Stereotactic Abative Body Radiotherapy (SABR) for Oligometastatic Prostate Cancer: A Prospective Clinical Trial.
Stereotactic ablative body radiotherapy (SABR) is an emerging treatment option for oligometastatic prostate cancer. However, limited prospective evidence is available. To determine the safety and feasibility of single fraction SABR for patients with oligometastatic prostate cancer. Secondary endpoints were local and distant progression-free survival (LPFS and DPFS), toxicity, quality of life (QoL), and prostate-specific antigen response. In a prospective clinical trial, patients were screened with computed tomography, bone scan, and sodium fluoride positron emission tomography scan and had one to three oligometastases. Kaplan-Meier methods were used to determine LPFS and DPFS. Toxicity was graded using Common Terminology Criteria for Adverse Event version 4.0. QoL was assessed using European Organization for Research and Treatment of Cancer QLQ-C30 and QLQ-BM22 at 1, 3,12, and 24 mo. A single fraction of 20-Gy SABR to each lesion. Between 2013 and 2014, 33 consecutive patients received SABR to a total of 50 oligometastases and were followed for 2 yr. The median age was 70 yr. The Gleason score was ≥8 in 15 patients (45%). Twenty patients had bone only, 12 had node only, and one had mixed disease. SABR was feasible and delivered as planned in 97% of cases. There was one grade 3 adverse event (3.0%, vertebral fracture). No patient died. The 1 and 2-yr LPFS was 97% (95% confidence interval [CI]: 91-100) and 93% (95% CI: 84-100), and DPFS was 58% (95% CI: 43-77) and 39% (95% CI: 25-60), respectively. In those not on androgen deprivation therapy (ADT; n=22), the 2-yr freedom from ADT was 48%. There was no significant difference from baseline QoL observed. Limitations include small sample size, limited duration of follow-up, and lack of a control arm. A single SABR session was feasible and associated with low morbidity in this cohort. Over one-third of patients did not progress and were free from ADT at 2-yr. QoL measures were maintained with this treatment strategy. This clinical trial investigated single treatment stereotactic radiotherapy for low volume advanced prostate cancer. The approach was found to be safe with avoidance of hormone therapy in almost half of the participants at 2 yr.


Problematic Alcohol Use and Associated Characteristics Following Bariatric Surgery.
Evidence suggests that a significant minority of individuals who undergo Roux-en-Y gastric bypass (RYGB) experience problematic alcohol and substance use following surgery. However, little research has examined characteristics, drinking patterns, and possible risk factors within this population. To provide descriptive information of a sample of adults with self-identified alcohol use problems following bariatric surgery, this study examined (1) alcohol and substance use symptoms using standardized assessments, (2) current and past psychiatric comorbidity, (3) subjective changes in alcohol sensitivity following surgery, and (4) specific patterns of alcohol use prior to and following bariatric surgery. Adult participants (N = 26) completed a series of structured diagnostic interviews and self-report assessments (e.g., Alcohol Use Disorders Identification Test [AUDIT], Michigan Alcohol Screening Test [MAST], Drug Abuse Screening Test [DAST]) by telephone 1 to 4 years following a RYGB or sleeve gastrectomy. All participants met objective criteria for current problematic alcohol use based on AUDIT and MAST cutoff scores, reported increased subjective sensitivity to alcohol following surgery, and evidenced significant current and past psychiatric comorbidities, most notably previous major depression (45.5%). Approximately one third of participants evidenced new-onset Diagnostic and Statistical Manual of Mental Disorders (DSM-IV) alcohol use or dependence following surgery. Preoperative drinking frequencies and quantities were similar to those reported during the period of the heaviest postoperative alcohol use. Findings have implications for pre- and postoperative prevention and intervention efforts. Additional research is needed to further elucidate risk factors for problematic alcohol use following bariatric surgery.


Can community health workers increase coverage of reproductive health services?
Health services were severely affected during the many years of instability and conflict in Afghanistan. In recent years, substantial increases in the coverage of reproductive health services have been achieved, yet absolute levels of coverage remain very low, especially in rural areas. One strategy for increasing use of reproductive health services is deploying community health workers (CHWs) to promote the use of services within the community and at health facilities. Using a multilevel model employing data from a cross-sectional survey of 8320 households in 29 provinces of Afghanistan conducted in 2006, this study determines whether presence of a CHW in the community leads to an increase in use of modern contraceptives, skilled antenatal care and skilled birth attendance. This study further examines whether the effect varies by the sex of the CHW. Results show that presence of a female CHW in the community is associated with higher use of modern contraception, antenatal care services and skilled birth attendants but presence of a male CHW is not. Community-level random effects were also significant. This study provides evidence that indicates that CHWs can contribute to increased use of reproductive health services and that context and CHW sex are important factors that need to be addressed in programme design.


Efficacy of manual versus free-weight training to improve maximal strength and performance for microgravity conditions.
We tested a simple and compact device designed for manual resistance training in conditions of microgravity (Self-Powered Rope Trainer Duo (SPoRT Duo)) to increase muscle performance. Twenty-four participants (20.8 ± 2.1 years) were randomly assigned to a manual resistance group (n = 12) and a free-weight group (n = 12). Participants performed eight exercises (three sets; 8-12 efforts) either with free weights or the SPoRT Duo twice a week for 6 weeks. Maximal isometric force of trunk flexion, back extension and chest press increased (P at least 0.01, d at least 0.52) both in the manual resistance group (18.4% ± 15.0%; 32.7% ± 22.7%; 15.3% ± 9.7%) and free-weight group (18.0% ± 13.9%; 26.6% ± 28.9%; 13.3% ± 7.6%). The change in maximal isometric force of wide grip row in both groups (d at best 0.38) did not reach statistical significance (P at best 0.08). The squat one-repetition-maximum increased in the manual resistance group (29.8% ± 22.1%) and the free-weight group (32.4% ± 26.6%). Jump height, determined by a jump-and-reach test, increased in the free-weight group (9.8% ± 13.2%) but not in the manual resistance group (2.0% ± 8.5%). Manual resistance training was equally effective in increasing strength as traditional resistance training with free weights. This apparatus is a useful addition to current in-flight exercise systems.


[The preparation for elective interventions--the spectrum and dissemination of somatic and psychological measures in Germany].
The positive effects of preoperative preparation on postoperative recovery in patients undergoing elective surgery have been demonstrated. On that account, we surveyed the practice procedures of preoperative preparation for elective surgery patients at German hospitals. During November 1994, we sent 1500 questionnaires to the directors of anaesthesiology departments in Germany. A total of 590 questionnaires (39.3%) were completed and returned. The participating hospitals range in size from 20 to 2600 beds (mean = 364; s = 334,97), totalling together more than 1.7 million surgical operations per year. The surveyed hospitals used one or more of the following procedures for pre-operative preparation: 573 (98.6%) of the replying hospitals used medical anxiolysis, 415 (71.3%) applied preoperative respiratory therapy. Furthermore, 222 (38.5%) of the studied hospitals trained their patients in postoperative relevant behaviour (respiratory therapy n = 167; physiotherapy n = 63 and patient-controlled analgesia n = 41). 74 (13%) offered psychological counselling, 29 (5%) made use of other psychological techniques (muscle relaxation; autogenic training, biofeedback) and 26 (4%) used other preparatory methods like video tapes (n = 13), music (n = 5), acupuncture (n = 4). Nearly all hospitals prepared their patients for surgery with a pre-op visit and anxiolytic medications. Further preparatory methods in most surveyed hospitals are only used on a case-by-case basis. At present psychological methods of preoperative preparation are not routinely used in clinical practice in Germany. Existing possibilities for optimising preoperative preparation in patients undergoing elective surgery are not used regularly. Preoperative preparation needs to be improved, especially in patients undergoing major surgery. Standardisation of management procedures and integration of several professional groups and regular application of known procedures for preoperative preparation may lead to cost-saving optimisation of the duration of hospital stay.


Problems with and a system to eliminate single-primer PCR product contamination in simple sequence repeat molecular marker-assisted selection in soybean.
Polymerase chain reaction (PCR) provides a foundation for simple sequence repeat molecular marker-assisted selection (SSR MAS) in soybean. This PCR system and its various conditions have been optimized by many researchers. However, current research on the optimization of the PCR system focuses on double-primer PCR products. We compared single- and double-SSR primer PCR products from 50 soybean samples and found that the use of single-PCR primers in the reaction system can lead to amplified fragments of portions of the SSR primers in the PCR process, resulting in both false-positives and fragment impurity of double-primer PCR amplification, inconvenient for subsequent analysis. We used "single-primer PCR correction" to eliminate interference caused by single-primer nonspecific PCR amplification and improve PCR quality. Using this method, the precision and success rates of SSR MAS in soybean can be increased.


Assessing Cochlear Implant Outcomes in Older Adults Using HERMES: A National Web-based Database.
Cochlear implant (CI) outcomes research has been largely limited to retrospective or single-institution studies in the United States. The objective is to demonstrate the feasibility of using a novel, national, web-based CI database through evaluating CI outcomes in older adults. Analysis of a prospective, national, web-based database designed for CI outcome tracking (HERMES; HIPAA-secure, Encrypted, Research Management and Evaluation Solution). Multi-centered at 18 private practice and academic US medical centers. Older (age ≥75, n = 47) or younger (age <75, n = 103) adult CI patients (n = 150 total, n = 160 ears). Arizona Biomedical (AzBio), CI usage, postoperative complications. Older adults had slightly lower performance on most recent AzBio (56.0%, n = 26, 12.2 ± 6.5 mo postoperatively) compared with younger adults (74.0%, n = 52, 12.6 ± 6.5 mo postoperatively; p < 0.01, Mann-Whitney). However, on multiple regression, age was not a significant predictor of AzBio scores after controlling for sex, hearing loss duration, use, and postoperative follow-up duration. Most recent CI use was similar but significantly different (11.4 h/d in older, n = 24 versus 13.0 in younger, n = 45; p = 0.01). Usage also did not decline over time (p = 0.81 in older versus p = 0.46, in younger). The most common complications were similar (vertigo 25 versus 20%, p = 0.45; tinnitus 4 versus 4%, p = 0.93) in older and younger adults, respectively. We demonstrate the feasibility of a novel user-friendly, web-based, national CI database to analyze CI outcomes. Older age was not a significant predictor of AzBio scores after adjusting for multiple factors. Additionally, CI use did not decline over time.


cAMP-dependent phosphorylation of betaig-h3 protein in human corneal endothelial cells.
To determine the identity of a major membrane/ extracellular matrix (ECM)-associated 66-kDa protein in human corneal endothelial cells and its phosphorylation in vivo. A membrane/ECM-associated 66 kDa protein was purified from human corneal endothelial cells of 50-80 year-old donors by a three-step procedure that included preparative SDS-PAGE, preparative isoelectric focusing (IEF) and HPLC using a C-18 column. The phosphorylation of the 66-kDa protein was determined by both endogenous kinases and exogenous protein kinase A in the presence of endogenous or added cAMP, or added Walsh inhibitor (a specific inhibitor of protein kinase A). The phosphorylated proteins were analyzed by SDS-PAGE followed by autoradiography. The phosphoamino acids were identified following hydrolysis of the purified phosphorylated 66-kDa protein and thin layer chromatography with standard phosphoamino acids. Following purification, the 66-kDa protein showed a single band on SDS-PAGE, a single species on two dimensional (2D)-gel electrophoresis and a single peak during C-18 HPLC. The partial N-terminal sequence of the 66-kDa protein matched with that of the 68-kDa beta ig-h3 protein (minus the signal peptide) of lung adenocarcinoma cells. Furthermore, on Western blot analysis, the 66-kDa protein immunoreacted with anti-beta ig-h3 past signal peptide (residue nos.24-32) antibody but not with the anti-beta ig-3 signal peptide (residue nos.1-9) antibody. During the phosphorylation of endothelial proteins by endogenous kinases or added protein kinase A in the presence of endogenous or added cAMP, the 66 kDa protein was the major substrate with its Ser residues phosphorylated in both cytosolic- and membrane/ECM-fractions. The human corneal endothelial 66-kDa protein is identical to the 68-kDa beta ig-h3 protein (minus signal peptide) from lung adenocarcinoma cells. The corneal protein exists in a phosphorylated state in vivo with its Ser residues phosphorylated by a cAMP-dependent protein kinase A.


Application of the tukey trend test procedure to assess developmental and reproductive toxicity. I. Measurement data.
Developmental and reproductive (DAR) toxicity studies typically include a series of increasing doses of a compound and a zero dose control. Given this framework, Tukey et al. (Biometrics, 41, 295-301, 1985) proposed a procedure (referred to as either the Tukey trend or TCH test procedure) for detecting a nonzero trend in response to increasing doses of the test compound. The procedure considers three candidate dosage scalings to ensure high power against relatively common dose-response patterns and appreciable power against most reasonable patterns. For toxicologic effects with near monotonic dose-response patterns, simulation studies have shown the TCH test to be overall more powerful than pairwise comparison procedures. The TCH test can be applied sequentially, eliminating the highest dose each time a statistically significant trend is observed, until a no-statistical-significance-of-trend dose is reached. This is the highest dose through which there is no statistically trustworthy evidence of the compound's impact on the response. Since DAR toxicity usually exhibits a progressive (monotonic) dose-response, we advocate routine use of Tukey's trend test for the evaluation of treatment effects in these studies. In this article, we discuss the procedure in detail and apply it to fetal body weight, a continuous measurement variable, from a developmental toxicity study.


The predictive value of C-reactive protein in end-stage renal disease: is it clinically significant?
Cardiovascular disease is the leading cause of death in patients with end-stage renal disease. Besides traditional risk factors, disturbances in mineral and bone metabolism and inflammation are thought to be responsible for the increased risk of death. In the last years C-reactive protein (CRP) has gained a lot of attention in the general population, especially with regard to its link with atherosclerosis. Although several studies suggest that CRP may be useful as a parameter in predicting future cardiovascular events in both the general population and in patients with end-stage renal disease, there is doubt about the clinical evidence of this assumption. A statistical association between CRP and cardiovascular disease was observed in various studies, but the predictive power of this association is markedly diminished when adjusted for other risk factors. The relative contributions of CRP as a marker, as a causative agent, or as a consequence of atherosclerotic vascular disease are unclear, both in the general population and in the dialysis population. The CRP levels are highly variable and influenced by intercurrent events in dialysis patients. In dialysis patients, it is possible to reduce the CRP levels by statins, although these agents do not reduce the cardiovascular mortality in diabetic dialysis patients.


Neural networks as a tool for utilizing laboratory information: comparison with linear discriminant analysis and with classification and regression trees.
Successful applications of neural network architecture have been described in various fields of science and technology. We have applied one such technique, error back-propagation, to a medical classification problem stemming from clinical chemistry, and we have compared the performance of two different neural networks with results obtained by conventional linear discriminant analysis or by the technique of classification and regression trees. The results obtained by the various models were tested for robustness by jackknife validation ("leave n out" method). Compared with the two other techniques, neural networks show a unique ability to detect features hidden in the input data which are not explicitly formulated as input. Thus, neural network techniques appear promising in the field of clinical chemistry, and their application, particularly in situations with complex data structures, should be investigated with more emphasis.


A microsphere-based assay for multiplexed single nucleotide polymorphism analysis using single base chain extension.
A rapid, high throughput readout for single-nucleotide polymorphism (SNP) analysis was developed employing single base chain extension and cytometric analysis of an array of fluorescent microspheres. An array of fluorescent microspheres was coupled with uniquely identifying sequences, termed complementary ZipCodes (cZipCodes), which allowed for multiplexing possibilities. For a given assay, querying a polymorphic base involved extending an oligonucleotide containing both a ZipCode and a SNP-specific sequence with a DNA polymerase and a pair of fluoresceinated dideoxynucleotides. To capture the reaction products for analysis, the ZipCode portion of the oligonucleotide was hybridized with its cZipCodes on the microsphere. Flow cytometry was used for microsphere decoding and SNP typing by detecting the fluorescein label captured on the microspheres. In addition to multiplexing capability, the ZipCode system allows multiple sets of SNPs to be analyzed by a limited set of cZipCode-attached microspheres. A standard set of non-cross reactive ZipCodes was established experimentally and the accuracy of the system was validated by comparison with genotypes determined by other technologies. From a total of 58 SNPs, 55 SNPs were successfully analyzed in the first pass using this assay format and all 181 genotypes across the 55 SNPs were correct. These data demonstrate that the microsphere-based single base chain extension (SBCE) method is a sensitive and reliable assay. It can be readily adapted to an automated, high-throughput genotyping system. [Primer sequences used in this study are available as online supplementary materials at www.genome.org.]


Postconcussion syndrome (PCS) in the emergency department: predicting and pre-empting persistent symptoms following a mild traumatic brain injury.
Head injuries across all age groups represent an extremely common emergency department (ED) presentation. The main focus of initial assessment and management rightly concentrates on the need to exclude significant pathology, that may or may not require neurosurgical intervention. Relatively little focus, however, is given to the potential for development of post-concussion syndrome (PCS), a constellation of symptoms of varying severity, which may bear little correlation to the nature or magnitude of the precipitating insult. This review aims to clarify the aetiology and terminology surrounding PCS and to examine the mechanisms for diagnosing and treating.


[Chromatic Swept-Source Laser Scanning - Concept for a Cell-Resolving Confocal Laser Slit Lamp?]
The in vivo characterisation of corneal epithelial tissue morphology is of considerable importance for diagnosis, disease prognosis, and the development of a treatment strategy for ocular surface diseases. In contrast to many alternative methods, in vivo corneal confocal microscopy (CCM) not only provides a macroscopic description of the corneal tissue but also allows its visualisation with cellular resolution. However, the translation of CCM from research to clinical practice is significantly limited by the complex and still largely manual operation of available CCM systems. In addition, for cross-sectional images, and analogously to conventional slit lamp microscopy, volume data must be acquired in time-consuming depth scans due to the frontal orientation of the image field in CCM, from which depth slices can subsequently be calculated. The pure acquisition time is already in the range of seconds, and additionally, motion artefacts have to be corrected in a sophisticated way. This paper presents the concept and optics simulation of a new imaging technique based on a swept-source laser in combination with special chromatic optics. Here, the laser periodically changes its wavelength and is focused at different depths due to the wavelength-dependent aberration of the chromatic optics. The optics simulation results promise good optical resolution at a total imaging depth of 145 µm. The long-term goal is cell-resolving in vivo corneal confocal microscopy in real time with differently oriented sectioning directions.


Phosphatidylethanolamine distribution and fluidity in outer and inner membranes of the gram-negative bacterium Erwinia carotovora.
1. The distribution of phosphatidylethanolamine, the major lipid of Erwinia carotovora, was investigated in intact bacteria, spheroplasts and outer- and inner-membrane preparations, with the amino-group reagent 2,4,6-trinitrobenzenesulphonic acid. Only 4% was found on the external surface of the outer membrane with 30% on the internal surface, whereas the inner membrane had 27 and 38% on its external and internal surfaces respectively. Some comparative studies were made with three other bacteria. 2. The fluidity of the membranes of E. carotovora was studied by using the fluorescent probe 1,6-diphenylhexa-1,3,5-triene. Results were consistent with the hydrocarbon region of the outer membrane bilayer being less fluid than that of the inner one. 3. On the basis of these and other results a model for the outer- and inner-membrane structures of E. carotovora is proposed.


Development and Standardization of Extended ChaeLee Korean Facial Expressions of Emotions.
In recent years there has been an enormous increase of neuroscience research using the facial expressions of emotion. This has led to a need for ethnically specific facial expressions data, due to differences of facial emotion processing among different ethnicities. FIFTY PROFESSIONAL ACTORS WERE ASKED TO POSE WITH EACH OF THE FOLLOWING FACIAL EXPRESSIONS IN TURN: happiness, sadness, fear, anger, disgust, surprise, and neutral. A total of 283 facial pictures of 40 actors were selected to be included in the validation study. Facial expression emotion identification was performed in a validation study by 104 healthy raters who provided emotion labeling, valence ratings, and arousal ratings. A total of 259 images of 37 actors were selected for inclusion in the Extended ChaeLee Korean Facial Expressions of Emotions tool, based on the analysis of results. In these images, the actors' mean age was 38±11.1 years (range 26-60 years), with 16 (43.2%) males and 21 (56.8%) females. The consistency varied by emotion type, showing the highest for happiness (95.5%) and the lowest for fear (49.0%). The mean scores for the valence ratings ranged from 4.0 (happiness) to 1.9 (sadness, anger, and disgust). The mean scores for the arousal ratings ranged from 3.7 (anger and fear) to 2.5 (neutral). We obtained facial expressions from individuals of Korean ethnicity and performed a study to validate them. Our results provide a tool for the affective neurosciences which could be used for the investigation of mechanisms of emotion processing in healthy individuals as well as in patients with various psychiatric disorders.


Effects of prolonged microscopic work on neck and back strain amongst male ENT clinicians and the benefits of a prototype postural support chair.
Musculoskeletal pain is a common occupational hazard experienced by surgeons. Ear, nose and throat (ENT) surgeons are predisposed to neck and back pain due to regular prolonged microscopic work. We conducted a prospective pilot study to investigate the effects of sustained microscopic work on the neck and back, its correlation to surgical experience and to assess the benefits of a prototype postural support chair (PSC) amongst 10 male, ENT clinicians. We used a subjective measure of time to fatigue and pain for the neck and back as well as objective readings from a surface electromyogram (sEMG). We found that an increase in surgical experience correlated with the time taken to experience fatigue and pain in the neck and back. This was corroborated by our sEMG findings. The PSC significantly delayed the sensations in the neck and also eliminated the difference seen amongst the varying seniority of clinicians.


Evaluation of transported dry and wet cervical exfoliated samples for detection of human papillomavirus infection.
We determined the feasibility of human papillomavirus (HPV) detection in cervical exfoliated cells collected as dry swab samples. Both dry cervical swab and specimen transport medium (STM) cervical swab samples were collected from 135 patients attending either colposcopy or women's clinics in Guayaquil, Ecuador, who had a cytology diagnosis within 6 months. HPV was detected by dot blot hybridization and genotyped by the liquid bead microarray assay (LBMA). Overall, 23.1% of dry samples were positive for any high-risk HPV types, and 24.6% of STM samples were positive for any high-risk HPV types. Of 125 paired samples, the type-specific high-risk HPV proportion positive agreement was 60.7% (kappa, 0.69; 95% confidence interval [CI], 0.53 to 0.82). Of six women with cytological evidence of invasive cervical cancer, high-risk HPV DNA was detected in three of their STM samples and in five of their dry samples. Dry samples were more likely to be insufficient for HPV testing than STM samples. Consistent with this observation, the amount of genomic DNA quantitated with the beta-actin gene was almost 20 times lower in dry samples than in STM samples when detected by the real-time TaqMan assay; however, HPV DNA viral loads in dry samples were only 1.6 times lower than those in matched STM samples. We concluded that exfoliated cervical cells could be collected as dry swab samples for HPV detection.


Incidence of advanced chronic renal failure and the need for end stage renal replacement treatment.
To determine the age related incidence of advanced chronic renal failure in two areas of England. Prospective study of patients newly identified as having advanced chronic renal failure within a two year period; subsequent monitoring of patients' clinical course for a further 26 months. Devon and Blackburn. Those patients in a population of 708,997 who developed advanced chronic renal failure (serum creatinine concentration greater than 500 mumol/l) for the first time during a two year period. 210 Patients (148 per million population per year) developed advanced chronic renal failure, 117 (51%) of whom were over 70. The age related incidence rose from 58 per million per year in those aged 20-49 to 588 per million per year in those aged 80 or over. Only 54% (113) of patients were referred to a nephrologist; 120 patients (57%) needed dialysis or died within three months of presenting without receiving dialysis, and 187 (89%) died or needed dialysis within three years. After those unsuitable for further treatment had been excluded, 78 patients per million population per year aged under 80 needed to start long term renal replacement treatment. Many patients suitable for renal replacement treatment are still not referred for nephrological opinion and are denied treatment. If the treatment rate in the United Kingdom rose from the 1988 rate of 55.1 per million per year to 78 per million per year then the number of patients receiving treatment would rise to about 800 per million. This is double the present number and has considerable but predictable resource implications for the NHS.


Vasorelaxation by amentoflavone isolated from Selaginella tamariscina.
In the courses of in vitro screening for the vasorelaxant effect of the various extracts from medicinal plants, an ethyl acetate-soluble extract of Selaginella tamariscina was found to exhibit distinctive vasorelaxant activity. Further purifications of the extract as guided by in vitro vasorelaxant assay afforded an active biflavonoid, amentoflavone. Amentoflavone induced concentration-dependent relaxation of the phenylephrine-precontracted aorta, which disappeared by removal of functional endothelium. Pretreatment of the aortic tissues with N(G)-nitro- L-arginine methyl ester (L-NAME), methylene blue, or 1 H- -oxadiazolo[4,3- a]quinoxalin-1-one (ODQ) inhibited the relaxation induced by amentoflavone. Amentoflavone-induced relaxations were also markedly attenuated by addition of tetraethylammonium (TEA) or verapamil. However, the relaxant effect of amentoflavone was not blocked by pretreatment with indomethacin, glibenclamide, atropine, or propranolol. Incubation of endothelium-intact aortic rings with amentoflavone increased the production of cGMP, but this effect was blocked by endothelium-denudation or pretreatment with L-NAME or ODQ. These results suggest that amentoflavone relaxes vascular smooth muscle via endothelium-dependent nitric oxide-cGMP signaling, with possible involvement of non-specific K (+) and Ca (2+) channels. Abbreviations. EDRF:endothelium-derived relaxing factor EDHF:endothelium-derived hyperpolarizing factor NO:nitric oxide cGMP:guanosine 3',5'-cyclic monophosphate DMSO:dimethyl sulfoxide L-NAME: N(G)-nitro- L-arginine methyl ester ODQ:1 H-[1,2,4]-oxadiazole-[4,3- a]-quinoxalin-1-one IBMX:3-isobutyl-1-methylxanthine K (Ca):Ca (2+)-dependent K (+) channel K (ATP):adenosine triphosphate (ATP)-sensitive K (+) channel TEA:tetraethylammonium


Comparison of the inter- and intra-observer repeatability of three gait-scoring scales for sows.
Most gait-scoring scales for pigs have a limited number of categories, supposedly to improve repeatability. However, reducing the number of categories could lead to loss of information if the observers' discriminative capacities are underused. With a recently estimated within-herd prevalence of sow lameness of 8.8% to 16.9% in the European Union and the associated losses, the availability of reliable tools for the timely detection of initial cases warrants attention. This study investigated the intra- and inter-observer repeatability (intra-OR and inter-OR) of three gait-scoring scales for sows: a continuous 'tagged' visual analogue scale (tVAS, measured in mm), a 5-point and a 2-point ordinal scale (5P and 2P), all with the same descriptors. Veterinary medicine students (n=108) were trained to use the scales and then asked to score 90 videos (30 per scale) of sows with normal and abnormal gait. Thirty-six videos were shown once and 18 were randomly shown three times, of which one mirrored horizontally. The students' opinions on the scales were also collected. Intra- and inter-OR were higher with the tVAS than the 2P scale (inter-OR: 0.73 v. 0.60; P<0.05. Intra-OR: 0.80 v. 0.67; P<0.05). Intra-OR was higher with the 5P (0.81) than the 2P scale (0.67; P<0.05). For all three scales, repeatabilities were lower (P<0.05) for non-lame sows (gait score of ⩽45 mm on the tVAS) than for sows showing some signs of lameness (gait score>45 mm). Video order (first 45 v. last 45 clips), mirroring, users' opinions on the scales, and previous declared experience in handling pigs or scoring lameness in other species had no effect on repeatabilities. Correlations between the students' and experts' scores were high (tVAS=0.92; 5P=0.91; 2P=0.88) but the association for the 2P was not linear and the frequency distribution showed lower correlations for a group of students. This study confirms recent evidence that it is possible to design high-resolution gait-scoring scales that do not reduce observer repeatability. Visual gait-scoring scales with fewer than five categories are likely to entail loss of information on lameness in individual sows.


Multimers of anionic amphiphiles mimic calmodulin stimulation of cyclic nucleotide phosphodiesterase.
Oleic acid, phosphatidylserine and pyrenedecanoic acid were found to activate calmodulin-deficient cyclic nucleotide phosphodiesterase at concentrations above their critical micellar concentration. In contrast with calmodulin these activators do not require the presence of Ca2+ for their action. It is shown that the size of phosphatidylserine vesicles is of crucial importance with respect to the activating potency of phosphatidylserine. Fluorescence measurements with the probe pyrenedecanoic acid revealed that micelles rather than monomers are the active species for stimulation of phosphodiesterase. There are indications that this result also may be applied to the other activators.


The crucial role of dendritic cells in rhinitis.
Dendritic cells generally play an important role as sentinels in the immune system. They are also very important in protecting the airways from invading pathogens and harmful particles and antigens. This review discusses current knowledge about dendritic cell function and the interaction between dendritic cells and their surroundings, the epithelium, during their presence in the nasal mucosa. There are some phenotypical differences between myeloid dendritic cells and plasmacytoid dendritic cells in different types of rhinitis. Since it has become possible to perform functional studies on purified dendritic cell populations obtained from the upper airway mucosa, a number of studies have appeared. Some confirm that thymic stromal lymphopoietin is present in the nasal mucosa and that it may influence dendritic cell-T-cell interaction in a pro-Th2 way. Epithelial cells share several characteristics with dendritic cells, but they cannot migrate and move antigens to draining lymph nodes. Several functional dendritic cell studies have been published indicating that there are disease-dependent dendritic cell differences in rhinitis. In addition to these differences, factors like epithelial cells and T cells influence dendritic cells. Several new therapeutic options are available targeting direct or indirect dendritic cell functions.


Tiered exposure-assessment strategy in the Semiconductor Health Study.
The Semiconductor Health Study included 14 U.S. companies with 82 silicon-based wafer-fabrication rooms and a complex array of processes and chemicals. Its epidemiologic components were a historical examination of spontaneous abortion rates among 902 women, a prospective evaluation of reproductive outcomes in 483 women, and a cross-sectional review of male fertility and of respiratory, ergonomic, neurologic, or gastrointestinal problems among 3,175 men and women. Designing an exposure assessment strategy presented unique problems, and multiple agents had to be evaluated. A three-tiered approach to exposure assessment was developed to reflect increasing specificity of exposures. At the first tier, employees were divided into fabrication and nonfabrication groups. At the second tier, work groups with qualitatively different exposures were determined. At the third tier, intensity of exposures to specific chemical, physical, and ergonomic agents was evaluated. Evaluations were based on worker reports of tasks performed, moderated by fabrication-specific factors observed by study industrial hygienists during site visits.


Spine growth precedes synapse formation in the adult neocortex in vivo.
Dendritic spines appear and disappear in an experience-dependent manner. Although some new spines have been shown to contain synapses, little is known about the relationship between spine addition and synapse formation, the relative time course of these events, or whether they are coupled to de novo growth of axonal boutons. We imaged dendrites in barrel cortex of adult mice over 1 month, tracking gains and losses of spines. Using serial section electron microscopy, we analyzed the ultrastructure of spines and associated boutons. Spines reconstructed shortly after they appeared often lacked synapses, whereas spines that persisted for 4 d or more always had synapses. New spines had a large surface-to-volume ratio and preferentially contacted boutons with other synapses. In some instances, two new spines contacted the same axon. Our data show that spine growth precedes synapse formation and that new synapses form preferentially onto existing boutons.


Combination spray washes of saponin with water or acetic acid to reduce aerobic and pathogenic bacteria on lean beef surfaces.
Saponins are naturally occurring compounds known as triterpenoid glycosides found in a variety of plant species. Saponins are approved for use in the food industry as foaming agents. When combined with water or organic acid in spray treatments, saponins' foaming property may improve carcass decontamination. In the first experiment of this study, lean beef carcass surfaces were experimentally inoculated with a fecal slurry containing antibiotic-resistant Escherichia coli O157:H7 and Salmonella Typhimurium. Spray-washing treatments with 1% saponin followed by a water wash, or 1% saponin followed by 2% acetic acid, were more effective for reducing aerobic bacteria than saponin, water, or 2% acetic acid washes alone. However, 1% saponin followed by a either a water or 2% acetic acid wash was no more effective than a 2% acetic acid wash for reducing populations of E. coli O157:H7 or Salmonella Typhimurium. In the second experiment, experimentally inoculated beef surfaces were subjected to spray treatments with water followed by another water wash, water followed by a 2% acetic acid wash, 1% saponin followed by a water wash, or 1% saponin followed by a 2% acetic acid wash. When examined for effectiveness against all bacterial populations, 1% saponin followed by a water wash and 1% saponin followed by a 2% acetic acid wash were as effective as two water washes or a water wash followed by 2% acetic acid for reducing aerobic bacteria, E. coli O157:H7, and Salmonella Typhimurium from beef surfaces. Under the conditions described, reductions associated with combination spray washes may be attributed to the physical removal of bacteria during the spraying process, not to any specific action of saponin.


[Biochemical markers of atherogenesis in a nephrotic form of glomerulonephritis in children].
A study was made of the regularities and changes in the fatty acid spectrum of total lipids of the blood serum, in interaction of carbohydrate lipid metabolism, and in the hypophyseal and thyroid system in 92 patients with the nephrotic form of acute and chronic glomerulonephritis. It has been discovered that the nephrotic form of glomerulonephritis is marked by a complex of changes in the fatty acid spectrum of the blood serum, in carbohydrate lipid metabolism at the sites of their conjunction, and in the hypophyseal and thyroid system which can be regarded as metabolic potentialities of atherosclerosis formation.


Hashimotos Thyroiditis with Coexistent Papillary Carcinoma and Non-hodgkin Lymphoma-thyroid.
Lymphocytic thyroiditis and hashimoto's thyroiditis (HT) are the two main forms of autoimmune thyroiditis among which the latter is most frequent. A vast majority of cases of papillary carcinoma and primary thyroid lymphoma (PTL) arise in the setting of HT. A case of 32-year-old female who presented with thyroid enlargement, post-thyroidectomy showed hashimoto's thyroiditis (HT) with coexistent papillary carcinoma and non-hodgkin lymphoma (NHL). The immunohistochemistry was positive for CD 20, CD 45, bcl 2 for lymphoma, low and high molecular cytokeratin for papillary carcinoma. The staging studies showed no evidence of metastasis. It is thus concluded that papillary carcinoma and NHL can coexist with HT. The thyroid lymphoma or papillary carcinoma has to be thought off, whenever patients presents with sudden enlargement in a known case of HT. A patient presenting with concomitant primary thyroid lymphoma and papillary thyroid carcinoma must be judiciously evaluated, since, the treatment has to prioritize the tumor with worst stage at the time of diagnosis.


Simulations of stable pores in membranes: system size dependence and line tension.
Amphiphilic bilayers with a pore were simulated using a coarse grained model. By stretching the bilayer to 70% beyond its equilibrium surface area, we established the phase diagram of pores, identifying regions where pores are stable, metastable, or unstable. A simple theoretical model is proposed to explain the phase diagram, and to calculate the critical and equilibrium relative stretches. Interestingly, these are found to scale with the inverse cubic root of the number of amphiphiles in the bilayer, thus explaining the order of magnitude difference between the simulated and the measured values. Three different methods are used to calculate a line tension coefficient of (3.5-4.0) x 10(-11) J/m, in good agreement with experimental data.


Diversity and community structure within anoxic sediment from marine salinity meromictic lakes and a coastal meromictic marine basin, Vestfold Hilds, Eastern Antarctica.
16S rDNA clone library analysis was used to examine the biodiversity and community structure within anoxic sediments of several marine-type salinity meromictic lakes and a coastal marine basin located in the Vestfolds Hills area of Eastern Antarctica. From 69 to 130 (555 total) 16S rDNA clones were analysed from each sediment sample, and restriction fragment length polymorphism (RFLP) and sequence analysis grouped the clones into 202 distinct phylotypes (a clone group with sequence similarity of >0.98). A number of phylotypes and phylotype groups predominated in all libraries, with a group of 10 phylotypes (31% of clones) forming a novel deep branch within the low G+C Gram-positive division. Other abundant phylotypes detected in several different clone libraries grouped with Prochlorococcus cyanobacteria, diatom chloroplasts, delta proteobacteria (Desulfosarcina group, Syntrophus and Geobacterl Pelobacter/Desulphuromonas group), order Chlamydiales (Parachlamydiaceae) and Spirochaetales (wall-less Antarctic spirochaetes). Most archaeal clones detected (3.1% of clones) belonged to a highly diverged group of Euryarchaeota clustering with clones previously detected in rice soil, aquifer sediments and hydrothermal vent material. Little similarity existed between the phylotypes detected in this study and other clone libraries based on marine sediment, suggesting that an enormous prokaryotic diversity occurs within marine and marine-derived sediments.


Direct monitoring of pulmonary disease treatment biomarkers using plasmonic gold nanorods with diffusion-sensitive OCT.
The solid concentration of pulmonary mucus (wt%) is critical to respiratory health. In patients with respiratory disease, such as Cystic Fibrosis (CF) and Chronic Obstructive Pulmonary Disorder (COPD), mucus hydration is impaired, resulting in high wt%. Mucus with high wt% is a hallmark of pulmonary disease that leads to obstructed airways, inflammation, and infection. Methods to measure mucus hydration in situ and in real-time are needed for drug development and personalized therapy. We employed plasmonic gold nanorod (GNR) biosensors that intermittently collide with macromolecules comprising the mucus mesh as they self-diffuse, such that GNR translational diffusion (DT) is sensitive to wt%. GNRs are attractive candidates for bioprobes due to their anisotropic optical scattering that makes them easily distinguishable from native tissue using polarization-sensitive OCT. Using principles of heterodyne dynamic light scattering, we developed diffusion-sensitive optical coherence tomography (DS-OCT) to spatially-resolve changing DT in real-time. DS-OCT enables, for the first time, direct monitoring of changes in nanoparticle diffusion rates that are sensitive to nanoporosity with spatial and temporal resolutions of 4.7 μm and 0.2 s. DS-OCT therefore enables us to measure spatially-resolved changes in mucus wt% over time. In this study, we demonstrate the applicability of DS-OCT on well-differentiated primary human bronchial epithelial cells during a clinical mucus-hydrating therapy, hypertonic saline treatment (HST), to reveal, for the first time, mucus mixing, cellular secretions, and mucus hydration on the micrometer scale that translate to long-term therapeutic effects.


Urinary free cortisol excretion in depression.
Urinary free cortisol (UFC) excretion was determined in 60 depressed inpatients and in 35 psychiatric inpatients with other disorders. The depressed patients had high daily UFC values, while the other patients excreted normal amounts. Over 40% of the depressed patients had UFC excretions in the range seen in Cushing's disease, while only 6% of the other patients excreted such high amounts of cortisol. Age and sex differences did not account for the results. Among the depressed patients those with depressive neuroses excreted less than unipolar or bipolar depressives. Following treatment, more normal UFC excretion was found in depressed patients. The estimation of UFC and its clinical utility are discussed in detail. UFC determination is a simple and informative indicator of adrenal cortical activation and its application to psychoendocrine studies is recommended.


Suicide in Bangla Movie and Drama: A Content Analysis.
We aimed to assess how suicidality has been depicted in Bangla movies and dramas. We conducted a search on YouTube by using search terms to identify movies and dramas with suicidal scripts. The search was performed between February and May 2022 resulting in 71 items consisting of 35 Bangla movies and 36 Bangla dramas. We scrutinized the contents of movies and dramas against our pre-designed instrument and we assessed their quality against World Health Organization guidelines. Among the 71 suicidal behaviors, 46.5% were suicides, 72% of the suicidal behavior was noted in young adults, 63.9% were unmarried, and 69% attempts were found in prominent characters. Hanging was found as the most prominent method (25.4%) and premarital and extramarital affairs and sexual harassment were the most prominent risk factors (60.6%). The potentially harmful characteristics were present in almost all events whereas potentially helpful contents were mentioned very minimally.


An integrated workflow for quantitative analysis of the newly synthesized proteome.
The analysis of proteins that are newly synthesized upon a cellular perturbation can provide detailed insight into the proteomic response that is elicited by specific cues. This can be investigated by pulse-labeling of cells with clickable and stable-isotope-coded amino acids for the enrichment and mass spectrometric characterization of newly synthesized proteins (NSPs), however convoluted protocols prohibit their routine application. Here we report the optimization of multiple steps in sample preparation, mass spectrometry and data analysis, and we integrate them into a semi-automated workflow for the quantitative analysis of the newly synthesized proteome (QuaNPA). Reduced input requirements and data-independent acquisition (DIA) enable the analysis of triple-SILAC-labeled NSP samples, with enhanced throughput while featuring high quantitative accuracy. We apply QuaNPA to investigate the time-resolved cellular response to interferon-gamma (IFNg), observing rapid induction of targets 2 h after IFNg treatment. QuaNPA provides a powerful approach for large-scale investigation of NSPs to gain insight into complex cellular processes.


Cloning and analysis of the promotor region of the human fibronectin gene.
Human fibronectin (FN) genomic clones were isolated by screening a human genomic library with a 75-base oligonucleotide. The sequence of the oligonucleotide corresponds to a region near the 5' end of the human FN cDNA clone pFH6 that contains the amino-terminal coding sequences but does not extend to the 5' end of the mRNA [Kornblihtt, A. R., Umezawa, K., Vibe-Pedersen, K. & Baralle, F. E. (1985) EMBO J. 4, 1755-1759]. The 5' end of the FN gene is found on a 3.7-kilobase-pair EcoRI fragment that contains about 2.7 kilobase pairs of flanking sequence. The first exon is 414 base pairs long, with a 5' untranslated region of 267 base pairs. As deduced on the basis of the position of the initiation codon, FN is synthesized with a 31-residue amino acid extension on the amino terminus that is not present in the mature polypeptide. This amino-terminal extension appears to contain both a signal peptide and a propeptide. The first 200 base pairs of 5'-flanking sequence is very G + C rich. Upstream of this the sequence becomes relatively A + T rich. The sequence ATATAA is found at -25 and the sequence CAAT is present at -150. The sequence GGGGCGGGGC at -102 exhibits homology to the binding site for the transcription factor SP1, and the sequence TGACGTCA at -173 exhibits homology to 5'-flanking sequences important for induction by cAMP.


Hospital Systems, Convenient Care Strategies, and Healthcare Reform.
Retail clinics (RCs) and urgent care centers (UCCs) are convenient care models that emerged on the healthcare scene in the past 10 to 15 years. Characterized as disruptive innovations, these models of healthcare delivery seem to follow a slightly different path from each other. Hospital systems, the very organizations that were originally threatened by convenient care models, are developing them and partnering with existing models. We posit that legislative changes such as the Affordable Care Act created challenges for hospital systems that accelerated their adoption of these models. In this study, we analyze 117 hospital systems in six states and report on their convenient care strategies. Our data suggest that UCCs are more prevalent than RCs among hospital systems, and that large and unexplained state-by-state variations exist in the adoption of these strategies. We also postulate about the future role of hospital systems in leading these innovations.


A novel fusion protein consisting of anti-ANGPTL3 antibody and interleukin-22 ameliorates diabetic nephropathy in mice.
The pathogenic mechanisms of diabetic nephropathy (DN) include podocyte injury, inflammatory responses and metabolic disorders. Although the antagonism of Angiopoietin-like protein 3 (ANGPTL3) can alleviate proteinuria symptoms by inhibiting the activation of integrin αvβ3 on the surface of podocytes, it can not impede other pathological processes, such as inflammatory responses and metabolic dysfunction of glucolipid. Interleukin-22 (IL-22) is considered to be a pivotal molecule involved in suppressing inflammatory responses, initiating regenerative repair, and regulating glucolipid metabolism. Genes encoding the mIL22IgG2aFc and two chains of anti-ANGPTL3 antibody and bifunctional protein were synthesized. Then, the DN mice were treated with intraperitoneal injection of normal saline, anti-ANGPTL3 (20 mg/kg), mIL22Fc (12 mg/kg) or anti-ANGPTL3 /IL22 (25.3 mg/kg) and irrigation of positive drug losartan (20mg/kg/d) twice a week for 8 weeks. In this research, a novel bifunctional fusion protein (anti-ANGPTL3/IL22) formed by the fusion of IL-22 with the C-terminus of anti-ANGPTL3 antibody exhibited favorable stability and maintained the biological activity of anti-ANGPTL3 and IL-22, respectively. The fusion protein showed a more pronounced attenuation of proteinuria and improved dysfunction of glucolipid metabolism compared with mIL22Fc or anti-ANGPTL3. Our results also indicated that anti-ANGPTL3/IL22 intervention significantly alleviated renal fibrosis via inhibiting the expression of the inflammatory response-related protein nuclear factor kappa light-chain enhancer of activated B cells (NF-κB) p65 and NOD-like receptor family pyrin domain-containing protein 3 (NLRP3) inflammasome. Moreover, transcriptome analysis revealed the downregulation of signaling pathways associated with injury and dysfunction of the renal parenchymal cell indicating the possible protective mechanisms of anti-ANGPTL3/IL22 in DN. Collectively, anti-ANGPTL3/IL22 bifunctional fusion protein can be a promising novel therapeutic strategy for DN by reducing podocyte injury, ameliorating inflammatory response, and enhancing renal tissue recovery.


SAR studies in the field of calcium(II) antagonists. Effect of modifications at the tetrasubstituted carbon of verapamil-like compounds.
A number of fluorenyl and diphenylmethane analogues of verapamil, chosen as having the same substituents arranged in different ways around the quaternary carbon, were synthesized in order to evaluate the importance of the stereoisomerism at that point of the molecule. The compounds were tested with the Langendorff technique and coronary perfusion pressure (CPP), left ventricular pressure (LVP), and heart rate (HR) were recorded. While most of the compounds were almost inactive on these parameters, three of them did show interesting cardiovascular action. In particular they produced a more pronounced decrease in CPP than verapamil, with a less marked negative inotropic effect. Structure-activity relationships and the mechanism of action of the compounds are discussed.


Rational Design of Asymmetric Polymethines to Attain NIR(II) Bioimaging at >1100 nm.
Organic molecules having emission in the NIR(II) region are emergent and receiving enormous attention. Unfortunately, attaining accountable organic emission intensity around the NIR(II) region is hampered by the dominant internal conversion operated by the energy gap law, where the emission energy gap and the associated internal reorganization energy λint play key roles. Up to the current stage, the majority of the reported organic NIR(II) emitters belong to those polymethines terminated by two symmetric chromophores. Such a design has proved to have a small λint that greatly suppresses the internal conversion. However, the imposition of symmetric chromophores is stringent, limiting further development of organic NIR(II) dyes in diversity and versatility. Here, we propose a new concept where as far as the emissive state of the any asymmetric polymethines contains more or less equally transition density between two terminated chromophores, λint can be as small as that of the symmetric polymethines. To prove the concept, we synthesize a series of new polymethines terminated by xanthen-9-yl-benzoic acid and 2,4-diphenylthiopyrylium derivatives, yielding AJBF1112 and AEBF1119 that reveal emission peak wavelength at 1112 and 1119 nm, respectively. The quantum yield is higher than all synthesized symmetric polymethines of 2,4-diphenylthiopyrylium derivatives (SC1162, 1182, 1185, and 1230) in this study. λint were calculated to be as small as 6.2 and 7.3 kcal/mol for AJBF1112 and AEBF1119, respectively, proving the concept. AEBF1119 was further prepared as a polymer dot to demonstrate its in vitro specific cellular imaging and in vivo tumor/bone targeting in the NIR(II) region.


Sialic Acid-Modified O-GlcNAc Transferase Inhibitor Liposome Presents Antitumor Effect in Hepatocellular Carcinoma.
O-linked-N-acetylglucosaminylation (O-GlcNAcylation) plays a key role in hepatocellular carcinoma (HCC) development, and the inhibition of O-GlcNAcylation has therapeutic potential. To decrease the systemic adverse events and increase targeting, we used sialic acid (SA)-decorated liposomes loaded with OSMI-1, an inhibitor of the O-GlcNAcylation, to further improve the anti-HCC effect. Fifty pairs of HCC tissue samples and the cancer genome atlas database were used to analyze the expression of O-GlcNAc transferase (OGT) and its effects on prognosis and immune cell infiltration. OSMI-1 cells were treated with SA and liposomes. Western blotting, immunofluorescence, cell proliferation assay, flow cytometry, enzyme-linked immunosorbent assay, immunohistochemistry, and tumorigenicity assays were used to investigate the antitumor effect of SA-modified OSMI-1 liposomes in vitro and in vivo. OGT was highly expressed in HCC tissues, negatively correlated with the degree of tumor infiltration of CD8+ and CD4+T cells and prognosis, and positively correlated with the degree of Treg cell infiltration. SA-modified OSMI-1 liposome (OSMI-1-SAL) was synthesized with stable hydrodynamic size distribution. Both in vitro and in vivo, OSMI-1-SAL exhibited satisfactory biosafety and rapid uptake by HCC cells. Compared to free OSMI-1, OSMI-1-SAL had a stronger capacity for suppressing the proliferation and promoting the apoptosis of HCC cells. Moreover, OSMI-1-SAL effectively inhibited tumor initiation and development in mice. OSMI-1-SAL also promoted the release of damage-associated molecular patterns, including anticalreticulin, high-mobility-group protein B1, and adenosine triphosphate, from HCC cells and further promoted the activation and proliferation of the CD8+ and CD4+T cells. In conclusion, the OSMI-1-SAL synthesized in this study can target HCC cells, inhibit tumor proliferation, induce tumor immunogenic cell death, enhance tumor immunogenicity, and promote antitumor immune responses, which has the potential for clinical application in the future.


Brief Report: Pilot Study of a Novel Interactive Digital Treatment to Improve Cognitive Control in Children with Autism Spectrum Disorder and Co-occurring ADHD Symptoms.
The presence of attention deficit/hyperactivity disorder (ADHD) symptoms in children with autism spectrum disorder (ASD) is associated with worse cognitive control. Children with ASD and ADHD often respond poorly to medications, thus we need alternative treatments. We examined the feasibility, acceptability, and preliminary efficacy of Project Evo-a digital treatment. Nineteen children with ASD and co-occurring ADHD symptoms completed this app-based treatment that targets multi-tasking through gameplay versus a comparison educational treatment. Children had a high engagement with both treatments, and parents and children reported high acceptability. Within-group analyses suggest the multi-tasking but not the educational treatment may improve cognitive control. This multi-tasking treatment is feasible, acceptable, and possibly efficacious for cognitive control impairments in children with ASD and ADHD.


Single-cell adhesion force mapping of a highly sticky bacterium in liquid.
The sticky bacterium Acinetobacter sp. Tol 5 adheres to various material surfaces via its cell surface nanofiber protein, AtaA. This adhesiveness has only been evaluated based on the amount of cells adhering to a surface. In this study, the adhesion force mapping of a single Tol 5 cell in liquid using the quantitative imaging mode of atomic force microscopy (AFM) revealed that the adhesion of Tol 5 was near 2 nN, which was 1-2 orders of magnitude higher than that of other adhesive bacteria. The adhesion force of a cell became stronger with the increase in AtaA molecules present on the cell surface. Many fibers of peritrichate AtaA molecules simultaneously interact with a surface, strongly attaching the cell to the surface. The adhesion force of a Tol 5 cell was drastically reduced in the presence of 1% casamino acids but not in deionized water (DW), although both liquids decrease the adhesiveness of Tol 5 cells, suggesting that DW and casamino acids inhibit the cell approaching step and the subsequent direct interaction step of AtaA with surfaces, respectively. Heterologous production of AtaA provided non-adhesive Acinetobacter baylyi ADP1 cells with a strong adhesion force to AFM tip surfaces of silicon and gold.


IgA-antigliadin antibodies in patients with IgA nephropathy: the secondary phenomenon?
Circulating IgA-antigliadin antibodies (IgA-AGA) are often found in patients with IgA nephropathy (NP). IgA-AGA are sensitive markers of an abnormal immune system reaction to gluten, seen particularly in patients with celiac disease. However, a lack of IgA-antireticulin and IgA-antiendomysium antibodies and often jejunal mucosal atrophy of patients with IgA NP suggest that most patients do not have latent celiac disease. To examine the relationship between IgA-AGA and clinical data, enzyme-linked immunosorbent assays for IgA-AGA were performed in 28 patients with IgA NP and in 50 healthy persons. The results were calculated in arbitrary units (AU). The cutoff level for a negative or a positive test was found to be 60 AU, calculated according to the AGA test result (mean + 3 SD) in 50 healthy persons. The following clinical data were assessed: age, gender, disease duration, daily proteinuria, blood pressure, serum creatinine, and creatinine clearance. Control sera were negative for IgA-AGA. Positive IgA-AGA tests were observed in 14 of the 28 patients (p < 0.0001 vs. controls) and high levels of IgA-AGA (AU >90) in 6 of the 28 patients (p < 0.001 vs. controls). The mean duration of the disease of the patients with positive IgA-AGA was significantly longer as compared with the patients who had a negative antibody test. IgA-AGA correlated with age (p < 0.05, r = 0. 56), disease duration (p < 0.05, r = 0.40), and blood pressure (p < 0.05, r = 0.48). Antireticulin and antiendomysium antibody tests were negative in all patient and control sera. We conclude that IgA-AGA are associated with the progression of IgA NP. Our findings support the current concept about the pathogenesis of IgA NP, where the defective IgA production itself may be the primary and intestinal lesions as well as the production of IgA-AGA the secondary phenomenon.


Regulation of IL-10 gene expression in Th2 cells by Jun proteins.
IL-10 is a key regulatory cytokine produced by T lymphocytes. Although Th2 cells are a major source of IL-10, little is known about IL-10 gene regulation in Th2 cells. High levels of IL-10 mRNA transcription are induced in the Th2 clone D10 after PMA plus ionomycin (P/I) stimulation; however we found that the IL-10 promoter was not inducible by P/I in D10 cells. We therefore sought regulatory regions in the IL-10 gene that could promote P/I-activated transcription in Th2 cells. Two strong DNase I-hypersensitive sites (DHSSs) were identified in the IL-10 gene in mouse T cells, and conserved noncoding sequences (CNSs) between the mouse and human IL-10 genes were also identified. One IL-10 DHSS maps within or next to a highly conserved CNS region, CNS-3. The CNS-3 region contains an AP-1 site that binds JunB and c-Jun proteins specifically in Th2 cells and not in Th1 cells. The CNS-3 element activates transcription from the IL-10 promoter after P/I stimulation and is responsive to c-Jun and JunB. Retroviral mediated-expression of either c-Jun or JunB in primary T cells led to a large increase in IL-10 expression, and inhibition of AP-1 activity by a dominant negative form of c-Jun in primary T cells strongly repressed IL-10 expression. IFN-gamma was relatively unaffected by modulations in AP-1 activity. These data indicate that we have identified a novel regulatory element that can specifically activate transcription of the IL-10 gene in Th2 cells via the AP-1/Jun pathway.


Glomangiosarcoma in the shoulder of a 51-year-old man.
Glomangiosarcoma is a very rare type of soft tissue neoplasm. Here we report such a case in the shoulder of a 51-year-old man.


A review of the effects of baclofen and of THC:CBD oromucosal spray on spasticity-related walking impairment in multiple sclerosis.
Multiple sclerosis (MS) is a complex disease with a heterogeneous and unpredictable clinical course. Mobility impairment after progressive paralyses and muscle tone spasticity is common. Areas covered: The prevalence, assessment, and pharmacological management of gait impairment and spasticity in MS and their effects on health-related quality of life (HRQoL) are discussed. The roles of oral and intrathecal baclofen and of delta-9-tetrahydrocannabinol/cannabidiol (THC:CBD) oromucosal spray in treating MS spasticity-related gait impairment are reviewed. Expert commentary: Mobility impairment and spasticity are experienced by approximately 90% and 80% of MS patients, respectively, during the disease course. Prevalence and severity of gait impairment and spasticity increase as disease progresses. The symptoms are related and both impact negatively on HRQoL. Oral baclofen and tizanidine are generally used for first-line treatment of MS spasticity but are ineffective in approximately 40% of cases. Second-line therapy includes add-on THC:CBD spray for patients with resistant MS spasticity. Results of studies evaluating baclofen for treating MS spasticity gait impairment are equivocal. In studies of patients with resistant MS spasticity, THC:CBD spray consistently improved the timed 10-meter walk test and significantly improved multiple spatial-temporal and kinematic gait parameters. THC:CBD oromucosal spray warrants further investigation as a treatment for MS spasticity-related gait impairment.


Internal dynamics of a supramolecular nanofibre.
A large variety of functional self-assembled supramolecular nanostructures have been reported over recent decades. The experimental approach to these systems initially focused on the design of molecules with specific interactions that lead to discrete geometric structures, and more recently on the kinetics and mechanistic pathways of self-assembly. However, there remains a major gap in our understanding of the internal conformational dynamics of these systems and of the links between their dynamics and function. Molecular dynamics simulations have yielded information on the molecular fluctuations of supramolecular assemblies, yet experimentally it has been difficult to obtain analogous data with subnanometre spatial resolution. Using site-directed spin labelling and electron paramagnetic resonance spectroscopy, we measured the conformational dynamics of a self-assembled nanofibre in water through its 6.7 nm cross-section. Our measurements provide unique insight for the design of supramolecular functional materials.


Evidence that DNA topoisomerase I is necessary for the cytotoxic effects of camptothecin.
The budding yeast Saccharomyces cerevisiae and the fission yeast Schizosaccharomyces pombe are both sensitive to camptothecin, an inhibitor of DNA topoisomerase I. An S. cerevisiae DNA repair mutant, rad52, is hypersensitive to the drug. In both species, topoisomerase I mutants totally lacking the enzyme are completely resistant to the drug. A strain with a mutation leading to a temperature-sensitive topoisomerase I exhibits temperature dependence in its in vivo response to camptothecin. A strain carrying a plasmid that overproduces topoisomerase I is hypersensitive to the drug. The rad52 mutant is killed by overproduction of the enzyme, even in the absence of the drug. The response of several of these strains to camptothecin analogs, to DNA topoisomerase II inhibitors, and to other drugs is reported. The cytotoxic effects of camptothecin are discussed in terms of the drug extending the lifetime of a topoisomerase I-DNA covalent intermediate, which is recognized as DNA damage by a DNA repair system.


HIV serostatus disclosure to sexual partners among HIV-positive methamphetamine-using gay, bisexual, and other men who have sex with men.
Nineteen semistructured interviews with HIV-positive methamphetamine-using gay, bisexual, or other MSM were analyzed qualitatively and using Critical Incident Measure (CIM). Among those who reported successful disclosure, themes regarding disclosure strategies and precontact disclosure emerged. Although few men spontaneously discussed the association between methamphetamine and disclosure, those who did reported less inclination to ascertain partner serostatus while under the influence. Men who reported disclosure to an HIV-seroconcordant partner were more likely to report not exchanging semen in this encounter compared with nondisclosers, whereas condom nonuse emerged as a theme across disclosure groups. Most of those who reported nondisclosure described their most recent encounter under the influence as occurring in the context of a public sex environment and/or with multiple partners. Understanding the interaction between methamphetamine use and serostatus disclosure practices, particularly among HIV-positive men, and how this interaction impacts sexual risk-taking has important implications for HIV prevention work.


Considerations for Membrane Protein Purification.
Isolating membrane proteins from their native cells while maintaining structural and functional integrity is challenging. Many detergents have been developed over the years that interact favorably with membrane proteins and mimic the physical properties of the lipid bilayer. Choosing the appropriate detergent is crucial for the successful extraction of a protein in its properly folded, active conformation.


Reduced E-cadherin and alpha-catenin expressions have no prognostic role in bladder carcinoma.
In various human cancers, dysfunction of the E-cadherin-catenin complex is associated with a decrease in cellular and tissue differentiation, and with higher invasive and metastatic potentials. The objective of this study was to investigate E-cadherin and alpha-catenin expression in superficial noninvasive papillary TCC and invasive TCC, and correlate these results with pathological and clinical parameters. We have used immunohistochemistry to localize Ecadherin and alpha-catenin in 56 formalin-fixed, paraffin-embedded tissue blocks from 41 patients with superficial bladder cancer and 15 with invasive bladder cancer. The 46 male and 10 female patients had a mean age of 67 years, with range of 40 to 82 years. The mean follow-up time was 33.4 (range 5-120) months. Tumor grade 1:2:3 ratios were 5:32:19. In superficial bladder tumor, abnormal expression of E-cadherin and alpha-catenin was demonstrated in 37 and 71% of the tumors, respectively. In advanced bladder tumor, abnormal expression of E-cadherin and alpha-catenin was demonstrated in 80 and 100% of the tumors, respectively. Differences in expression of E-cadherin and alpha-catenin could be discerned between superficial and advanced bladder tumors (p=0.004, p=0.024, respectively). However, the association between E-cadherin and alpha-catenin expression and tumor grade was not statistically significant (p>0.05). In addition, the expression of E-cadherin and alpha-catenin did not correlate with tumor number and size (p>0.05). We have demonstrated that abnormal expression of E-cadherin and/or alpha-catenin occurs in more than 85% of bladder carcinomas and correlates significantly only with advanced stage. Nevertheless, these observations need to be confirmed in larger prospective clinical studies.


Older adults as adaptive decision makers: evidence from the Iowa Gambling Task.
Older adults process emotional information differently than younger adults and may demonstrate less of a negativity bias on cognitive tasks. The Iowa Gambling Task designed by A. Bechara, H. Damasio, D. Tranel, and A. R. Damasio (1997) has been used to examine the integration of emotion and cognition in a risky-choice decision task and may give insight into differences in the decision-making strategies in younger and older adults. Eighty-eight younger adults (18-34 years) and 67 older adults (65-88 years) completed the Iowa Gambling Task. Using a theoretical decomposition of the task designed by J. R. Busemeyer and J. C. Stout (2002), the authors found that both groups were successful at solving the task but used very different strategies that reflected each group's strength. For younger adults, that strength was learning and memory. For older adults, that strength was an accurate representation of wins and losses (valence).


Decay of SARS-CoV-2 RNA along the wastewater treatment outfitted with Upflow Anaerobic Sludge Blanket (UASB) system evaluated through two sample concentration techniques.
For the first time, we present, i) an account of decay in the genetic material loading of SARS-CoV-2 during Upflow Anaerobic Sludge Blanket (UASB) treatment of wastewater, and ii) comparative evaluation of polyethylene glycol (PEG), and ultrafiltration as virus concentration methods from wastewater for the quantification of SARS-CoV-2 genes. The objectives were achieved through tracking of SARS-CoV-2 genetic loadings i.e. ORF1ab, N and S protein genes on 8th and 27th May 2020 along the wastewater treatment plant (106000 m3 million liters per day) equipped with UASB system in Ahmedabad, India. PEG method performed better in removing materials inhibiting RT-qPCR for SARS-CoV-2 gene detection from the samples, as evident from constant and lower CT values of control (MS2). Using the PEG method, we found a reduction >1.3 log10 reduction in SARS-CoV-2 RNA abundance during UASB treatment, and the RNA was not detected at all in the final effluent. The study implies that i) conventional wastewater treatment systems is effective in SARS-CoV-2 RNA removal, and ii) UASB system significantly reduces SARS-CoV-2 genetic loadings. Finally, PEG method is recommended for better sensitivity and inhibition removal during SARS-CoV-2 RNA quantification in wastewater.


Highly Sensitive Detection of Ionizing Radiations by a Photoluminescent Uranyl Organic Framework.
Precise detection of low-dose X- and γ-radiations remains a challenge and is particularly important for studying biological effects under low-dose ionizing radiation, safety control in medical radiation treatment, survey of environmental radiation background, and monitoring cosmic radiations. We report here a photoluminescent uranium organic framework, whose photoluminescence intensity can be accurately correlated with the exposure dose of X- or γ-radiations. This allows for precise and instant detection of ionizing radiations down to the level of 10-4 Gy, representing a significant improvement on the detection limit of approximately two orders of magnitude, compared to other chemical dosimeters reported up to now. The electron paramagnetic resonance analysis suggests that with the exposure to radiations, the carbonyl double bonds break affording oxo-radicals that can be stabilized within the conjugated uranium oxalate-carboxylate sheet. This gives rise to a substantially enhanced equatorial bonding of the uranyl(VI) ions as elucidated by the single-crystal structure of the γ-ray irradiated material, and subsequently leads to a very effective photoluminescence quenching through phonon-assisted relaxation. The quenched sample can be easily recovered by heating, enabling recycled detection for multiple runs.


Stapled gastro/duodenojejunostomy shortens reconstruction time during pylorus-preserving pancreaticoduodenectomy.
To investigate whether a stapled technique is superior to the conventional hand-sewn technique for gastro/duodenojejunostomy during pylorus-preserving pancreaticoduodenectomy (PpPD). In October 2010, we introduced a mechanical anastomotic technique of gastro- or duodenojejunostomy using staplers during PpPD. We compared clinical outcomes between 19 patients who underwent PpPD with a stapled gastro/duodenojejunostomy (stapled anastomosis group) and 19 patients who underwent PpPD with a conventional hand-sewn duodenojejunostomy (hand-sewn anastomosis group). The time required for reconstruction was significantly shorter in the stapled anastomosis group than in the hand-sewn anastomosis group (186.0 ± 29.4 min vs 219.7 ± 50.0 min, P = 0.02). In addition, intraoperative blood loss was significantly less (391.0 ± 212.0 mL vs 647.1 ± 482.1 mL, P = 0.03) and the time to oral intake was significantly shorter (5.4 ± 1.7 d vs 11.3 ± 7.9 d, P = 0.002) in the stapled anastomosis group than in the hand-sewn anastomosis group. There were no differences in the incidences of delayed gastric emptying and other postoperative complications between the groups. These results suggest that stapled gastro/duodenojejunostomy shortens reconstruction time during PpPD without affecting the incidence of delayed gastric emptying.


Altered ultrastructural morphology of self-aggregated low density lipoproteins: coalescence of lipid domains forming droplets and vesicles.
Lipid droplets and vesicles can presumably be formed directly from lipoproteins in the extracellular space in atherosclerosis, but an in vitro demonstration of the phenomenon in the absence of cellular pathways has been lacking. Low density lipoproteins (LDL) are known to undergo self-aggregation after brief vortexing in vitro. In the present study, LDL aggregates were examined by electron microscopy, using new mordant techniques for lipid visualization, and by chemical analysis. Aggregation of LDL by vortexing is regularly accompanied by the formation of comparatively large lipid droplets (up to 600 nm diameter) and vesicles. Aggregates containing droplets and vesicles were formed after as little as 5 sec of vortexing, and LDL protein and cholesteryl ester were almost completely (95%) incorporated into aggregates after 4 min vortexing. Substantial fractions of phospholipid and unesterified cholesterol from the original LDL remained in solution even after 4 min vortexing, forming large multilamellar vesicles that did not adhere to the aggregated material. Spontaneous aggregates retrieved from LDL solutions after prolonged storage were also examined by electron microscopy, revealing similar lipid droplets and vesicles. The ultrastructural appearance of LDL aggregated in vitro is remarkably similar to the appearance of extracellular lipid deposits in atherosclerosis, lending credence to the hypothesis of direct extracellular formation of these deposits from lipoproteins.


[Gastritis, dyspepsia and Helicobacter pylori].
Helicobacter pylori is a major culprit in chronic gastritis. Interactions between the infected host, the bacteria and the environment, influence the type of gastritis and the occurrence of specific diseases (peptic ulcer, gastric lymphoma and adenocarcinoma) associated with the infection in a minority of cases. The benefit of H. pylori eradication is not proved in isolated gastritis, except for rare forms such as hypertrophic or lymphocytic gastritis. In spite of a high prevalence in functional dyspepsia with normal gastroduodenal endoscopy, H. pylori is not the main cause of symptoms in dyspeptic patients, but the gastritis could be involved in the appearance of ulcer-like symptoms. Strategies for non-invasive H. pylori detection for primary eradication are developed to reduce the endoscopy workload in the management of uninvestigated dyspepsia.


A noncontrastive model for assessment of phonology.
The contrastive/noncontrastive model developed by Seymour and Seymour (1977) can be applied to the development of a dialect-sensitive phonological assessment that uses a single scoring and test format, regardless of a child's dialect. Through extensive field research, stimulus items were found that respect the phonotactics of African American English (AAE) (i.e., no targets are final consonants or final consonant clusters), yet are sufficiently demanding to show development in the age range from 4 to 9 years. Consonant clusters of varying levels of phonological difficulty are shown to discriminate between typically developing and phonologically impaired children of different dialect groups, Mainstream American English (MAE) and non-MAE. Implications for further diagnosis and remediation are presented.


Opportunities for antimicrobial stewardship among carbapenem-treated patients in 18 North American hospitals.
Here we describe the characteristics of carbapenem use at 18 hospitals across North America. Adult inpatients treated with a carbapenem for ≥24 h were included in this multicentre, retrospective, cross-sectional study. Outcomes evaluated included classification of therapy as empirical or definitive, discharge disposition and 30-day re-admission. A total of 621 patients were included in this study. Of these, 467 patients (75.2%) received a carbapenem empirically, among whom negative cultures occurred in 313 (67.0%) and 93% were eligible for de-escalation of therapy. In-hospital mortality occurred in 72 patients (11.6%) and 549 patients (88.4%) were discharged. Of the 549 patients who were discharged, 349 patients (63.6%) went home and 30-day infection-related re-admission occurred in 95 patients (17.3%). This population represents a significant need for carbapenem stewardship. Institutional guidelines should focus on four common disease states (respiratory, genitourinary, intra-abdominal and bloodstream), and diagnostic stewardship should be employed to aid in rapid de-escalation of carbapenem therapy. Additional studies aiming to identify antimicrobial stewardship techniques that may help to optimise carbapenem therapy and increase education about the importance of utilising carbapenem-sparing regimens are required.


Noncovalent associations of glutathione S-transferase and ligands: a study using electrospray quadrupole/time-of-flight mass spectrometry.
Human glutathione S-transferase A1-1 was observed predominantly as dimeric ions (51 kDa) during electrospray mass spectrometric analysis from aqueous solution at pH 7.4, in keeping with the known dimeric structure in solution. When analyses were performed on solutions of the enzyme containing glutathione (GSH), noncovalent adducts of protein dimer and one or two ligand molecules were observed; each mass increment, which exceeded the mass of GSH alone, was provisionally interpreted to indicate concomitant association of two water molecules per bound GSH. Noncovalent adducts of ligand and protein dimer were similarly observed for oxidized glutathione and for two glutathione inhibitors, both incorporating substituted thiol structures. In these instances, the mass increments exactly matched the ligand masses, suggesting that the apparent concomitant binding of water was associated with the presence in the ligand of a free thiol group. Collisionally activated decomposition during tandem mass spectrometry analyses of noncovalent adducts incorporating protein dimer and ligands yielded initially the denuded dimer; at higher collision energies the monomer and a protein fragment were formed.


Immunochemical investigations of Rh0(D) activity detected in Band 3 of red cell membrane.
Stroma from D positive red cell was solubilized with 1% deoxycholic acid (DOC) and was applied onto Sepharose 4B and 6B, followed by fractionation with activated thiol Sepharose 4B. By this method Band 3 containing no carbohydrate was prepared. The Band 3 was incubated with human anti-D, then immune complex soluble in DOC solution was produced. This complex was not prepared from red cell components other than Band 3, nor from Band 3 from D negative red cell. The complex agglutinated papainized D positive red cells. Molecular weight of the complex was estimated to be 500,000 to 700,000 by the gel filtration method. The molecular weight increased according to the time of incubation. The complex gave a precipitation line in counter immunoelectrophoresis (IEP), a single peak in crossed IEP against anti-Band 3, and a line at gamma-globulin region in IEP against anti-human serum. SDS-polyacrylamide gel disc electrophoresis of the complex showed the pattern consisting of fragments of immunoglobulin G and Band 3 which was not stained with periodic acid Schiff reagent. These support our results that Rh-Hr blood type activities are located in Band 3 almost free of carbohydrate.


[Multicenter clinical trial of the antitumor drug Dioxadet (phase II)].
A stage II of the joint clinical study of Dioxadet, an ethylene imine drug, was carried out to evaluate its therapeutic effect in 229 patients and side-effects in 239 patients with malignancies of various sites. Marked therapeutic effect was observed in ascitic malignancies of the ovary and a moderate one--in disseminated breast cancer. The most frequent side-effect was reversible myelodepression, often delayed.


Effects of neck exercise on high-school students' neck-shoulder posture.
[Purpose] This study examined the effects of deep flexor muscle-strengthening exercise on the neck-shoulder posture, and the strength and endurance of the deep flexor muscles of high-school students. [Subjects] The subjects were 30 seventeen-year-old female high-school students who complained about bad posture and chronic neck-shoulder pain. They were randomly divided into an experimental group of 15 subjects, who performed a deep flexor muscle-strengthening exercise and a control group of 15 subjects, who performed a basic stretching exercise. [Methods] The experimental group of 15 subjects performed a deep flexor muscle-strengthening exercise consisting of low-load training of the cranio-cervical flexor muscle, and the control group of 15 subjects performed a basic stretching exercise consisting of seven motions. [Results] The experimental group showed statistically significant changes in head tilt angle, neck flexion angle, forward shoulder angle, and the result of the cranio-cervical flexion test after the training. In contrast, the control group showed no statistically significant changes in these measures following the training. When the results of the groups were compared, statistically significant differences were found for all items between the experimental group and the control group. [Conclusion] Strengthening cranio-cervical flexor muscles is important for the adjustment of neck posture, and maintaining their stability is required to improve neck-shoulder posture.


High-risk HPV and survival in patients with oral and oropharyngeal squamous cell carcinoma - 5-year follow up of a population-based study.
No statistically significant 5-year survival difference was seen in patients with oral and oropharyngeal squamous cell carcinoma (OOPSCC) between high-risk HPV-positive and -negative groups in this population-based study. To see if the formerly observed higher risk for recurrence or second primary tumour (SPT) in high-risk HPV-positive patients with OOPSCC corresponds to worse survival. A total of 128 consecutive, previously untreated patients with OOPSCC, who were part of a population-based case-control study in southern Sweden during 2000-2004, were included. A mouthwash sample was collected and exfoliated cells were collected with cotton-tipped swabs from the tonsillar fossa and the tumour. Specimens were analysed for HPV DNA using nested polymerase chain reaction (PCR). Disease-specific survival (DSS) and DSS difference between HPV-negative and HPV-positive patients were calculated. The relationship between age, stage, high-risk HPV status and DSS was assessed. Oral and oropharyngeal tumours were assessed separately. Mean DSS in months was 80.7/68.6 (high-risk HPV-negative/high-risk HPV-positive) for oral cavity tumours (p = 0.18) and 67.6/78.3 (high-risk HPV-negative/high-risk HPV-positive) for oropharyngeal tumours (p = 0.47). For oral cavity tumours, age, T status, N status and stage all showed significant differences in DSS. For oropharyngeal tumours, no significant difference regarding DSS was found.


Protein-energy Malnutrition in Goitrous Schoolchildren of Isfahan, Iran.
Some studies have shown the possible role of protein-energy malnutrition (PEM) in persistence of endemic goiter in iodine replenished areas. The present study was conducted to assess the association between PEM and goiter in schoolchildren of Isfahan, Iran. In a cross-sectional study using multistage cluster random-sampling, 2331 schoolchildren with age ranged from 6-13 years old with a female to male ratio of 1.60 were enrolled. Thyroid size was examined by two endocrinologists for goiter detection. Children were considered goitrous if they had palpable or visible goiters according to World Health Organization (WHO)/United Nations children's Fund/International Council for the Control of Iodine Deficiency criteria. Weight and standing height were measured using the standard tools and anthropometric indices were calculated using the WHO AnthroPlus software developed by the World Health Organization. Height-for-age Z-scores (HAZ), weight-for-age Z-scores (WAZ) and body mass index (BMI) for age were calculated for each child. Children with a HAZ, WAZ or BMI-for-age of Z-score < -2.0 were classified as stunted, underweight or thin, respectively. Blood samples were drowned to measure serum thyroid hormones. Overall, 32.9% of subjects were classified as goitrous. Weight, height, BMI, WAZ and BMI-for-age Z-score were significantly lower in children with goiter than in children who did not have goiter (P < 0.05). The prevalence of goiter in thin children was higher than that in non-thin ones (48.4 vs. 31.6%, odds ratio [OR]: 2.02, 95% confidence interval [CI]: 1.52-2.69, P < 0.001). Although 33.4% of non-stunted children were goitrous, 31% of stunted ones had goiter (P = 0.5). According to the logistic regression model taking sex and age as covariates, the only significant parameter affecting palpable goiter detection was thinness (OR = 2.13, 95% CI: 1.22-3.69, P < 0.001). In the present study, we found a high prevalence of goiter in children who were malnourished. It seems that PEM may play a role in the still high prevalence of goiter in this region.


Identification of human gene research articles with wrongly identified nucleotide sequences.
Nucleotide sequence reagents underpin molecular techniques that have been applied across hundreds of thousands of publications. We have previously reported wrongly identified nucleotide sequence reagents in human research publications and described a semi-automated screening tool Seek & Blastn to fact-check their claimed status. We applied Seek & Blastn to screen >11,700 publications across five literature corpora, including all original publications in Gene from 2007 to 2018 and all original open-access publications in Oncology Reports from 2014 to 2018. After manually checking Seek & Blastn outputs for >3,400 human research articles, we identified 712 articles across 78 journals that described at least one wrongly identified nucleotide sequence. Verifying the claimed identities of >13,700 sequences highlighted 1,535 wrongly identified sequences, most of which were claimed targeting reagents for the analysis of 365 human protein-coding genes and 120 non-coding RNAs. The 712 problematic articles have received >17,000 citations, including citations by human clinical trials. Given our estimate that approximately one-quarter of problematic articles may misinform the future development of human therapies, urgent measures are required to address unreliable gene research articles.


Enhanced anti-tumor effect of an IFN-gamma-EGF fusion protein.
The novel fusion proteins harboring human or mouse interferon combined with epidermal growth factor receptor binding domain were constructed using methods of genetic and protein engineering. The fusion proteins were assayed to retain complete antiviral activities. The EGF receptor binding moiety of the fusion proteins exhibited competitive binding against 125I-EGF for EGF receptors on A431 cells. The fusion proteins were shown to be more potent in inhibiting the growth of cultured target carcinoma cells than interferon-gamma alone. Experimental data derived from mouse B16 malignant melanoma models indicates that the weight of tumor in mice treated with IFN fusion proteins was significantly smaller than that of mice treated with interferon-gamma alone. The work here is unprecedented in the world and provides a reliable evidence to support the upcoming clinical employment of a class of interferons that specifically target tumor cell.


RhoA and RhoC are both required for the ROCK II-dependent promotion of centrosome duplication.
CDK2-cyclin E triggers centrosome duplication, and nucleophosmin (NPM/B23) is found to be one of its targets. NPM/B23 phosphorylated by CDK2-cyclin E acquires a high binding affinity to Rho-associated kinase (ROCK II), and physically associates with ROCK II. The NPM/B23-binding results in superactivation of ROCK II, which is a critical event for initiation of centrosome duplication. The activation of ROCK II also requires the binding of Rho small GTPase to the auto-inhibitory region; hence the availability of the active Rho protein is an important aspect of the centrosomally localized ROCK II to properly initiate centrosome duplication. There are three isoforms of Rho (RhoA, B and C), all of which are capable of binding to and priming the activation of ROCK II. Here, we investigated which Rho isoform(s) are involved in the activation of ROCK II in respect to the initiation of centrosome duplication. We found that both RhoA and RhoC, but not RhoB, were required for initiation of centrosome duplication, and overactivation of RhoA, as well as RhoC, but not RhoB, promoted centrosome duplication and centrosome amplification.


Binding affinities of insulin-like growth factor-I (IGF-I) fusion proteins to IGF binding protein 1 and IGF-I receptor are not correlated with mitogenic activity.
In this report, comparisons between molecular affinities and cellular proliferation activities have been made for insulin-like growth factor-I (IGF-I) and two IGF-I fusion proteins in order to evaluate fusion proteins as tools for receptor binding studies. Binding affinities and growth promoting effects of the N-terminal fusion Z-IGF-I and the C-terminal fusion IGF-I-Z, and native recombinant human IGF-I, were analyzed. Binding kinetic properties of the three IGF-I variants were analyzed using BIAcore kinetic interaction analysis testing for binding to both human IGF binding protein 1 (IGFBP-1) and a soluble form of the human IGF type I receptor extracellular domains (sIGF-IR). The growth promoting effects on SaOS-2 human osteosarcoma cells of the different fusion proteins were analyzed. A comparison of receptor binding affinities and growth promoting effects shows that the fusion protein receptor affinity does not correlate with proliferative potential. The IGF-I-Z fusion, with the lowest receptor affinity, shows similar proliferative potential to native IGF-I. However, the Z-IGF-I fusion protein, with twice the receptor affinity of IGF-I-Z, displays only about 70% of the IGF-I-Z growth promoting activity. Both IGF-I fusion proteins possess similar affinity to IGFBP-1. These results indicate that determinants other than the receptor affinity could be involved in the regulation of IGF-I proliferative action. This study demonstrates that ligand fusion proteins may be useful to study mechanisms of ligand induced receptor activation.


[Molecular diagnostics for vitreoretinal lymphoma].
Primary vitreoretinal lymphoma (PVRL) represents a subtype of intraocular lymphomas, which are a subgroup of malignant lymphomas of the eye. PVRL is considered a special form of primary diffuse large cell lymphoma (DLBCL) of the CNS (central nervous system) (PCNSL) and arises primary or secondary to PCNSL. According to the cell of origin (COO) classification of DLBCL, PVRL largely belongs to the activated B‑cell (ABC) type of DLBCL. Based on a recently established genetic-biological classification of DLBCL, PCNSL and thus also PVRL belong to a group of DLBCL of the MYD88/CD79B-mutated (MCD) or cluster 5 subtype, which often shows extranodal manifestations and MYD88 and CD79A mutations as well as CDKN2A deletions.PVRL diagnostics is often complicated as it represents a classic masquerade syndrome. Due to the usually limited material with often large numbers of reactive lymphocytes and/or degenerative changes in the cells, the results of diagnostic tests are difficult to interpret. Classic diagnostic tests include cytology on vitreous aspirates, immunocytochemistry, and clonality analysis.New insights into the spectrum of genetic alterations of vitreoretinal lymphomas (VRL) confirm the close relationship to PCNSL and could significantly improve pathological diagnosis. Next-generation sequencing panel-based diagnostics allow VRL diagnosis confirmation with little DNA in almost 100% of patients in cases with insufficient cytological evidence or lack of clonality detection. PVRL, as well as secondary vitreoretinal lymphomas after PCNSL or extracerebral DLBCL, have high mutation frequencies in characteristically mutated genes in PCNSL or MCD/cluster 5 type DLBCL. Supporting diagnostics, mutation detection can also be performed on cell-free DNA from the vitreous supernatant.


[Chromatin diminution, mitotic anomalies and cell death in early mouse embryos].
The Feulgen-positive particles, similar to those in the nuclei, fragments of chromosomes and the whole chromosomes, and micronuclei have been found in the cytoplasm of early mouse embryos. Some morphological peculiarities of the interphase nuclei (extrusion of nucleoli-like structures from the nuclei, so called "partial" pycnoses) and mitoses (loss of chromosomes, the pattern of distribution of condensed chromatin similar to that with C-mitosis) are described. Dead cells were found in early blastocysts. The dynamics of these phenomena was traced in the course of development starting from the 4-cell stage to the blastocyst. The high frequency of enumerated findings enables the author propose the existence of the process of chromatin degradation in normally developing mammalian embryos, which causes genetic abnormalities in some cells of the embryo. This may constitute a step of differentiation for at least provisional structures of the embryo.