Myroides pelagicus sp. nov., isolated from seawater in Thailand.
An aerobic, Gram-negative, non-motile, yellow-to-orange pigmented and rod-shaped bacterium, designated strain SM1(T), was isolated from seawater in Thailand and subjected to a polyphasic taxonomic study. Phylogenetic analyses based on 16S rRNA gene sequences revealed that the novel isolate shared 93-95 % sequence similarity with species of the genus Myroides. The DNA-DNA relatedness values of strain SM1(T) with Myroides odoratimimus JCM 7460(T) and Myroides odoratus JCM 7458(T) were below 70 %. The DNA G+C content of strain SM1(T) was 33.6 mol%, the major menaquinone was MK-6 and the major fatty acids were C(15 : 0) iso, C(17 : 1)omega9c iso and C(17 : 0) iso 3-OH. Based on its phenotypic and chemotaxonomic characteristics, strain SM1(T) was classified as representing a novel species of the genus Myroides, for which the name Myroides pelagicus sp. nov. is proposed. The type strain is SM1(T) (=IAM 15337(T)=KCTC 12661(T)).


Dehydration stress and Mayaro virus vector competence in Aedes aegypti.
Relative humidity (RH) is an environmental variable that affects mosquito physiology and can impact pathogen transmission. Low RH can induce dehydration in mosquitoes, leading to alterations in physiological and behavioral responses such as blood-feeding and host-seeking behavior. We evaluated the effects of a temporal drop in RH (RH shock) on mortality and Mayaro virus vector competence in Ae. aegypti. While dehydration induced by humidity shock did not impact virus infection, we detected a significant effect of dehydration on mosquito mortality and blood-feeding frequency, which could significantly impact transmission dynamics.


Lysosomes, caspase-mediated apoptosis, and cytoplasmic activation of P21, but not cell senescence, participate in a redundant fashion in embryonic morphogenetic cell death.
Micromass cultures of embryonic limb skeletal progenitors replicate the tissue remodelling processes observed during digit morphogenesis. Here, we have employed micromass cultures in an in vitro assay to study the nature of cell degeneration events associated with skeletogenesis. In the assay, "naive" progenitors obtained from the autopod aggregate to form chondrogenic nodules and those occupying the internodular spaces exhibit intense apoptosis and progressive accumulation of larger cells, showing intense SA-β-Gal histochemical labelling that strictly overlaps with the distribution of neutral red vital staining. qPCR analysis detected intense upregulation of the p21 gene, but P21 immunolabelling showed cytoplasmic rather than the nuclear distribution expected in senescent cells. Semithin sections and transmission electron microscopy confirmed the presence of canonical apoptotic cells, degenerated cell fragments in the process of phagocytic internalization by the neighbouring cells, and large vacuolated cells containing phagosomes. The immunohistochemical distribution of active caspase 3, cathepsin D, and β-galactosidase together with the reduction in cell death by chemical inhibition of caspases (Q-VAD) and lysosomal cathepsin D (Pepstatin A) supported a redundant implication of both pathways in the dying process. Chemical inhibition of P21 (UC2288) revealed a complementary role of this factor in the dying process. In contrast, treatment with the senolytic drug Navitoclax increased cell death without changing the number of cells positive for SA-β-Gal. We propose that this model of tissue remodelling involves the cooperative activation of multiple degradation routes and, most importantly, that positivity for SA-β-Gal reflects the occurrence of phagocytosis, supporting the rejection of cell senescence as a defining component of developmental tissue remodelling.


[Integrated skills laboratory concept for undergraduate training in internal medicine].
An amendment to the German medical curriculum in April 2002 will place basic practical skills at the centre of medical training. We report here on the implementation and evaluation of an obligatory, tutor-guided, and integrated skills laboratory concept in the field of internal medicine. To test the effectiveness of a skills laboratory training on OSCE performance a pilot study was carried out. The experimental group, of 77 students, participated in seven sessions of communication training, skills laboratory training, and bedside teaching, each lasting one and a half hours. The control group of 66 students had as many sessions but was only offered bedside-teaching. The evaluation of acceptance of skills' training as well as the related increase in individual competence is on-going (summer term 2004: n = 176 students). The integrated skills laboratory concept was rated at 3.5 (SD = 1.2) on a 5-point scale and was acknowledged as practice-oriented (M = 4.2; SD = 1.0) and relevant for doctors' everyday lives (M = 3.6; SD = 1.1). Increased levels of competence according to individual self-evaluations proved to be highly significant (p<.001), and results of the pilot study showed that the experimental group had a significantly better OSCE performance than the control group (p<.001). This pilot study shows that curriculum changes promoting basic clinical skills are effective and lead to an improved practical education of tomorrow's physicians. The integrated skills laboratory concept is well accepted and leads to a relevant increase in competence in the practice of internal medical. The presented skills laboratory concept in internal medicine is proving to be a viable and efficient learning tool.


Ultra-short stems with proximal load transfer: Clinical and radiographic results at five-year follow-up.
Bone stock preservation is crucial when performing THR in young patients. Following this philosophy, an original ultra-short custom-made implant with extensive proximal load transfer was developed. It presents three very innovative features: absence of the diaphyseal portion of the stem, a well defined lateral flare with load transfer on the lateral column of the femur and a very high femoral neck cut. These innovations resulted in a very conservative implant both of the bone stock and the soft tissues. In this study we reviewed the X-rays of 111 patients with 131 primary total hip replacements performed with this implant. The average pain score using the Harris Hip Score system, at an average of five years after surgery, was 42 of 44 points; 95 per cent (124) of the patients had no or slight pain. We had no cases of thigh pain. None of the patients required a femoral stem revision. Two cases had to be re-operated for polyethylene liner exchange but the femoral implant was stable and left in place. At five years, all implants appeared radiographically stable with well maintained proximal bone stock. It was concluded that the geometry of this implant provides significant initial stability, which seems to be preserved throughout a long follow-up period. This study validates the assumption that torsional loads can be controlled even without the diaphyseal portion of the stem and that neck preservation combined with lateral flare support guarantees a more natural loading of the femur.


Rod photoreceptors protect from cone degeneration-induced retinal remodeling and restore visual responses in zebrafish.
Humans are largely dependent upon cone-mediated vision. However, death or dysfunction of rods, the predominant photoreceptor subtype, results in secondary loss of cones, remodeling of retinal circuitry, and blindness. The changes in circuitry may contribute to the vision deficit and undermine attempts at restoring sight. We exploit zebrafish larvae as a genetic model to specifically characterize changes associated with photoreceptor degenerations in a cone-dominated retina. Photoreceptors form synapses with two types of second-order neurons, bipolar cells, and horizontal cells. Using cell-specific reporter gene expression and immunolabeling for postsynaptic glutamate receptors, significant remodeling is observed following cone degeneration in the pde6c(w59) larval retina but not rod degeneration in the Xops:mCFP(q13) line. In adults, rods and cones are present in approximately equal numbers, and in pde6c(w59) mutants glutamate receptor expression and synaptic structures in the outer plexiform layer are preserved, and visual responses are gained in these once blind fish. We propose that the abundance of rods in the adult protects the retina from cone degeneration-induced remodeling. We test this hypothesis by genetically manipulating the number of rods in larvae. We show that an increased number and uniform distribution of rods in lor/tbx2b(p25bbtl) or six7 morpholino-injected larvae protect from pde6c(w59)-induced secondary changes. The observations that remodeling is a common consequence of photoreceptor death across species, and that in zebrafish a small number of surviving photoreceptors afford protection from degeneration-induced changes, provides a model for systematic analysis of factors that slow or even prevent the secondary deteriorations associated with neural degenerative disease.


Behavior of attenuated mycobacteria in organs of neonatal and adult mice.
The consequences of Calmette-Guérin bacillus (BCG) vaccination were followed in newborn and adult mice. BCG failed to multiply in the organs of adult mice when administered peritoneally. In contrast, extensive multiplication of the vaccine occurred in both splenic and pulmonary tissue after its peritoneal administration to newborn mice. This absence of tuberculostasis occurred during the period when the animal's spleen, lung, and thymus were rapidly growing. Animals achieved similar levels of resistance to virulent respiratory challenge 10 wk after vaccination irrespective of whether the vaccine had been administered when the mice were newly born and BCG had multiplied in vivo or administered when the animals were fully grown and the BCG had remained in the dormant state. Although neonatal infection with BCG was severe, as shown by the large numbers of organisms recovered from the animals' tissues, the animals suffered no mortality or overt signs of disease. Neonatal vaccination did not significantly affect either the animal's growth rate or the gross development of its organs.


Death rates for acquired hypothyroidism and thyrotoxicosis in English populations (1979-2010): comparison of underlying cause and all certified causes.
Overt hypothyroidism and thyrotoxicosis have widespread systemic effects and are associated with increased mortality. Most death certificates that include them do not have the thyroid disease coded as the underlying cause of death. To describe regional (1979-2010) and national (1995-2010) trends in mortality rates for acquired hypothyroidism and thyrotoxicosis, analysing all certified causes of death (termed 'mentions') and not just the underlying cause. Analysis of death registration data. Analysis of data for the Oxford region (mentions available from 1979) and English national data (mentions available from 1995). The data were grouped in periods defined by different national rules for selecting the underlying cause of death (1979-83, 1984-92, 1993-2000 and 2001-10) and were also analysed as single calendar years. Mentions mortality for acquired hypothyroidism in the Oxford region declined significantly from 1979 to 2010: the average annual percentage change (AAPC) was -2.6% (95% confidence intervals -3.5, -1.8). Most of the decrease occurred during the 1980s. The AAPC in rates for later years in England (1995-2010) was non-significant at 0.2% (-0.7, 1.0). Mortality rates for thyrotoxicosis decreased significantly: the AAPC was -2.8% (-4.1, -1.5) in the Oxford region and -3.8% (-4.7, -3.0) in England. In England, between 2001 and 2010, hypothyroidism or thyrotoxicosis was coded as the underlying cause of death on, respectively, 17 and 24% of death certificates that included them. Mortality rates for hypothyroidism and thyrotoxicosis have fallen substantially. The fall is probably wholly or mainly a result of improved care.


Quantum noise measurements for fourier multiaperture emission tomography.
The mean and variance of an image reconstructed by Fourier multiaperture emission tomography are estimated by a physical experiment and by simulations. Results correspond closely to those obtained from a stochastic model of the noise process. A new scheme to enhance the statistical performance is proposed which improves the signal-to-noise ratio by 35 percent.


Motor impairment in rats exposed to PCBs and methylmercury during early development.
Epidemiological and laboratory studies indicate that polychlorinated biphenyls (PCBs) and methyl mercury (MeHg) may have additive or interactive adverse effects on nervous system function. Prior studies have shown that high doses of MeHg target the cerebellum and impair balance and coordination, but the effects of PCBs on cerebellar function were unknown. In addition, the combined effects of PCBs and MeHg on cerebellar function have not been studied previously. Therefore, we investigated the effects of developmental exposure to PCBs, MeHg, or PCBs + MeHg on three motor tasks that involve cerebellar functions. Female Long-Evans rats were exposed to MeHg (0.5 ppm in drinking water), PCBs (6-mg/kg/d Aroclor 1254), PCBs + MeHg, or vehicle only beginning 4 weeks prior to breeding, through pregnancy, and continuing through postnatal day (PND) 16. Starting at approximately PND 60, one male and one female from each litter were tested on three motor tasks that involve cerebellar function. PCB + MeHg-exposed rats were impaired relative to the controls on a task requiring them to traverse a rotating rod. Rats exposed to PCBs alone were also somewhat impaired relative to the controls, whereas MeHg-exposed rats were not significantly different from the controls. There were no statistically significant deficits related to PCB or MeHg exposure on a vertical rope-climbing test or a parallel bar test. Our results demonstrate that the possibility of additive neurotoxic effects of PCBs and MeHg needs to be seriously considered.


Forecasting phenology under global warming.
As a consequence of warming temperatures around the world, spring and autumn phenologies have been shifting, with corresponding changes in the length of the growing season. Our understanding of the spatial and interspecific variation of these changes, however, is limited. Not all species are responding similarly, and there is significant spatial variation in responses even within species. This spatial and interspecific variation complicates efforts to predict phenological responses to ongoing climate change, but must be incorporated in order to build reliable forecasts. Here, we use a long-term dataset (1953-2005) of plant phenological events in spring (flowering and leaf out) and autumn (leaf colouring and leaf fall) throughout Japan and South Korea to build forecasts that account for these sources of variability. Specifically, we used hierarchical models to incorporate the spatial variability in phenological responses to temperature to then forecast species' overall and site-specific responses to global warming. We found that for most species, spring phenology is advancing and autumn phenology is getting later, with the timing of events changing more quickly in autumn compared with the spring. Temporal trends and phenological responses to temperature in East Asia contrasted with results from comparable studies in Europe, where spring events are changing more rapidly than are autumn events. Our results emphasize the need to study multiple species at many sites to understand and forecast regional changes in phenology.


Laser remote sensing of hydrazine, MMH, and UDMH using a differential-absorption CO2 lidar.
A dual mini-TEA CO(2) laser differential-absorption lidar system has been used to test the remote sensing of hydrazine, unsymmetrical dimethylhydrazine (UDMH), and monomethylhydrazine (MMH) in atmospheric conditions. Average concentrations of these compounds were measured using backscattered laser radiation from a target located at a range of 2.7 km. The experimental results indicate that average atmospheric concentration levels of the hydrazine compounds of the order of 40-100 ppb can be detected over ranges between 0.5 and 5 km. The level of concentration sensitivity over this interval was found to be limited primarily by atmospheric fluctuations. An investigation of the effect of these fluctuations on measurement uncertainties indicated that the fluctuations reduce the benefits of signal averaging over N pulses significantly below the expected square root of N improvement. It is also shown that uncertainties due to long-term atmospheric drifts can be effectively reduced through use of dual-laser lidar return ratios.


Characteristics of patients with shingles admitted to a district general hospital.
Little is known about why some patients with shingles are admitted to hospital. We reviewed 72 case notes from a list of 80 patients admitted to hospital with shingles over a six-year period. Pain was the main complaint of the patients admitted, most of whom were elderly and lived alone. The commonest site of involvement in hospital admissions was the eye (herpes zoster ophthalmicus). Diagnosis of shingles was made after admission in 12 patients, eight of whom had originally been diagnosed as having an acute medical or surgical condition. We conclude that the prodromal phase of shingles may lead to misdiagnosis.


Autonomic and adrenocortical reactivity and buccal cell telomere length in kindergarten children.
To examine associations between autonomic nervous system and adrenocortical reactivity to laboratory stressors and buccal cell telomere length (BTL) in children. The study sample comprised 78 children, aged 5 to 6 years, from a longitudinal cohort study of kindergarten social hierarchies, biologic responses to adversity, and child health. Buccal cell samples and reactivity measures were collected in the spring of the kindergarten year. BTL was measured by real-time polymerase chain reaction, as the telomere-to-single-copy gene ratio. Parents provided demographic information; parents and teachers reported children's internalizing and externalizing behavior problems. Components of children's autonomic (heart rate, respiratory sinus arrhythmia [RSA], and preejection period [PEP]) and adrenocortical (salivary cortisol) responses were monitored during standardized laboratory challenges. We examined relationships between reactivity, internalizing and externalizing behaviors, and BTL, adjusted for age, race, and sex. Heart rate and cortisol reactivity were inversely related to BTL, PEP was positively related to BTL, and RSA was unrelated to BTL. Internalizing behaviors were also inversely related to BTL (standardized β = -0.33, p = .004). Split at the median of reactivity parameters, children with high sympathetic activation (decreasing PEP), and parasympathetic withdrawal (decreasing RSA) did not differ with regard to BTL. However, children with both this profile and high cortisol reactivity (n = 12) had significantly shorter BTL (0.80 versus 1.00; χ² = 7.6, p = .006), compared with other children. The combination of autonomic and adrenocortical reactivity was associated with shorter BTL in children. These data suggest that psychophysiological processes may influence, and that BTL may be a useful marker of, early biologic aging.


Role of porin proteins in acquisition of transferrin iron by enteropathogens.
Acquisition of iron from key innate immune defence proteins such as transferrin (Tf) and lactoferrin is an important mechanism by which pathogenic bacteria obtain essential iron for growth within their host. Bacterial species that do not produce siderophores often use specific Tf-binding proteins, the best characterized being the Neisseriaceae-type Tf-binding proteins TbpA and TbpB. Previous work from our laboratory has shown that siderophore-producing enteric species such as Escherichia coli also readily bind Tf, although no genomic evidence exists for Tbp-like Tf-binding proteins. Application of proteomic analyses and molecular mutagenesis strategies to an enteropathogenic E. coli identified the OmpA and OmpC porins as Tf-binding proteins. Mutagenesis of the ompA or ompC genes affected E. coli Tf binding and, furthermore, compromised the ability of the ompA mutant to respond to growth promotion by certain catecholamine stress hormones. Evidence was also found to implicate the OmpA porin as an entry point for catecholamine stress hormones. Further proteomic analyses in other bacterial pathogens revealed wide-scale involvement of porins in Tf binding: Salmonella typhimurium (OmpC), and Shigella sonnei, Shigella flexneri and Shigella boydii (OmpC and/or OmpA). This study shows that in addition to their existing housekeeping functions, the Gram-negative porin family of proteins can also act as Tf-capture proteins for those bacteria that lack the classical Neisseriaceae-type Tf-binding proteins.


Light trapping design for low band-gap polymer solar cells.
We demonstrate numerically a 2-D nanostructured design for light trapping in a low band-gap polymer solar cell. Finite element method simulations are used to study the effect of varying nanostructure periodicity, height, and shape on active layer absorption. Maintaining a constant active layer thickness of 100 nm we observe an enhancement in solar absorption of almost 40% relative to a planar cell. Improvements of this magnitude enable single-junction, low-band-gap cells to achieve power conversion efficiencies of 11.2% and perform competitively with even state-of-the-art tandem cells. Our design is also shown to significantly outperform tandem cells at off-normal angles of incidence.


Results from 15years of quality surveillance for a National Indigenous Point-of-Care Testing Program for diabetes.
Diabetes is a major health problem for Australia's Aboriginal and Torres Strait Islander peoples. Point-of-care testing for haemoglobin A1c (HbA1c) has been the cornerstone of a long-standing program (QAAMS) to manage glycaemic control in Indigenous people with diabetes and recently, to diagnose diabetes. The QAAMS quality management framework includes monthly testing of quality control (QC) and external quality assurance (EQA) samples. Key performance indicators of quality include imprecision (coefficient of variation [CV%]) and percentage acceptable results. This paper reports on the past 15years of quality testing in QAAMS and examines the performance of HbA1c POC testing at the 6.5% cut-off recommended for diagnosis. The total number of HbA1c EQA results submitted from 2002 to 2016 was 29,093. The median imprecision for EQA testing by QAAMS device operators averaged 2.81% (SD 0.50; range 2.2 to 3.9%) from 2002 to 2016 and 2.44% (SD 0.22; range 2.2 to 2.9%) from 2009 to 2016. No significant difference was observed between the median imprecision achieved in QAAMS and by Australasian laboratories from 2002 to 2016 (p=0.05; two-tailed paired t-test) and from 2009 to 2016 (p=0.17; two-tailed paired t-test). For QC testing from 2009 to 2016, imprecision averaged 2.5% and 3.0% for the two levels of QC tested. Percentage acceptable results averaged 90% for QA testing from 2002 to 2016 and 96% for QC testing from 2009 to 2016. The DCA Vantage was able to measure a patient and an EQA sample with an HbA1c value close to 6.5% both accurately and precisely. HbA1c POC testing in QAAMS has remained analytically sound, matched the quality achieved by Australasian laboratories and met profession-derived analytical goals for 15years.


The effects of body-weight supported treadmill training on cardiovascular regulation in individuals with motor-complete SCI.
Four-month longitudinal within-subject exercise training study. Although body-weight supported treadmill training (BWSTT) has not shown promise as a means of improving ambulation in individuals with motor-complete spinal cord injury (SCI), it may still improve cardiovascular health and function in this population. The purpose of this study was to (i) investigate the effects of BWSTT on peripheral muscular and elastic artery dimension and function and measures of heart rate variability (HRV) and blood pressure variability (BPV) in individuals with motor-complete SCI, and (ii) to make a preliminary examination of what factors may predict favourable cardiovascular outcomes following BWSTT in this population. Centre for Health Promotion and Rehabilitation, McMaster University, Hamilton, Ontario, Canada. Six individuals (four male, two female; age 37.7+/-15.4 years) with chronic SCI (C4-T12; ASIA A-B; 7.6+/-9.4 years post-injury) were included in the present investigation. Doppler ultrasound was used to determine femoral (exercising; muscular), carotid (elastic) and brachial (non-exercising control; muscular) artery dimension and function before and after 4 months of BWSTT. Continuous heart rate and blood pressure were also recorded before and after 4-months of BWSTT to determine frequency domain measures of HRV and BPV; clinically valuable indices of neurocardiac and neurovascular control, respectively. Two-way ANOVA (vessel x time) revealed no exercise-induced change in femoral or carotid artery cross-sectional area, blood flow or resistance and no change in carotid artery compliance following the 4 months of BWSTT compared to the non-exercising control brachial artery. However, there was a significant exercise-induced increase in femoral artery compliance. There were no exercise-induced changes in HRV or BPV when all participants were considered together. However, the results suggest that the subgroup of individuals who had a substantial heart rate response to BWSTT (n=3), experienced exercise-training induced changes in HRV reflective of a relative shift toward cardiac vagal predominance and reductions in BPV. BWSTT may cause an increase in femoral artery compliance in individuals with motor-complete SCI and therefore, should be encouraged as a means of improving cardiovascular health in this population. BWSTT may also cause modest improvements in measures of HRV and BPV in a select subgroup of individuals who respond to ambulation with moderate to large increases in HR. In the present study, factors associated with a substantial HR response to BWSTT were a propensity to orthostatic intolerance and muscular spasticity.


Gait recognition across various walking speeds using higher order shape configuration based on a differential composition model.
Gait has been known as an effective biometric feature to identify a person at a distance. However, variation of walking speeds may lead to significant changes to human walking patterns. It causes many difficulties for gait recognition. A comprehensive analysis has been carried out in this paper to identify such effects. Based on the analysis, Procrustes shape analysis is adopted for gait signature description and relevant similarity measurement. To tackle the challenges raised by speed change, this paper proposes a higher order shape configuration for gait shape description, which deliberately conserves discriminative information in the gait signatures and is still able to tolerate the varying walking speed. Instead of simply measuring the similarity between two gaits by treating them as two unified objects, a differential composition model (DCM) is constructed. The DCM differentiates the different effects caused by walking speed changes on various human body parts. In the meantime, it also balances well the different discriminabilities of each body part on the overall gait similarity measurements. In this model, the Fisher discriminant ratio is adopted to calculate weights for each body part. Comprehensive experiments based on widely adopted gait databases demonstrate that our proposed method is efficient for cross-speed gait recognition and outperforms other state-of-the-art methods.


Asbestos Exposure and Malignant Mesothelioma in Construction Workers-Epidemiological Remarks by the Italian National Mesothelioma Registry (ReNaM).
Notwithstanding the ban in 1992, asbestos exposure for workers in the construction sector in Italy remains a concern. The purpose of this study is to describe the characteristics of malignant mesothelioma (MM) cases recorded by the Italian registry (ReNaM) among construction workers. Incident mesothelioma cases with a definite asbestos exposure have been analyzed. Characteristics of cases and territorial clusters of crude rates of MM in construction workers have been described, as well as the relation between asbestos use before the ban and the historical trend of workforce in the construction sector in Italy. ReNaM has collected 31,572 incident MM cases in the period from 1993 to 2018 and asbestos exposure has been assessed for 24,864 (78.2%) cases. An occupational exposure has been reported for 17,191 MM cases (69.1% of subjects with a definite asbestos exposure). Among them, 3574 had worked in the construction sector, with an increasing trend from 15.8% in the 1993-98 period to 23.9% in 2014-2018 and a ubiquitous territorial distribution. The large use of asbestos in construction sector before the ban makes probability of exposure for workers a real concern still today, particularly for those working in maintenance and removal of old buildings. There is a clear need to assess, inform, and prevent asbestos exposure in this sector.


DNA measurement by image analysis of paraffin-embedded breast carcinoma tissue. A comparative investigation.
Measurement of cellular DNA content may provide useful prognostic information in several human neoplasms. The DNA content by image analysis of fresh tissue has been compared with flow cytometry with good correlation, but the use of paraffin-embedded tissue for image analysis has not been studied widely. This study reports the DNA content of 54 breast carcinomas and compares the results of image analysis of paraffin-embedded tissue and results of image analysis and flow cytometry of corresponding fresh tumors. Image analysis of paraffin blocks and fresh tumors showed comparable results in 51 tumors (94%), whereas 47 tumors (87%) were similar by all three methods. The DNA indices from image analysis of fresh and paraffin-embedded tumors showed significant correlation by linear regression analysis (r = 0.96; P less than 0.001). Discordances between image analysis of fresh and paraffin-embedded tumors are the result of technical problems, such as staining and tissue preservation. Discordances between image analysis and flow cytometry reflect the advantages and pitfalls of the two techniques. These data suggest that image analysis of paraffin-embedded tissue is a viable technique that will permit the performance of retrospective studies with long-term follow-up data for the evaluation of the prognostic significance of DNA content.


Surgical and transcatheter (Amplatzer) closure of atrial septal defects: a prospective comparison of results and cost.
To compare effectiveness, complications, and cost of Amplatzer with surgical atrial septal defect (ASD) closure. Prospective study. Tertiary cardiac referral centre. 43 consecutive patients (excluding non-UK residents) aged between 2.1 and 56.8 years (median 7) undergoing ASD closure. Procedural success, complications, regression of right ventricular dilatation (up to one year postprocedure), cost, inpatient stay, and home convalescent time. Amplatzer ASD closure was successful in 24 of 27 (89%) patients. Surgical closure was successful in all 19 cases. Cardiac complications affecting management occurred in three (11%) of the Amplatzer group (two procedural failures, one device embolisation) and 4 of 19 (21%) surgical patients (one pericardial pain, one global pericardial effusion requiring drainage, and one patient with anaemia requiring haematinics in addition to an incidental pericardial effusion and one further incidental pericardial effusion) (p = NS). There were complications that did not affect management in a further 5 of 19 surgical patients. There was no significant difference in regression of right ventricular dilatation by six months postprocedure (median right ventricular end diastolic diameter decrease: Amplatzer group 17.5%, surgical group 15.1%; median cardiothoracic ratio decrease: Amplatzer 7.9%, surgical 7.5%). Both hospital stay and home convalescent times were significantly shorter after Amplatzer closure (median hospital stay: Amplatzer one day, surgery six days; median convalescent time: Amplatzer two weeks, surgery 5.5 weeks). Median cost was similar for both groups (Amplatzer pound5375, surgical pound5412). Amplatzer ASD closure has a lower chance of success with a single procedure than surgery. Overall, there were more complications in the surgical group but the majority of these were minor and did not require any change in management. Resolution of right ventricular dilatation over the study period was similar for both techniques. Time spent in hospital and away from work or school was shorter for the Amplatzer group. The cost of both techniques was similar.


Genetic and Histopathologic Intertumor Heterogeneity in Primary Aldosteronism.
Whether primary aldosteronism (PA) is the consequence of a monoclonal or multiclonal process is unclear. A 48-year-old man with severe bilateral PA refractory to medical therapy underwent unilateral adrenalectomy of the dominant adrenal. Although computed tomography showed three left-sided cortical nodules, postsurgical histopathology and genetic analysis revealed five different adrenocortical adenomas. Two zona fasciculata (ZF)-like aldosterone-producing adenomas (APAs) each harbored distinct known somatic KCNJ5 mutations (L168R and T158A). A zona glomerulosa-like APA harbored a known CACNA1D G403R somatic mutation, whereas a zona reticularis-like adenoma, which was grossly black in pigmentation with histologic characteristics more associated with cortisol-producing adenomas, expressed CYP11B2, CYP17, and DHEA-ST by immunohistochemistry (IHC) and harbored no known somatic mutations. The fifth adenoma was ZF-type, negative for CYP11B2 and CYP17 IHC, and harbored no known somatic mutations. This case highlights complex intertumor heterogeneity in histology, steroidogenesis, and somatic mutations in multiple adrenocortical adenomas arising in a single patient with PA. These findings suggest that the syndrome of PA can involve heterogeneous and multiclonal functional adrenal adenomas.


Comparison of the Onset of Action, Maintenance, and Recovery of Three Weight-based Dosing of Cisatracurium in Patients with Morbid Obesity in Laparoscopic Bariatric Surgery: A Randomized Clinical Trial.
For patients with morbid obesity, different cisatracurium dosage regimens are recommended. This study aimed to compare the onset of action, the sufficiency of neuromuscular blockade during infusion, and the recovery of the three distinct cisatracurium dosage scalars in patients with morbid obesity undergoing laparoscopic bariatric surgery. In this randomized clinical trial, 55 patients were scheduled for bariatric surgery at Firoozgar Hospital from March 2020 to August 2021. Using a block randomization method, they were randomly divided into three groups, based on total body weight (TBW group), fat-free mass (FFM group), or ideal body weight (IBW group), to receive a bolus of cisatracurium 0.2 mg/Kg, followed by an infusion of 2 µg/Kg, to maintain a train-of-four (TOF) count≤2. Data were analyzed using SPSS software. P<0.05 was considered statistically significant. The mean time (seconds) to reach TOF0 in the TBW group was significantly shorter (201.89, 95%CI=192.99-210.79; P=0.004) than the IBW group (233.53, 95%CI=218.71-248.34; P=0.01). However, this difference was not statistically significant between TBW and FFM groups (220.83, 95%CI=199.73-241.94; P=0.81) or between FFM and the IBW groups (P=0.23). The rescue dose and increments of cisatracurium infusion were not required in the TBW group, whereas their probability was 4.81 times higher in the IBW group than the FFM group. Furthermore, the TBW and FFM groups had higher mean surgical condition scores than the IBW group (P<0.001, and P=0.006, respectively). Cisatracurium loading and infusion dosing based on FFM provide a comparable onset of action and surgical field condition to the TBW-based dosing with a shorter recovery time. However, IBW-based dosing of cisatracurium was insufficient for laparoscopic bariatric surgery.Trial Registration Number: IRCT20151107024909N9.A preprint of this study was published at . doi: .


Morphological and genetic characterization of a new cytoplasmic male sterility system (oxa CMS) in stem mustard (Brassica juncea).
KEY MESSAGE: oxa CMS is a new cytoplasmic male sterility type in Brassica juncea. oxa CMS is a cytoplasmic male sterility (CMS) line that has been widely used in the production and cultivation of stem mustard in the southwestern China. In this study, different CMS-type specific mitochondrial markers were used to confirm that oxa CMS is distinct from the pol CMS, ogu CMS, nap CMS, hau CMS, tour CMS, Moricandia arvensis CMS, orf220-type CMS, etc., that have been previously reported in Brassica crops. Pollen grains of the oxa CMS line are sterile with a self-fertility rate of almost 0% and the sterility strain rate and sterility degree of oxa CMS is 100% due to a specific flower structure and flowering habit. Scanning electron microscopy revealed that most pollen grains in mature anthers of the oxa CMS line are empty, flat and deflated. Semi-thin section further showed that the abortive stage of anther development in oxa CMS is initiated at the late uninucleate stage. Abnormally vacuolated microspores caused male sterility in the oxa CMS line. This cytological study combined with marker-assisted selection showed that oxa CMS is a novel CMS type in stem mustard (Brassica juncea). Interestingly, the abortive stage of oxa CMS is later than those in other CMS types reported in Brassica crops, and there is no negative effect on the oxa CMS line growth period. This study demonstrated that this novel oxa CMS has a unique flower structure with sterile pollen grains at the late uninucleate stage. Our results may help to uncover the mechanism of oxa CMS in Brassica juncea.


Macular pigment optical density is related to cognitive function in older people.
the xanthophylls lutein (L) and zeaxanthin (Z) exist in relatively high concentration in multiple central nervous tissues (e.g. cortex and neural retina). L + Z in macula (i.e. macular pigment, MP) are thought to serve multiple functions, including protection and improvement of visual performance. Also, L + Z in the macula are related to L + Z in the cortex. to determine whether macular pigment optical density (MPOD, L + Z in the macula) is related to cognitive function in older adults. participants were older adults (n = 108, 77.6 ± 2.7 years) sampled from the age-related maculopathy ancillary study of the Health Aging and Body Composition Study (Memphis, TN, USA). Serum carotenoids were measured using high performance liquid chromatography. MPOD was assessed using heterochromatic flicker photometry. Eight cognitive tests designed to evaluate several cognitive domains including memory and processing speed were administered. Partial correlation coefficients were computed to determine whether cognitive measures were related to serum L + Z and MPOD. MPOD levels were significantly associated with better global cognition, verbal learning and fluency, recall, processing speed and perceptual speed, whereas serum L + Z was significantly related to only verbal fluency. MPOD is related to cognitive function in older people. Its role as a potential biomarker of cognitive function deserves further study.


Molecular dissection of membrane-transport proteins: mass spectrometry and sequence determination of the galactose-H+ symport protein, GalP, of Escherichia coli and quantitative assay of the incorporation of [ring-2-13C]histidine and (15)NH(3).
The molecular mass of the galactose-H(+) symport protein GalP, as its histidine-tagged derivative GalP(His)(6), has been determined by electrospray MS (ESI-MS) with an error of <0.02%. One methionine residue, predicted to be present from the DNA sequence, was deduced to be absent. This is a significant advance on the estimation of the molecular masses of membrane-transport proteins by SDS/PAGE, where there is a consistent under-estimation of the true molecular mass due to anomalous electrophoretic migration. Addition of a size-exclusion chromatography step after Ni(2+)-nitrilotriacetate affinity purification was essential to obtain GalP(His)(6) suitable for ESI-MS. Controlled trypsin, trypsin+chymotrypsin and CNBr digestion of the protein yielded peptide fragments suitable for ESI-MS and tandem MS analysis, and accurate mass determination of the derived fragments resulted in identification of 82% of the GalP(His)(6) protein. Tandem MS analysis of selected peptides then afforded 49% of the actual amino acid sequence of the protein; the absence of the N-terminal methionine was confirmed. Matrix-assisted laser-desorption ionization MS allowed identification of one peptide that was not detected by ESI-MS. All the protein/peptide mass and sequence determinations were in accord with the predictions of amino acid sequence deduced from the DNA sequence of the galP gene. [ring-2-(13)C]Histidine was incorporated into GalP(His)(6) in vivo, and ESI-MS analysis enabled the measurement of a high (80%) and specific incorporation of label into the histidine residues in the protein. MS could also be used to confirm the labelling of the protein by (15)NH(3) (93% enrichment) and [(19)F]tryptophan (83% enrichment). Such MS measurements will serve in the future analysis of the structures of membrane-transport proteins by NMR, and of their topology by indirect techniques.


Inferring the Regulatory Network of miRNAs on Terpene Trilactone Biosynthesis Affected by Environmental Conditions.
As a medicinal tree species, ginkgo (Ginkgo biloba L.) and terpene trilactones (TTLs) extracted from its leaves are the main pharmacologic activity constituents and important economic indicators of its value. The accumulation of TTLs is known to be affected by environmental stress, while the regulatory mechanism of environmental response mediated by microRNAs (miRNAs) at the post-transcriptional levels remains unclear. Here, we focused on grafted ginkgo grown in northwestern, southwestern, and eastern-central China and integrally analyzed RNA-seq and small RNA-seq high-throughput sequencing data as well as metabolomics data from leaf samples of ginkgo clones grown in natural environments. The content of bilobalide was highest among detected TTLs, and there was more than a twofold variation in the accumulation of bilobalide between growth conditions. Meanwhile, transcriptome analysis found significant differences in the expression of 19 TTL-related genes among ginkgo leaves from different environments. Small RNA sequencing and analysis showed that 62 of the 521 miRNAs identified were differentially expressed among different samples, especially the expression of miRN50, miR169h/i, and miR169e was susceptible to environmental changes. Further, we found that transcription factors (ERF, MYB, C3H, HD-ZIP, HSF, and NAC) and miRNAs (miR319e/f, miRN2, miRN54, miR157, miR185, and miRN188) could activate or inhibit the expression of TTL-related genes to participate in the regulation of terpene trilactones biosynthesis in ginkgo leaves by weighted gene co-regulatory network analysis. Our findings provide new insights into the understanding of the regulatory mechanism of TTL biosynthesis but also lay the foundation for ginkgo leaves' medicinal value improvement under global change.


Israel's rapid rollout of vaccinations for COVID-19.
As of the end of 2020, the State of Israel, with a population of 9.3 million, had administered more COVID-19 vaccine doses than all countries aside from China, the US, and the UK. Moreover, Israel had administered almost 11.0 doses per 100 population, while the next highest rates were 3.5 (in Bahrain) and 1.4 (in the United Kingdom). All other countries had administered less than 1 dose per 100 population.While Israel's rollout of COVID-19 vaccinations was not problem-free, its initial phase had clearly been rapid and effective. A large number of factors contributed to this early success, and they can be divided into three major groups.The first group of factors consists of long-standing characteristics of Israel which are extrinsic to health care. They include: Israel's small size (in terms of both area and population), a relatively young population, relatively warm weather in December 2020, a centralized national system of government, and well-developed infrastructure for implementing prompt responses to large-scale national emergencies.The second group of factors are also long-standing, but they are health-system specific. They include: the organizational, IT and logistical capacities of Israel's community-based health care providers, the availability of a cadre of well-trained, salaried, community-based nurses who are directly employed by those providers, a tradition of effective cooperation between government, health plans, hospitals, and emergency care providers - particularly during national emergencies; and support tools and decisionmaking frameworks to support vaccination campaigns.The third group consists of factors that are more recent and are specific to the COVID-19 vaccination effort. They include: the mobilization of special government funding for vaccine purchase and distribution, timely contracting for a large amount of vaccines relative to Israel's population, the use of simple, clear and easily implementable criteria for determining who had priority for receiving vaccines in the early phases of the distribution process, a creative technical response that addressed the demanding cold storage requirements of the Pfizer-BioNTech COVID-19 vaccine, and well-tailored outreach efforts to encourage Israelis to sign up for vaccinations and then show up to get vaccinated.While many of these facilitating factors are not unique to Israel, part of what made the Israeli rollout successful was its combination of facilitating factors (as opposed to each factor being unique separately) and the synergies it created among them. Moreover, some high-income countries (including the US, the UK, and Canada) are lacking several of these facilitating factors, apparently contributing to the slower pace of the rollout in those countries.


Individualised antimicrobial dosing in critically ill patients undergoing continuous renal replacement therapy: focus on total drug clearance.
Continuous renal replacement therapy (CRRT) is common practice in critical care patients with acute renal failure. To evaluate the adequacy of antimicrobial doses calculated based on the total drug clearance and dose recommended by different guides in critically ill patients undergoing CRRT. Retrospective observational study. Patients admitted to a critical care unit during May 2014 to May 2016 and subjected to CRRT were included. The recommended dose was established as the product of the usual dose of the drug by total drug clearance. 177 antimicrobial agents, used in 64 patients were analysed; 45 (25.4%) antimicrobials were given in an insufficient dose (<20%) according to the theoretical calculation. Following the recommendations in the revised guidelines, between 10% and 20% of antimicrobials were given in insufficient doses. A higher success rate of treatment in those patients not receiving a low drug dosage was seen (35.2% vs 24.0%). There is a great disparity between the antimicrobial dose prescribed, recommended and calculated based on drug clearance in critically ill patients undergoing CRRT.


Cost-effectiveness of colorectal cancer screening protocols in urban Chinese populations.
Colorectal cancer (CRC) takes a second and fourth position in the incidence and mortality lists respectively among all malignant tumors in urban populations in China. This study was designed to evaluate the cost-effectiveness of two different CRC screening protocols: faecal occult blood test (FOBT) alone, and FOBT plus a high-risk factor questionnaire (HRFQ) as the respective initial screens, followed by colonoscopy. We developed a Markov model to simulate the progression of a cohort of 100,000 average risk asymptomatic individuals moving through a defined series of states between the ages of 40 to 74 years. The parameters used for the modeling came from the CESP (Comparison and Evaluation of Screening Programs for Colorectal Cancer in Urban Communities in China) study and published literature. Eight CRC screening scenarios were tested in the Markov model. The cost-effectiveness of CRC screening under each scenario was measured by an incremental cost-effectiveness ratio (ICER) compared with a scenario without CRC screening. The study revealed that a combined use of FOBT and HRFQ is preferable in CRC screening programs as an initial screening instrument. Annual FOBT+HRFQ screening is recommended for those who have a negative initial result and those who have a positive result but have failed to continue to colonoscopic examination. Repeated colonoscopy (for those with a positive result in initial screening but a negative colonoscopy result) should be performed at a ten-year interval instead of one-year. Such a protocol would cost 7732 Yuan per life year saved, which is the most cost-effective option. In conclusion, the current Chinese Trial Version for CRC Screening Strategy should be revised in line with the most cost-effective protocol identified in this study.


Acute decreases in cerebrospinal fluid glutathione levels after intracerebroventricular morphine for cancer pain.
Intracerebroventricular (ICV) morphine administration is effective for the management of refractory cancer pain. Recent preclinical observations of acute depletion of the major endogenous intracellular antioxidant glutathione (GSH) in brain and peripheral organs after ICV morphine in rodents led us to apply microchemical methods to profile the neurochemical effects of ICV morphine in three patients treated for intractable cancer pain. Assessment of morphine, morphine-6-glucuronide, and a panel of endogenous compounds and metabolites in ventricular and cisternal cerebrospinal fluid (CSF) demonstrated transient, postdose increases in morphine and morphine-6-glucuronide in ventricular and cistemal CSF, accompanied by acute decreases in CSF GSH levels. Significant changes were also observed in the CSF levels of 4-hydroxybenzoic acid, homovanillic acid, 5-hydroxyphenyllactic acid, and uric acid. These pilot clinical observations of acute central GSH depletion after ICV morphine suggest a novel mechanism for neuropsychiatric toxicity or preclinical findings, such as hyperalgesia or increased motoric activity observed in nonhuman species after central morphine administration. Because ICV morphine is a mainstay of treatment for refractory cancer pain, elucidation of a mechanism's (or mechanisms') mediating a potential pro-oxidant state in the central nervous system induced by ICV morphine is important. We observed acute decreases in glutathione levels in cerebrospinal fluid sampled from patients after intracerebroventricular doses of morphine for intractable cancer pain. Such doses may, by depleting the antioxidant glutathione, render the central nervous system vulnerable to damage from oxidative stress.


Eye contact detection in humans from birth.
Making eye contact is the most powerful mode of establishing a communicative link between humans. During their first year of life, infants learn rapidly that the looking behaviors of others conveys significant information. Two experiments were carried out to demonstrate special sensitivity to direct eye contact from birth. The first experiment tested the ability of 2- to 5-day-old newborns to discriminate between direct and averted gaze. In the second experiment, we measured 4-month-old infants' brain electric activity to assess neural processing of faces when accompanied by direct (as opposed to averted) eye gaze. The results show that, from birth, human infants prefer to look at faces that engage them in mutual gaze and that, from an early age, healthy babies show enhanced neural processing of direct gaze. The exceptionally early sensitivity to mutual gaze demonstrated in these studies is arguably the major foundation for the later development of social skills.


In silico study directed towards identification of the key structural features of GyrB inhibitors targeting MTB DNA gyrase: HQSAR, CoMSIA and molecular dynamics simulations.
Mycobacterium tuberculosis DNA gyrase subunit B (GyrB) has been identified as a promising target for rational drug design against fluoroquinolone drug-resistant tuberculosis. In this study, we attempted to identify the key structural feature for highly potent GyrB inhibitors through 2D-QSAR using HQSAR, 3D-QSAR using CoMSIA and molecular dynamics (MD) simulations approaches on a series of thiazole urea core derivatives. The best HQSAR and CoMSIA models based on IC50 and MIC displayed the structural basis required for good activity against both GyrB enzyme and mycobacterial cell. MD simulations and binding free energy analysis using MM-GBSA and waterswap calculations revealed that the urea core of inhibitors has the strongest interaction with Asp79 via hydrogen bond interactions. In addition, cation-pi interaction and hydrophobic interactions of the R2 substituent with Arg82 and Arg141 help to enhance the binding affinity in the GyrB ATPase binding site. Thus, the present study provides crucial structural features and a structural concept for rational design of novel DNA gyrase inhibitors with improved biological activities against both enzyme and mycobacterial cell, and with good pharmacokinetic properties and drug safety profiles.


Dissociation of β-amyloid from lipoprotein in cerebrospinal fluid from Alzheimer's disease accelerates β-amyloid-42 assembly.
Monoclonal 2C3 specific to β-amyloid (Aβ) oligomers (AβOs) enabled us to test our hypothesis that the alteration of lipoprotein-Aβ interaction in the central nervous system (CNS) initiates and/or accelerates the cascade favoring Aβ assembly. Immunoprecipitation of frontal cortex employing 2C3 unequivocally detected soluble 4-, 8-, and 12-mers in Alzheimer's disease (AD) brains. Immunoblot analysis of the entorhinal cortex employing 2C3 revealed that the accumulation of soluble 12-mers precedes the appearance of neuronal loss or cognitive impairment and is enhanced as the Braak neurofibrially tangle (NFT) stages progress. The dissociation of soluble Aβ from lipoprotein particles occurs in cerebrospinal fluid (CSF), and the presence of lipoprotein-free oligomeric 2C3 conformers (4- to 35-mers) was evident, which mimic CNS environments. Such CNS environments may strongly affect conformation of soluble Aβ peptides, resulting in the conversion of soluble Aβ(42) monomers into soluble Aβ(42) assembly. The findings suggest that functionally declined lipoproteins may accelerate the generation of metabolic conditions leading to higher levels of soluble Aβ(42) assembly in the CNS.


Recruitment of Mcm10 to Sites of Replication Initiation Requires Direct Binding to the Minichromosome Maintenance (MCM) Complex.
Mcm10 is required for the initiation of eukaryotic DNA replication and contributes in some unknown way to the activation of the Cdc45-MCM-GINS (CMG) helicase. How Mcm10 is localized to sites of replication initiation is unclear, as current models indicate that direct binding to minichromosome maintenance (MCM) plays a role, but the details and functional importance of this interaction have not been determined. Here, we show that purified Mcm10 can bind both DNA-bound double hexamers and soluble single hexamers of MCM. The binding of Mcm10 to MCM requires the Mcm10 C terminus. Moreover, the binding site for Mcm10 on MCM includes the Mcm2 and Mcm6 subunits and overlaps that for the loading factor Cdt1. Whether Mcm10 recruitment to replication origins depends on CMG helicase assembly has been unclear. We show that Mcm10 recruitment occurs via two modes: low affinity recruitment in the absence of CMG assembly ("G1-like") and high affinity recruitment when CMG assembly takes place ("S-phase-like"). Mcm10 that cannot bind directly to MCM is defective in both modes of recruitment and is unable to support DNA replication. These findings indicate that Mcm10 is localized to replication initiation sites by directly binding MCM through the Mcm10 C terminus.


[Effects of irrigation with treated wastewater on nutrient distribution in cucumber and tomato plants and their fruit quality].
A field experiment was conducted to investigate the effects of irrigation with treated wastewater on the nutrient distribution in cucumber and tomato plants and their fruit quality. Irrigation with treated wastewater promoted tomato growth significantly, but had definite inhibition effect on cucumber growth. After the irrigation with treated wastewater, the nitrogen in plants had the characteristics of upward translocation, potassium was easily to be accumulated in cucumber leaf but not accumulated in tomato root, and sodium was mostly accumulated in root but less enriched in leaf, not giving damage to the plants. No significant effects were observed on the distribution of calcium, magnesium, and chlorine in plants. Under the irrigation with treated wastewater, the overall quality of cucumber and tomato fruits was less affected. The nitrate concentration in cucumber and tomato fruits was increased by 5.3% and 32.9%, respectively, but still lower than the state food safety standard of China.


Facile Synthesis of Different Morphologies of Cu2SnS3 for High-Performance Supercapacitors.
Cu2SnS3 is considered as an emerging potential candidate for electrode materials due to considerable interlayer spaces and tunnels in its crystal structures and excellent conducting ability. Ternary Cu2SnS3 as anode in lithium ion batteries has already been reported, but it is rarely mentioned to be applied in supercapacitors which is considered to be a complementary energy storage device for lithium ion batteries. It is an effective method to improve the electrochemical performance of materials by adjusting the morphology and microstructure of materials. In present study, ternary nanosheet-assembled Cu2SnS3 microspheres (M-CTS) and nanoparticles-like Cu2SnS3 (N-CTS) are synthesized via a facile solvothermal route. The results suggest that Cu2SnS3 microspheres (M-CTS) exhibit better capacitive performance compared with Cu2SnS3 (N-CTS) nanoparticles, which means that morphology does have a significant effect on the electrochemical reaction. M-CTS presents excellent supercapacitor performances with the high specific capacity of about 406 C g-1 at a current density of 1 A g-1 and achieves a high energy density of 85.6 W h kg-1 and power density of 720 W kg-1. The remarkable electrochemical performance of Cu2SnS3 can be attributed to the large specific surface area, smaller average pore size, and improved electrical conductivity. Our research indicates that it is very suitable for large-scale production and has enormous potential in the practical application of high-performance supercapacitors.


Crm1p mediates regulated nuclear export of a yeast AP-1-like transcription factor.
The yeast AP-1-like transcription factor, Yap1p, activates genes required for the response to oxidative stress. Yap1p is normally cytoplasmic and inactive, but will activate by nuclear translocation if cells are placed in an oxidative environment. Here we show that Yap1p is a target of the beta-karyopherin-like nuclear exporter, Crm1p. Yap1p is constitutively nuclear in a crm1 mutant, and Crm1p binds to a nuclear export sequence (NES)-like sequence in Yap1p in the presence of RanGTP. Recognition of Yap1p by Crm1p is inhibited by oxidation, and this inhibition requires at least one of the three cysteine residues flanking the NES. These results suggest that Yap1p localization is largely regulated at the level of nuclear export, and that the oxidation state affects the accessibility of the Yap1p NES to Crm1p directly. We also show that a mutation in RanGAP (rna1-1) is synthetically lethal with crm1 mutants. Yap1p export is inhibited in both rna1-1 and prp20 (RanGNRF) mutant strains, but Yap1p rapidly accumulates at the nuclear periphery after shifting rna1-1, but not other mutant cells to the non-permissive temperature. Thus, disassembly of export complexes in response to RanGTP hydrolysis may be required for release of substrate from a terminal binding site at the nuclear pore complex (NPC).


Effects of nitroglycerin versus labetalol on peripheral perfusion during deliberate hypotension for sinus endoscopic surgery: a randomized, controlled, double-blinded trial.
Deliberate hypotension is used to provide a bloodless field during functional endoscopic sinus surgery; however, the impact of controlled hypotension during anesthesia on peripheral tissue perfusion has not been extensively evaluated. The aim of this study was to compare the impact of nitroglycerin- versus labetalol-induced hypotension on peripheral perfusion. The present randomized, double-blinded, controlled trial included adult patients undergoing endoscopic sinus surgery. Patients were allocated to one of two groups according to the drug received for induction of deliberate hypotension: nitroglycerin (n = 20) or labetalol (n = 20). Mean arterial pressure was maintained at 55-65 mmHg in both groups. Both study groups were compared according to pulse oximeter-derived peripheral perfusion index (primary outcome), serum lactate level, mean arterial pressure, heart rate, surgical field score, and intraoperative blood loss. Forty patients were included in the final analysis. The nitroglycerin group exhibited a higher peripheral perfusion index at nearly all records (p < 0.0001) and lower postoperative serum lactate levels (1.3 ± 0.2 mmol/L vs. 1.7 ± 0.4 mmol/L; p = 0.001) than the labetalol group. The peripheral perfusion index was higher in the nitroglycerin group than at baseline at most intraoperative readings. The median surgical field score was modestly lower in the labetalol group than in the nitroglycerin group in the first 20 min (2 [interquartile range (IQR) 2-2.5] versus 1.5 [IQR 1-2]; p = 0.001). Both groups demonstrated comparable and acceptable surgical field scores in all subsequent readings. Nitroglycerin-induced deliberate hypotension was accompanied by higher peripheral perfusion index and lower serum lactate levels than labetalol-induced deliberate hypotension during sinus endoscopic surgery. The study was registered at clinicaltrials registry system with trial number: NCT03809065. Registered at 19 January 2019. This study adheres to CONSORT guidelines.


Characteristics and function of sulfur dioxygenase in Echiuran worm Urechis unicinctus.
Sulfide is a common toxin to animals and is abundant in coastal and aquatic sediments. Sulfur dioxygenase (SDO) is thought to be the key enzyme involved in sulfide oxidation in some organisms. The echiuran worm, Urechis unicinctus, inhabits coastal sediment and tolerates high concentrations of sulfide. The SDO is presumably important for sulfide tolerance in U. unicinctus. The full-length cDNA of SDO from the echiuran worm U. unicinctus, proven to be located in the mitochondria, was cloned and the analysis of its sequence suggests that it belongs to the metallo-β-lactamase superfamily. The enzyme was produced using an E. coli expression system and the measured activity is approximately 0.80 U mg protein(-1). Furthermore, the expression of four sub-segments of the U. unicinctus SDO was accomplished leading to preliminary identification of functional domains of the enzyme. The identification of the conserved metal I (H113, H115, H169 and D188), metal II (D117, H118, H169 and H229) as well as the potential glutathione (GSH) (R197, Y231, M279 and I283) binding sites was determined by enzyme activity and GSH affinity measurements. The key residues responsible for SDO activity were identified by analysis of simultaneous mutations of residues D117 and H118 located close to the metal II binding site. The recombinant SDO from U. unicinctus was produced, purified and characterized. The metal binding sites in the SDO were identified and Y231 recognized as the mostly important amino acid residue for GSH binding. Our results show that SDO is located in the mitochondria where it plays an important role in sulfide detoxification of U. unicinctus.


Dual-specificity tyrosine phosphorylation-regulated kinase 1A (DYRK1A) inhibitors: a survey of recent patent literature.
Dual-specificity tyrosine phosphorylation-regulated kinase 1A (DYRK1A) is a eukaryotic serine-threonine protein kinase belonging to the CMGC group. DYRK1A hyperactivity appears to contribute to the development of a number of human malignancies and to cognitive deficits observed in Down syndrome and Alzheimer's disease. As a result, the DYRK1A kinase represents an attractive target for the synthesis and optimization of pharmacological inhibitors of potential therapeutic interest. Like most tyrosine kinase inhibitors developed up to the market, DYRK1A inhibitors are essentially acting by competing with ATP for binding at the catalytic site of the kinase. Areas covered: This paper reviews patent activity associated with the discovery of synthetic novel heterocyclic molecules inhibiting the catalytic activity of DYRK1A. Expert opinion: Despite the important role of DYRK1A in biological processes and the growing interest in the design of new therapeutic drugs, there are only few patented synthetic DYRK1A inhibitors and most of them were and are still developed by academic research groups, sometimes with industrial partners.


Following movement of the L1 stalk between three functional states in single ribosomes.
The L1 stalk is a mobile domain of the large ribosomal subunit E site that interacts with the elbow of deacylated tRNA during protein synthesis. Here, by using single-molecule FRET, we follow the real-time dynamics of the L1 stalk and observe its movement relative to the body of the large subunit between at least 3 distinct conformational states: open, half-closed, and fully closed. Pretranslocation ribosomes undergo spontaneous fluctuations between the open and fully closed states. In contrast, posttranslocation ribosomes containing peptidyl-tRNA and deacylated tRNA in the classical P/P and E/E states, respectively, are fixed in the half-closed conformation. In ribosomes with a vacant E site, the L1 stalk is observed either in the fully closed or fully open conformation. Several lines of evidence show that the L1 stalk can move independently of intersubunit rotation. Our findings support a model in which the mobility of the L1 stalk facilitates binding, movement, and release of deacylated tRNA by remodeling the structure of the 50S subunit E site between 3 distinct conformations, corresponding to the E/E vacant, P/E hybrid, and classical states.


Role of Th1/Th2 cells and related cytokines in autoimmune hepatitis.
Dysregulation of T cell response is thought to play an important role in the immunopathogenesis of autoimmune hepatitis. However, no consensus has yet been reached regarding the implications of a distinct T cell subset in the pathogenesis of this progressive liver disease. Therefore, T-bet and GATA-3 expression was examined in patients with autoimmune hepatitis (AIH) and in healthy controls. Moreover, the profile of Th1 (IFN-γ) and Th2 (IL-4) cytokine gene expression was analyzed. Levels of mRNA transcripts were measured in peripheral blood mononuclear cells (PBMCs) using a two-step reverse transcription quantitative real-time polymerase chain reaction with SYBR Green. T-bet and IFN-γ mRNA expression was significantly higher in AIH patients compared to healthy controls (p<0.05), whereas no differences were observed for either GATA-3 or IL-4 mRNA expression (p>0.05). Alterations in the Th1/Th2 cell balance may be responsible for both disease progression and the resulting complications.


Oligonol, a lychee fruit-derived low-molecular form of polyphenol mixture, suppresses inflammatory cytokine production from human monocytes.
Monocytes produce high levels of inflammatory cytokines including IL-6 and TNF-α that are involved in autoimmunity, inflammatory diseases, cardiovascular disease and obesity. Therapies targeting IL-6 and TNF-α have been utilized in treating chronic inflammatory diseases. Oligonol is a lychee fruit-derived low-molecular form of polyphenol mixture, typically catechin-type monomers and oligomers of proanthocyanidins, which are produced by an oligomerization process. Although previous studies reported anti-inflammatory properties of Oligonol, it is unknown whether and how Oligonol suppresses IL-6 and TNF-α production in human monocytes. The results of our study demonstrate that Oligonol (25μg/ml) decreases the production of IL-6 and TNF-α from human primary monocytes as measured by flow cytometry and ELISA. Such an anti-cytokine effect was likely mediated by the suppression of NF-κB activation without inducing cell death. Our findings raise the possibility of exploring the benefits of Oligonol in controlling inflammatory conditions, especially those associated with monocytes, in humans.


[Preparation of films and fibers containing proteolytic enzymes].
Immobilization of the proteolytic enzymes trypsin and terrylitin in fluorplastic-42 films and fibers as well as in water-soluble polymeric (polyvinyl alcohol and carboxymethyl cellulose) films was being studied during preparation of the films. The properties of the proteolytic enzymes immobilized in the polymers are dependent on the conditions of preparation of fibers and films.


[Pulmonary blood flow in bronchiectasis patients before and after lung resection].
The analysis of data obtained in the investigation of functions of the pulmonary blood flow in 44 patients prior to and following segmentary and combined resections of the lungs fulfilled for bronchiectasis has shown that almost all the patients with bronchiectasis have the disturbed hemodynamics of the small circulation circuit. The direct relation was shown between the duration of the disease, spread of the pathological process and the degree of lung hypertension. It has been found that a resection within the limits of 6 lung segments fails to exert a negative influence on hemodynamics of the small blood circulation circuit and allows prevention of progressing pulmonary hypertension in most of the patients.


Early palliative care and the opioid crisis: ten pragmatic steps towards a more rational use of opioids.
In palliative care, opioids and other controlled drugs are among the most commonly used and important medications. Opioids are associated with significant risk of dependence and misuse. In many developed countries, there is an epidemic of prescription opioid misuse and overdose deaths. Palliative care has a critical role educating patients about the safe use of opioids, providing universal screening and close monitoring, and prescribing opioids appropriately balancing the risks and benefits. This is particularly important in the era of early palliative care, when patients have much longer survival and potentially greater risk of misuse while on chronic opioid therapy. Here, we provided a critical appraisal of opioid use in the context of opioid crisis and early palliative care. We also present a pragmatic 10-step approach for the judicious use of opioids.


Analytical Method Development for Sodium Valproate through Chemical Derivatization.
Sodium valproate has anticonvulsant activity and is structurally different to conventional antiepileptic drugs. The problem with valproic acid is the lack of a chromophore, which means that gas chromatography is the sole assay methodology. The introduction of benzoyl and phenyl groups to the molecule is a useful derivatisation, which enables the creation of detectable chromophores for HPLC analysis for pharmaceutical dosages as well as biological systems. Methodology. Sodium valproate was derivatised by the addition of a chromophore to its structure by introducing a methyl benzoyl or a phenyl group. Trichlorophenol and 2-hydroxyacetophenone were used to introduce phenyl and benzoyl groups to valproic acid, respectively. The reaction used was estrification reaction using coupling agents. An analytical method was then developed and validated using reverse-phase HPLC. The method was validated for parameters like linearity, range, accuracy precision, and robustness. The developed method was easy and feasible and can be applied to both routine analysis and bioanalysis. The method was very sensitive and could quantify valproic acid at a very low concentration of 0.75 × 10-5 mg/ml. The developed method was found to be linear (R 2 = 0.997), accurate, precise, and robust. The proposed chemical derivatisation and the developed analytical method are novel. The developed analytical procedure is the first of its kind; it is easy and feasible and can be used to quantify and detect sodium valproate at very low concentrations compared to other available methods in the literature.


ESR1 and PGR polymorphisms are associated with estrogen and progesterone receptor expression in breast tumors.
Hormone receptor-positive (HR+) breast cancers express the estrogen (ERα) and/or progesterone (PgR) receptors. Inherited single nucleotide polymorphisms (SNPs) in ESR1, the gene encoding ERα, have been reported to predict tamoxifen effectiveness. We hypothesized that these associations could be attributed to altered tumor gene/protein expression of ESR1/ERα and that SNPs in the PGR gene predict tumor PGR/PgR expression. Formalin-fixed paraffin-embedded breast cancer tumor specimens were analyzed for ESR1 and PGR gene transcript expression by the reverse transcription polymerase chain reaction based Oncotype DX assay and for ERα and PgR protein expression by immunohistochemistry (IHC) and an automated quantitative immunofluorescence assay (AQUA). Germline genotypes for SNPs in ESR1 (n = 41) and PGR (n = 8) were determined by allele-specific TaqMan assays. One SNP in ESR1 (rs9322336) was significantly associated with ESR1 gene transcript expression (P = 0.006) but not ERα protein expression (P > 0.05). A PGR SNP (rs518162) was associated with decreased PGR gene transcript expression (P = 0.003) and PgR protein expression measured by IHC (P = 0.016), but not AQUA (P = 0.054). There were modest, but statistically significant correlations between gene and protein expression for ESR1/ERα and PGR/PgR and for protein expression measured by IHC and AQUA (Pearson correlation = 0.32-0.64, all P < 0.001). Inherited ESR1 and PGR genotypes may affect tumor ESR1/ERα and PGR/PgR expression, respectively, which are moderately correlated. This work supports further research into germline predictors of tumor characteristics and treatment effectiveness, which may someday inform selection of hormonal treatments for patients with HR+ breast cancer.


Methodological and practical issues related to shiftwork research.
This paper cites inconsistencies in the current literature regarding the health impact of shiftwork and suggests that these disparities may be attributed, in part, to shortcomings in the nature and reporting of cross-sectional field research. Selected methodological (e.g., differential accretion, cross-over) and practical (e.g., definition of independent and dependent variables) problems in the design and communication of shiftwork investigations are discussed and strategies are provided for their solution.


Transcription factors lhx1/5-1 and pitx are required for the maintenance and regeneration of serotonergic neurons in planarians.
In contrast to most adult organisms, freshwater planarians can regenerate any injured body part, including their entire nervous system. This allows for the analysis of genes required for both the maintenance and regeneration of specific neural subtypes. In addition, the loss of specific neural subtypes may uncover previously unknown behavioral roles for that neural population in the context of the adult animal. Here we show that two homeodomain transcription factor homologs, Smed-lhx1/5-1 and Smed-pitx, are required for the maintenance and regeneration of serotonergic neurons in planarians. When either lhx1/5-1 or pitx was knocked down by RNA interference, the expression of multiple canonical markers for serotonergic neurons was lost. Surprisingly, the loss of serotonergic function uncovered a role for these neurons in the coordination of motile cilia on the ventral epidermis of planarians that are required for their nonmuscular gliding locomotion. Finally, we show that in addition to its requirement in serotonergic neurons, Smed-pitx is required for proper midline patterning during regeneration, when it is required for the expression of the midline-organizing molecules Smed-slit in the anterior and Smed-wnt1 in the posterior.


[Allergy to protein hydrolysates. Report of 30 cases].
To describe the population of children allergic to cow's milk protein and their usual substitutes made of protein hydrolysates, and who are efficiently taken care of by using Neocate, an amino acid based formula. Allergy to protein hydrolysates was diagnosed in 30 infants, aged from 15 d to 13 months (median = 3 months), who remained symptomatic while receiving a protein hydrolysate for 15 d to 12 months (median = 1.8 months). After a complete clinical and biological evaluation, an amino acid based diet using Neocate was attempted, followed after one month by an oral challenge test with the protein hydrolysate. Symptoms occurred mainly in the digestive tract (25 cases) in the form of regurgitations (18 cases), diarrhea (17 cases) and colicky pain (10 cases); failure to thrive was seen in ten cases. Neocate improved the clinical condition rapidly (within 3-10 d), and allowed the children to gain weight (27.5 +/- 10.8 g/d). Skin prick tests with cow's milk hydrolysates were positive in 13 out of 26 children; total IgE was raised in cases of cutaneous symptoms (six cases), and associated with increased specific IgE for cow's milk. During the intestinal permeability test, the lactitol/mannitol ratio which initially increased, decreased following Neocate (6.58 +/- 2.92% vs. 3.47 +/- 1.58%, P = 0.0004). Multiple food allergies were present in 22 cases. During challenge with a hydrolysate, the clinical reaction was immediate in 13 cases, partially delayed in 13 cases and delayed in four cases. Allergy to cow's milk hydrolysates may occur and has to be considered in the presence of anaphylaxis and also chronic digestive symptoms such as regurgitations, diarrhea and colicky pain, when these symptoms persist during cow's milk free diet.


Compound heterozygous mutation of Rag1 leading to Omenn syndrome.
Omenn syndrome is a primary immunodeficiency disorder, featuring susceptibility to infections and autoreactive T cells and resulting from defective genomic rearrangement of genes for the T cell and B cell receptors. The most frequent etiologies are hypomorphic mutations in "non-core" regions of the Rag1 or Rag2 genes, the protein products of which are critical members of the cellular apparatus for V(D)J recombination. In this report, we describe an infant with Omenn syndrome with a previously unreported termination mutation (p.R142*) in Rag1 on one allele and a partially characterized substitution mutation (p.V779M) in a "core" region of the other Rag1 allele. Using a cellular recombination assay, we found that while the p.R142* mutation completely abolished V(D)J recombination activity, the p.V779M mutation conferred a severe, but not total, loss of V(D)J recombination activity. The recombination defect of the V779 mutant was not due to overall misfolding of Rag1, however, as this mutant supported wild-type levels of V(D)J cleavage. These findings provide insight into the role of this poorly understood region of Rag1 and support the role of Rag1 in a post-cleavage stage of recombination.


Behavioral Interventions on Surgeons' Preoperative Decision-Making: A Scoping Review and Critical Analysis.
Consensus is lacking regarding the optimal strategy to influence surgeons' behaviors to reduce low-value surgical care. Comprehensively describing the existing body of literature that seeks to intervene on surgeons' preoperative decision-making may aid in structuring future behavior change strategies. We performed a scoping review using four databases (Medical Literature Analysis and Retrieval System Online, Embase, Web of Science, and Cumulated Index to Nursing and Allied Health Literature) for articles that tested the effect of behavioral-based interventions on any aspect of surgeons' decision-making in the preoperative setting. Abstracted data were characterized by summative descriptions and analyzed using the Tailored Implementation for Chronic Disease framework, mapping aspects of deimplementation strategies in the studies onto the determinant(s) that they altered. Data abstraction and mapping tools were piloted and iteratively revised before two researchers independently assessing studies and categorizing determinants, and then meeting to discuss their decisions. There were 1460 articles identified from the initial search, with 17 full text articles ultimately included in the scoping review. Eight studies relied on a multidisciplinary preoperative conference to accomplish their aims, while five were multifaceted in their approach to deimplementation, and four studies used only a clinical decision support tool to accomplish their aims. Mapping determinants addressed in these studies onto the Tailored Implementation for Chronic Disease framework demonstrated that most strategies attempted to close knowledge gaps, leverage communication between providers, and broadcast institutional prioritization of change. There is a small but growing field of implementation and deimplementation strategies in preoperative surgical decision-making, and different approaches may be equally effective in varied clinical contexts. Deliberate measurement and comparison of outcomes, as well as selection of control groups, are areas for improvement in future work.


An ECVAG trial on assessment of oxidative damage to DNA measured by the comet assay.
The increasing use of single cell gel electrophoresis (the comet assay) highlights its popularity as a method for detecting DNA damage, including the use of enzymes for assessment of oxidatively damaged DNA. However, comparison of DNA damage levels between laboratories can be difficult due to differences in assay protocols (e.g. lysis conditions, enzyme treatment, the duration of the alkaline treatment and electrophoresis) and in the end points used for reporting results (e.g. %DNA in tail, arbitrary units, tail moment and tail length). One way to facilitate comparisons is to convert primary comet assay end points to number of lesions/10(6) bp by calibration with ionizing radiation. The aim of this study was to investigate the inter-laboratory variation in assessment of oxidatively damaged DNA by the comet assay in terms of oxidized purines converted to strand breaks with formamidopyrimidine DNA glycosylase (FPG). Coded samples with DNA oxidation damage induced by treatment with different concentrations of photosensitizer (Ro 19-8022) plus light and calibration samples irradiated with ionizing radiation were distributed to the 10 participating laboratories to measure DNA damage using their own comet assay protocols. Nine of 10 laboratories reported the same ranking of the level of damage in the coded samples. The variation in assessment of oxidatively damaged DNA was largely due to differences in protocols. After conversion of the data to lesions/10(6) bp using laboratory-specific calibration curves, the variation between the laboratories was reduced. The contribution of the concentration of photosensitizer to the variation in net FPG-sensitive sites increased from 49 to 73%, whereas the inter-laboratory variation decreased. The participating laboratories were successful in finding a dose-response of oxidatively damaged DNA in coded samples, but there remains a need to standardize the protocols to enable direct comparisons between laboratories.


Loss of Control as a Discriminating Factor Between Different Latent Classes of Disordered Gambling Severity.
Analyses of disordered gambling assessment data have indicated that commonly used screens appear to measure latent categories. This stands in contrast to the oft-held assumption that problem gambling is at the extreme of a continuum. To explore this further, we report a series of latent class analyses of a number of prevalent problem gambling assessments (PGSI, SOGS, DSM-IV Pathological Gambling based assessments) in nationally representative British surveys between 1999 and 2012, analysing data from nearly fifty thousand individuals. The analyses converged on a three class model in which the classes differed by problem gambling severity. This identified an initial class of gamblers showing minimal problems, a additional class predominantly endorsing indicators of preoccupation and loss chasing, and a third endorsing a range of disordered gambling criteria. However, there was considerable evidence to suggest that classes of intermediate and high severity disordered gamblers differed systematically in their responses to items related to loss of control, and not simply on the most 'difficult' items. It appeared that these differences were similar between assessments. An important exception to this was one set of DSM-IV criteria based analyses using a specific cutoff, which was also used in an analysis that identified an increase in UK problem gambling prevalence between 2007 and 2010. The results suggest that disordered gambling has a mixed latent structure, and that present assessments of problem gambling appear to converge on a broadly similar construct.


Do participants in a physical activity program from a Care Sport Connector become healthier? An explorative study from the Netherlands.
Care Sport Connectors (CSCs) have been appointed to create a connection between the primary care and physical activity (PA) sectors to stimulate residents who are inactive to become more physically active to gain health benefits. The objective of this explorative study was to find out whether CSCs achieve these goals by testing the hypothesis that more residents become physically active, and score higher for health-related fitness and health-related quality of life. We conducted a longitudinal study design whereby participants (n = 402) were measured at three time points: at the start of their PA program (T0); after 6 months (T1); and after 1 year (T2). Participants conducted a fitness test to measure their health-related physical fitness and filled in questionnaires to assess PA level (PA-, Fit-, Combi-, and sport norm), health-related quality of life, motivation for PA, and personal information. We used a multi-level analysis to test whether outcomes of participants differ over time. Participants who dropped out and maintainers were compared with a chi-square test and a one-way ANOVA. This study showed that one-third of the participants dropped out (n = 139). Participants who dropped out were, compared with maintainers, less physically active (P = 0.004) and were more often reached in bigger municipalities, by an integral approach. More participants meet the PA norm (P = 0.007) and sport norm (P<0.001) at T2 then at T0. Scores in health-related physical fitness and quality of life were significant but not a meaningful gain in health-related fitness. More residents become physically active and participate in sport because they took part in a PA programs or activity organized by a CSC. Lifestyle interventions should be offered with a higher frequency, intensity, and focus on behavior change. It is necessary to invest in combined lifestyle interventions offered by a collaboration of primary care, welfare, and PA professionals.


Enabling Oxidation Protection and Carrier-Type Switching for Bismuth Telluride Nanoribbons via in Situ Organic Molecule Coating.
Thermoelectric materials with high electrical conductivity and low thermal conductivity (e.g., Bi2Te3) can efficiently convert waste heat into electricity; however, in spite of favorable theoretical predictions, individual Bi2Te3 nanostructures tend to perform less efficiently than bulk Bi2Te3. We report a greater-than-order-of-magnitude enhancement in the thermoelectric properties of suspended Bi2Te3 nanoribbons, coated in situ to form a Bi2Te3/F4-TCNQ core-shell nanoribbon without oxidizing the core-shell interface. The shell serves as an oxidation barrier but also directly functions as a strong electron acceptor and p-type carrier donor, switching the majority carriers from a dominant n-type carrier concentration (∼1021 cm-3) to a dominant p-type carrier concentration (∼1020 cm-3). Compared to uncoated Bi2Te3 nanoribbons, our Bi2Te3/F4-TCNQ core-shell nanoribbon demonstrates an effective chemical potential dramatically shifted toward the valence band (by 300-640 meV), robustly increased Seebeck coefficient (∼6× at 250 K), and improved thermoelectric performance (10-20× at 250 K).


Co-localization of apoptosis-regulating proteins in mouse mammary epithelial HC11 cells exposed to TGF-beta1.
TGF-beta1 is an apoptogenic agent for mammary epithelial cells (MEC). The molecular mechanism of the TGF-beta1-induced apoptosis remains, however, obscure. In the present study we used laser scanning cytometry, confocal microscopy and immunogold electron microscopy to analyze the expression, aggregation and co-localization of caspase-8, Bid, Bax and VDAC-1. These proteins are regarded as the most important factors involved in the regulatory phase of TGF-beta1-induced apoptosis. Apoptosis in HC11 mouse MEC manifested with a simultaneous increase in expression and subcellular aggregation of caspase-8, Bid, Bax and VDAC-1. Confocal microscopy revealed a strong pattern of co-localization of examined proteins during both early and late apoptosis. Experiments with double- and triple-staining immunoelectron microscopy showed a co-localization of Bax/Bid, caspase-8/Bax/Bid, and Bax/VDAC-1, on the membranes of mitochondria, Golgi apparatus, rough endoplasmic reticulum, nuclear envelope, nuclear pore, and within the nucleus. In conclusion, the observed pattern of changes in aggregation and subcellular localization of caspase-8, Bid, Bax and VDAC-1 during TGF-beta1-induced apoptosis in HC11 mouse MEC suggests an interaction between these proteins and formation of multimeric complexes on organellar membranes, thus controlling their permeability for intracellular mediators of apoptosis.


[Women and alcohol: From risky use to addiction, the issues at stake].
Present in all social circles with proven effects on health, alcohol is the second leading cause of avoidable death after tobacco. Like men, women can be affected by alcohol misuse, or even addiction. As the female organism is more vulnerable to the deleterious effects of alcohol, women are particularly exposed in terms of risks and complications for their physical and mental health.


Perturbations in nucleosome structure from heavy metal association.
Heavy metals have the potential to engage in strong bonding interactions and can thus function in essential as well as toxic or therapeutic capacities. We conducted crystallographic analyses of heavy cation binding to the nucleosome core particle and found that Co(2+) and Ni(2+) preferentially associate with the DNA major groove, in a sequence- and conformation-dependent manner. Conversely, Rb(+) and Cs(+) are found to bind only opportunistically to minor groove elements of the DNA, in particular at narrow AT dinucleotide sites. Furthermore, relative to Mn(2+) the aggressive coordination of Co(2+) and Ni(2+) to guanine bases is observed to induce a shift in histone-DNA register around the nucleosome center by stabilizing DNA stretching over one region accompanied by expulsion of two bases at an opposing location. These 'softer' transition metals also associate with multiple histone protein sites, including inter-nucleosomal cross-linking, and display a proclivity for coordination to histidine. Sustained binding and the ability to induce structural perturbations at specific locations in the nucleosome may contribute to genetic and epigenetic mechanisms of carcinogenesis mediated by Co(2+) and Ni(2+).


[Duration of splint therapy in patients with temporomandibular joint disorders].
Studying the duration of treatment in patients with temporomandibular joint pain dysfunction syndrome, and the relationship of the duration of treatment with the age of the patient at the beginning of therapy. The study was carried out using information from medical records of dental patients, information from additional examinations of patients who were treated at the National Medical Research Centre for Dentistry and Maxillofacial Surgery of the Ministry of Health of Russia from 2016 to 2022. Statistical research methods: to evaluate the normality of the distribution, graphical methods were used, as well as the Shapiro-Wilk criterion. The duration of splint therapy in patients with temporomandibular joint pain dysfunction syndrome varied from 4 to 27 months. The average duration of treatment of patients using occlusive splints was 10.5±5.3 months. Without abnormal observations, the average duration of splint therapy in patients with TMJ pain syndrome was 9.6±4.1 months. The obtained data allow stating the absence of a correlation between the age of patients and the duration of splint-therapy. The majority of patients (68.4%) complete the splint therapy stage within 1 year, and a very small part (1.8%) are treated for more than 1.5 years. The duration of treatment of patients with temporomandibular joint pain dysfunction syndrome does not depend on age or gender.


Comparative Study of Adeno-associated Virus, Adenovirus, Bacu lovirus and Lentivirus Vectors for Gene Therapy of the Eyes.
The eye possesses unique anatomical features that make it a valuable target for gene therapy applications. The aim of the current study was to compare transduction efficiency, safety and biodistribution of four viral vectors following intravitreal injection. Adenovirus (AdV), Adeno-Associated Virus (AAV), Baculovirus (BV) and Lentivirus (LV) vectors encoding Green Fluorescent Protein (GFP) were injected bilaterally intravitreally into adult C57BL/6OlaHsd mice. Control mice received saline. Eyes and other organs were studied at multiple time points from 3 days to 6 months. Immunohistochemical stainings with retinal cell markers were performed to verify GFP-positive cells. Biodistribution in retina and various non-target tissues was studied using a qPCR method. Inflammatory responses and toxicity were investigated from cryostat eye sections and serum samples. AAV-injected eyes showed GFP expression both in inner and outer retinal cells from 7 days up to 6 months. LV eyes showed long lasting transgene expression mostly in retinal pigment epithelium whereas AdV transiently transduced mainly cells in the anterior chamber. In BV-injected eyes, GFP positivity was very low. qPCR results showed that AdV, AAV and LV spread into the optic nerve, but were below the detection limit in other organs. The strongest immune responses were evoked by intravitreal injections of AdV and BV. The highest concentration of anti-GFP IgG was detected in the AdV-treated group, whereas the AAV group showed the lowest concentration. Neither blood chemistry screen nor the number of apoptotic cells showed any differences between the viral vector and saline injected groups. Our findings show that intravitreal gene delivery is safe and feasible with AAV, AdV and lentivirus vectors.


Chitosan and Its Derivatives - Biomaterials with Diverse Biological Activity for Manifold Applications.
Derived from chitin, chitosan is a natural polycationic linear polysaccharide being the second most abundant polymer next to cellulose. The main obstacle in the wide use of chitosan is its almost complete lack of solubility in water and alkaline solutions. To break this obstacle, the structure of chitosan is subjected to modification, improving its physic-chemical properties and facilitating application as components of composites or hydrogels. Derivatives of chitosan are biomaterials useful for different purposes because of their lack of toxicity, low allergenicity, biocompatibility and biodegradability. This review presents the methods of chemical modifications of chitosan which allow to obtain tailor- made properties required for a variety of biomedical applications. Selected pharmaceutical and biomedical applications of chitosan derivatives are also highlighted. Possibility to manage waste from arthropod and crab processing is also emphasized.


Posterolateral approaches to the craniovertebral junction.
The indications and operative technique for a number of posterolateral approaches to the craniovertebral junction (CVJ) are reviewed. The literature addressing posterolateral approaches to the CVJ is reviewed, and illustrative cases are presented. The far lateral approach and its variants, including the transcondylar, supracondylar, and paracondylar approaches, are an effective means of addressing intradural anterior and anterolateral CVJ lesions. These approaches provide exposure of the lower third of the clivus, the foramen magnum, and the upper cervical spine; do not cross contaminated regions; and enable a watertight dural closure to be performed. They are associated with minimal morbidity and usually do not significantly decrease the stability of the CVJ. All surgeons treating lesions of the CVJ should be familiar with the posterolateral approach and its modifications.