Direct costs and cost-effectiveness of dual-source computed tomography and invasive coronary angiography in patients with an intermediate pretest likelihood for coronary artery disease.
The study aims to determine the direct costs and comparative cost-effectiveness of latest-generation dual-source computed tomography (DSCT) and invasive coronary angiography for diagnosing coronary artery disease (CAD) in patients suspected of having this disease. The study was based on a previously elaborated cohort with an intermediate pretest likelihood for CAD and on complementary clinical data. Cost calculations were based on a detailed analysis of direct costs, and generally accepted accounting principles were applied. Based on Bayes' theorem, a mathematical model was used to compare the cost-effectiveness of both diagnostic approaches. Total costs included direct costs, induced costs and costs of complications. Effectiveness was defined as the ability of a diagnostic test to accurately identify a patient with CAD. Direct costs amounted to €98.60 for DSCT and to €317.75 for invasive coronary angiography. Analysis of model calculations indicated that cost-effectiveness grew hyperbolically with increasing prevalence of CAD. Given the prevalence of CAD in the study cohort (24%), DSCT was found to be more cost-effective than invasive coronary angiography (€970 vs €1354 for one patient correctly diagnosed as having CAD). At a disease prevalence of 49%, DSCT and invasive angiography were equally effective with costs of €633. Above a threshold value of disease prevalence of 55%, proceeding directly to invasive coronary angiography was more cost-effective than DSCT. With proper patient selection and consideration of disease prevalence, DSCT coronary angiography is cost-effective for diagnosing CAD in patients with an intermediate pretest likelihood for it. However, the range of eligible patients may be smaller than previously reported.


The attractive female body weight and female body dissatisfaction in 26 countries across 10 world regions: results of the international body project I.
This study reports results from the first International Body Project (IBP-I), which surveyed 7,434 individuals in 10 major world regions about body weight ideals and body dissatisfaction. Participants completed the female Contour Drawing Figure Rating Scale (CDFRS) and self-reported their exposure to Western and local media. Results indicated there were significant cross-regional differences in the ideal female figure and body dissatisfaction, but effect sizes were small across high-socioeconomic-status (SES) sites. Within cultures, heavier bodies were preferred in low-SES sites compared to high-SES sites in Malaysia and South Africa (ds = 1.94-2.49) but not in Austria. Participant age, body mass index (BMI), and Western media exposure predicted body weight ideals. BMI and Western media exposure predicted body dissatisfaction among women. Our results show that body dissatisfaction and desire for thinness is commonplace in high-SES settings across world regions, highlighting the need for international attention to this problem.


Does systemic lidocaine reduce ketamine requirements for endotracheal intubation in calves?
To investigate whether an intravenous (IV) lidocaine bolus in calves premedicated with xylazine-butorphanol reduces the amount of ketamine required to allow endotracheal intubation. Randomized, prospective clinical study. In total, 41 calves scheduled for elective umbilical surgery. Calves were randomly assigned to one of two groups (L: lidocaine or S: saline). The calves were administered xylazine (0.07 mg kg-1) and butorphanol (0.1 mg kg-1) intramuscularly and 10 minutes later lidocaine (2 mg kg-1; group L) or saline (group S) IV over 1 minute. After 2 minutes, ketamine (2.5 mg kg-1) was injected IV. If the depth of anaesthesia was insufficient for intubation, additional ketamine (1 mg kg-1) was administered every minute until intubation was successful. The amount of ketamine required for intubation, respiratory rate, pulse rate, arterial pressures, the depth of sedation and conditions of endotracheal intubation after induction of anaesthesia were compared between the two groups. The calves in group L were sedated more deeply than those in group S; however, neither the median (range) amount of ketamine required for intubation, 3.5 (2.5-4.5) mg kg-1 and 3.5 (2.5-3.5) mg kg-1, respectively, nor the induction quality differed significantly between the groups. A bolus of lidocaine (2 mg kg-1) administered 10 minutes after xylazine-butorphanol in calves deepened the degree of sedation but did not decrease the requirement of ketamine for endotracheal intubation. No adverse effects were recorded in the physiological variables measured.


The role of proximal circumstances and child behaviour in toddlers' risk for minor unintentional injuries.
Much of the research on child injury risk has focused on trait-like factors (eg, hyperactivity, child gender) that influence injury risk rather than state-like factors (eg, environmental circumstances, child behaviour). Additional research is needed to better identify the proximal risk factors for children's risk for unintentional injury. The present study examined the antecedents to minor unintentional injury events and whether unusual circumstances and child behaviour predicted injury risk. The study used archival data that were collected via biweekly in-person interviews with 170 mothers of toddlers (15-36 months) for 6 months. A case crossover design was used to predict children's risk for injury from proximal risk factors. Children were at a higher risk for injury when circumstances were unusual and when they were engaging in an unusual behaviour. When a child was engaging in an unusual behaviour, higher levels of maternal supervision predicted lower injury risk. Children were more likely to be injured in a new environment, in an environment with animals or other people, in an environment with hazards or when engaging in a new activity or in a familiar activity performed in an unfamiliar way. The results indicate that toddlers may be at a greater risk for minor unintentional injury when environmental circumstances are outside of the norm or when a child is engaging in unusual behaviours. The findings also indicate that higher levels of caregiver supervision may be especially beneficial when children are engaging in new or unfamiliar activities.


Combination treatment with cetuximab in advanced nasopharyngeal carcinoma patients: a meta-analysis.
Cetuximab, an anti-epidermal growth factor receptor monoclonal antibody, carries the potential for combination treatment against nasopharyngeal carcinoma (NPC). We conducted a meta-analysis to assess the possible benefits and safety between the combination treatment with cetuximab and conventional treatment in NPC patients. Skin toxicity (ST) associated with additional cetuximab was evaluated as well. We performed a systematic search (PubMed, Embase, Cochrane library, China National Knowledge Infrastructure, and WanFang Data) for studies comparing combination treatment with cetuximab versus conventional treatment in NPC patients. The selected studies included completely or partly reported clinical outcomes including survivals, complete and partial responses, and adverse reactions (ST). The pooled HR, relative risk (RR), and respective 95% CI were estimated by using fixed effects model or random effects model. A total of 23 relevant studies with available data were included in the final analysis. According to the pooled data, combination treatment with cetuximab showed improved efficacy on increased objective response rate (studies with cetuximab treatment: RR: 1.39, 95% CI: 1.29-1.50; concurrent chemoradiotherapy with or without cetuximab: RR: 1.39, 95% CI: 1.25-1.54) and prolonged survival (studies with cetuximab treatment: the pooled HR for OS was 0.70, 95% CI: 0.55-0.89; concurrent chemoradiotherapy with or without cetuximab: the pooled HR for OS was 0.64, 95% CI: 0.49-0.84) compared with conventional treatment. Moreover, the improved efficacy was invariably accompanied by an increased occurrence of ST (studies with cetuximab treatment: RR: 2.46, 95% CI: 1.81-3.34; concurrent chemoradiotherapy with or without cetuximab: RR: 1.84, 95% CI: 1.02-3.31). However, the majority of adverse reactions exhibited similar occurrence rates between the different treatments. Patients with NPC receiving additional cetuximab treatment can benefit more from this systemic comprehensive therapy, while the efficiency of conventional treatment for NPC is limited. ST associated with cetuximab may be used as a potential on-treatment marker to guide treatment with cetuximab against NPC.


Racing pigeons: a reservoir for nitro-imidazole-resistant Trichomonas gallinae.
Trichomonas gallinae , the cause of avian trichomonosis, is most commonly found in the order Columbiformes. Racing pigeons are often treated preventively with nitro-imidazoles, which could result in the emergence of resistant isolates, and these isolates can be a threat to wildlife when exchanges occur. The sequence type of 16 T. gallinae isolates obtained from racing pigeons and 15 isolates from wild pigeons was determined based on the ITS1/5.8S rRNA/ITS2 region sequence. In addition, the resistance profiles of these isolates against 5 different nitro-imidazoles (metronidazole, dimetridazole, ronidazole, tinidazole, and carnidazole) were determined. Two different Trichomonas sequence types were isolated. Sequence type A isolates were recovered from racing and wild pigeons, in contrast to sequence type B, which was only isolated from wild pigeons. Isolates with sequence type B were all susceptible to the tested nitro-imidazoles, except for tinidazole resistance in 3 isolates. Resistance to the nitro-imidazoles was observed more frequently in isolates obtained from racing pigeons than from wild pigeons, with most isolates belonging to sequence type A. A higher percentage of the sequence type A isolated from racing pigeons, in comparison with those isolated from the wild pigeons, were resistant to the nitro-imidazoles and displayed higher mean lethal concentration (MLC) values. Two isolates belonging to sequence type A, 1 recovered from a racing pigeon and 1 from a wild pigeon, displayed a similar resistance pattern, suggesting a potential exchange of resistant isolates between racing pigeons and wild pigeons.


Exploiting the Generative Adversarial Network Approach to Create a Synthetic Topography Corneal Image.
Corneal diseases are the most common eye disorders. Deep learning techniques are used to perform automated diagnoses of cornea. Deep learning networks require large-scale annotated datasets, which is conceded as a weakness of deep learning. In this work, a method for synthesizing medical images using conditional generative adversarial networks (CGANs), is presented. It also illustrates how produced medical images may be utilized to enrich medical data, improve clinical decisions, and boost the performance of the conventional neural network (CNN) for medical image diagnosis. The study includes using corneal topography captured using a Pentacam device from patients with corneal diseases. The dataset contained 3448 different corneal images. Furthermore, it shows how an unbalanced dataset affects the performance of classifiers, where the data are balanced using the resampling approach. Finally, the results obtained from CNN networks trained on the balanced dataset are compared to those obtained from CNN networks trained on the imbalanced dataset. For performance, the system estimated the diagnosis accuracy, precision, and F1-score metrics. Lastly, some generated images were shown to an expert for evaluation and to see how well experts could identify the type of image and its condition. The expert recognized the image as useful for medical diagnosis and for determining the severity class according to the shape and values, by generating images based on real cases that could be used as new different stages of illness between healthy and unhealthy patients.


Hyperresponsiveness of monoaminergic mechanisms in DOCA/NaCl hypertensive rats.
To examine the role of the sympathetic nervous system in the development of deoxycorticosterone acetate (DOCA)/NaCl hypertension and to test the hypothesis that the responsiveness of the sympathetic nervous system to stress is enhanced during the developmental phase of hypertension in this model before resting sympathetic activity becomes increased, DOCA/NaCl-treated rats and uninephrectomized control animals were studied after 3, 7, 14, and 28 days of treatment. Basal plasma norepinephrine and epinephrine in conscious, unrestrained resting DOCA/NaCl-treated rats were the same as in controls at 3, 7, and 14 days but were significantly elevated at 28 days of treatment. Ganglionic blockade resulted in a significantly greater decrease in mean arterial pressure in DOCA/NaCl rats than in controls at 14 and 28 days of treatment. At 14 days, DOCA/NaCl rats exhibited significantly greater increments in plasma norepinephrine and epinephrine following cold stress than did H2O controls. Basal plasma prolactin levels were elevated and release of dopamine from isolated superfused mediobasal hypothalami reduced in 28-day DOCA/NaCl hypertensive rats. These results indicate that sympathetic nervous system activity increases progressively during the development of DOCA/NaCl hypertension and that the sympathoadrenal system is hyperresponsive to environmental stress even early in the course of DOCA/NaCl treatment and suggest that hypothalamo-hypophyseal function is altered in this model of hypertension.


Sexual dysfunction in patients with multiple sclerosis: a multidisciplinary approach to evaluation and management.
Multiple sclerosis (MS) is the most common cause of progressive neurological disability in young adults. In addition to spasticity, tremors, weakness, sensory disturbances, depression, cognitive problems, and bladder or bowel dysfunction, sexual dysfunction (SD) is also a prevalent and destructive manifestation of the disease that severely affects quality of life. Evaluation of this disorder requires insight into the primary (changes that directly affect libido, sexual response and orgasm due to direct damage to the nervous system), secondary (complaints which are related to the physical disability of MS, such as fatigue, muscle rigidity, weakness and spasms), and tertiary (emotional, social and cultural aspects of MS) components of MS-associated SD. Given the complexity and multifactorial nature of SD, a multidisciplinary approach is necessary when treating patients with MS. The aim of this Review is to provide a holistic approach to the evaluation and management of SD in patients with MS, incorporating the latest data from the fields of urology, neurology, nursing, social work, and psychology. What is currently known regarding the evaluation and management of SD in patients with MS will be presented from the perspective of these specialties.


A pilot study of the acceptability, efficacy, and iatrogenic effects of a brief dynamic norms intervention for reducing young adult alcohol use.
Alcohol use and its related consequences are a public health problem among young adults. Building upon efficacious personalized normative feedback interventions, dynamic norms can be used to highlight the decreasing prevalence of alcohol use over time among young adults' peers, thereby increasing their motivation to change drinking consistent with the trend. Because limited research has examined dynamic norms feedback interventions for alcohol use, we examined the acceptability and initial efficacy of such an intervention, and potential iatrogenic effects of showing norms feedback about drinking to light drinkers and nondrinkers. Participants were 546 unvaccinated young adults ages 18-24 who completed a baseline survey, intervention, and 1-month follow-up assessment. Participants were block randomized to receive a brief web-based dynamic norms intervention, with feedback content focused on either (a) alcohol-related behaviors (intervention) or (b) COVID-19 vaccine behaviors (the attention-matched control for the present study). On average, participants who received the alcohol intervention rated it as generally engaging, helpful, and acceptable, with the majority (90.8%) indicating that they would recommend it to a friend. Supporting initial efficacy, in generalized linear models controlling for demographics and baseline alcohol outcomes, at 1-month follow-up the alcohol intervention was associated with statistically and clinically significant reductions in all indices of perceived drinking norms, drinking quantity, drinking frequency, and driving after drinking occasions. Lighter drinkers showed no adverse iatrogenic effects. Presenting alcohol-related personalized normative feedback using dynamic trends is a promising intervention for reducing alcohol use in a community sample of young adults. Further research clarifying the optimal presentation of dynamic norms is needed.


The anatomy of supercoiled loops in the Drosophila 7F locus.
The genome in eucaryotes is organized into a series of supercoiled loops, topologically anchored at their bases by components of the nuclear matrix. Previous studies have shown that active genes are associated with the nuclear matrix. We wished to know whether loops in general were solely organized by active genes. We therefore examined a locus of the Drosophila X-chromosome comprising 163,000 bp of continuous DNA sequences and devoid of known active genes. Of the 52 EcoRI restriction fragments comprising this region, we found 5 anchored fragments which non-randomly organized this region into 4 DNA loops. Each of the 5 anchored fragments contained a transcribed sequence. These results strongly suggest that supercoiled loops are organized in a specific fashion with respect to DNA sequence, with the anchorage points exclusively demarcated by transcriptionally active genes.


[Cryptococcosis in AIDS patients: case study from 1996 to 2006 in Paroissien Hospital].
Clinic, epidemiological and microbiological characteristics of 128 episodes of cryptococcosis were retrospectively evaluated in 106 positive HIV patients hospitalized at the Paroissien Hospital on period 1996-2007. There were 75 male and 31 female patients, with a median age of 34 years, ranging from 20 to 68 years. Addiction to intravenous drugs was the main cause of HIV infection in 55 patients (51.9%). Cryptococcosis was detected as single episode in 85 patients (80.2%) and as relapse in 19 (17.9%). It appeared as the first marking disease in 36 patients (34.0%). Diagnosis was established in 116 episodes by CSF study (Indian ink, culture, antigen detection), in 9 cases by Cryptococcus sp. recovery from blood cultures, and in 3 cases by antigen detection in patient's serum with a latex reactive. Neurological symptomatology, with headache and fever as the most common signs, was presented by 89.6% of patients. Induction treatment was done in all cases with amphotericin B, and maintenance treatment was carried out with fluconazole. Mortality rate was 35.8%, being higher in those patients who had suffered relapses (41.3%) compared to those who presented a first episode of the mycosis (33.3%).


Diagnosis of an intact hydatidiform mole with coexistent fetus by amniography.
A case hydatidiform mole with coexistent living fetus is presented and the steps by which the diagnosis was reached are given prominence. The diagnosis in utero of an intact hydatidiform mole with associated fetus is extremely difficult, and the author thinks that the most useful diagnostic means for this rare situation is amniography.


Behavioral Plasticity and the Origins of Novelty: The Evolution of the Rattlesnake Rattle.
Environmentally induced behavior (behavioral plasticity) has long been hypothesized to promote the origins of novel morphological traits, but this idea remains controversial. One context in which this hypothesis can be evaluated is animal communication, where behavior and morphology are often linked. Here, we examined the evolution of one of nature's most spectacular communication signals: the rattlesnake rattle. We specifically evaluated whether rattlesnake rattling behavior-and, hence, the rattle-originated from a simple behavior: vibrating the tail when threatened. By reconstructing the ancestral state of defensive tail vibration, we show that this behavior is nearly ubiquitous in the Viperidae (the family that includes rattlesnakes) and widespread in the Colubridae (the largest snake family, nearly all of which are nonvenomous), suggesting a shared origin for the behavior between these families. After measuring tail vibration in 56 species of Viperidae and Colubridae, we show that the more closely related a species was to rattlesnakes, the more similar it was to rattlesnakes in duration and rate of tail vibration. Thus, the rattlesnake rattle might have evolved via elaboration of a simple behavior. These data thereby support the long-standing hypothesis that behavioral plasticity often precedes-and possibly instigates-the evolution of morphological novelty.


Psychometric evaluation of the Worst Pruritus Numerical Rating Scale (NRS), Atopic Dermatitis Symptom Scale (ADerm-SS), and Atopic Dermatitis Impact Scale (ADerm-IS).
Atopic dermatitis (AD) is a chronic inflammatory skin disease characterized by pruritus, skin pain, and sleep impacts, which are only reportable by patients themselves. The goal of this research is to evaluate the reliability, validity, and interpretability of the scores from three patient-reported outcome measures within the context of a clinical trial for adolescents and adults with moderate to severe AD. Data from a Phase 3 randomized, double-blind, placebo-controlled, multinational clinical trial for individuals 12-75 years of age with moderate to severe AD (AD Up [ClinicalTrials.gov NCT03568318]) were used to assess the reliability, validity, and interpretability of scores on the Worst Pruritus Numerical Rating Scale (NRS) and the Atopic Dermatitis Symptom and Impact Scales (ADerm-SS and ADerm-IS). Analyses were conducted separately for the adult and adolescent subgroups. Of the 882 participants included in the psychometric analyses, the majority were adults (n = 769, 87.2%), male (n = 536, 60.8%), and white (n = 630, 71.4%). Multi-item scores from the ADerm-SS and ADerm-IS had good internal consistency reliability, and most scores demonstrated acceptable test-retest reliability. Scores from the three questionnaires demonstrated adequate validity, exhibiting correlations with other conceptually related outcome assessments and score differences between clinically distinct subgroups. Finally, the score interpretation analyses provide estimates for meaningful within-person change and between-groups difference thresholds that may be useful for future research in adults and adolescents with moderate to severe AD. These results provide evidence that the scores produced by the Worst Pruritus NRS, ADerm-SS, and ADerm-IS are reliable and construct-valid when completed by adults and adolescents with moderate to severe AD in a clinical trial setting. The results presented here expand upon the previous qualitative evidence of these tools and provide further support for their use in future clinical studies. While results are specific to clinical trials, next steps would be to evaluate the use of these questionnaires in clinical practice. This can provide clinicians and dermatologists a window into the patient's disease experience outside of the clinic, aid in shared decision making, and support a patient-centric approach to management of moderate to severe AD.


Impact of elapsed treatment time on outcome of external-beam radiation therapy for localized carcinoma of the prostate.
The purpose of this study was to evaluate the impact of elapsed treatment time in external-beam radiation therapy for localized prostate carcinoma. The medical records of 1083 patients with localized prostate carcinoma treated between 1970 and December 1999 with external irradiation alone were reviewed. Median follow-up was 6 years (range, 4-24 years). Since 1987, prostate-specific antigen levels were obtained in 687 patients before the initiation of radiation therapy, and all patients seen in follow-up had prostate-specific antigen determinations. There were 344 patients with T1c, 496 with T2, and 243 with T3 tumors. The elapsed treatment time was divided into < or = 7, 7.1-9, or > 9 weeks. Local tumor control was determined by rectal examination and cause-specific survival or prostate-specific antigen failure according to American Society of Therapeutic Radiology and Oncology consensus criteria. Because of dose-escalation studies, tumor dose levels ranged from 66-73.8 Gy, given in 1.8- to 2-Gy fractions. In patients with stage T1c, local failure ranged from 0% to 10% with doses < or = 72 Gy with; elapsed treatment time had no impact. No pelvic failures were detected in 88 patients receiving doses > 72 Gy. In patients with T2 who received < or = 70 Gy, overall pelvic failure rate was 4% (12/306) in those with an elapsed treatment time of < or = 9 weeks, in contrast to 27% (12/44) for those with an elapsed treatment time > 9 weeks; at 10 years, patients with T2 tumors treated in > 9 weeks had a higher actuarial pelvic failure rate (35%), in contrast to 5% to 18% with shorter treatment times. For patients with T2 tumors who received 70-72 Gy, pelvic failure rate ranged from 0% to 32%, and there were no failures in 37 patients treated to higher doses. In patients with prostate-specific antigen values whose tumors were stage T1c, the chemical failure rate was 41% (60/147) with a tumor dose < 70 Gy, compared with 17% (4/24) in those who received higher doses. In patients with stage 2 disease who were treated with < 70 Gy, the chemical failure rate was 31%, and the rate was 12%-18% in those who received higher doses. In stage T3, the clinical pelvic failure rate ranged from 25% to 32% in the three elapsed time groups, and the chemical failure rate ranged from 48% to 69%, and there was no significant correlation with elapsed time or total irradiation dose. Cause-specific survival without chemical failure in patients with stage T1c disease at 10 years was 85%-90% in the three elapsed treatment time groups. In patients with stage T2 disease, the corresponding values were 80% and 90% for elapsed treatment times < 9 weeks, in contrast to 65% for patients treated > 9 weeks. In patients with stage T3 disease, cause-specific survival was about 60% in all elapsed treatment groups. There was no significant correlation of elapsed treatment time with urinary or rectal morbidity. Patients treated with radiation therapy for stage T2 localized prostate carcinoma showed a greater incidence of pelvic and chemical failures and a lower cause-specific survival when elapsed treatment time was > 9 weeks in comparison with the failure and survival rates occurring with shorter times. Higher doses of irradiation (> 72 Gy) eliminate the influence of prolongation of treatment time on outcome.


[Fragment-based screening by X-ray structure analysis].
The first step of FBDD is fragment-based screening (FBS). There are various analytical methods used to perform FBS: NMR, X-rays, SPR, etc. The advantage of X-ray structure analysis in FBS is that it can cope with relatively large structural changes upon the binding of a fragment and unexpected events such as multiple binding. The first thing needed to perform FBS by X-rays is a fragment library. The library currently employed at our laboratory was designed especially for X-ray structure analysis and consists of 384 compounds. This library was built based on Ro3 and special attention was given to eliminating ambiguity when interpreting electron density. FBS by X-rays requires more than 100 times more data collection and structure analysis. Recent development of critical technologies dramatically reduced the time required for X-ray structure analysis. Additionally, development of software and the incorporation of an industrial robot to a laboratory system has enabled us to construct a fully automated structure analysis system. However, there still are some limitations to X-ray structure analysis. It demands hundreds of high quality crystals and those crystals must not only survive soaking of compounds dissolved in DMSO but also be resistant to X-ray damage. In this article, a practical example of FBS by X-rays will be presented using HSP90 along with facts on the limitations of X-rays.


[The incidence of the antiphospholipid syndrome in the clinical and biological manifestations of systemic lupus erythematosus].
In the present investigation a study was made on the incidence of antiphospholipid syndrome (APS) on clinical and biological manifestations in a series or 32 patients (28 females and 4 males with a mean age of 25 years) diagnosed of SLE (ARA criteria) and APS (Harris criteria) compared with a group of 25 patients (19 females and 6 males with a mean age of 38 years) diagnosed of SLE without APS. This entails a selection from 124 patients diagnosed of SLE, and an incidence of 25.8% and 9.7% for ACA and LA, respectively. After a clinical protocol was filled, a complete immunological profile was obtained, with lymphocyte subsets, IL-2 receptor, coagulation study, isotype determination for anticardiolipin antibody (ACA), lupus anticoagulant (LA), serology for syphilis and imaging diagnostic techniques. Comparative results, with an statistic assessment, are shown in tables. It is concluded that SLE + APS population can be considered as definite for a peculiar SLE subtype.


pKa does not predict pH potentiation of sodium channel blockade by lidocaine and W6211 in guinea pig ventricular myocardium.
During diastole, tertiary amine local anesthetic molecules may exit cardiac sodium channels quickly through the membrane if they are neutral, or more slowly through the aqueous channel pore if they are charged. Extracellular acidosis potentiates sodium channel blockade by these drugs, and drug pKa should be a potent predictor of the degree of response of drug dissociation kinetics to changes in extracellular pH. To test this hypothesis, we measured kinetics of recovery from drug-induced channel blockade in guinea pig papillary muscle exposed to lidocaine (pKa 7.86) and to W6211 (pKa 6.29) using Vmax as a measure of peak sodium current. Both compounds, which are physicochemically very similar in respects other than pKa, delayed Vmax recovery in a pH-dependent fashion. As pH was lowered from 7.9 to 6.5, the recovery time constant rose from 86 to 230 msec for lidocaine and from 53 to 154 msec for W6211. We revised an earlier kinetic scheme of drug-channel interaction to incorporate newer concepts of drug trapping and ionization within the channel, and the resulting analytic expressions fit the data well. Important implications of the new scheme are that the pKa of the drug-receptor complex may differ from the drug pKa, and that deprotonation of channel-bound charged drug molecules may be a rate-limiting process.


Genetic diversity of populations of Monilinia fructicola (Fungi, Ascomycota, Helotiales) from China.
The genetic variation among 128 isolates of Monilinia fructicola (Fungi, Ascomycota, Helotiales) from China was analyzed using Inter-Simple Sequence Repeat (ISSR) markers and compared with those of samples from California, USA and New Zealand. A total of 72 reproducible DNA fragments were scored, of which 87.5% (63/72) were polymorphic. The Nei's gene diversity and Shannon's diversity indices of three Chinese regional populations were very similar to that from California. However, several differences were observed among geographic populations of M. fructicola from both within China and between China and California. The analysis of molecular variance (AMOVA) of isolates from different geographic locations suggested that most of the observed genetic variation was found within populations. Results of this study are inconsistent with the hypothesis that the Chinese populations of M. fructicola were derived from a single or few recent migrants from other countries. Instead, our results suggest that M. fructicola has been in China long before its first official recording in 2003.


A Two-Component regulatory system with opposite effects on glycopeptide antibiotic biosynthesis and resistance.
The glycopeptide A40926, produced by the actinomycete Nonomuraea gerenzanensis, is the precursor of dalbavancin, a second-generation glycopeptide antibiotic approved for clinical use in the USA and Europe in 2014 and 2015, respectively. The final product of the biosynthetic pathway is an O-acetylated form of A40926 (acA40926). Glycopeptide biosynthesis in N. gerenzanensis is dependent upon the dbv gene cluster that encodes, in addition to the two essential positive regulators Dbv3 and Dbv4, the putative members of a two-component signal transduction system, specifically the response regulator Dbv6 and the sensor kinase Dbv22. The aim of this work was to assign a role to these two genes. Our results demonstrate that deletion of dbv22 leads to an increased antibiotic production with a concomitant reduction in glycopeptide resistance. Deletion of dbv6 results in a similar phenotype, although the effects are not as strong as in the Δdbv22 mutant. Consistently, quantitative RT-PCR analysis showed that Dbv6 and Dbv22 negatively regulate the regulatory genes (dbv3 and dbv4), as well as some dbv biosynthetic genes (dbv23 and dbv24), whereas Dbv6 and Dbv22 positively regulate transcription of the single, cluster-associated resistance gene. Finally, we demonstrate that exogenously added acA40926 and its precursor A40926 can modulate transcription of dbv genes but with an opposite extent: A40926 strongly stimulates transcription of the Dbv6/Dbv22 target genes while acA40926 has a neutral or negative effect on transcription of those genes. We propose a model in which glycopeptide biosynthesis in N. gerenzanensis is modulated through a positive feedback by the biosynthetic precursor A40926 and a negative feedback by the final product acA40926. In addition to previously reported control systems, this sophisticated control loop might help the producing strain cope with the toxicity of its own product. This work, besides leading to improved glycopeptide producing strains, enlarges our knowledge on the regulation of glycopeptide biosynthesis in actinomycetes, setting N. gerenzanensis and its two-component system Dbv6-Dbv22 apart from other glycopeptide producers.


Lead to hit ruthenium-cyclopentadienyl anticancer compounds: Cytotoxicity against breast cancer cells, metabolic stability and metabolite profiling.
The successful choice of hit compounds during drug development programs involves the integration of structure-activity relationship (SAR) studies with pharmacokinetic determinations, including metabolic stability assays and metabolite profiling. A panel of nine ruthenium-cyclopentadienyl (RuCp) compounds with the general formula [Ru(η5-C5H4R)(PPh3)(bipyR')]+ (with R = H, CHO, CH2OH; R' = H, CH3, CH2OH, CH2Biotin) has been tested against hormone-dependent MCF-7 and triple negative MDA-MB-231 breast cancer cells. In general, all compounds showed important cytotoxicity against both cancer cell lines and were able to inhibit the formation of MDA-MB-231 colonies in a dose-dependent manner, while showing selectivity for cancer cells over normal fibroblasts. Among them, four compounds stood out as lead structures to be further studied. Cell distribution assays revealed their preference for the accumulation at cell membrane (Ru quantification by ICP-MS) and the mechanism of cell death seemed to be mediated by apoptosis. Potential structural liabilities of lead compounds were subsequently flagged upon in vitro metabolic stability assays and metabolite profiling. The implementation of this integrated strategy led to the selection of RT151 as a promising hit compound.


What is the unit of visual attention? Object for selection, but Boolean map for access.
In the past 20 years, numerous theories and findings have suggested that the unit of visual attention is the object. In this study, I first clarify 2 different meanings of unit of visual attention, namely the unit of access in the sense of measurement and the unit of selection in the sense of division. In accordance with this distinction, I argue that an object, as commonly described, is only the unit of selection. The unit of access is better characterized as a Boolean map (Huang & Pashler, 2007), that is, the linkage of a single feature value per dimension associated with a map (i.e., a set of locations). The experiments in this study demonstrated the following: (a) Grouping items into a single object (by connecting them) does not improve the perception of these items (Experiment 1); (b) same-object advantage exists only when the features to be perceived are different dimensions of a single Boolean map and not when they belong to different parts of an object (Experiments 2 and 3); (c) cuing the relevant feature does not help perception when the features to be perceived are different dimensions of a single Boolean map but does help significantly when these features belong to different parts of an object (Experiment 4); and (d) connection, as used in Experiments 1-4, is effective in affecting object structure (i.e., affecting the mechanism of selection) in both an enumeration and a tracking task (Experiments 5 and 6). The results of these experiments, together with data available in the literature, demonstrate that the unit of access is a Boolean map, not an object.


Sizes and distributions of intrinsic neurons incorporating tritiated GABA in monkey sensory-motor cortex.
Neurons accumulating [3H]gamma-aminobutyric acid (GABA) were identified autoradiographically in frozen and plastic sections after injection of the material in the monkey somatic sensory (areas 3, 1, and 2), motor (area 4), and parietal (area 5) cortex following intravenous administration of a GABA transaminase inhibitor, amino-oxyacetic acid. Two general forms of labeled cells are recognized: those with large diameter (15- to 20-micrometer) somata, probably corresponding to the basket cells of Golgi studies, and those with small diameter (6- to 12-micrometer) somata, probably corresponding to several other types. The sizes and laminar distributions of labeled cells in the cytoarchitectonic fields of the first somatic sensory are (SI) are similar. In these areas, the greatest concentrations of labeled cells are in layer II, in layer IV, and in the superficial stratum of layer IV. In area 5, there are many fewer labeled cells in layer IV. In area 4, the labeled cells are larger and distributed more homogeneously, though there are fewer in layer VI. The large labeled cell type is concentrated in layers III and V of all areas. The smaller labeled types are found in all other layers, including layer I. In layer IV, they form approximately 40% of the total small-celled neuronal population.


Beware of TIPS in liver transplant candidates. Transjugular Intrahepatic Portosystemic Shunt.
Transjugular intrahepatic portosystemic shunt (TIPS) reduce portal pressure and prevent bleeding from esophageal varices in cirrhotic patients. The method is often used in liver transplant candidates. Two cases of TIPS malpositioning in liver transplantation candidates are reported. In the first patient, the caudal end of the TIPS was situated distally in the portal trunk and during transplantation it was necessary to isolate the spleno-portal confluence in order to ensure anastomosis in an area of the wall without endothelial lesions. In the second case, still on the waiting list, the cephalead end of the stent is situated in the right atrium and in this case a more complex trans-diaphragmatic and probably trans-atrial approach is foreseen to allow extraction of the stent. In cirrhotic patients who may be possible transplant candidates, shorter TIPS must be used and positioned with care intrahepatically. Careful radiological evaluation is recommended, together with a CT scan and possibly angiography, in patients with TIPS before liver transplantation is performed, to avoid surprises with detrimental effects during the transplant.


Green innovations and environmentally friendly technologies: examining the role of digital finance on green technology innovation.
The digital finance created by technological empowerment has a significant impact on the inventive behavior of micro-enterprises. This paper uses a correlation analysis that combines the fixed effect model (FE) and the panel threshold model (PTM) to evaluate the impact of digital financing on the quantity and quality of innovation in green technology. In addition, its process is dissected in this work with respect to resource limitations and financial expenditures. The empirical evidence demonstrates that the use of digital financing considerably increases both the rate and quality of innovation in environmentally friendly technologies. Further, the effect of user engagement on green innovation is dynamically overlaid and accumulates over time, as opposed to the coverage of digital finance and digital services. In terms of ownership, growth cycle, and company size, digital finance may assist remedy the misallocation of financial resources and further drive inclusive green innovation. Based on the examination of underlying mechanisms, it is clear that digital finance may play a significant role in fostering innovation in environmentally friendly technologies by easing financial limitations and decreasing associated costs. Depending on the context, "quantitative change before qualitative change" describes the dynamic development process of green innovation fueled by digital finance. This paper proposes that the combination of technological innovation and digital financial services should focus on establishing an inclusive digital financial service system, fostering diverse financial forms, and enhancing the market environment for digital financial services.


Fatigue, muscle strength and vitamin D status in women with systemic lupus erythematosus compared with healthy controls.
Recent studies have demonstrated an inverse relationship between vitamin D levels and fatigue in systemic lupus erythematosus (SLE). The aims of this study were to evaluate proximal muscle strength, fatigue and vitamin D levels in women with SLE compared with healthy controls and to investigate relationships between these factors in a cohort of women with SLE. Forty-five women (24 SLE, 21 healthy controls) participated. Primary outcome measures were the fatigue severity scale (FSS), isometric muscle strength of dominant limbs using hand held dynamometry, two functional tests--the 30-second chair stand test and the 1-kg arm lift test, with vitamin D status measured using 25(OH)D. Overall 25(OH)D levels were 68.4 (22.4) nmol/L with no difference between SLE and control groups. There was a statistically and clinically significant difference in fatigue, 1-kg arm lift, 30-second sit to stand, knee extension, hip flexion, hip abduction, shoulder flexion and grip strength in the SLE group compared with the control group (p < 0.05). In the SLE group FSS was moderately correlated with both functional measures (1-kg arm lift r = -0.42, 30-second chair stand r = -0.44, p < 0.05). However, no statistically significant correlation between dynamometry measures and fatigue was evident. There was no association between fatigue and 25(OH)D level (r = -0.12). In summary, women with SLE were weaker and demonstrated reduced physical function and higher fatigue levels than healthy controls. Fatigue was related to physical function but not vitamin D status or maximal isometric strength in vitamin D replete individuals with SLE.


Sleep Disturbances and Associated Factors in Chinese Children with Autism Spectrum Disorder: A Retrospective and Cross-Sectional Study.
Sleep disturbances in children with autism spectrum disorder (ASD) exist worldwide, but little is known about this issue in non-Western cultures. This study aimed to characterize sleep disturbances in Chinese children with ASD and to examine associated sociodemographic factors and emotional/behavioral problems. Parents of 60 Chinese children with ASD (aged 6-17 years) from Shenzhen, China completed the Children's Sleep Habits Questionnaire (CSHQ), and the Strengths and Difficulties Questionnaire (SDQ). Sleep disturbances were severe and common, with rates of 70.0% for overall disturbances and 15.0% (daytime sleepiness) to 40.0% (sleep duration) for specific domains. The severity and rate of sleep disturbances were higher compared to previous studies in typically developing children from the same region of China and American children with ASD, respectively. Further, there were significant correlations between most CSHQ and SDQ domains. Female gender, older parental age, higher hyperactivity, and poorer prosocial behavior were associated with increased overall sleep disturbances.


Bi-phasic expression of Heterochromatin Protein 1 (HP1) during breast cancer progression: Potential roles of HP1 and chromatin structure in tumorigenesis.
Epigenetics in cancer prognosis and therapy is gaining recognition in recent years. Breast cancer is a genetic disease harboring numerous genetic mutations, including tumor suppressor BRCA1 and BRCA2 mutations. However, the functions of BRCA1 in cancer cells are also altered by non-genetic mechanisms, including DNA methylation and chromatin structure. Therefore, identification of epigenetic markers for breast cancer is very important for early diagnosis and effective therapy. This review focuses on recent findings on the roles of Heterochromatin protein 1 (HP1) in BRCA1 functions and breast cancer progression. We previously showed that BRCA1 function and breast cancer progression are frequently associated with HP1 expression level and potentially with chromatin structure. Herein, we suggest that bi-phasic expression of HP1 during breast cancer progression indicates dual roles of HP1 in tumorigenesis. Exploiting differential HP1 expression in tumors could lead to effective cancer therapy. Re-setting the chromatin structure may be a critical step for high-efficiency cancer therapy for many breast cancer patients.


[The development of the scientific and methodological basis of Russian forensic biology].
The development of the scientific and methodological basis of Russian forensic biology is retraced since the time of its formation as a self-contained branch of forensic medical examination. Comprehensive analysis of scientific publications and technical documents revealed principal directions along which forensic examination of evidential objects of biological origin developed. The wealth of research data obtained by forensic biologists during a long period (from the 1930s to the 2000s) is for the first time summarized and thoroughly analysed.


Cellular alterations in the alveolar wall in bleomycin-induced pulmonary fibrosis in rats. An ultrastructural morphometric study.
This study was designed to measure quantitatively changes in populations of major alveolar cell types, with a particular focus on nonmuscle, contractile interstitial cells, in lungs of rats 14 and 28 days after a single intratracheal instillation of bleomycin sulfate. Standard ultrastructural morphometric techniques, including stratified random sampling, were applied to airway-perfused lungs from bleomycin-instilled and control (saline-instilled) rats. Cell numbers and volumes were calculated for each major cell type populating the pulmonary parenchyma. At 14 days postinstillation, the number and volume of fibroblasts, contractile interstitial cells, Type II epithelial cells, and macrophages were increased significantly. At 28 days, the largest increase was in contractile interstitial cells, approximately tenfold in number and fourfold in volume over control cells. The number and volume of non-filament-containing fibroblasts did not differ between control and treated groups at this time. These data suggest proliferating fibroblasts undergo "differentiation" into cell types with increasing contractile capability during the development of fibrosis in this animal model. This study represents the first quantitative description of increases in contractile interstitial cells related to pulmonary fibrosis.


Improved outcomes with home-based administration of palivizumab: results from the 2000-2004 Palivizumab Outcomes Registry.
Palivizumab Outcomes Registry data collected during 4 years were examined to assess compliance and respiratory syncytial virus (RSV) hospitalization rates in high-risk children receiving palivizumab prophylaxis at home compared with an outpatient setting. Prospective observational registry enrolling high-risk infants who received > or = 1 dose of palivizumab throughout the 2000-2001 to 2003-2004 RSV seasons at participating U.S. pediatric sites. Registry data were analyzed for compliance and RSV hospitalization outcomes in 19,548 infants receiving doses at home versus an outpatient setting. Compliance with the injection regimen was determined by comparing the number of palivizumab injections received versus the projected number of anticipated doses and by comparing infants receiving all injections within a 35-day interval. Compliance was significantly greater for infants who received palivizumab at home (n = 1226) as compared with those who received palivizumab in a clinic or office (n = 17,641), whether measured by the number of doses received (88% versus 81%, P < 0.0001) or by the timing of doses (73% versus 66%, P < 0.0001). Infants who received palivizumab at home also had fewer RSV-associated hospitalizations compared with those who received palivizumab in a clinic or office [0.4% (5/1226) versus 1.2% (207/17,641), P = 0.0139]. Home administration of palivizumab was associated with a significantly higher rate of compliance and lower hospitalization rate for RSV illness in high-risk infants.


Reliability of a graded exercise test for assessing recovery from concussion.
To evaluate a graded treadmill test for retest reliability (RTR) and interrater reliability (IRR) in the evaluation of the physiologic effects of symptom exacerbation from concussion. Prospective case series (RTR) and blinded rater assessment of 10 actors portraying patients with and without symptom exacerbation (IRR). University Sports Medicine Concussion Clinic. For RTR, 21 refractory concussed patients (11 athletes and 10 nonathletes) and 10 healthy subjects; for IRR, 32 raters representing a variety of health care disciplines. For RTR, a Balke protocol treadmill test to symptom exacerbation before and after 2 to 3 weeks. For IRR, video recordings of actors during the treadmill test viewed by raters blinded to condition. For RTR, agreement of the tests for maximal heart rate (HR), systolic blood pressure, diastolic blood pressure, and rating of perceived exertion. For IRR, presence or absence of symptom exacerbation and the symptom exacerbation HR. Raters achieved a sensitivity of 99% for identifying actors with symptom exacerbation and a specificity of 89% for ruling out concussion symptoms and agreed on 304 of 320 observations (accuracy of 95%). The intraclass correlation coefficient for the symptom exacerbation HR was large at 0.90 (95% confidence interval, 0.78-0.98). The treadmill test had good RTR for maximum HR (intraclass correlation coefficient, 0.79) but not for systolic blood pressure, diastolic blood pressure, or rating of perceived exertion. The Balke exercise treadmill protocol has very good IRR and sufficient RTR for identifying patients with symptom exacerbation from concussion.


Exploring the mechanical properties of 3D-printed multilayer lattice structures for use in accommodative insoles.
Full-contact insoles fabricated from multilayer foams are the standard of care (SoC) for offloading and redistributing high plantar pressures in individuals with diabetes at risk of plantar ulceration and subsequent lower limb amputation. These devices have regional variations in total thickness and layer thickness to create conformity with a patient's foot. Recent work has demonstrated that metamaterials can be tuned to match the mechanical properties of SoC insole foams. However, for devices fabricated using a multilayer lattice structure, having regional variations in total thickness and layer thickness may result in regional differences in mechanical properties that have yet to be investigated. Three lattices, two dual-layer and one uniform-layer lattice structure, designed to model the mechanical properties of SoC insoles, were 3D-printed at three structure/puck thicknesses representing typical regions seen in accommodative insoles. The pucks underwent cyclic compression testing, and the stiffness profiles were assessed. Three pucks at three structure/puck thicknesses fabricated from SoC foams were also tested. Initial evaluations suggested that for the latticed pucks, structure thickness and density inversely impacted puck stiffness. Behaving most like the SoC pucks, a dual-layer lattice that increased in density as structure thickness increased demonstrated consistent stiffness profiles across puck thicknesses. Identifying a lattice with constant mechanical properties at various structure thicknesses may be important to produce a conforming insole that emulates the standard of care from which patient-specific/regional lattice modulations can be made.


Chromosome and DNA methylation dynamics during meiosis in the autotetraploid Arabidopsis arenosa.
Variation in chromosome number due to polyploidy can seriously compromise meiotic stability. In autopolyploids, the presence of more than two homologous chromosomes may result in complex pairing patterns and subsequent anomalous chromosome segregation. In this context, chromocenter, centromeric, telomeric and ribosomal DNA locus topology and DNA methylation patterns were investigated in the natural autotetraploid, Arabidopsis arenosa. The data show that homologous chromosome recognition and association initiates at telomeric domains in premeiotic interphase, followed by quadrivalent pairing of ribosomal 45S RNA gene loci (known as NORs) at leptotene. On the other hand, centromeric regions at early leptotene show pairwise associations rather than associations in fours. These pairwise associations are maintained throughout prophase I, and therefore likely to be related to the diploid-like behavior of A. arenosa chromosomes at metaphase I, where only bivalents are observed. In anthers, both cells at somatic interphase as well as at premeiotic interphase show 5-methylcytosine (5-mC) dispersed throughout the nucleus, contrasting with a preferential co-localization with chromocenters observed in vegetative nuclei. These results show for the first time that nuclear distribution patterns of 5-mC are simultaneously reshuffled in meiocytes and anther somatic cells. During prophase I, 5-mC is detected in extended chromatin fibers and chromocenters but interestingly is excluded from the NORs what correlates with the pairing pattern.


Detection of frameshift mutations of RIZ in gastric cancers with microsatellite instability.
To study the frameshift mutations of the retinoblastoma protein-interacting zinc finger gene RIZ in gastric cancer with microsatellite instability, and to identify two coding polyadenosine tracts of RIZ. Frameshift mutations at (A)8 and (A)9 tracts of RIZ were detected in 70 human gastric cancer (HGC) specimens by DHPLC and DNA sequencing. Microsatellite instability (MSI) status was assessed by two mononucleotide markers, BAT26 and BAT25, by means of denaturing high-performance liquid chromatography (DHPLC). In 70 HGC samples, 8 (11.4%) were found positive for instabilities at BAT26 and BAT25. In 7 of the 8 cases with instabilities at both BAT26 and BAT25 (MSI-H), 1 was unstable at BAT26 but stable at BAT25. Frameshift mutations were identified in 4 (57.1%) of the 7 samples with MSI-H in the (A)9 tract of RIZ without mutations in the (A)8 tract. In contrast, frameshift mutations were found in neither of the polyadenosine tracts in 63 samples of MSI-L or MSI stable tumors. Pro704 LOH detection in 4 cases with frameshift mutations did not find LOH in these cases. Frameshift mutations of RIZ may play an important role in gastric cancers with MSI.


Single polymer dynamics in an elongational flow.
The stretching of individual polymers in a spatially homogeneous velocity gradient was observed through use of fluorescently labeled DNA molecules. The probability distribution of molecular extension was determined as a function of time and strain rate. Although some molecules reached steady state, the average extension did not, even after a approximately 300-fold distortion of the underlying fluid element. At the highest strain rates, distinct conformational shapes with differing dynamics were observed. There was considerable variation in the onset of stretching, and chains with a dumbbell shape stretched more rapidly than folded ones. As the strain rate was increased, chains did not deform with the fluid element. The steady-state extension can be described by a model consisting of two beads connected by a spring representing the entropic elasticity of a worm-like chain, but the average dynamics cannot.


Effect of individual counseling on physical activity fitness and health: a randomized controlled trial in a workplace setting.
Physical inactivity and obesity are major public health problems. Our objective was to investigate the effectiveness of an individual counseling intervention at the workplace on physical activity fitness and health. Counseling content derived from the Patient-centered Assessment and Counseling for Exercise and Nutrition (PACE) program. A total of 299 employees of three municipal services in the Dutch town of Enschede were randomly allocated into intervention (n =131) and control group (n =168). Over a 9-month period, intervention group subjects were offered seven counseling sessions. Counseling was based on the individual's stage of behavioral change using PACE physical activity and nutrition protocols. Subjects in both the intervention and control group received written information about several lifestyle factors. Primary outcome measures were physical activity (total energy expenditure, during sports activities, during physical activity leisure time other than sports, and meeting the moderate-intensity public health recommendations); cardiorespiratory fitness; and prevalence of musculoskeletal symptoms. Secondary outcome measures were body composition (body mass index [BMI], and percentage of body fat measured via skinfold thicknesses); blood pressure; and blood cholesterol. There were significant positive effects on total energy expenditure, physical activity during sports, cardiorespiratory fitness, percentage of body fat, and blood cholesterol. No effects were found for the proportion of subjects meeting the public health recommendation of moderate-intensity physical activity, physical activity during leisure time other than sports, prevalence of musculoskeletal symptoms, body mass index, and blood pressure. Individual face-to-face counseling at the workplace based on PACE protocols positively influenced physical activity levels and some components of physical fitness. The implementation of workplace counseling programs for individuals should therefore be promoted.


Evaluation of anion influence on the formation and extraction capacity of ionic-liquid-based aqueous biphasic systems.
Extractive fermentation using aqueous biphasic systems (ABS) is a promising separation process since it provides a nondenaturing environment for biomolecules and improves the stability of cells. Due to environmental concerns and toxicity issues related with common volatile organic solvents, ionic liquids (ILs), a new class of nonvolatile alternative solvents, are being currently investigated for extraction purposes. In this work, a wide range of imidazolium-based ILs was studied aiming at obtaining new insights regarding their ability toward the formation of ABS and their capacity to the extraction of biomolecules. On the basis of the IL cations 1-ethyl-3-methylimidazolium and 1-butyl-3-methylimidazolium, the IL anion influence on ABS formation was assessed through their combination with chloride, bromide, acetate, hydrogensulfate, methanesulfonate, methylsulfate, ethylsulfate, trifluomethanesulfonate, trifluoroacetate, and dicyanamide. Ternary phase diagrams (and respective tie-lines) formed by these hydrophilic ILs, water, and the inorganic salt K(3)PO(4), were measured and are reported. The results indicate that the ability of an IL to induce ABS closely follows the decrease in the hydrogen bond accepting strength or the increase in the hydrogen bond acidity of the IL anion. In addition, the extraction capacity of the studied ABS was evaluated through their application to the extraction of an essential amino acid, L-tryptophan. It is shown that the partition coefficients obtained between the IL and the K(3)PO(4)-aqueous rich phases were substantially larger than those typically obtained with polymers-inorganic salts or polymers-polysaccharides aqueous systems.


Seeded aggregation and toxicity of {alpha}-synuclein and tau: cellular models of neurodegenerative diseases.
The deposition of amyloid-like filaments in the brain is the central event in the pathogenesis of neurodegenerative diseases. Here we report cellular models of intracytoplasmic inclusions of α-synuclein, generated by introducing nucleation seeds into SH-SY5Y cells with a transfection reagent. Upon introduction of preformed seeds into cells overexpressing α-synuclein, abundant, highly filamentous α-synuclein-positive inclusions, which are extensively phosphorylated and ubiquitinated and partially thioflavin-positive, were formed within the cells. SH-SY5Y cells that formed such inclusions underwent cell death, which was blocked by small molecular compounds that inhibit β-sheet formation. Similar seed-dependent aggregation was observed in cells expressing four-repeat Tau by introducing four-repeat Tau fibrils but not three-repeat Tau fibrils or α-synuclein fibrils. No aggregate formation was observed in cells overexpressing three-repeat Tau upon treatment with four-repeat Tau fibrils. Our cellular models thus provide evidence of nucleation-dependent and protein-specific polymerization of intracellular amyloid-like proteins in cultured cells.


Development and analysis of a nationwide cost database of acute-care hospitals in Japan.
Understanding of hospital cost is crucial to achieve an ideal balance between the assurance and improvement of patient safety and quality, and efficient use of finite resources. However, neither a standardized calculation methodology nor a large-scale database of costs in acute-care hospitals exists in Japan. This study aims to develop a standardized methodology, construct a nationwide cost database in Japan, analyse the characteristics of the database and examine the relationship between the cost and the charge from the viewpoint of an appropriate reflection of the cost to the price. We designed the costing framework, gathered the data for patients discharged from 139 acute-care hospitals in Japan between July 2004 and October 2004 and constructed a database containing information on 284,730 patients. The characteristics of the database and the relationship between the cost and the charge were investigated. In the nationwide database we constructed, a wide range in the average cost per hospitalization and average cost per diem was observed. A wide variation of cost components was seen across major diagnostic categories. Moreover, there was a high correlation between the cost and the charge (Correlation coefficient = 0.94). After designing a costing framework, a nationwide database comprised of individual case-level costs with components for acute-care hospitals in Japan was successfully developed. We hope this study contributes to appropriate decision making and helps motivate further research geared towards efficient hospital management and a rational payment system in Japan.


Synthesis of 3'-deoxy-3'-difluoromethyl azanucleosides from trans-4-hydroxy-l-proline.
[reaction: see text] Two strategies were tried to synthesize 3'-deoxy-3'-difluoromethyl azanucleosides. After the failure of the first route, the key intermediate 12 from trans-4-hydroxyproline 7 in 8 steps was stereoselectively prepared. The alcohol 12 was subjected to selective protection, oxidation, and difluoromethylenation to afford the fluorinated compound 18, whose hydrogenation was then systematically investigated. After a series of transformations of protecting groups, the resultant compounds 22 and 23 were oxidized to the desired lactams 24 and 25, which were successfully utilized to synthesize our target molecules, 3'-deoxy-3'-difluoromethyl azanucleosides 33, 34a, 34b, and 35.


Linkage between ATP consumption and mechanical unfolding during the protein processing reactions of an AAA+ degradation machine.
Proteolytic machines powered by ATP hydrolysis bind proteins with specific peptide tags, denature these substrates, and translocate them into a sequestered compartment for degradation. To determine how ATP is used during individual reaction steps, we assayed ClpXP degradation of ssrA-tagged titin variants with different stabilities in native and denatured forms. The rate of ATP turnover was 4-fold slower during denaturation than translocation. Importantly, this reduced turnover rate was constant during denaturation of native variants with different stabilities, but total ATP consumption increased with substrate stability, suggesting an iterative application of a uniform, mechanical unfolding force. Destabilization of substrate structure near the degradation tag accelerated degradation and dramatically reduced ATP consumption, revealing an important role for local protein stability in resisting denaturation. The ability to denature more stable proteins simply by using more ATP endows ClpX with a robust unfolding activity required for its biological roles in degradation and complex disassembly.


Enzymatic outside-in cross-linking enables single-step microcapsule production for high-throughput three-dimensional cell microaggregate formation.
Cell-laden hydrogel microcapsules enable the high-throughput production of cell aggregates, which are relevant for three-dimensional tissue engineering and drug screening applications. However, current microcapsule production strategies are limited by their throughput, multistep protocols, and limited amount of compatible biomaterials. We here present a single-step process for the controlled microfluidic production of single-core microcapsules using enzymatic outside-in cross-linking of tyramine-conjugated polymers. It was hypothesized that a physically, instead of the conventionally explored biochemically, controlled enzymatic cross-linking process would improve the reproducibility, operational window, and throughput of shell formation. Droplets were flown through a silicone delay line, which allowed for highly controlled diffusion of the enzymatic cross-linking initiator. The microcapsules' cross-linking density and shell thickness is strictly depended on the droplet's retention time in the delay line, which is predictably controlled by flow rate. The here presented hydrogel cross-linking method allows for facile and cytocompatible production of cell-laden microcapsules compatible with the formation and biorthogonal isolation of long-term viable cellular spheroids for tissue engineering and drug screening applications.


Herpesviruses assimilate kinesin to produce motorized viral particles.
Neurotropic alphaherpesviruses initiate infection in exposed mucosal tissues and, unlike most viruses, spread rapidly to sensory and autonomic nerves where life-long latency is established1. Recurrent infections arise sporadically from the peripheral nervous system throughout the life of the host, and invasion of the central nervous system may occur, with severe outcomes2. These viruses directly recruit cellular motors for transport along microtubules in nerve axons, but how the motors are manipulated to deliver the virus to neuronal nuclei is not understood. Here, using herpes simplex virus type I and pseudorabies virus as model alphaherpesviruses, we show that a cellular kinesin motor is captured by virions in epithelial cells, carried between cells, and subsequently used in neurons to traffic to nuclei. Viruses assembled in the absence of kinesin are not neuroinvasive. The findings explain a critical component of the alphaherpesvirus neuroinvasive mechanism and demonstrate that these viruses assimilate a cellular protein as an essential proviral structural component. This principle of viral assimilation may prove relevant to other virus families and offers new strategies to combat infection.


Intracranial pressure before and after cranioplasty: insights into intracranial physiology.
Decompressive craniectomy (DC) is an emergency neurosurgical procedure used in cases of severe intracranial hypertension or impending intracranial herniation. The procedure is often lifesaving, but it exposes the brain to atmospheric pressure in the subsequent rehabilitation period, which changes intracranial physiology and probably leads to complications such as hydrocephalus, hygromas, and "syndrome of the trephined." The objective of the study was to study the effect of cranioplasty on intracranial pressure (ICP), postural ICP changes, and intracranial pulse wave amplitude (PWA). The authors performed a prospective observational study including patients who underwent DC during a 12-month period. Telemetric ICP sensors were implanted in all patients at the time of DC. ICP was evaluated before and after cranioplasty during weekly measurement sessions including a standardized postural change program. Twelve of the 17 patients enrolled in the study had cranioplasty performed and were included in the present investigation. Their mean ICP in the supine position increased from -0.5 ± 4.8 mm Hg the week before cranioplasty to 6.3 ± 2.5 mm Hg the week after cranioplasty (p < 0.0001), whereas the mean ICP in the sitting position was unchanged (-1.2 ± 4.8 vs -1.1 ± 3.6 mm Hg, p = 0.90). The difference in ICP between the supine and sitting positions was minimal before cranioplasty (1.1 ± 1.8 mm Hg) and increased to 7.4 ± 3.6 mm Hg in the week following cranioplasty (p < 0.0001). During the succeeding 2 weeks of the follow-up period, the mean ICP in the supine and sitting positions decreased in parallel to, respectively, 4.6 ± 3.0 mm Hg (p = 0.0003) and -3.9 ± 2.7 mm Hg (p = 0.040), meaning that the postural ICP difference remained constant at around 8 mm Hg. The mean intracranial PWA increased from 0.7 ± 0.7 mm Hg to 2.9 ± 0.8 mm Hg after cranioplasty (p < 0.0001) and remained around 3 mm Hg throughout the following weeks. Cranioplasty restores normal intracranial physiology regarding postural ICP changes and intracranial PWA. These findings complement those of previous investigations on cerebral blood flow and cerebral metabolism in patients after decompressive craniectomy.


Contemporary pacemakers: what the primary care physician needs to know.
Pacemaker therapy is most commonly initiated because of symptomatic bradycardia, usually resulting from sinus node disease. Randomized multicenter trials assessing the relative benefits of different pacing modes have made possible an evidence-based approach to the treatment of bradyarrhythmias. During the past several decades, major advances in technology and in our understanding of cardiac pathophysiology have led to the development of new pacing techniques for the treatment of heart failure in the absence of bradycardia. Left ventricular or biventricular pacing may improve symptoms of heart failure and objective measurements of left ventricular systolic dysfunction by resynchronizing cardiac contraction. However, emerging clinical data suggest that long-term right ventricular apical pacing may have harmful effects. As the complexity of cardiac pacing devices continues to grow, physicians need to have a basic understanding of device indications, device function, and common problems encountered by patients with devices in the medical and home environment.


Strain-Induced Magnetic Transitions in SrMO2.5 (M = Mn, Fe) Thin Films with Ordered Oxygen Vacancies.
We examine the epitaxial-strain-induced phase transitions in thin films of perovskite-derived SrMnO2.5 and SrFeO2.5 exhibiting ordered oxygen vacancies (OOVs). We find that SrMnO2.5 hosts multiple magnetic transitions to other ordered states, including antiferromagnetic (AFM) and ferromagnetic (FM) orders of E*-AFM, C-AFM, and FM types depending on the compressive or tensile strain state. In contrast, no magnetic transitions occur in thin-film SrFeO2.5 (G-AFM to FM), whereas its bulk phase exhibits a hydrostatic pressure-induced AFM-to-FM transition. We explain the origin of these dependencies on the transition-metal configuration, that is, d4 Mn versus d5 Fe, and the relative orientation of the OOVs in the Ca2Mn2O5-type structure with respect to the epitaxial interface. We find that the magnetic phase stability can be predicted by using exchange striction arguments, with FM (AFM) spin interactions preferring longer (shorter) Mn-O bonds in the square pyramidal MnO5 unit comprising SrMnO2.5. Because the Mn-O bond lengths directly shrink or elongate to accommodate the applied stress without considerable polyhedral rotations, we show that compressive and tensile strain tune the unit cell structure to favor different combinations of exchange interactions that stabilize the various magnetic spin orders. Our study shows that the strong coupling between the OOV structure and spin orders with epitaxial strain is a promising route to achieve picoscale control of functional electronic and magnetic responses in complex oxide thin films.


Targeting V600EB-Raf and Akt3 using nanoliposomal-small interfering RNA inhibits cutaneous melanocytic lesion development.
Most events promoting early melanoma development are yet to be identified, but deregulation of the B-Raf and Akt3 signaling cascades is an important regulator of this process. Approximately 90% of normal moles and approximately 60% of early invasive cutaneous melanomas contain a T1799A B-Raf mutation ((V600E)B-Raf), leading to 10 times higher enzyme activity and constitutive activation of the mitogen-activated protein kinase pathway. Furthermore, approximately 70% of melanomas have elevated Akt3 signaling due to increased gene copy number and PTEN loss. Therefore, targeting (V600E)B-Raf and Akt3 signaling is necessary to prevent or treat cutaneous melanocytic lesions. Agents specifically targeting these proteins are needed, having fewer side effects than those inhibiting both normal and mutant B-Raf protein or targeting all three Akt isoforms. In this study, a unique nanoliposomal-ultrasound-mediated approach has been developed for delivering small interfering RNA (siRNA) specifically targeting (V600E)B-Raf and Akt3 into melanocytic tumors present in skin to retard melanoma development. Novel cationic nanoliposomes stably encapsulate siRNA targeting (V600E)B-Raf or Akt3, providing protection from degradation and facilitating entry into melanoma cells to decrease expression of these proteins. Low-frequency ultrasound using a lightweight four-cymbal transducer array enables penetration of nanoliposomal-siRNA complex throughout the epidermal and dermal layers of laboratory-generated or animal skin. Nanoliposomal-mediated siRNA targeting of (V600E)B-Raf and Akt3 led to a cooperatively acting approximately 65% decrease in early or invasive cutaneous melanoma compared with inhibition of each singly with negligible associated systemic toxicity. Thus, cationic nanoliposomes loaded with siRNA targeting (V600E)B-Raf and Akt3 provide an effective approach for targeted inhibition of early or invasive cutaneous melanomas.


A psychometric comparison of two carer quality of life questionnaires in Huntington's disease: implications for neurodegenerative disorders.
The carer impact of neurodegenerative disorders such as Huntington's disease (HD) is vast. Attempts to measure carer QoL in neurodegenerative disorders include the three-dimensional (Practical aspects of Caregiving, PC; Satisfaction with Life, SL; Feelings about Living with Huntington's disease, FL) Huntington's Disease Quality of Life Battery for Carers (HDQoL-C) and the unidimensional Alzheimer's Carer's Quality of Life Inventory (ACQLI). However, evidence regarding their psychometric properties is sparse. To test and compare the psychometric properties of the HDQoL-C, its short-form, and the ACQLI among HD carers. Data from 61 HD carers (36 women; mean age, 55) were analysed using traditional psychometric methods. Data completeness was good (>95% computable scale scores) but compromised for the PC and total HDQoL-C scales (≤80% computable scale scores). Scaling assumptions were supported for the SL and ACQLI scales (corrected item-total correlations ≥0.38; scaling success rates, 94-100%) but not for the PC, FL or total HDQoL-C scales (corrected item-total correlations ≥0.08; scaling success rates, 39-62%). Floor/ceiling effects were ≤9.8%. Reliabilities were ≥0.84, except for the PC scale (0.62). The HDQoL-C failed to exhibit suitability as a HD carer outcome measure, as two of its three scales did not meet basic psychometric criteria. The third scale (SL) did not outperform the ACQLI. This suggests that carer impact is not disease specific across neurodegenerative disorders.


Association of Cigarette Smoking with Risk of Chronic Musculoskeletal Pain: A Meta-Analysis.
Chronic musculoskeletal pain (CMP) management is a major global public health goal owing to increased social and economic burdens. However, the risk of CMP in smokers compared with nonsmokers remains uncertain. This study aims to determine the magnitude and importance of the relationship between cigarette smoking and risk of CMP. A meta-analysis of the CMP risk of cigarette smokers. We systematically searched PubMed, Embase, and Cochrane library databases from inception to August 2020. Data extraction and quality assessment were performed by 2 independent reviewers using a standardized extraction checklist. Data were pooled using a random-effects model. In this meta-analysis of 32 studies involving 296,109 participants, current smoking was associated with increased CMP risk (OR: 1.23, 95% CI: 1.09-1.40), whereas ever and past smoking did not show such an association (OR: 1.14, 95% CI: 0.95-1.37; OR: 1.06, 95% CI: 0.83-1.35, respectively). Stratified analyses showed that there was a marked significance in almost all strata of current smokers compared with non-smokers, except for mean age (>= 50 years), location of pain (neck pain, sacral pain, and knee pain), smoking frequency (occasionally), study design (cross-sectional), mean follow-up (< 10 years), and adjustment for confounding factors (>= 6). Interestingly, there was statistically negative association between cigarette smoking and knee pain risk in current smokers, ever smokers, and past smokers. The major limitation of this meta-analysis relates to the heterogeneities across included studies. Cigarette smoking was associated with increased risk of CMP. In view of the high prevalence of smoking in many countries and the increasing number of CMP patients worldwide, reducing tobacco use should be an important public health strategy to prevent and control the global epidemic of CMP. Future research should attempt to establish whether this association is causal and clarify its mechanisms.


Increased lodging resistance in long-culm, low-lignin gh2 rice for improved feed and bioenergy production.
Lignin modification has been a breeding target for the improvements of forage digestibility and energy yields in forage and bioenergy crops, but decreased lignin levels are often accompanied by reduced lodging resistance. The rice mutant gold hull and internode2 (gh2) has been identified to be lignin deficient. GH2 has been mapped to the short arm of chromosome 2 and encodes cinnamyl-alcohol dehydrogenase (CAD). We developed a long-culm variety, 'Leaf Star', with superior lodging resistance and a gh phenotype similar to one of its parents, 'Chugoku 117'. The gh loci in Leaf Star and Chugoku 117 were localized to the same region of chromosome 2 as the gh2 mutant. Leaf Star had culms with low lignin concentrations due to a natural mutation in OsCAD2 that was not present in Chugoku 117. However, this variety had high culm strength due to its strong, thick culms. Additionally, this variety had a thick layer of cortical fiber tissue with well-developed secondary cell walls. Our results suggest that rice can be improved for forage and bioenergy production by combining superior lodging resistance, which can be obtained by introducing thick and stiff culm traits, with low lignin concentrations, which can be obtained using the gh2 variety.


T cells and aging.
Deterioration of the immune system with aging ("immunosenescence") is believed to contribute to morbidity and mortality in man due to the greater incidence of infection, as well as possibly autoimmune phenomena and cancer in the aged. Dysregulation of T cell function is thought to play a critical part in these processes. Factors contributing to T cell immunosenescence may include a) stem cell defects, b) thymus involution, c) defects in antigen presenting cells (APC), d) aging of resting immune cells, e) disrupted activation pathways in immune cells, f) replicative senescence of clonally expanding cells. This review aims to consider the current state of knowledge on the scientific basis for and potential clinical relevance of those factors in immunosenescence.


Involvement of the telomeric protein Pin2/TRF1 in the regulation of the mitotic spindle.
Pin2/TRF1 was independently identified as a telomeric DNA-binding protein (TRF1) that regulates telomere length, and as a protein (Pin2) that can bind the mitotic kinase NIMA and suppress its lethal phenotype. We have previously demonstrated that Pin2/TRF1 levels are cell cycle-regulated and its overexpression induces mitotic arrest and then apoptosis. This Pin2/TRF1 activity can be potentiated by microtubule-disrupting agents, but suppressed by phosphorylation of Pin2/TRF1 by ATM; this negative regulation is critical in mediating for many, but not all, ATM-dependent phenotypes. Interestingly, Pin2/TRF1 specifically localizes to mitotic spindles in mitotic cells and affects the microtubule polymerization in vitro. These results suggest a role of Pin2/TRF1 in mitosis. However, nothing is known about whether Pin2/TRF1 affects the spindle function in mitotic progression. Here we characterized a new Pin2/TRF1-interacting protein, EB1, that was originally identified in our yeast two-hybrid screen. Pin2/TRF1 bound EB1 both in vitro and in vivo and they also co-localize at the mitotic spindle in cells. Furthermore, EB1 inhibits the ability of Pin2/TRF1 to promote microtubule polymerization in vitro. Given that EB1 is a microtubule plus end-binding protein, these results further confirm a specific interaction between Pin2/TRF1 and the mitotic spindle. More importantly, we have shown that inhibition of Pin2/TRF1 in ataxia-telangiectasia cells is able to fully restore their mitotic spindle defect in response to microtubule disruption, demonstrating for the first time a functional involvement of Pin2/TRF1 in mitotic spindle regulation.


Synthetic double-stranded RNA induces retinoic acid-inducible gene-I in mouse osteoblastic cells.
Retinoic acid inducible gene-I (RIG-I) is a member of the DExH box family of proteins. RIG-I acts as a sensor of viral infections through the recognition of viral double-stranded RNA (dsRNA). Recently, it was demonstrated that polyinosinic acid:polycytidylic acid [poly(I):poly(C)], a synthetic dsRNA analogue, induced the expression of RIG-I in various cell types, such as vascular endothelial cells and gingival fibroblasts. However, it remains unclear whether RIG-I is induced in osteoblasts in response to poly(I):poly(C). In the present study, we investigated the effects of poly(I):poly(C) on the expression of RIG-I in mouse osteoblastic MC3T3-E1(E1) cells. We found that poly(I):poly(C) increased the expression level of RIG-I in E1 cells, and that recombinant interferon-β (IFN-β) induced the expression of RIG-I mRNA in E1 cells. An anti-IFN-β neu-tralizing antibody partially inhibited poly(I):poly(C)-induced RIG-I expression. These results indicate that RIG-I production is induced by poly(I):poly(C)-provoked IFN-β in mouse osteoblastic E1 cells. We suggest that osteoblasts are involved in antiviral defense as well as in bone metabolism. Results of further studies will provide more clues regarding the molecular function of osteoblasts in viral infection.


Combining affinity purification and mass spectrometry to define the network of the nuclear proteins interacting with the N-terminal region of FMRP.
Fragile X-Syndrome (FXS) represents the most common inherited form of intellectual disability and the leading monogenic cause of Autism Spectrum Disorders. In most cases, this disease results from the absence of expression of the protein FMRP encoded by the FMR1 gene (Fragile X messenger ribonucleoprotein 1). FMRP is mainly defined as a cytoplasmic RNA-binding protein regulating the local translation of thousands of target mRNAs. Interestingly, FMRP is also able to shuttle between the nucleus and the cytoplasm. However, to date, its roles in the nucleus of mammalian neurons are just emerging. To broaden our insight into the contribution of nuclear FMRP in mammalian neuronal physiology, we identified here a nuclear interactome of the protein by combining subcellular fractionation of rat forebrains with pull- down affinity purification and mass spectrometry analysis. By this approach, we listed 55 candidate nuclear partners. This interactome includes known nuclear FMRP-binding proteins as Adar or Rbm14 as well as several novel candidates, notably Ddx41, Poldip3, or Hnrnpa3 that we further validated by target-specific approaches. Through our approach, we identified factors involved in different steps of mRNA biogenesis, as transcription, splicing, editing or nuclear export, revealing a potential central regulatory function of FMRP in the biogenesis of its target mRNAs. Therefore, our work considerably enlarges the nuclear proteins interaction network of FMRP in mammalian neurons and lays the basis for exciting future mechanistic studies deepening the roles of nuclear FMRP in neuronal physiology and the etiology of the FXS.


Two-Stage Revision Total Hip Arthroplasty With a Specific Articulating Antibiotic Spacer Design: Reliable Periprosthetic Joint Infection Eradication and Functional Improvement.
Two-stage revision utilizing spacers loaded with high-dose antibiotic cement prior to reimplantation remains the gold standard for treatment of periprosthetic joint infections (PJI) in total hip arthroplasty (THA) in North America, but there is a paucity of data on mid-term outcomes. We sought to analyze the survivorship free of infection, clinical outcomes, and complications of a specific articulating spacer utilized during 2-stage revision. One hundred thirty-five hips (131 patients) undergoing a 2-stage revision THA for PJI with a specific articulating antibiotic spacer design from 2005 to 2013 were retrospectively reviewed. Infections were classified according to the Musculoskeletal Infection Society criteria. Mean age at resection was 65 years and mean follow-up was 5 years (rang, 2-10). Survivorship free of any infection after reimplantation was 92% and 88% at 2 and 5 years, respectively. Patients with a host-extremity grade of C3 compared to all patients with a host grade of A [hazard ratio (HR) 4.1, P = .05] were significant risk factors for poorer infection-free survivorship after reimplantation. Harris hip scores improved from a mean of 58 to a mean of 71 in the spacer phase (P = .002) and a mean of 81 post-reimplantation (P = .001). Fourteen (10%) patients dislocated after reimplantation, 9 (7%) of which required re-revision. Trochanteric deficiency (HR 19, P < .0001), dislocation of the articulating spacer prior to reimplantation [which occurred in 7 (5%) patients, 5 of whom subsequently dislocated the definitive implant] (HR 16, P < .0001), and female gender (HR 5, P = .002) were significant risk factors for post-reimplantation dislocation. Insertion of an articulating antibiotic spacer during a 2-stage revision THA for PJI demonstrates reliable infection eradication and improvement in clinical function, including the spacer phase. Patients with trochanteric deficiency and an articulating spacer dislocation are at high risk of post-reimplantation dislocation; judicial use of a dual-mobility or constrained device should be considered in these patients.


On the ATP-dependent activation of the radical enzyme (R)-2-hydroxyisocaproyl-CoA dehydratase.
Members of the 2-hydroxyacyl-CoA dehydratase enzyme family catalyze the β,α-dehydration of various CoA-esters in the fermentation of amino acids by clostridia. Abstraction of the nonacidic β-proton of the 2-hydroxyacyl-CoA compounds is achieved by the reductive generation of ketyl radicals on the substrate, which is initiated by the transfer of an electron at low redox potentials. The highly energetic electron needed on the dehydratase is donated by a [4Fe-4S] cluster containing ATPase, termed activator. We investigated the activator of the 2-hydroxyisocaproyl-CoA dehydratase from Clostridium difficile. The activator is a homodimeric protein structurally related to acetate and sugar kinases, Hsc70 and actin, and has a [4Fe-4S] cluster bound in the dimer interface. The crystal structures of the Mg-ADP, Mg-ADPNP, and nucleotide-free states of the reduced activator have been solved at 1.6-3.0 Å resolution, allowing us to define the position of Mg(2+) and water molecules in the vicinity of the nucleotides and the [4Fe-4S] cluster. The structures reveal redox- and nucleotide dependent changes agreeing with the modulation of the reduction potential of the [4Fe-4S] cluster by conformational changes. We also investigated the propensity of the activator to form a complex with its cognate dehydratase in the presence of Mg-ADP and Mg-ADPNP and together with the structural data present a refined mechanistic scheme for the ATP-dependent electron transfer between activator and dehydratase.


Boll weevil (Coleoptera: Curculionidae) survival through cotton gin trash fans.
There is concern that cotton gins may serve as loci for reintroduction of boll weevils, Anthonomus grandis grandis Boheman, to eradicated or suppressed zones when processing weevil-infested cotton from neighboring zones. Previous work has shown that virtually all weevils entering the gin in the seed cotton will be removed before they reach the gin stand. Those not killed by the seed cotton cleaning machinery will be shunted alive into the trash fraction, which passes through a centrifugal trash fan before exiting the gin. The objective of this study was to determine survival potential of boll weevils passed through a trash fan. Marked adult weevils were distributed in gin trash and fed through a 82.6-cm (32.5-in.) diameter centrifugal fan operated across a range of fan-tip speeds. A small number of boll weevils were recovered alive immediately after passage through the fan, but all were severely injured and did not survive 24 h. In another experiment, green bolls infested with both adult- and larval-stage weevils were fed through the fan. Several teneral adults survived 24 h, and there was no evidence that fan-tip speed affected either initial survival of weevils, or the number of unbroken boll locks that could harbor an infesting weevil. Thus, designating a minimum fan-tip speed for ensuring complete kill is not possible for the boll weevil. Experiments suggest that a device installed in a gin that partially crushes or cracks bolls open before entering a trash fan will increase mortality, possibly enough that further precautions would be unnecessary.


Should a drain be placed in early breast cancer surgery?
The current surgical debate has led to a reduction in the extent of surgery performed and thereby to a reduced occurrence of surgical trauma and, over the recent years, reduced seroma formation. This reduction in surgical procedures calls the need for a drain into question. Using Google Scholar and the National Library of Medicine (PubMed), a literature review was performed on systematic reviews and meta-analyses regarding breast cancer surgery ± axillary dissection. Additionally, randomized trials for the time period after the last systematic review were included and evaluated according to the Jadad score. The search returned 5 systematic reviews, in which a total of 1,075 patients were included (537 cases and 538 controls). Since the last review, no prospective randomized trial meeting the inclusion criteria has been published. The current reviews conclude that insertion of a drain is associated with a longer hospital stay and reduced seroma formation. The data regarding wound infection and drain insertion is inconclusive. The omission of a drain is associated with early discharge, reduced postsurgical pain, and early mobilization, but also with an increase in outpatient seroma aspirations. The omission of a drain is possible in early breast cancer surgery (wide local excision and sentinel node biopsy) with adequate surgical techniques and instruments.


Materials for phantoms for terahertz pulsed imaging.
Phantoms are commonly used in medical imaging for quality assurance, calibration, research and teaching. They may include test patterns or simulations of organs, but in either case a tissue substitute medium is an important component of the phantom. The aim of this work was to identify materials suitable for use as tissue substitutes for the relatively new medical imaging modality terahertz pulsed imaging. Samples of different concentrations of the candidate materials TX151 and napthol green dye were prepared, and measurements made of the frequency-dependent absorption coefficient (0.5 to 1.5 THz) and refractive index (0.5 to 1.0 THz). These results were compared qualitatively with measurements made in a similar way on samples of excised human tissue (skin, adipose tissue and striated muscle). Both materials would be suitable for phantoms where the dominant mechanism to be simulated is absorption (approximately 100 cm(-1) at 1 THz) and where simulation of the strength of reflections from boundaries is not important; for example, test patterns for spatial resolution measurements. Only TX151 had a frequency-dependent refractive index close to that of tissue, and could therefore be used to simulate the layered structure of skin, the complexity of microvasculature or to investigate frequency-dependent interference effects that have been noted in terahertz images.


Effect of forage to concentrate ratio with sorghum silage as a source of forage on rumen fermentation, N balance, and purine derivative excretion in limit-fed dairy heifers.
Sorghum silage has been shown to be a good alternative to corn silage for dairy cows; however, studies regarding heifers are insufficiently explored. Therefore, the objective of this study was to evaluate effects of changing forage to concentrate ratio (FOR:CON) in diets based on sorghum silage on N digestibility, rumen fermentation, N balance, C excretion, and microbial N yield in limit-fed dairy heifers. A split-plot 4×4 Latin square design with 19-d periods (15 d of adaptation and 4 d of sampling) was conducted with 8 rumen cannulated dairy heifers (age 13.7±0.6mo and weight 364.8±17.6kg). Heifers were fed sorghum silage-based diets with 4 FOR:CON (85:15, 75:25, 65:35, and 55:45) balanced for similar metabolizable energy intake per unit of body weight and crude protein concentration. Diets were fed to allow 900 to 1,000g/d body weight gain and were fed once daily. Total collection of feces and urine was completed on d 15 to 19 to determine N, C, urea N, allantoin, uric acid, and creatinine excretion. Rumen contents were sampled on d 19 at 0, 1.5, 3, 4.5, 6, 9, 13, 17, 21, and 23h after feeding to measure pH, volatile fatty acid (VFA), ammonia-N, and free AA concentrations. The pH decreased linearly while ammonia-N and free AA levels increased linearly with decreasing FOR:CON of diets. Although mean total VFA did not differ among treatment diets, molar proportions of VFA did. Acetate proportion decreased while propionate and butyrate increased with decreasing FOR:CON. Intake of N and urea N excretion decreased with decreasing forage proportion in diets while total N excretion, apparent N digestibility, and N retention were not different. Intake of C and excretion in feces (g/d) decreased linearly with decreasing FOR:CON in diets. Creatinine, allantoin, and uric acid excretion were not affected by FOR:CON; however, microbial N yield tended to increase linearly with greater concentrate in diets. Heifers limit fed diets based on sorghum silage demonstrated the effect of available ammonia-N and readily fermentable carbohydrates with subsequent effects on nutrient utilization when different FOR:CON were applied. Based on the presented results, FOR:CON 65:35 had the most suitable balance of available ammonia-N and readily fermentable carbohydrates for the most optimal results.