nomograms. However, a small study size limits the power to detect clinically relevant differences. The results warrant larger prospective studies of UFH monitoring in children with thromboembolic disease.


Quantitative analysis of the age-related fragmentation of hamster 24-h activity rhythms.
The continuous monitoring of spontaneous locomotor activity has emerged as one of the most widely used metrics in rodent circadian research. This behavioral measure is also extremely useful for the description of the effects of aging on circadian rhythms. The present study describes the successful use of a log-survivorship approach to identify discrete bouts of hamster wheel-running activity and provides a detailed description of the age-related fragmentation in the 24-h profile of this behavioral variable. In addition, stepwise discriminant analysis identified the most important quantitative measures for distinguishing between the individual patterns of wheel-running activity of young (3 mo) and old (17-18 mo) golden hamsters. The results suggest that this method of bout analysis can be a valuable tool for the study of genetic, developmental, neurochemical, physiological, and environmental factors involved in the temporal control of rodent locomotor behavior.


Prospective outcomes evaluation after decompression with or without instrumented fusion for lumbar stenosis and degenerative Grade I spondylolisthesis.
There is considerable debate among spine surgeons regarding whether fusion should be used to augment decompressive surgery in patients with symptomatic lumbar spinal stenosis involving Grade I degenerative spondylolisthesis. The authors prospectively evaluated the outcomes of patients treated between 2000 and 2002 at two institutions to determine whether fusion improves functional outcome 1 year after surgery. Patients ranged in age from 50 to 81 years. They presented with degenerative Grade I (3- to 14-mm) spondylolisthesis and lumbar stenosis without gross instability (< 3 mm of motion at the level of subluxation). Those in whom previous surgery had been performed at the level of subluxation were excluded. Each patient completed Oswestry Disability Index (ODI) and Short Form-36 (SF-36) questionnaires preoperatively and at 6 to 12 months postoperatively. Some patients underwent decompression alone (20 cases), whereas others underwent decompression and posterolateral instrumentation-assisted fusion (14 cases), at the treating surgeon's discretion. Baseline demographic data, radiographic features, and ODI and SF-36 scores were similar in both groups. The 1-year fusion rate was 93%. Both forms of surgery independently improved outcome compared with baseline status, based on ODI and SF-36 physical component summary (PCS) results (p < 0.001). Decompression combined with fusion led to an improvement in ODI scores of 27.5 points, whereas decompression alone was associated with a 13.6-point increase (p = 0.02). Analysis of the SF-36 PCS data also demonstrated a significant intergroup difference (p = 0.003). Surgery substantially improved 1-year outcomes based on established outcomes instruments in patients with Grade I spondylolisthesis and stenosis. Fusion was associated with greater functional improvement.


Long-term Patterns of Regional Failure for Nasopharyngeal Carcinoma following Intensity-Modulated Radiation Therapy.
To analyze the long-term patterns of regional failure following intensity-modulated radiotherapy (IMRT) for nasopharyngeal carcinoma (NPC). From January 2005 to December 2010, 275 non-metastatic NPC patients treated with IMRT were retrospectively enrolled. Patients staged as II (lymph nodes measuring 4 or more cm in diameter), III or IV also received chemotherapy. Failures were assessed as in-field or out-field relative to the pretreatment planning computed tomography data sets. Univariate and multivariate analyses were performed with Cox proportional hazards model to analyze the effect of various prognostic factors on regional failure-free survival (RFFS) and overall survival (OS). During a median follow-up of 71 months, the RFFS and OS rates were 94.3% and 83.9%, respectively. Seventeen patients developed regional failures, of which 16 were in-field; one patient showed an out-field failure in the parotid gland, and no recurrences were seen for level Ib. Failures in level II and in the retropharyngeal area accounted for 70.6% (12/17) and 52.9% (9/17) of all failures, respectively. The 5-year RFFS rates for patients with classifications of N0-1 and N2-3 were 98.5% and 90.2%, respectively (p = 0.001). Multivariate analysis showed that N stage was the only independent prognostic predictor of RFFS (HR 7.363, 95% CI 1.516-35.756, p = 0.013). The regional failure of NPC after treatment with IMRT is uncommon but is significantly higher in N2-3 patients than in N0-1 patients. In-field failures represent the main pattern of regional recurrence and are most often detected in level II and in the retropharyngeal area, while out-field failure is rare. Close attention should be directed to NPC patients with advanced N stages.


Myocardial perfusion imaging in ischemic heart disease anno 1990.
The field of radionuclide myocardial perfusion imaging is in a rapid state of change. Stress-rest myocardial imaging is important not only for the detection of coronary artery disease but also for prognostic stratification of patients. In particular, assessment of myocardial viability in patients with left ventricular dysfunction is a recent focus of investigation. Single-photon emission computed tomography has become widely accepted as the preferred (albeit challenging) imaging modality for myocardial perfusion imaging. Silent myocardial ischemia and its clinical significance continues to be an intriguing aspect of the clinical manifestation of coronary artery disease. Myocardial perfusion imaging is an invaluable independent method to unravel this problem. Dipyridamole was approved for pharmacologic vasodilation in conjunction with myocardial perfusion imaging. At the same time, direct infusion of adenosine was proposed as an alternative method of effecting vasodilatory stress. In 1990, several new technetium-99m-labeled myocardial perfusion imaging agents have been introduced (teboroxime and hexakis-2-methoxyisobutyl-isonitrile [sestaMIBI]) that may have a profound impact on imaging techniques and applications of myocardial perfusion imaging.


Taste function in mice with a targeted mutation of the pkd1l3 gene.
Recent studies, both in vitro and in vivo, have suggested the involvement of the polycystic kidney disease-1 and -2 like genes, Pkd1l3 and Pkd2l1, in acid taste transduction. In mice, disruption of taste cells expressing PKD2L1 eliminates gustatory neural responses to acids. However, no previous data exist on taste responses in the absence of PKD1L3 or on behavioral responses in mice lacking either of these proteins. In order to assess the function of PKD1L3, we genetically engineered mice with a targeted mutation of the Pkd1l3 gene. We then examined taste responsiveness of mutant and wild-type mice using several different approaches. In separate groups of mice, we measured preference scores in 48-h 2-bottle tests, determined NaCl or citric acid taste thresholds using a conditioned taste aversion technique, and conducted electrophysiological recordings of activity in the chorda tympani and glossopharyngeal nerves. Multiple taste compounds representing all major taste qualities were used in the preference tests and nerve-recording experiments. We found no significant reduction in taste responsiveness in Pkd1l3 mutant mice in behavioral or electrophysiological tests when compared with wild-type controls. Therefore, further studies are needed to elucidate the function of PKD1L3 in taste bud cells.


Association between length of stay and readmission for COPD.
Recent financial penalties for high risk-adjusted chronic obstructive pulmonary disease (COPD) readmissions are causing hospitals to search for ways to reduce COPD readmissions. Although some have advocated for increasing the length of stay (LOS) as a method to decrease readmissions, the association between LOS and readmission is unclear. Our primary objective was to examine the association between LOS and readmission among patients admitted for COPD. We conducted an observational study of 33,558 veterans admitted to 130 Veterans Affairs hospitals for COPD from October 1, 2008, to September 30, 2011. We used multivariable regression to separately examine the associations of patient and hospital LOS with 30-day all-cause readmission. At the patient level, compared with short LOS (<3 days), a longer LOS was associated with increased risk for readmission. The adjusted odds ratio was 1.39 (95% confidence interval [CI], 1.18-1.63) for medium LOS (3-4 days) and 2.03 (95% CI, 1.72-2.40) for long LOS (>4 days). On the hospital level, there was no association between LOS and readmission. On a patient level, a longer LOS for COPD hospitalizations was associated with higher risk for readmission, which is likely confounded by the severity of the illness. On a hospital level, LOS was not associated with readmission. These findings imply that, independent of other transitional care practices, altering the hospital LOS may not influence the risk of readmission.


An Analysis of Computer Game Addiction in Primary School Children and Its Affecting Factors.
Computer games are included in next-generation technologies in today's developed visual media world. They are attractive to all ages, but the dramatic increase in the use of computer games in children and adolescents is remarkable. This study aims to determine the computer game addiction in primary school children and its affecting factors. The study sample consisted of 476 students among 952 students enrolled in three primary schools in Fethiye, Muğla. Data were collected from the students using the "Child Information Form" and the "Computer Game Addiction Scale for Children." The data were analyzed using numbers, percentages, independent samples, one-way analysis of variance, and regression analysis. This study found that there was a statistically significant difference between gender, class grade, level of income, mothers' educational level, the presence of a game console/computer at home, and the computer game addiction scale scores (p < .05). It was also found that students who spend more time on the Internet and playing computer game constitute the most at-risk group for computer game addiction (p < .05). Some interventions can be planned to decrease computer game addiction particularly in male students, children and families with low income and educational level, and students who have computers and game consoles at home with longer duration of gaming and Internet use with the cooperation of schools, school nurses, teachers, and parents.


Effects of antiepileptic drugs on refractory seizures in the intact immature corticohippocampal formation in vitro.
We developed a new in vitro preparation of immature rats, in which intact corticohippocampal formations (CHFs) depleted in magnesium ions become progressively epileptic. The better to characterize this model, we examined the effects of 14 antiepileptic drugs (AEDs) currently used in clinical practice. Recurrent ictal-like seizures (ILEs, four per hour) were generated in intact CHFs of P7-8 rats, and extracellular recordings were performed in the hippocampus and neocortex. AEDs were applied at clinically relevant concentrations (at least two), during 30 min after the third ILE. Their ability to prevent or to delay the next ILE was examined. Valproic acid and benzodiazepines (clobazam and midazolam) but also phenobarbital and levetiracetam prevent the occurrence of seizures. In contrast, usual concentrations of carbamazepine (CBZ), phenytoin, vigabatrin, tiagabine, gabapentin, lamotrigine (LTG), topiramate, felbamate, and ethosuximide did not suppress ILEs. In addition, LTG and CBZ aggravate seizures in one third of the cases. This intact in vitro preparation in immature animals appears to be quite resistant to most AEDs. Blockade of seizures was achieved with drugs acting mainly at the gamma-aminobutyric acid (GABA)A-receptor site but not with those that increase the amount of GABA. Drugs with a broad spectrum of activity are efficient but not those preferentially used in partial seizures or absences. We suggest that this preparation may correspond to a model of epilepsy with generalized convulsive seizures and could be helpful to develop new AEDs for refractory infantile epilepsies.


Central corneal thickness and anterior chamber depth measurement by Sirius(®) Scheimpflug tomography and ultrasound.
The purpose of this study was to compare the accuracy of the new Sirius(®) Scheimpflug anterior segment examination device for measurement of central corneal thickness (CCT) and anterior chamber depth (ACD) with that of CCT measurements obtained by ultrasound pachymetry and ACD measurements obtained by ultrasound biometry, respectively. CCT and ACD was measured in 50 right eyes from 50 healthy subjects using a Sirius Scheimpflug camera, SP100 ultrasound pachymetry, and US800 ultrasound biometry. CCT measured with the Sirius was 546 ± 39 μm and 541 ± 35 μm with SP100 ultrasound pachymetry (P = 0.003). The difference was statistically significant (mean difference 4.68 ± 10.5 μm; limits of agreement -15.8 to 25.20 μm). ACD measured with the Sirius was 2.96 ± 0.3 mm compared with 3.36 ± 0.29 mm using US800 ultrasound biometry (P < 0.001). The difference was statistically significant (mean difference -0.40 ± 0.16 mm; limits of agreement -0.72 to 0.07 mm). When the ACD values obtained using ultrasound biometry were corrected according to the values for CCT measured by ultrasound, the agreement increased significantly between both technologies for ACD measurements (mean difference 0.15 ± 0.16 mm; limits of agreement -0.16 to 0.45 mm). CCT and ACD measured by Sirius and ultrasound methods showing good agreement between repeated measurements obtained in the same subjects (repeatability) with either instrument. However, CCT and ACD values, even after correcting ultrasound ACD by subtracting the CCT value obtained with either technology should not be used interchangeably.


Comparison of perceived and actual times spent by residents performing ultrasound examinations on patients.
Emergency medicine (EM) ultrasonography (US) has become a part of residency education. More residents will be taking time during their shifts to perform bedside US examinations for educational purposes, thus further challenging time resources. The authors' hypothesis was that EM residents could accurately estimate the amount of time spent on an US examination of a patient, thus being able to determine when too much time is being taken away from other duties. The authors performed a prospective, single-blind observational study over a four-month period of EM residents at a large community hospital emergency department (ED) with an EM residency program. When a study physician was present, residents were observed and timed from plugging in the machine to turning it off. The residents, who were not aware of the study, were then asked how long the examination had taken. The true scan time, perceived scan time by the resident, and whether the department was busy or not were recorded on standardized quality assurance data sheets by the study physician. When all beds in the department were occupied, the ED was considered to be busy; otherwise, it was noted as not busy. Statistical analysis included descriptive statistics, paired Student's t-test to compare perceived and actual scan times, and correlation analysis to evaluate for any effect of how busy the ED was on accuracy of perceived times. Ninety-three observations were made on 17 different residents. The mean perceived time of examinations was 8 min 12 sec (95% CI = 6:42 to 9:42); the mean actual time was 9 min 53 sec (95% CI = 8:29 to 11:18). The difference of 1 min 42 sec (95% CI = 0:37 to 2:47) yielded a p = 0.003. Residents underestimated the time spent on the scan 64 of 93 (69%) times. How busy the ED was did not affect accuracy. In this study, residents underestimated the amount of time spent performing an ultrasound examination. The small difference between the actual and perceived scan times was statistically significant; the clinical significance of this time difference is not known.


NK cell recruitment in salivary glands provides early viral control but is dispensable for tertiary lymphoid structure formation.
Salivary glands (SGs) represent a permissive site for several sialotropic viruses whose persistence is linked to the development of autoimmunity. Natural Killer (NK) cells play a key role in viral clearance but their involvement in viral infection control and in tertiary lymphoid structures (TLS) development within SGs is unknown. By using an inducible model of TLS in the SGs of wild-type C57BL/6 mice, induced by the local delivery of a replication-defective adenovirus (AdV), we demonstrated that circulating NK cells are rapidly recruited to SGs and highly enrich the early inflammatory infiltrate prior to TLS development. NK cells migrating to SGs in response to AdV infection up-regulate NKp46, undergo proliferation, acquire cytotoxic potential, produce Granzyme-B and IFN-γ, and reduce viral load in the acute phase of the infection. Nonetheless, the selective depletion of both circulating and infiltrating NK cells in AdV-infected mice neither affect the development and frequency of TLS nor the onset of autoimmunity. These data demonstrate that, upon local viral delivery of AdV, peripheral NK cells homing to SGs can exert an early control of the viral infection but are dispensable for the formation of TLS and breach of immunologic tolerance.


Stem cell mobilization with plerixafor and healing of diabetic ischemic wounds: A phase IIa, randomized, double-blind, placebo-controlled trial.
Bone marrow-derived cells contribute to tissue repair, but traffic of hematopoietic stem/progenitor cells (HSPCs) is impaired in diabetes. We therefore tested whether HSPC mobilization with the CXCR4 antagonist plerixafor improved healing of ischemic diabetic wounds. This was a pilot, phase IIa, double-blind, randomized, placebo-controlled trial (NCT02790957). Patients with diabetes with ischemic wounds were randomized to receive a single subcutaneous injection of plerixafor or saline on top of standard medical and surgical therapy. The primary endpoint was complete healing at 6 months. Secondary endpoints were wound size, transcutaneous oxygen tension (TcO2 ), ankle-brachial index (ABI), amputations, and HSPC mobilization. Twenty-six patients were enrolled: 13 received plerixafor and 13 received placebo. Patients were 84.6% males, with a mean age of 69 years. HSPC mobilization was successful in all patients who received plerixafor. The trial was terminated after a preplanned interim analysis of 50% of the target population showed a significantly lower healing rate in the plerixafor vs the placebo group. In the final analysis data set, the rate of complete healing was 38.5% in the plerixafor group vs 69.2% in the placebo group (chi-square P = .115). Wound size tended to be larger in the plerixafor group for the entire duration of observation. No significant difference was noted for the change in TcO2 and ABI or in amputation rates. No other safety concern emerged. In conclusion, successful HSPC mobilization with plerixafor did not improve healing of ischemic diabetic wounds. Contrary to what was expected, outside the context of hematological disorders, mobilization of diabetic HSPCs might exert adverse effects on wound healing.


Infliximab's influence on anastomotic strength and degree of inflammation in intestinal surgery in a rabbit model.
Infliximab, a TNF-α inhibitor, is a potent anti-inflammatory drug in the treatment of inflammatory bowel diseases. Recent studies have investigated the effect of infliximab treatment on postoperative complications such as anastomotic leakage, however, with conflicting results and conclusions. The purpose of this study was to investigate whether a single dose infliximab has an adverse effect on the anastomotic healing process, observed as reduced anastomotic breaking strength and histopathologically verified lower grade of inflammatory response, in the small intestine of a rabbit. Thirty New Zealand rabbits (median weight 2.5 kg) were allocated to treatment with an intravenous bolus of either 10 mg/kg infliximab (n = 15) or placebo (n = 15). One week later all rabbits underwent two separate end-to-end anastomoses in the jejunum under general anesthesia. At postoperative day three, the anastomotic breaking strength was determined and histopathological changes were examined. The mean value of anastomotic breaking strength in the placebo group was 1.89 ± 0.36 N and the corresponding value was 1.81 ± 0.33 N in the infliximab treated rabbits. There was no statistically significant difference between the groups (p = 0.51). The infliximab-treated rabbits had a significant lower degree of inflammatory infiltration response compared to the placebo group (p = 0.047). Our conclusion, limited by the small sample sizes in both groups, is that a single dose of infliximab, given one week prior to surgery, does not have an impact on the anastomotic breaking strength on the third postoperative day in the small intestine of rabbits.


Potter's notion of bioethics.
In 1970 Van Rensselaer Potter was the first to use the term "bioethics" in a publication to advocate the development of a new discipline to address the basic problems of human flourishing. This article analyzes Potter's notion of bioethics in order to understand its origins, sources, and substance. In early publications, Potter conceptualized bioethics as a bridge: between present and future, nature and culture, science and values, and finally between humankind and nature. In later publications, disappointed by a predominant focus on individual and medical issues, and with a wish to underscore the need for a broader perspective, Potter introduced the new term "global bioethics," meant to transcend ethics specialties and integrate them into a new interdisciplinary endeavor to address global problems. A growing interest in global bioethics today means that Potter's original insights are more timely than ever.


Regulation of agouti-related protein messenger ribonucleic acid transcription and peptide secretion by acute and chronic inflammation.
Agouti-related protein (AgRP) is an orexigenic neuropeptide produced by neurons in the hypothalamic arcuate nucleus (ARC) that is a key component of central neural circuits that control food intake and energy expenditure. Disorders in energy homeostasis, characterized by hypophagia and increased metabolic rate, frequently develop in animals with either acute or chronic diseases. Recently, studies have demonstrated that proopiomelanocortin-expressing neurons in the ARC are activated by the proinflammatory cytokine IL-1beta. In the current study, we sought to determine whether inflammatory processes regulate the expression of AgRP mRNA and to characterize the response of AgRP neurons to IL-1beta. Here, we show by real-time RT-PCR and in situ hybridization analysis that AgRP mRNA expression in rodents is increased in models of acute and chronic inflammation. AgRP neurons were found to express the type I IL-1 receptor, and the percentage of expression was significantly increased after peripheral administration of lipopolysaccharide. Furthermore, we demonstrate that IL-1beta inhibits the release of AgRP from hypothalamic explants. Collectively, these data indicate that proinflammatory signals decrease the secretion of AgRP while increasing the transcription of the AgRP gene. These observations suggest that AgRP neurons may participate with ARC proopiomelanocortin neurons in mediating the anorexic and metabolic responses to acute and chronic disease processes.


Somatostatin receptors on peripheral primary afferent terminals: inhibition of sensitized nociceptors.
Somatostatin (SST) is in primary afferent neurons and reduces vascular and nociceptive components of inflammation. SST receptor (SSTR) agonists provide analgesia following intrathecal or epidural administration in humans, but neurotoxicity in the central nervous system (CNS) has been reported in experimental animals. With the rationale that targeting peripheral SSTRs would provide effective analgesia while avoiding CNS side effects, the goals of the present study are to investigate the presence of SSTRs on peripheral primary afferent fibers and determine the behavioral and physiological effects of the SST agonist octreotide (OCT) on formalin-induced nociception and bradykinin-induced primary afferent excitation and sensitization in the rat. The results demonstrate that: (1) SSTR2as are present on 11% of peripheral primary afferent sensory fibers in rat glabrous skin; (2) intraplantar injection of OCT reduces formalin-induced nociceptive behaviors; (3) OCT reduces, in a dose-dependent fashion, responses to thermal stimulation in C-mechanoheat sensitive fibers; and (4) OCT reduces the responses of C-mechanoheat fibers to bradykinin-induced excitation and sensitization to heat. Each of these actions can be reversed following co-injection of OCT with the SSTR antagonist cyclo-somatostatin (c-SOM). Thus, activation of peripheral SSTRs reduces both inflammatory pain and the activity of sensitized nociceptors, avoids deleterious CNS side effects and may be clinically useful in the treatment of pain of peripheral origin.


Systems Pharmacology Approach for Prediction of Pulmonary and Systemic Pharmacokinetics and Receptor Occupancy of Inhaled Drugs.
Pulmonary drug disposition after inhalation is complex involving mechanisms, such as regional drug deposition, dissolution, and mucociliary clearance. This study aimed to develop a systems pharmacology approach to mechanistically describe lung disposition in rats and thereby provide an integrated understanding of the system. When drug- and formulation-specific properties for the poorly soluble drug fluticasone propionate were fed into the model, it proved predictive of the pharmacokinetics and receptor occupancy after intravenous administration and nose-only inhalation. As the model clearly distinguishes among drug-specific, formulation-specific, and system-specific properties, it was possible to identify key determinants of pulmonary selectivity of receptor occupancy of inhaled drugs: slow particle dissolution and slow drug-receptor dissociation. Hence, it enables assessment of factors for lung targeting, including molecular properties, formulation, as well as the physiology of the animal species, thereby providing a general framework for rational drug design and facilitated translation of lung targeting from animal to man.


Haplotype phasing and inheritance of copy number variants in nuclear families.
DNA copy number variants (CNVs) that alter the copy number of a particular DNA segment in the genome play an important role in human phenotypic variability and disease susceptibility. A number of CNVs overlapping with genes have been shown to confer risk to a variety of human diseases thus highlighting the relevance of addressing the variability of CNVs at a higher resolution. So far, it has not been possible to deterministically infer the allelic composition of different haplotypes present within the CNV regions. We have developed a novel computational method, called PiCNV, which enables to resolve the haplotype sequence composition within CNV regions in nuclear families based on SNP genotyping microarray data. The algorithm allows to i) phase normal and CNV-carrying haplotypes in the copy number variable regions, ii) resolve the allelic copies of rearranged DNA sequence within the haplotypes and iii) infer the heritability of identified haplotypes in trios or larger nuclear families. To our knowledge this is the first program available that can deterministically phase null, mono-, di-, tri- and tetraploid genotypes in CNV loci. We applied our method to study the composition and inheritance of haplotypes in CNV regions of 30 HapMap Yoruban trios and 34 Estonian families. For 93.6% of the CNV loci, PiCNV enabled to unambiguously phase normal and CNV-carrying haplotypes and follow their transmission in the corresponding families. Furthermore, allelic composition analysis identified the co-occurrence of alternative allelic copies within 66.7% of haplotypes carrying copy number gains. We also observed less frequent transmission of CNV-carrying haplotypes from parents to children compared to normal haplotypes and identified an emergence of several de novo deletions and duplications in the offspring.


Successful Treatment of a Case of Late-onset Colitis after Umbilical Cord Transplantation with Metronidazole: A Case Report and Literature Review.
Diarrhea after hematopoietic stem cell transplantation (HSCT) can be life-threatening, and its etiology includes conditioning regimens, graft-versus-host disease (GVHD), infections, and transplantation-associated microangiopathy (iTAM). Cord colitis syndrome (CCS) has been described as a syndrome of culture-negative and antibiotic-responsive persistent watery and non-bloody diarrhea of uncertain pathogenesis and occurs in umbilical cord blood transplantation (UCBT) recipients. We encountered a case similar to CCS that developed severe watery diarrhea after UCBT without any signs of GVHD or infection and responded well to metronidazole (MNZ) treatment. Since CCS is very rare, we herein describe a case of MNZ-effective diarrhea after UCBT.


Surgeon volume is associated with cost and variation in surgical treatment of proximal humeral fractures.
The issue of rising costs will likely dominate the healthcare debate in the forthcoming years. We assessed factors including surgeon volume that were associated with lower hospital costs and variations in surgical treatment for proximal humeral fractures. We used national databases for 2001 to 2008 to extract information on 25,731 patients undergoing surgery for proximal humeral fractures. We calculated hospital cost by converting hospital charges based on the hospital accounting reports collected by the Centers for Medicare & Medicaid Services. In a multivariate linear regression analysis, higher surgeon volume, open reduction and internal fixation (versus hemiarthroplasty), and lower burden of comorbidities were associated with lower hospital cost. Higher surgeon volume was linearly associated with lower hospital costs such that, on average, adjusting for all other factors, a surgeon performing 20 shoulder arthroplasties per year saves a hospital approximately US $1800 per surgery. Factors associated with higher utilization of hemiarthroplasty included high surgeon volume (odds ratio [OR] = 1.46; 95% CI = 1.43, 1.97; as compared with low surgeon volume) and earlier years of our study period (OR = 0.61; 95% CI = 0.56, 0.66; for hemiarthroplasty in 2007-2008 versus 2001-2002). Higher surgeon volume was associated with lower hospital costs for proximal humeral fractures. Therefore, policies on minimum volume requirements by hospitals may result in substantial cost savings. There is provider-based practice variation in the surgical treatment of proximal humeral fractures and evidence-based guidelines in this area are needed. Level III, economic analysis. See Instructions for Authors for a complete description of levels of evidence.


Detection of cervical intraepithelial neoplasias and cancers in cervical tissue by in vivo light scattering.
OBJECTIVE: To examine the utility of in vivo elastic light scattering measurements to identify cervical intraepithelial neoplasias (CIN) 2/3 and cancers in women undergoing colposcopy and to determine the effects of patient characteristics such as menstrual status on the elastic light scattering spectroscopic measurements. MATERIALS AND METHODS: A fiber optic probe was used to measure light transport in the cervical epithelium of patients undergoing colposcopy. Spectroscopic results from 151 patients were compared with histopathology of the measured and biopsied sites. A method of classifying the measured sites into two clinically relevant categories was developed and tested using five-fold cross-validation. RESULTS: Statistically significant effects by age at diagnosis, menopausal status, timing of the menstrual cycle, and oral contraceptive use were identified, and adjustments based upon these measurements were incorporated in the classification algorithm. A sensitivity of 77±5% and a specificity of 62±2% were obtained for separating CIN 2/3 and cancer from other pathologies and normal tissue. CONCLUSIONS: The effects of both menstrual status and age should be taken into account in the algorithm for classifying tissue sites based on elastic light scattering spectroscopy. When this is done, elastic light scattering spectroscopy shows good potential for real-time diagnosis of cervical tissue at colposcopy. Guiding biopsy location is one potential near-term clinical application area, while facilitating "see and treat" protocols is a longer term goal. Improvements in accuracy are essential.


Transcriptomic analysis of polyketide synthases in a highly ciguatoxic dinoflagellate, Gambierdiscus polynesiensis and low toxicity Gambierdiscus pacificus, from French Polynesia.
Marine dinoflagellates produce a diversity of polyketide toxins that are accumulated in marine food webs and are responsible for a variety of seafood poisonings. Reef-associated dinoflagellates of the genus Gambierdiscus produce toxins responsible for ciguatera poisoning (CP), which causes over 50,000 cases of illness annually worldwide. The biosynthetic machinery for dinoflagellate polyketides remains poorly understood. Recent transcriptomic and genomic sequencing projects have revealed the presence of Type I modular polyketide synthases in dinoflagellates, as well as a plethora of single domain transcripts with Type I sequence homology. The current transcriptome analysis compares polyketide synthase (PKS) gene transcripts expressed in two species of Gambierdiscus from French Polynesia: a highly toxic ciguatoxin producer, G. polynesiensis, versus a non-ciguatoxic species G. pacificus, each assembled from approximately 180 million Illumina 125 nt reads using Trinity, and compares their PKS content with previously published data from other Gambierdiscus species and more distantly related dinoflagellates. Both modular and single-domain PKS transcripts were present. Single domain β-ketoacyl synthase (KS) transcripts were highly amplified in both species (98 in G. polynesiensis, 99 in G. pacificus), with smaller numbers of standalone acyl transferase (AT), ketoacyl reductase (KR), dehydratase (DH), enoyl reductase (ER), and thioesterase (TE) domains. G. polynesiensis expressed both a larger number of multidomain PKSs, and larger numbers of modules per transcript, than the non-ciguatoxic G. pacificus. The largest PKS transcript in G. polynesiensis encoded a 10,516 aa, 7 module protein, predicted to synthesize part of the polyether backbone. Transcripts and gene models representing portions of this PKS are present in other species, suggesting that its function may be performed in those species by multiple interacting proteins. This study contributes to the building consensus that dinoflagellates utilize a combination of Type I modular and single domain PKS proteins, in an as yet undefined manner, to synthesize polyketides.


Cloning of NruI and Sbo13I restriction and modification sstems in E. coli and amino acid sequence comparison of M.NruI and M.Sbo13I with other amino-methyltransferases.
NruI and Sbo13I are restriction enzyme isoschizomers with the same recognition sequence 5' TCG downward arrowCGA 3' (cleavage as indicated downward arrow). Here we report the cloning of NruI and Sbo13I restriction-modification (R-M) systems in E. coli. The NruI restriction endonuclease gene (nruIR) was cloned by PCR and inverse PCR using primers designed from the N-terminal amino acid sequence. The NruI methylase gene (nruIM) was derived by inverse PCR walking. The amino acid sequences of NruI endonuclease and methylase are very similar to the Sbo13I R-M system which has been cloned and expressed in E. coli by phage selection of a plasmid DNA library. Dot blot analysis using rabbit polyclonal antibodies to N6mA- or N4mC-modified DNA indicated that M.NruI is possibly a N6mA-type amino-methyltransferase that most likely modifies the external A in the 5' TCGCGA 3' sequence. M.Sbo13I, however, is implicated as a probable N4mC-type methylase since plasmid carrying sbo13IM gene is not restricted by Mrr endonuclease and Sbo13I digestion is not blocked by Dam methylation of the overlapping site. The amino acid sequence of M.NruI and M.Sbo13I did not show significant sequence similarity to many known amino-methyltransferases in the alpha, beta, and gamma groups, except to a few putative methylases in sequenced microbial genomes. The order of the conserved amino acid motifs (blocks) in M.NruI/M.Sbo13I is similar to the gamma. group amino-methyltranferases, but with two distinct features: In motif IV, the sequence is DPPY instead of NPPY; there are two additional conserved motifs, IVa and Xa as extension of motifs IV and X, in this family of enzymes. We propose that M.NruI and M.Sbo13I form a subgroup in the gamma group of amino-methyltransferases.


The allometric propagation of COVID-19 is explained by human travel.
We analyzed the number of cumulative positive cases of COVID-19 as a function of time in countries around the World. We tracked the increase in cases from the onset of the pandemic in each region for up to 150 days. We found that in 81 out of 146 regions the trajectory was described with a power-law function for up to 30 days. We also detected scale-free properties in the majority of sub-regions in Australia, Canada, China, and the United States (US). We developed an allometric model that was capable of fitting the initial phase of the pandemic and was the best predictor for the propagation of the illness for up to 100 days. We then determined that the power-law COVID-19 exponent correlated with measurements of human mobility. The COVID-19 exponent correlated with the magnitude of air passengers per country. This correlation persisted when we analyzed the number of air passengers per US states, and even per US metropolitan areas. Furthermore, the COVID-19 exponent correlated with the number of vehicle miles traveled in the US. Together, air and vehicular travel explained 70% of the variability of the COVID-19 exponent. Taken together, our results suggest that the scale-free propagation of the virus is present at multiple geographical scales and is correlated with human mobility. We conclude that models of disease transmission should integrate scale-free dynamics as part of the modeling strategy and not only as an emergent phenomenological property.


Pharmacokinetic drug interactions between oral contraceptives and second-generation anticonvulsants.
Drug interactions between oral contraceptives (OCs) and traditional anticonvulsants have been well described. However, in the past decade, a number of new anticonvulsants have been developed, as well as modifications made in the composition of the OC preparations themselves. Additionally, anticonvulsants are increasingly employed in the therapy of nonseizure-related disorders, placing more women at risk of potential drug interactions that may lead to contraceptive failure. Second-generation anticonvulsants include felbamate, gabapentin, lamotrigine, oxcarbazepine, tiagabine, topiramate, vigabatrin and zonisamide. Most have been approved for adjunctive management of seizures refractory to therapy with traditional anticonvulsants. On the basis of available study data in women receiving concomitant OC preparations, gabapentin, lamotrigine, tiagabine and vigabatrin may be administered without significant pharmacokinetic interactions that potentially diminish contraceptive efficacy. However, additional or alternative contraceptive measures, including using OCs with higher estrogen content, are recommended when using felbamate, oxcarbazepine and topiramate, as these agents have demonstrated enzyme-inducing activity leading to reduced plasma steroid concentrations. The effects of zonisamide in women receiving OCs have yet to be reported. It is important to characterise the properties [e.g. substrate and enzyme activity (particularly cytochrome P450 3A4 induction)] of new anticonvulsants and recognise their potential to interfere with OCs. However, a pharmacokinetic interaction does not in itself indicate loss of OC efficacy. Contraceptive failure should be measured by changes in ovarian hormone concentrations, maturation of ovarian follicle(s) or ovulation.


Telemedicine Use in Orthopaedic Surgery Varies by Race, Ethnicity, Primary Language, and Insurance Status.
Healthcare disparities are well documented across multiple subspecialties in orthopaedics. The widespread implementation of telemedicine risks worsening these disparities if not carefully executed, despite original assumptions that telemedicine improves overall access to care. Telemedicine also poses unique challenges such as potential language or technological barriers that may alter previously described patterns in orthopaedic disparities. Are the proportions of patients who use telemedicine across orthopaedic services different among (1) racial and ethnic minorities, (2) non-English speakers, and (3) patients insured through Medicaid during a 10-week period after the implementation of telemedicine in our healthcare system compared with in-person visits during a similar time period in 2019? This was a retrospective comparative study using electronic medical record data to compare new patients establishing orthopaedic care via outpatient telemedicine at two academic urban medical centers between March 2020 and May 2020 with new orthopaedic patients during the same 10-week period in 2019. A total of 11,056 patients were included for analysis, with 1760 in the virtual group and 9296 in the control group. Unadjusted analyses demonstrated patients in the virtual group were younger (median age 57 years versus 59 years; p < 0.001), but there were no differences with regard to gender (56% female versus 56% female; p = 0.66). We used self-reported race or ethnicity as our primary independent variable, with primary language and insurance status considered secondarily. Unadjusted and multivariable adjusted analyses were performed for our primary and secondary predictors using logistic regression. We also assessed interactions between race or ethnicity, primary language, and insurance type. After adjusting for age, gender, subspecialty, insurance, and median household income, we found that patients who were Hispanic (odds ratio 0.59 [95% confidence interval 0.39 to 0.91]; p = 0.02) or Asian were less likely (OR 0.73 [95% CI 0.53 to 0.99]; p = 0.04) to be seen through telemedicine than were patients who were white. After controlling for confounding variables, we also found that speakers of languages other than English or Spanish were less likely to have a telemedicine visit than were people whose primary language was English (OR 0.34 [95% CI 0.18 to 0.65]; p = 0.001), and that patients insured through Medicaid were less likely to be seen via telemedicine than were patients who were privately insured (OR 0.83 [95% CI 0.69 to 0.98]; p = 0.03). Despite initial promises that telemedicine would help to bridge gaps in healthcare, our results demonstrate disparities in orthopaedic telemedicine use based on race or ethnicity, language, and insurance type. The telemedicine group was slightly younger, which we do not believe undermines the findings. As healthcare moves toward increased telemedicine use, we suggest several approaches to ensure that patients of certain racial, ethnic, or language groups do not experience disparate barriers to care. These might include individual patient- or provider-level approaches like expanded telemedicine schedules to accommodate weekends and evenings, institutional investment in culturally conscious outreach materials such as advertisements on community transport systems, or government-level provisions such as reimbursement for telephone-only encounters. Level III, therapeutic study.


[2 newborn infants with severe arrhythmia caused by hyperkalemia].
Two newborn infants with ventricular arrhythmias secondary to hyperkalaemia are presented. One child also showed a decreased serum calcium concentration. There is scanty literature concerning the often life threatening cardiac arrhythmias due to hyperkalaemia in the newborn infants. Treatment of the cardiac arrhythmias require intravenous calcium gluconat and sodium bicarbonate infusion beside lowering the serum potassium level in the usual way.


[Effect of transurethral resection of the prostate on the state of patients with ischemic heart disease].
The water-salt equilibrium and the degree of endogenous intoxication (albumin fluorescence test) were examined in 60 patients with transurethral resection of the prostate (TURP), 40 of whom had ischemic heart disease (IHD). Body temperature, ECG, ST segment and echocardioscopy were daily monitored perioperatively in all patients. The results showed a lack of any pronounced changes in the water-salt equilibrium in TURP that lasted up to 1.5 hours and included big volumes of 5% glucose; they were also indicative of a lower postoperative binding albumin ability, which is normally most pronounced in patients with hyperthermia. As for the IHD patients, hyperthermia was found to be concurrent in them with the onset of ischemia of the myocardium and with its low contractive ability, which can be referred to as a significant factor in case of the above patients.


Managing clinical trials.
Managing clinical trials, of whatever size and complexity, requires efficient trial management. Trials fail because tried and tested systems handed down through apprenticeships have not been documented, evaluated or published to guide new trialists starting out in this important field. For the past three decades, trialists have invented and reinvented the trial management wheel. We suggest that to improve the successful, timely delivery of important clinical trials for patient benefit, it is time to produce standard trial management guidelines and develop robust methods of evaluation.


Transgenic tobacco and Arabidopsis plants expressing the two multifunctional sorghum cytochrome P450 enzymes, CYP79A1 and CYP71E1, are cyanogenic and accumulate metabolites derived from intermediates in Dhurrin biosynthesis.
Novel cyanogenic plants have been generated by the simultaneous expression of the two multifunctional sorghum (Sorghum bicolor [L.] Moench) cytochrome P450 enzymes CYP79A1 and CYP71E1 in tobacco (Nicotiana tabacum cv Xanthi) and Arabidopsis under the regulation of the constitutive 35S promoter. CYP79A1 and CYP71E1 catalyze the conversion of the parent amino acid tyrosine to p-hydroxymandelonitrile, the aglycone of the cyanogenic glucoside dhurrin. CYP79A1 catalyzes the conversion of tyrosine to p-hydroxyphenylacetaldoxime and CYP71E1, the subsequent conversion to p-hydroxymandelonitrile. p-Hydroxymandelonitrile is labile and dissociates into p-hydroxybenzaldehyde and hydrogen cyanide, the same products released from dhurrin upon cell disruption as a result of pest or herbivore attack. In transgenic plants expressing CYP79A1 as well as CYP71E1, the activity of CYP79A1 is higher than that of CYP71E1, resulting in the accumulation of several p-hydroxyphenylacetaldoxime-derived products in the addition to those derived from p-hydroxymandelonitrile. Transgenic tobacco and Arabidopsis plants expressing only CYP79A1 accumulate the same p-hydroxyphenylacetaldoxime-derived products as transgenic plants expressing both sorghum cytochrome P450 enzymes. In addition, the transgenic CYP79A1 Arabidopsis plants accumulate large amounts of p-hydroxybenzylglucosinolate. In transgenic Arabidopsis expressing CYP71E1, this enzyme and the enzymes of the pre-existing glucosinolate pathway compete for the p-hydroxyphenylacetaldoxime as substrate, resulting in the formation of small amounts of p-hydroxybenzylglucosinolate. Cyanogenic glucosides are phytoanticipins, and the present study demonstrates the feasibility of expressing cyanogenic compounds in new plant species by gene transfer technology to improve pest and disease resistance.


Remarkable frequency of a history of liver disease in dogs fed homemade diets with buckwheat.
In our nutrition consultation service we observed liver disease in 2 dogs of one owner who was feeding buckwheat. This led to the hypothesis that buckwheat may cause problems. The present retrospective study in a German and a Russian nutrition consultation service was carried out to see whether there is an increased incidence of liver disease in dogs fed buckwheat. A retrospective study was carried out on the nutrition consultation cases of the Chair of Animal Nutrition and Dietetics, LMU Munich and a Russian nutrition consultant. All cases of dogs with buckwheat in their nutritional history were evaluated and compared with randomly selected dogs that had not been fed buckwheat from the same case set. Two German and 1 Russian buckwheat samples were compared (appearance, nutrient content, starch gelatinization, flavonoids, fagopyrin) as well as cooking methods. In the years 2007-2017, 34 cases of dogs fed buckwheat were identified in Germany and 57 in Russia. Eighty-five control cases in Germany and 48 in Russia were evaluated. In Germany, the incidence of liver disease in dogs fed buckwheat was 32 %, while that of the control group was 3.5 %. However, in Russia there was no significant difference between dogs fed buckwheat and control dogs. The appearance of the German and Russian buckwheat differed, with smaller seeds and more greenish colour in the German specimens while the Russian buckwheat presented larger and more brownish seeds. There was no difference in the analyses of the 3 buckwheat samples in crude nutrient and rutin content. Quercetin, quercitrin and fagopyrin were not detectable in all three samples. The degree of starch gelatinization in the Russian sample was higher than in the German. In Russia it is recommended to remove the reddish scum during boiling whereas this is rarely mentioned in Germany. German buckwheat may represent a risk in canine diets. With the difference remaining unclear, it is recommended to refrain from feeding buckwheat to dogs. In dogs fed homemade diets and suffering from liver disease, buckwheat should be considered in the nutrition history.


Furosemide Stress Test as a Predicting Biomarker for Delayed Graft Function in Kidney Transplantation.
Delayed graft function (DGF) could worsen early and long-term outcomes of kidney transplantation (KT). DGF is caused by several pre-transplantation and perioperative factors in both donors and recipients. At present, there are no biomarkers or tests during the immediate post-KT period that can accurately predict the development of DGF. This prospective study was conducted in deceased donor KT (DDKT) at King Chulalongkorn Memorial Hospital, Thailand. All recipients underwent furosemide stress test (FST) by receiving a single dose of intravenous furosemide, 1.5 mg/kg at 3 h after allograft reperfusion. We determined the correlations between DGF (requiring dialysis within the first week after transplantation) and the values of urine volume recorded hourly after FST until 6 h, the parameters of postoperative dynamic tests, including resistive index (RI) of renal arteries and effective renal plasma flow (ERPF), and urine neutrophil gelatinase-associated lipocalin (NGAL). Of the 59 total DDKT recipients enrolled, 24 developed DGF. The FST is a more accurate biomarker than urine NGAL, RI of renal arteries, and ERPF in the prediction of DGF. The 4-h urine volume less than 350 mL (FST non-responsive) was the best cut-off value in predicting DGF with 87.5% sensitivity, 82.9% specificity, and 82.5% accuracy. Multiple logistic regression analyses showed an odds ratio of 0.993 (0.986-0.999, p = 0.035) for the 4-h urine volume to predict DGF. The FST is a simple and accurate biomarker for predicting DGF in early post-KT period. Close monitoring and well prepared dialysis are suggested in patients with urine volume < 350 mL after 4 h of FST. The FST non-responsive patients could be the target for further DGF preventive intervention. ClinicalTrials.gov identifier: NCT03071536.


Photoassisted Electrolysis of Water by Visible Irradiation of a p-Type Gallium Phosphide Electrode.
Photoelectrolytic decomposition of water with visible irradiation is demonstrated in a cell made of p-type gallium phosphide as the cathode and platinum as the anode. A maximum energy conversion efficiency of 0.1 percent is achieved with an external bias of 1.3 volts. The stability of the electrode is demonstrated, and the results are discussed in terms of a composite energy diagram.


Clinical course and outcome of severe head injury in Australia.
The records of 159 severely head-injured patients (all in coma for longer than 6 hours) from Sydney, Australia, were studied. The clinical course, charted over a 2-week period, indicated that 60% of deaths occur by Day 3 and that 12% of patients remain in coma (Glagow Coma Scale (GCS) score less than 7) for more than 2 weeks. Overall, at long-term follow-up review more than 2 years after injury, 51% of patients were dead, 7% were severely disabled or vegetative, and 42% had a good to moderate recovery. Outcome of the patients in prolonged coma was assessed separately, with only one-third making a good or moderate recovery; two-thirds of the severely disabled patients came from this group. The high proportion of poor outcomes associated with prolonged coma suggests that this group of patients should be specifically targeted in research. One appropriate intervention with this group would be the restructuring and intensification of early rehabilitation. However, the GCS score lacks the precision needed for this type of study, and a better measure of recovery should be developed.


When less is better: a comparison of Bach Flower Remedies and homeopathy.
Homeopathy and Bach Flower Remedies (BFRs), historically-related complementary healing modalities classified as prescription/nonprescription drugs and over-the-counter homeopathic nutritional supplements, respectively, are compared with respect to indications, dosage philosophies, associated procedures, reported outcomes, safety profiles, and the possible operation of the placebo effect. Original data and published research reports, including case studies, retrospective meta-analyses, and double-blind clinical trials are compiled and evaluated for both healing systems. Homeopathy and BFR therapy both feature highly diluted natural medicinal substances, flexible dosage schedules tailored to individual patients, and energy-based healing action. They differ with respect to practitioner training and certification, number and types of medicinal source materials, remedy combinations and applications, and potential toxicity or other side-effects. Extensive testing has produced mixed or equivocal results regarding the efficacy of both of these health care systems. While a variety of positive outcomes have been frequently recorded with Homoeopathy and BFR treatments, it is likely that the placebo effect operates to a significant extent in both approaches.


Duration of treatment for urinary tract infections in children.
In a double-blind trial 45 children aged 6 months to 14 years with Escherichia coli infections of the urinary tract were given co-trimoxazole for two weeks and then allotted at random to one of two treatment groups for the remainder of six months; one continued with the active drug and the other with dummy tablets of identical appearance. Of the 24 children who took co-trimoxazole for two weeks and the 21 who took it for six months, 11 and 10, respectively, remained without further infections for at least a year. Over 90% of the reinfections occurred within five months of stopping the antibiotics, and the longer treatment did not cause any delay in their appearance. Thus probably a six-month course of treatment is no more likely to achieve a cure than a two-week course; nevertheless, no infection occurred during treatment, and there may be an advantage in continuing with antibiotics in small dosage.


A Tiered Approach to Evaluating Salinity Sources in Water at Oil and Gas Production Sites.
A suspected increase in the salinity of fresh water resources can trigger a site investigation to identify the source(s) of salinity and the extent of any impacts. These investigations can be complicated by the presence of naturally elevated total dissolved solids or chlorides concentrations, multiple potential sources of salinity, and incomplete data and information on both naturally occurring conditions and the characteristics of potential sources. As a result, data evaluation techniques that are effective at one site may not be effective at another. In order to match the complexity of the evaluation effort to the complexity of the specific site, this paper presents a strategic tiered approach that utilizes established techniques for evaluating and identifying the source(s) of salinity in an efficient step-by-step manner. The tiered approach includes: (1) a simple screening process to evaluate whether an impact has occurred and if the source is readily apparent; (2) basic geochemical characterization of the impacted water resource(s) and potential salinity sources coupled with simple visual and statistical data evaluation methods to determine the source(s); and (3) advanced laboratory analyses (e.g., isotopes) and data evaluation methods to identify the source(s) and the extent of salinity impacts where it was not otherwise conclusive. A case study from the U.S. Gulf Coast is presented to illustrate the application of this tiered approach.


Sheep model in orthopedic research: a literature review.
The aim of the study reported here was to provide some basic and general information on the suitability of an experimental sheep model for conducting in vivo orthopedic studies. The authors have classified the fundamental aspects that should be carefully evaluated when using sheep as an experimental model in orthopedic research: factors strictly related to bone anatomy and formation; and factors strictly affecting bone physiology, such as gastrointestinal mineral and vitamin absorption, and reproductive cycle. Future investigations should address all of the aspects highlighted, since there is no animal with the same anatomic, biochemical, physiologic, and biological characteristics as those of human beings. Moreover, useful data for treating orthopedic patients are based not only on good planning and study design, but also on perfect knowledge of the animal used and the differences between the model and the human being. The authors hope that this report will contribute to extrapolation of reliable data for use of sheep in the orthopedics field.


Biotechnological applications of mobile group II introns and their reverse transcriptases: gene targeting, RNA-seq, and non-coding RNA analysis.
Mobile group II introns are bacterial retrotransposons that combine the activities of an autocatalytic intron RNA (a ribozyme) and an intron-encoded reverse transcriptase to insert site-specifically into DNA. They recognize DNA target sites largely by base pairing of sequences within the intron RNA and achieve high DNA target specificity by using the ribozyme active site to couple correct base pairing to RNA-catalyzed intron integration. Algorithms have been developed to program the DNA target site specificity of several mobile group II introns, allowing them to be made into 'targetrons.' Targetrons function for gene targeting in a wide variety of bacteria and typically integrate at efficiencies high enough to be screened easily by colony PCR, without the need for selectable markers. Targetrons have found wide application in microbiological research, enabling gene targeting and genetic engineering of bacteria that had been intractable to other methods. Recently, a thermostable targetron has been developed for use in bacterial thermophiles, and new methods have been developed for using targetrons to position recombinase recognition sites, enabling large-scale genome-editing operations, such as deletions, inversions, insertions, and 'cut-and-pastes' (that is, translocation of large DNA segments), in a wide range of bacteria at high efficiency. Using targetrons in eukaryotes presents challenges due to the difficulties of nuclear localization and sub-optimal magnesium concentrations, although supplementation with magnesium can increase integration efficiency, and directed evolution is being employed to overcome these barriers. Finally, spurred by new methods for expressing group II intron reverse transcriptases that yield large amounts of highly active protein, thermostable group II intron reverse transcriptases from bacterial thermophiles are being used as research tools for a variety of applications, including qRT-PCR and next-generation RNA sequencing (RNA-seq). The high processivity and fidelity of group II intron reverse transcriptases along with their novel template-switching activity, which can directly link RNA-seq adaptor sequences to cDNAs during reverse transcription, open new approaches for RNA-seq and the identification and profiling of non-coding RNAs, with potentially wide applications in research and biotechnology.


Updated Cost-effectiveness Assessments of PCSK9 Inhibitors From the Perspectives of the Health System and Private Payers: Insights Derived From the FOURIER Trial.
Preliminary cost-effectiveness analyses of proprotein convertase subtilisin/kexin type 9 inhibitor (PCSK9i) were based on benefits estimated from reductions in low-density lipoprotein cholesterol that occurred in PCSK9i trials with variable results. The recent Further Cardiovascular Outcomes Research with PCSK9 Inhibition in Subjects with Elevated Risk (FOURIER) trial provides better information about the effectiveness of the drug. To use the trial results to determine the cost-effectiveness of a PCSK9i and statin treatment strategy compared with a statin alone strategy. We derived observed rates of events, outcomes, cost of care, and health insurance from existing literature for a theoretical cohort of patients designed to resemble the FOURIER PCSK9i trial population and created a Markov model during the time horizon of a full lifetime. We evaluated the incremental cost-effectiveness ratio from a health system perspective, and the return on investment from a private payer perspective. For both measures, we assumed an annual PCSK9i drug price of $14 300, with a lapse in US patent protection that would reduce the price by 43% in year 12. Costs were reported in 2016 US dollars. This study modeled 1000 hypothetical patients with attributes similar to those of the FOURIER trial cohort. At the current price, the incremental cost-effectiveness ratio of statin plus PCSK9i therapy was $337 729 per quality-adjusted life-year. Our probabilistic sensitivity analysis found that a statin plus PCSK9i strategy had a low probability (<1%) of being cost effective at the commonly accepted societal threshold of $100 000 per quality-adjusted life-year. Furthermore, PCSK9i produced a negative return on investment of 86% for private payers. In our threshold analysis, the price of PCSK9i would need to drop 62%, to $5459 per year, to reach $100 000 per quality-adjusted life year. At current prices, the addition of PCSK9i to statin therapy is estimated to provide an additional quality-adjusted life year for $337 729 . Significant discounts are necessary to meet conventional cost-effectiveness standards.


[Multiple cause of death in cardio- and cerebrovascular diseases].
In order to validate mortality studies, we described the role of hypertension at the time of death of people who died during April and May 1985. This analysis take into account all the causes listed in death certificates and not only the underlying cause. A proportional mortality design was used to evaluate the relationship of hypertension, cerebrovascular and cardiovascular disease. The number of deaths associated to hypertension was larger by multiple cause than by underlying cause. The odds ratio for hypertension and ischemic heart disease was 4.1 adjusted by age and sex, and 21.1 for cerebrovascular disease.


The relationship between hypertension and salt intake in Turkish population: SALTURK study.
This population-based epidemiological study was aimed to evaluate the daily salt intake and its relation to blood pressure in a representative group of Turkish population. The enrolled normotensive and hypertensive individuals (n = 1970) completed a questionnaire including demographics, dietary habits, hypertension awareness and drug usage. Blood pressure was measured and to estimate salt consumption, 24-h urine samples were collected. The daily urinary sodium excretion was 308.3 ± 143.1 mmol/day, equal to a salt intake of 18.01 g/day. Salt intake was higher in obese participants, rural residents, participants with lower education levels and elderly. A positive linear correlation between salt intake and systolic and diastolic blood pressures was demonstrated (r = 0.450, p = 0.020; r = 0.406, p = 0.041; respectively), and each 100 mmol/day of salt intake resulted in 5.8 and 3.8 mmHg increase in systolic and diastolic blood pressures, respectively. Salt intake and systolic blood pressure was significantly correlated in normal weight individuals (r = 0.257, p < 0.01). The Turkish population consumes a great amount of salt; salt intake and blood pressure was positively correlated. Efforts in sodium restriction are therefore crucial in the management of hypertension as part of national and global health policies.


Halothane-induced liver injury in outbred guinea pigs: role of trifluoroacetylated protein adducts in animal susceptibility.
Halothane causes a mild form of liver injury in guinea pigs that appears to model the hepatotoxicity seen in approximately 20% of patients treated with this drug. In previous studies, it was concluded that the increased susceptibility of some outbred guinea pigs to halothane-induced liver injury is not caused by their inherent ability to metabolize halothane to form toxic levels of trifluoroacetylated protein adducts in the liver. In this study, we reevaluated the role of trifluoroacetylated protein adducts in halothane-induced liver injury in guinea pigs. Male outbred Hartley guinea pigs were treated with halothane intraperitoneally. On the basis of serum alanine aminotransferase levels and liver histology, treated animals were designated as being susceptible, mildly susceptible, or resistant to halothane. Immunoblot studies with the use of anti-trifluoroacetylated antibodies showed that susceptible guinea pigs for the most part had higher levels of trifluoroacetylated protein adducts in the liver 48 h after treatment with halothane than did less susceptible animals. In support of this finding, the level of trifluoroacetylated protein adducts detected immunochemically in the sera of treated guinea pigs correlated with sera levels of alanine aminotransferase activity. In addition, the levels of cytochrome P450 2A-related protein but not those of other cytochrome P450 isoforms, measured by immunoblot analysis with isoform-specific antibodies, correlated with the amount of trifluoroacetylated protein adducts detected in the livers of guinea pigs 8 h after halothane administration. The results of this study indicate that the susceptibility of outbred guinea pigs to halothane-induced liver injury is related to an enhanced ability to metabolize halothane in the liver to form relatively high levels of trifluoroacetylated protein adducts. They also suggest that cytochrome P450 2A-related protein might have a major role in catalyzing the formation of trifluoroacetylated protein adducts in the liver of susceptible guinea pigs. Similar mechanisms may be important in humans.


DIACYLGLYCEROL ACYLTRANSFERASE and DIACYLGLYCEROL KINASE Modulate Triacylglycerol and Phosphatidic Acid Production in the Plant Response to Freezing Stress.
Plants accumulate the lipids phosphatidic acid (PA), diacylglycerol (DAG), and triacylglycerol (TAG) during cold stress, but how plants balance the levels of these lipids to mediate cold responses remains unknown. The enzymes ACYL-COENZYME A:DIACYLGLYCEROL ACYLTRANSFERASE (DGAT) and DIACYLGLYCEROL KINASE (DGK) catalyze the conversion of DAG to TAG and PA, respectively. Here, we show that DGAT1, DGK2, DGK3, and DGK5 contribute to the response to cold in Arabidopsis (Arabidopsis thaliana). With or without cold acclimation, the dgat1 mutants exhibited higher sensitivity upon freezing exposure compared with the wild type. Under cold conditions, the dgat1 mutants showed reduced expression of C-REPEAT/DRE BINDING FACTOR2 and its regulons, which are essential for the acquisition of cold tolerance. Lipid profiling revealed that freezing significantly increased the levels of PA and DAG while decreasing TAG in the rosettes of dgat1 mutant plants. During freezing stress, the accumulation of PA in dgat1 plants stimulated NADPH oxidase activity and enhanced RbohD-dependent hydrogen peroxide production compared with the wild type. Moreover, the cold-inducible transcripts of DGK2, DGK3, and DGK5 were significantly more up-regulated in the dgat1 mutants than in the wild type during cold stress. Consistent with this observation, dgk2, dgk3, and dgk5 knockout mutants showed improved tolerance and attenuated PA production in response to freezing temperatures. Our findings demonstrate that the conversion of DAG to TAG by DGAT1 is critical for plant freezing tolerance, acting by balancing TAG and PA production in Arabidopsis.


Contemporary trends of in-hospital complications and mortality for radical cystectomy.
What's known on the subject? and What does the study add? Radical cystectomy (RC) carries significant risks of morbidity and mortality. Little is known whether in-hospital outcomes are improving for RC. Using a contemporary population-based cohort, the present study suggests minimal improvement in postoperative complications and mortality overall or by hospital-volume category from 2001 to 2008. About 29% and 2% of patients undergoing RC will experience a postoperative complication or die during hospitalisation, respectively. To characterise the contemporary trends of in-hospital complications and mortality for radical cystectomy (RC) from a contemporary population-based cohort, as patients undergoing RC for bladder cancer are at significant risk for complications and mortality and the degree to which in-hospital outcomes have changed over time is unknown. We identified 50 625 individuals who underwent RC for bladder cancer between 2001 and 2008 from the Nationwide Inpatient Sample. Multivariable regression models were used to identify hospital and patient covariates associated with in-hospital complications and mortality and to estimate predicted probabilities of each outcome. Temporal trends of in-hospital mortality and complications were assessed by Wilcoxon rank-sum test. The proportion of patients with in-hospital complications remained stable at 28.3% in 2001-2002 compared with 28.0% in 2007-2008 (P = 0.81 for trend). In-hospital mortality was also unchanged from 2.4% in 2001-2002 compared with 2.3% in 2007-2008 (P = 0.87 for trend). While high-volume hospitals were associated with lower odds of in-hospital complications (odds ratio [OR] 0.77, P = 0.01) and mortality (OR 0.60, P = 0.02) compared with low-volume hospitals, the predicted probabilities of in-hospital complications or mortality were unchanged within each volume category between 2001 and 2008. In-hospital complications and mortality for RC remain unchanged from 2001 to 2008. While high-volume hospitals continue to have better outcomes, there is little evidence that postoperative mortality and morbidity are improving among low-, medium- and high-volume hospitals. Increased attention is needed to identify the modifiable aspects of postoperative care to improve in-hospital outcomes and safety for patients undergoing RC.


Association between kidney measurements and cognitive performance in patients with ischemic stroke.
Individuals with chronic kidney disease (CKD) are at a substantially higher risk for stroke, which may predispose individuals to cognitive impairment. However, the association of low estimated glomerular filtration rate (eGFR) and albuminuria with poorer cognitive performance in patients with stroke is not fully understood, and the current evidence for this association is contradictory. Our aim was to retrospectively investigate whether low eGFR and albuminuria, as indicated by the urine albumin-creatinine ratio (UACR), are independently or jointly associated with worse cognitive performance in patients with ischemic stroke. This retrospective study included 608 patients with acute ischemic stroke. Their UACR and eGFR values were obtained from inpatient medical records. Global cognitive function was assessed with the mini-mental state exam (MMSE) and Montreal Cognitive Assessment (MoCA) one month after hospital discharge. The relationship between renal measures and cognitive performance was assessed using univariate and multiple linear regression analyses. Potential confounders included age, gender, BMI, education, diabetes and hypertension history, NIHSS score, smoking and alcohol consumption status, serum total cholesterol, triglyceride, fasting glucose, uric acid, homocysteine, systolic blood pressure, and either eGFR or UACR. Patients had an average age of 66.6±4.1 years, and 48% were females. Average eGFR and UACR were 88.4±12.9 ml/min/1.73m2 and 83.6±314.2 mg/g, respectively. The number of patients with eGFR ≥90, 60-89, and <60 ml/min/1.73 m2 was 371 (61%), 207 (34%), and 30 (5%), respectively, and the percentage of patients with UACR <30 mg/g, 30-300 mg/g, and >300 mg/g was 56%, 39%, and 5%, respectively. Multivariate adjusted models showed that eGFR was independently associated with MMSE (β = -0.4; 95% CI = -0.5,-0.4; p <0.001) and MoCA (β = -0.6; 95% CI = -0.7,-0.5; p <0.001). However, UACR was not significantly correlated with MMSE or MoCA. In patients with ischemic stroke, reduced eGFR but not albuminuria was associated with lower cognitive performance. These results show that the eGFR decline could be an effective indicator of cognitive impairment after a stroke. Therefore, regular monitoring and early detection of mild renal dysfunction in patients with acute ischemic stroke might be needed.


Induction of mammary gland carcinomas by the subcutaneous injection of 1-methyl-1-nitrosourea.
The efficacy of s.c. administration of 1-methyl-1-nitrosourea (MNU) for the induction of mammary carcinomas was compared with the i.v. method of carcinogen injection in female Sprague-Dawley rats. Group-housed animals fed a laboratory chow diet and distilled water ad libitum throughout the study were injected at 50 days of age with 50 mg MNU per kg body weight. The carcinogen was given either s.c. or i.v., via the jugular vein, to one of the two groups of 20 rats each. Animals were palpated for tumor detection weekly and necropsied 180 days after injection with the carcinogen. At the termination of the study, 180 days postcarcinogen, cancer incidences were similar, 95 versus 90% in animals given MNU either s.c. or i.v. with an average of 3.9 and 3.9 cancers per rat, respectively. Time of tumor appearances were essentially identical under both treatment conditions. Using either method of carcinogen administration resulted in the induction of approximately 2.4 times more carcinomas in the cervical-thoracic mammary glands than in the abdominal-inguinal glands with no differences observed in cancer occurrence in the left versus the right mammary gland chains. The data indicate that s.c. administration of MNU is as effective and specific in the induction of mammary carcinomas as is i.v. administration. The s.c. method has the advantage of being easier and faster to perform and permits reproducible treatment of large numbers of rats by a small technical staff.


One-pot multi-segment condensation strategies for chemical protein synthesis.
With the growing requirement for otherwise-difficult-to-obtain proteins, it is necessary to develop more efficient chemical protein synthesis methods for rapid access to designed protein samples. In particular, a one-pot multi-segment condensation method, with only one purification step to obtain the final product, is expected to demonstrate unique benefits in chemical protein synthesis, such as the requirement of fewer handling procedures and the higher efficiency in obtaining aimed protein samples. The utilization of the one-pot multi-segment condensation strategy is demonstrated via the synthesis of a series of post-translational modification (PTM) or disease-associated peptides or proteins for basic and advanced scientific research. This review summarizes the recent one-pot multi-segment condensation methods utilized in chemical protein synthesis, in which two aspects of drive-strategies will be mainly included: a kinetically controlled strategy and a protecting group-removal strategy, respectively. On one hand, the activities of peptides in N-terminal thiol amino acids or C-terminal acyl donors can be largely different based on the differences in properties, such as steric hindrance, migration rates, electrophilicity, and introduction of active elements such as selenium, etc. Using the different activities, regio-selective peptide ligation can be performed in a kinetically controlled manner. On the other hand, the protecting group-removal strategy involves various moieties, which can block the activity of functional groups arising from N-terminal thiol amino acids or C-terminal acyl donors, and they can be removed by using additives, and pH- or photo-stimulation conditions with further achievement of chemical protein synthesis by the one-pot strategy.


Cooperative regulation of c-myc expression in differentiation of human promyelocytic leukemia induced by recombinant gamma-interferon and 1,25-dihydroxyvitamin D3.
The biological activity of recombinant human gamma-interferon (IFN-gamma) to induce the phenotypic differentiation of human promyelocytic leukemia cell line HL-60 and to regulate c-myc expression was evaluated. Treatment with IFN-gamma increased monocyte-associated cell surface antigens detected by monocyte-specific monoclonal antibodies in a dose- and time-dependent manner. These antigenic changes were accompanied by a functional differentiation, determined by the increase of phagocytic capability and superoxide generation. IFN-gamma was also found to suppress the growth of HL-60 cells and reduce expression of a c-myc oncogene. These phenotypic and morphological changes to macrophage-like cells induced by IFN-gamma were similar to those by 1,25-dihydroxyvitamin D3, whereas the plasma membrane antigenic changes were different. Moreover, the combination of IFN-gamma and suboptimal doses of 1,25-dihydroxyvitamin D3 have synergistic effects in augmenting mature monocyte specific antigens (Mo2, 63D3, OKM1, and OKM5). In the reduction of c-myc expression by these drugs, a cooperative effect was observed with the inhibition of transferrin receptor expression and cell growth. These results indicate that human recombinant IFN-gamma induces a monocyte phenotype in the HL-60 cells via a mechanism different from the action of 1,25-dihydroxyvitamin D3.


Adolescent Attentional Bias toward Real-world Flavored E-cigarette Marketing.
E-cigarettes are now the most commonly-used tobacco product among adolescents; yet, little work has examined how the appealing food and flavor cues used in their marketing might attract adolescents' attention, thereby increasing willingness to try these products. In the present study, we tested whether advertisements for fruit/sweet/savory-flavored ("flavored") e-cigarettes attracted adolescent attention in real-world scenes more than tobacco flavored ("unflavored") e-cigarettes. Additionally, we examined the relationship between adolescent attentional bias and willingness to try flavored e-cigarettes. Participants were 46 adolescents (age range: 16-18 years). All participants took part in an eye-tracking paradigm that examined attentional bias to flavored and unflavored e-cigarette advertisements embedded in pictures of real-world storefront scenes. Afterwards, participants' willingness to try flavored and unflavored e-cigarettes was assessed. In support of our primary hypothesis, adolescents looked longer and fixated more frequently on flavored (vs unflavored) e-cigarette advertisements. Moreover, this attentional bias towards flavored e-cigarette advertisements predicted a greater willingness to try flavored vs unflavored e-cigarettes. These findings suggest that flavored e-cigarette marketing attracts the attention of adolescents, in- creases their willingness to try flavored e-cigarette products, and could, therefore, put them at greater risk for tobacco initiation.


Profibrogenic phenotype in caveolin-1 deficiency via differential regulation of STAT-1/3 proteins.
Fibrosis underlies the pathogenesis of several human diseases, which can lead to severe injury of vital organs. We previously demonstrated that caveolin-1 expression is reduced in experimental fibrosis and that caveolin-1 exerts antiproliferative and antifibrotic effects in lung fibrosis models. The signal transducers and activators of transcription (STAT) proteins, STAT1 and STAT3, can be activated simultaneously. STAT1 can inhibit cell growth and promote apoptosis while STAT3 inhibits apoptosis. Here, we show that caveolin-1-deficient (cav-1(-/-)) lung fibroblasts display dramatically upregulated STAT3 activation in response to platelet-derived growth factor-BB and transforming growth factor-β stimuli, whereas STAT1 activation is undetectable. Downregulation of protein tyrosine phosphatase-1B played a role in the preferential activation of STAT3 in cav-1(-/-) fibroblasts. Genetic deletion of STAT3 by siRNA modulated the expression of genes involved in cell proliferation and fibrogenesis. Basal expression of α-smooth muscle actin was prominent in cav-1(-/-) liver and kidney, consistent with deposition of collagen in these organs. Collectively, we demonstrate that the antiproliferative and antifibrogenic properties of caveolin-1 in vitro are mediated by the balance between STAT1 and STAT3 activation. Deregulated STAT signaling associated with caveolin-1 deficiency may be relevant to proliferative disorders such as tissue fibrosis.


Effect monitoring in dual-task performance.
Actions aim to produce effects in the environment. To accomplish this properly, we not only have to recruit the appropriate motor patterns, but also we must be able to monitor whether an intended effect has ultimately been realized. Here, we investigated the impact of such effect monitoring on performance in multitasking situations: Multitasking basically means to produce and monitor multiple actions and effects in fast succession. We show that effect monitoring cannot run in parallel, without causing processing decrements, with a second task. Also, monitoring of effects that are spatially incompatible to a response seems to take longer than the monitoring of spatially compatible action effects (Experiments 1 through 4). We further argue that effect monitoring is essential toward learning of response-effect associations, as it captures not only anticipated action effects, but also unpredictable occurrences in the environment (Experiments 5a and 5b). (PsycINFO Database Record


Running interfacial waves in a two-layer fluid system subject to longitudinal vibrations.
We study the waves at the interface between two thin horizontal layers of immiscible fluids subject to high-frequency horizontal vibrations. Previously, the variational principle for energy functional, which can be adopted for treatment of quasistationary states of free interface in fluid dynamical systems subject to vibrations, revealed the existence of standing periodic waves and solitons in this system. However, this approach does not provide regular means for dealing with evolutionary problems: neither stability problems nor ones associated with propagating waves. In this work, we rigorously derive the evolution equations for long waves in the system, which turn out to be identical to the plus (or good) Boussinesq equation. With these equations one can find all the time-independent-profile solitary waves (standing solitons are a specific case of these propagating waves), which exist below the linear instability threshold; the standing and slow solitons are always unstable while fast solitons are stable. Depending on initial perturbations, unstable solitons either grow in an explosive manner, which means layer rupture in a finite time, or falls apart into stable solitons. The results are derived within the long-wave approximation as the linear stability analysis for the flat-interface state [D.V. Lyubimov and A.A. Cherepanov, Fluid Dynamics 21, 849 (1986)] reveals the instabilities of thin layers to be long wavelength.