Direct evidence for RNA-RNA interactions at the 3' end of the Hepatitis C virus genome using surface plasmon resonance.
Surface plasmon resonance was used to investigate two previously described interactions analyzed by reverse genetics and complementation mutation experiments, involving 5BSL3.2, a stem-loop located in the NS5B coding region of HCV. 5BSL3.2 was immobilized on a sensor chip by streptavidin-biotin coupling, and its interaction either with the SL2 stem-loop of the 3' end or with an upstream sequence centered on nucleotide 9110 (referred to as Seq9110) was monitored in real-time. In contrast with previous results obtained by NMR assays with the same short RNA sequences that we used or SHAPE analysis with longer RNAs, we demonstrate that recognition between 5BSL3.2 and SL2 can occur in solution through a kissing-loop interaction. We show that recognition between Seq9110 and the internal loop of 5BSL3.2 does not prevent binding of SL2 on the apical loop of 5BSL3.2 and does not influence the rate constants of the SL2-5BSL3.2 complex. Therefore, the two binding sites of 5BSL3.2, the apical and internal loops, are structurally independent and both interactions can coexist. We finally show that the stem-loop SL2 is a highly dynamic RNA motif that fluctuates between at least two conformations: One is able to hybridize with 5BSL3.2 through loop-loop interaction, and the other one is capable of self-associating in the absence of protein, reinforcing the hypothesis of SL2 being a dimerization sequence. This result suggests also that the conformational dynamics of SL2 could play a crucial role for controlling the destiny of the genomic RNA.


Circadian rhythm of heart rate variability is reversibly abolished in ischemic stroke.
Acute brain infarction significantly decreases heart rate variability as a result of cardiovascular autonomic dysregulation. However, information regarding circadian rhythms of heart rate and heart rate variability is limited. In this prospective study, we analyzed 24-hour circadian rhythm of heart rate and the time and frequency domain measures of heart rate variability in 24 patients with hemispheric brain infarction, 8 patients with medullary brainstem infarction, and 32 age- and sex-matched healthy control subjects. ECG data were obtained from the patients in the acute phase and at 6 months after the infarction. In the acute phase of stroke, all the components of heart rate variability, ie, standard deviation of RR intervals, total power, high-frequency power, low-frequency power, and very-low-frequency power, were similar at night (from midnight to 6 AM) and during the day (from 9 AM to 9 PM), indicating that the circadian oscillation of heart rate variability had been abolished. At 6 months after brain infarction, the circadian rhythm had returned and, as in the control subjects, the values at night were significantly higher than those in the daytime. The values in hemispheric and in brainstem infarction did not differ significantly from each other. These results suggest that circadian fluctuation of heart rate variability is reversibly abolished in the acute phase of ischemic stroke and that it returns during the subsequent 6 months. The loss of the relative vagal nocturnal dominance may contribute to the incidence of cardiac arrhythmias and other cardiovascular complications after acute stroke.


On modelling large deformations of heterogeneous biological tissues using a mixed finite element formulation.
This study addresses the issue of modelling material heterogeneity of incompressible bodies. It is seen that when using a mixed (displacement-pressure) finite element formulation, the basis functions used for pressure field may not be able to capture the nonlinearity of material parameters, resulting in pseudo-residual stresses. This problem can be resolved by modifying the constitutive relation using Flory's decomposition of the deformation gradient. A two-parameter Mooney-Rivlin constitutive relation is used to demonstrate the methodology. It is shown that for incompressible materials, the modification does not alter the mechanical behaviour described by the original constitutive model. In fact, the modified constitutive equation shows a better predictability when compared against analytical solutions. Two strategies of describing the material variation (i.e. linear and step change) are explained, and their solutions are evaluated for an ideal two-material interfacing problem. When compared with the standard tied coupling approach, the step change method exhibited a much better agreement because of its ability to capture abrupt changes of the material properties. The modified equation in conjunction with integration point-based material heterogeneity is then used to simulate the deformations of heterogeneous biological structures to illustrate its applications.


Serum cholesterol response to dietary cholesterol.
Serum cholesterol change, delta Chol mg/dl, from change only in dietary cholesterol, mg/1,000 kcal, x, was analyzed with data from 39 controlled experiments on 64 adults with the model delta Chol = b(x2k-x1k). Values of b were found by iteration for exponents k = 0.3 to k = i.0 yielding eight regression equations to check predicted against observed. The smallest root mean square deviation regression was 7 mg/dl given by delta Chol = 1.5(x2(1/2)-x1(1/2] which predicted an average serum cholesterol change of 18.7 versus 19.9 observed. The relationship pertains to calorie balance on a wide range of nutritionally acceptable diets. A prediction equation proposed from experiments in Massachusetts that took k = 1.0 overpredicted the effect of dietary cholesterol in these experiments by 300%.


Selective utilization of pre-operative endoscopic ultrasound to exclude choledocholithiasis prior to laparoscopic cholecystectomy: a retrospective study.
The objective of this study was to define the clinical, biochemical and ultrasonographic criteria correlating with a likelihood of a positive preoperative endoscopic ultrasound (EUS) in patients presenting with acute gallstone-related pancreatobiliary disease. All patients who underwent EUS prior to elective laparoscopic cholecystectomy were analyzed at the Gastroenterology Unit, Kaplan Medical Center, following acute admission with cholecystitis, cholelithiasis, cholangitis, acute pancreatitis and obstructive jaundice. One hundred and seventy four patients met the inclusion criteria. EUS showed choledocholithiasis in 37 (21.3%) with 35/37 undergoing a preoperative ERCP (24/35 cases - 64.9% positive yield). Twenty seven of the positive EUS studies (73%) were performed during the acute illness, with 50 of the 137 negative studies (36.4%) performed during the acute phase of the illness (p=0.0001). On multivariate analysis, a positive EUS was more commonly found during the acute phase of the illness [OR: 3.445; 95% CI: 1.48-8.008, p=0.004] or if there was transient jaundice [OR: 1.167; 95% CI: 1.002-1.36, p=0.047]. The timing of the examination influences the detection rate of CBD stones by EUS prior to surgery although it may be appropriate to more selectively use EUS in those patients with hyperbilirubinaemia and/or where the CBD is dilated.


Measuring student motivation on the use of a mobile assisted grammar learning tool.
Language learning is an emerging research area where researchers have done significant contributions by incorporating technological assistantship (i.e., computer- and mobile-assistant learning). However, it has been revealed from the recent empirical studies that little attention is given on grammar learning with the proper instructional materials design and the motivational framework for designing an efficient mobile-assisted grammar learning tool. This paper hence, reports a preliminary study that investigated learner motivation when a mobile-assisted tool for tense learning was used. This study applied the Attention-Relevance-Confidence-Satisfaction (ARCS) model. It was hypothesized that with the use of the designed mobile- assisted tense learning tool students would be motivated to learn grammar (English tense). In addition, with the increase of motivation, performance outcome in paper- based test would also be improved. With the purpose to investigate the impact of the tool, a sequential mixed-method research design was employed with the use of three research instruments; Instructional Materials Motivation Survey (IMMS), a paper-based test and an interview protocol using a semi-structured interview. Participants were 115 undergraduate students, who were enrolled in a remedial English course. The findings showed that with the effective design of instructional materials, students were motivated to learn grammar, where they were positive at improving their attitude towards learning (male 86%, female 80%). The IMMS findings revealed that students' motivation increased after using the tool. Moreover, students improved their performance level that was revealed from the outcome of paper-based instrument. Therefore, it is confirmed that the study contributed to designing an effective multimedia based instructions for a mobile-assisted tool that increased learners' motivational attitude which resulted in an improved learning performance.


Effects of the T/L-type calcium channel blocker benidipine on albuminuria and plasma aldosterone concentration. A pilot study involving switching from L-type calcium channel blockers to benidipine.
Albuminuria and a high plasma aldosterone concentration (PAC) are prognosis factors predicting a poor outcome for cardiovascular disease. We examined here the effects of benidipine, a T/L-type calcium channel blocker (CCB), on albuminuria and PAC.Thirty-one patients with essential hypertension who received an L-type CCB and achieved the target blood pressure (BP) indicated by the Treatment Guidelines of the Japan Society of Hypertension (JSH2009) were investigated. The Ltype CCB under treatment was switched to benidipine at a dose in which equivalent BP reduction was expected. BP and estimated glomerular filtration rate at 6 months after switching to benidipine were not significantly different from those at baseline. The urinary-albumin-creatinine ratio (UACR) decreased significantly by 36.9% (P = 0.001). No significant change was observed in plasma renin activity (P = 0.063). The PAC of all patients decreased significantly by 11.8% (P = 0.002). When analyzed by daily doses of benidipine, the PAC appeared to have decreased in patients who received 4 mg per day of benidipine (n = 14), although statistical significance was not reached (P = 0.096). The PAC in patients who received 8 mg per day of benidipine (n =17) was significantly reduced by 13.2% (P = 0.017).In hypertensive patients whose BP is controlled by L-type CCB, switching to the T/L-type CCB benidipine maintained BP control and reduced UACR. In addition, the high dose of benidipine reduced the PAC independent of BP control. These results suggest the T/L-type CCB benidipine may contribute to cardio-renal protection in addition to lowering BP.


Case-controlled comparison of video-assisted and conventional minimally invasive parathyroidectomy.
Video-assisted parathyroidectomy (VAP) is a new approach to parathyroid exploration for primary hyperparathyroidism (PH). We examined the VAP learning curve and hypothesized that compared with conventional minimally invasive parathyroidectomy (MIS), VAP has similar complication rates and the added benefit of a shorter hospital length of stay. Using a case-control study design, patients with PH with single-focus imaging results undergoing VAP or MIS were compared during a 5-year VAP implementation period. VAP was possible in 18 per cent of patients undergoing initial parathyroid exploration. In comparing 125 VAP cases with 95 MIS control subjects, patients undergoing MIS had higher mean preoperative levels of calcium (P = 0.007) and parathyroid hormone (P = 0.008), greater mean adenoma weight (P < 0.001), and increased long-term mortality (4% MIS vs 0% VAP, P = 0.03). Mean operative time, in-house analgesia use, and operative complications did not differ. The rate of conversion from VAP to MIS was 14 per cent. Patients undergoing VAP were less likely to require an overnight hospital stay (P = 0.01). VAP is a safe surgical option for selected patients with PH, offering improved cosmesis with operative times comparable to conventional MIS. VAP can be done with a low conversion rate even during implementation and allows the added benefit of shorter hospital stay.


[Monitoring of winter wheat stripe rust based on the spectral knowledge base for TM images].
In most cases, the reversion model for monitoring the severity degree of stripe rust based on the hyperspectral information can not be directly applied by the satellite images with relatively broad bandwidth, while the airborne hyperspectral images can not be applied for large-scale monitoring either, due to the scale limitation of its data and high cost. For resolving this dilemma, we developed a monitoring method based on PHI images, which relies on the construction of spectral knowledge base of winter wheat stripe rust. Three PHI images corresponding to the winter wheat experimental field that included different severity degree of stripe rust were used as a medium to establish the spectral knowledge base of relationships between disease index (DI) and the simulated reflectance of TM bands by using the empirical reversion model of DI(%) and the relative spectral response (RSR) function of TM-5 sensor. Based on this, we can monitor and identify the winter wheat stripe rust by matching the spectral information of an untested pixel to the spectral knowledge base via Mahalanobis distance or spectral angle mapping (SAM). The precision of monitoring was validated by simulated TM pixels, while the effectiveness of identification was tested by pixels from TM images. The results showed that the method can provide high precision for monitoring and reasonable accuracy for identification in some certain growth stages of winter wheat. Based on the simulated TM pixels, the model performed best in the pustulation period, yielded a coefficient of determination R2 = 0.93, while the precision of estimates dropped in the milk stage, and performed worst in the jointing stage, which is basically inappropriate for monitoring. Moreover, by using the pixels from TM images, the infected pixels could be identified accurately in pustulation and milk stages, while failed to be identified in jointing stage. For matching algorithms, the Mahalanobis distance method produced a slightly better result than SAM method.


Retention of a co-translational translocated mutant protein of carboxypeptidase Y of Saccharomyces cerevisiae in endoplasmic reticulum.
Co-translational translocation of Saccharomyces cerevisiae vacuolar glycoprotein carboxypeptidase Y (CpY) was highly efficient when studied with an in vivo and in vitro homologous system, comparison of limited proteolytic cleavage of immunoprecipitated translational products of CpY and subcellular localisation of a mutant CpY. The efficient segregation of CpY mRNA in highly purified fractions of rough microsomes was characterised. CpY1 mutant showed retention of core glycosylated material (proCpY1) in the rough and smooth endoplasmic reticulum fractions. It is suggested that the presence of structures that are incompatible with intercompartmental transport of vacuolar protein leads to retention of the mutated CpY by the endoplasmic reticulum.


Force-induced recruitment of cten along keratin network in epithelial cells.
The cytoskeleton provides structural integrity to cells and serves as a key component in mechanotransduction. Tensins are thought to provide a force-bearing linkage between integrins and the actin cytoskeleton; yet, direct evidence of tensin's role in mechanotransduction is lacking. We here report that local force application to epithelial cells using a micrometer-sized needle leads to rapid accumulation of cten (tensin 4), but not tensin 1, along a fibrous intracellular network. Surprisingly, cten-positive fibers are not actin fibers; instead, these fibers are keratin intermediate filaments. The dissociation of cten from tension-free keratin fibers depends on the duration of cell stretch, demonstrating that the external force favors maturation of cten-keratin network interactions over time and that keratin fibers retain remarkable structural memory of a cell's force-bearing state. These results establish the keratin network as an integral part of force-sensing elements recruiting distinct proteins like cten and suggest the existence of a mechanotransduction pathway via keratin network.


Season of birth and eating disorders.
A statistically significant season of birth variation is found in an unselected nationwide sample of 1,939 eating disorders patients, with peak season of birth occurring in May. However, among younger patients (n = 882), peak season of birth is in March, which is statistically significantly different to that expected from the general population season of birth cycle. This finding may imply links between etiology of earlier-onset eating disorders and the psychoses; similar first quarter peak seasonal patterns of birth have been found in schizophrenic and affective psychoses--with birth peaks in January and February. In contrast, for the neuroses and personality disorders, birth peaks have been found to be in June and August, similar to the June birth peak found in this study for later-onset eating disorders (n = 1,057), which was not statistically significantly different to season of birth peaks expected from general population data.


Mental Health Services in Texas Jails.
Jails and prisons in the United States have become the places where people with mental illness go. Texas jails were surveyed in 2012 to learn how they screened inmates for mental illness. Of these jails, 13% responded. Most screened for suicidal ideation and whether or not an inmate took a medicine. About half the jails offered in-house care, and the other half referred inmates to the local mental health authority. Most jails had a formal jail diversion program, and most thought that mental health illness was increasing. About half had an annual 4-hour training program for staff. Recommendations are made for future care in jails.


Patients with acute myocardial infarction and interfering factors when seeking emergency care: implications for health education.
With the objective of estimating the time elapsed between the beginning of the signs and symptoms of a heart attack until the arrival at the cardiology emergency service (Delta T) and the factors that influence in this process, 112 patients were studied, with an infarction diagnosis with supraunleveling ST segment. The delta T was on average of 3h59+/-2h55min; 99(88%) of those patients sought out an emergency service within 1h after the beginning of the event. Unmarried patients presented a delta T smaller in relation to the others (P=0,006), as well as those that recognized the symptoms as a heart event; thoracic pain with burning symptoms, were described by 25 (24%) of patients, and the first attitude taken, in view those symptoms, was self-medication 37(33%). The recognition of the signs and symptoms of Acute Myocardium Infarction, is a decisive factor for the seeking of a specialized service.


Per Artursson's Major Contributions to the Caco-2 Cell Literature in Pharmaceutical Sciences.
This edition of the Journal of Pharmaceutical Sciences is dedicated to the wonderful career of Per Artursson from the Uppsala University. My Commentary focusses on Per's major contributions to the Caco-2 cell literature over the past 30 years. Two especially influential papers have been cited more than 1000 times out of a total citation count of almost 30,000 and a h-index of 93 (Google Scholar), making Per one of the most cited and influential Pharmaceutical scientists of his generation. The Caco-2 field to which Per contributed so many advances has informed the community on key areas including predictive drug fluxes across the intestine, metabolism by intestinal epithelia, the role of transporters during flux, enantiomer-selective flux, excipient interaction with tight junctions, and nanoparticle uptake by enterocytes. In this pioneering work, Per has been careful to emphasise that Caco-2 monolayers have limitations and are a model of the human small intestine where observations must be backed up with in vitro tissue and in vivo work. Throughout, he has paid great attention to detail in methodology, as reflected by co-authorship of two Nature Protocols on Caco-2 assays. The article briefly assesses some of the most important milestones in Per's published Caco-2 research.


Adherence to and Dropout from Liraglutide 3.0 mg Obesity Treatment in a Real-World Setting.
The factors associated with non-adherence to obesity treatment using liraglutide 3.0 mg in a real-world setting remain elusive. We performed a secondary data analysis of 769 participants treated with liraglutide 3.0 mg from December 2017 to June 2020 at nine Korean hospitals. Data were collected 2, 4, and 6 months after treatment initiation. Adherence groups were defined as <2, 2-4, 4-6, and ≥6 months. Among the 769 patients, 539 (70.1%) were lost to follow-up within 6 months because of unknown reasons (54.2%), adverse events (14.8%), change of treatment (13.7%), or discontinuation due to poor weight loss (9.3%). Dropout at 6 months was significantly associated with the presence of diabetes mellitus in step 1 and the presence of diabetes mellitus with regular exercise in step 2 of the logistic regression analysis using the forward stepwise selection method. After adjusting for covariates, the presence of diabetes mellitus (odds ratio [OR], 0.25; 95% confidence interval [CI], 0.10-0.63; OR, 0.47; 95% CI, 0.31-0.73; and OR, 0.52; 95% CI, 0.34-0.80) and regular exercise (OR, 2.86; 95% CI, 1.31-6.23; OR, 2.09; 95% CI, 1.26-3.48; and OR, 2.99; 95% CI, 1.81-4.92) showed significant associations in the <2, 2-4, and 4-6 groups compared with the highest adherence group (≥6 months). Non-adherence to obesity treatment with liraglutide is related to regular exercise and absence of diabetes mellitus. Further prospective studies are warranted to increase medication adherence in those groups.


Plantazolicin, a novel microcin B17/streptolysin S-like natural product from Bacillus amyloliquefaciens FZB42.
Here we report on a novel thiazole/oxazole-modified microcin (TOMM) from Bacillus amyloliquefaciens FZB42, a Gram-positive soil bacterium. This organism is well known for stimulating plant growth and biosynthesizing complex small molecules that suppress the growth of bacterial and fungal plant pathogens. Like microcin B17 and streptolysin S, the TOMM from B. amyloliquefaciens FZB42 undergoes extensive posttranslational modification to become a bioactive natural product. Our data show that the modified peptide bears a molecular mass of 1,335 Da and displays antibacterial activity toward closely related Gram-positive bacteria. A cluster of 12 genes that covers ∼10 kb is essential for the production, modification, export, and self-immunity of this natural product. We have named this compound plantazolicin (PZN), based on the association of several producing organisms with plants and the incorporation of azole heterocycles, which derive from Cys, Ser, and Thr residues of the precursor peptide.


Bone abnormalities in the surgical treatment of patients with rheumatoid arthritis.
Profound abnormalities of bone are an important component of the morbidity of rheumatoid arthritis and partly determine the orthopaedic treatment of patients. The principal bone abnormality, which is osteoporosis associated with rapid remodeling, degrades the mechanical properties of the skeleton in juxtaarticular bone, in the diaphyses of long bones, in the pelvis, and in the base of skull. Abnormalities of bone affect clinical decision making in the surgical treatment of patients with rheumatoid arthritis and must be considered for the optimal treatment of these patients. Increased fracture risk and compromised bony fixation complicate fracture treatment. Techniques of surgery must be modified to protect bone from intraoperative fracture. Methods of fixation of implants and other devices must be appropriate to the biomechanics of the bone. Bone healing usually is rapid if not compromised by mechanical instability. Specific patterns of bone deformation and failure can be identified in the evolution of rheumatoid arthritis. Bone graft used in reconstruction of the protruded acetabulum is incorporated rapidly. Bone resorption with joint instability is a common feature of rheumatoid arthritis; however, the opposite pattern of bony ankylosis with stiffness is observed in a smaller percentage of patients. Recognition of the tissue type is necessary for the individualization of surgical procedures to achieve optimal joint stability and mobility. Based on growing understanding of the pathophysiology of bone in patients with rheumatoid arthritis, new pharmacologic therapies may become available for the prevention and treatment of bone abnormalities in patients with rheumatoid arthritis.


Gonadal dysgenesis in a 46,XY female mosaic for double autosomal trisomies 8 and 21.
The proband was evaluated at 19 years of age because of primary amenorrhoea and, on chromosomal analysis, was found to have a 46,XY karyotype in 75% of her cells and 48,XY, +8, +21 in 25% of her cells. She appeared normal at birth and exhibited normal intellectual and physical development until puberty when secondary sexual differentiation failed. This young women showed none of the dysmorphic features associated with either trisomy 8 or trisomy 21. Her XY gonadal dysgenesis was manifested by late developmental problems of amenorrhoea, sexual infantilism, and gonadal neoplasia.


Catastrophic Health Expenditure Among Colorectal Cancer Patients and Families: A Case of Malaysia.
This study aimed to estimate the cost of colorectal cancer (CRC) management and to explore the prevalence and determinants of catastrophic health expenditure (CHE) among CRC patients and their families arising from the costs of CRC management. Data were collected prospectively from 138 CRC patients. Patients were interviewed by using a structured questionnaire at the time of the diagnosis, then at 6 months and 12 months following diagnosis. Simple descriptive methods and multivariate binary logistic regression were used in the analysis. The mean cost of managing CRC was RM8306.9 (US$2595.9), and 47.8% of patients' families experienced CHE. The main determinants of CHE were the economic status of the family and the likelihood of the patient undergoing surgery. The results of this study strongly suggest that stakeholders and policy makers should provide individuals with financial protection against the consequences of cancer, a costly illness that often requires prolonged treatment.


Epithelial barrier function of the filtering bleb conjunctiva and the cornea after trabeculectomy with mitomycin C.
To evaluate epithelial barrier function of the filtering bleb conjunctiva and the cornea after trabeculectomy with adjunct, intraoperative mitomycin C, by using a newly developed slit-lamp fluorophotometer. Thirteen patients with normal-tension glaucoma with a cystic filtering bleb who had undergone unilateral trabeculectomy with mitomycin C only once at least 1 year previously were subjected to the study. Epithelial barrier function of the bleb conjunctiva and the cornea in the operated eye was evaluated using an Anterior Fluorometer FL-500 to measure the uptake of topically applied fluorescein. Fluorescein uptake (nanograms per milliliter, mean +/- standard error) by the bleb conjunctiva (1,857 +/- 380) was not significantly different from either the uptake by the superior bulbar conjunctiva in the operated eyes, located approximately 90 degrees C away from the filtering bleb (1,974 +/- 258), or the uptake by the conjunctiva in the intact eyes, located symmetrically against the bleb (1,913 +/- 248). The corneal uptake in the operated eye (56.1 +/- 10.2) was not significantly different from that in the intact eye (53.5 +/- 8.3). Epithelial barrier function of the cystic filtering bleb conjunctiva and the cornea long after trabeculectomy with mitomycin C was demonstrated to be almost identical to that of the control.


The Target of Rapamycin pathway antagonizes pha-4/FoxA to control development and aging.
FoxA factors are critical regulators of embryonic development and postembryonic life, but little is know about the upstream pathways that modulate their activity. C. elegans pha-4 encodes a FoxA transcription factor that is required to establish the foregut in embryos and to control growth and longevity after birth. We previously identified the AAA+ ATPase homolog ruvb-1 as a potent suppressor of pha-4 mutations. Here we show that ruvb-1 is a component of the Target of Rapamycin (TOR) pathway in C. elegans (CeTOR). Both ruvb-1 and let-363/TOR control nucleolar size and promote localization of box C/D snoRNPs to nucleoli, suggesting a role in rRNA maturation. Inactivation of let-363/TOR or ruvb-1 suppresses the lethality associated with reduced pha-4 activity. The CeTOR pathway controls protein homeostasis and also contributes to adult longevity. We find that pha-4 is required to extend adult lifespan in response to reduced CeTOR signaling. Mutations in the predicted CeTOR target rsks-1/S6 kinase or in ife-2/eIF4E also reduce protein biosynthesis and extend lifespan, but only rsks-1 mutations require pha-4 for adult longevity. In addition, rsks-1, but not ife-2, can suppress the larval lethality associated with pha-4 loss-of-function mutations. The data suggest that pha-4 and the CeTOR pathway antagonize one another to regulate postembryonic development and adult longevity. We suggest a model in which nutrients promote TOR and S6 kinase signaling, which represses pha-4/FoxA, leading to a shorter lifespan. A similar regulatory hierarchy may function in other animals to modulate metabolism, longevity, or disease.


[Kooy's explanation of the drop in the marriage age in the Netherlands between 1958 and 1968].
A critique of an article by G. A. Kooy on reasons for the declining age at marriage in the Netherlands is presented. The author argues first "that a normative change does not in itself constitute the cause of a behavioural change: both changes ought to be related to a common determinant; second, that patrism and matrism, being two alternative ways of managing an Oedipal tension, do not constitute a social process and hence cannot explain social change." (summary in ENG)


The Early Infancy Temperament Questionnaire.
Although there are several scales routinely used for assessment of temperament during the first year of life, none of them is well suited to the infant younger than 4 months old. The Early Infancy Temperament Questionnaire (EITQ) was designed to meet this need. The EITQ is a 76-item parent questionnaire for assessing the nine New York Longitudinal Study temperament characteristics in 1- to 4-month-old infants. The majority of the items were adapted from the Revised Infant Temperament Questionnaire to be developmentally appropriate for the very young infant. The standardization population consisted of 404 infants from one pediatric practice. Means for the nine categories were calculated separately for infants from 1 to 2 months and 3 to 4 months old. Internal consistency for the nine categories ranged from .42 to .76. Test-retest scores, completed between 2 to 3 weeks after the first rating, ranged from .43 to .87, with generally increasing retest levels in the older age group. None of the categories showed significant differences between male and female infants. This newly developed instrument should enhance the ability of both researchers and clinicians to assess temperament reliably and to understand better its contribution to clinical problems in the very young child.


Successful Treatment of a 19-Month-Old Boy with Hepatitis Associated Aplastic Anemia by Infusion of Umbilical Cord-Derived Mesenchymal Stromal Cells: A Case Report.
Here we presented a case of a 19-month-old boy who developed severe aplastic anemia postacute hepatitis. He was treated successfully with the umbilical cord-derived mesenchymal stromal cells (UC-MSCs) infusion and cyclosporine A (CsA). The boy achieved both hematopoietic recovery and normal lymphocyte proportion. So far, his condition still remains stable. To our knowledge, there is a rare previous report on the utility of MSCs infusion for the treatment of hepatitis-associated aplastic anemia (HAAA). Considering the efficacy, safety, and strong operability, particularly for pediatric patient, the infusion of UC-MSCs combined with CsA could be an effective alternative for the treatment of HAAA.


[A new handpiece attachment for Nd:YAG laser ablation of emphysematous bullae].
Recently, attention has been drawn to lung volume reduction for bullous emphysema by wedge excision or laser ablation. We used a neodymium: yttrium-aluminum garnet (Nd:YAG) laser with a free-beam mode to ablate peripheral emphysematous areas of lung tissue. To aid us in this procedure, we developed an attachment consisting of a stainless steel rod 3 cm in effective length, with a 1.8-cm diameter ring attached to its tip, and installed it on the tip of the laser handpiece used for free-beam laser ablation. This device has the following distinct advantages: 1) the distance between the tip of the laser and the pleura can easily be held constant, 2) the angle of irradiation can easily be maintained at 90 degrees. These result in 3) a constant area of irradiation (one square centimeter), and 4) an irradiated area that is clearly distinguished by the ring. In addition, the attachment is 5) simple and easy to construct. Three patients with bullous emphysema underwent unilateral laser ablation of the visceral pleura with the new device. All patients recovered completely. We found this rod-and-ring attachment to be useful for free-beam laser ablation.


Fock-Space Coupled Cluster Theory: Systematic Study of Partial Fourth Order Triples Schemes for Ionization Potential and Comparison with Bondonic Formalism.
In this paper, we have made a systematic study of partial fourth order perturbative schemes due to triples to compute the ionization potential within Fock-space multi-reference coupled-cluster theory. In particular, we have obtained computationally less expensive correlation schemes due to fourth order triples. Prototype examples have been considered to explore the efficacy of the approximate methods mentioned, while the bondonic formalism supporting the bonding phenomenology is also respectively for the first time here advanced.


Analysis of advanced lung cancer patients diagnosed following emergency admission.
Data on prognosis and predictors of overall survival in advanced lung cancer patients diagnosed following emergency admission (DFEA) are currently lacking. We retrospectively analysed data from 771 patients with advanced nonsmall cell lung cancer between April 2004 and April 2012. Of the 771 patients, 103 (13%) were DFEA. DFEA was not an independent predictor of overall survival by multivariate Cox proportional hazard models, whereas good performance status (PS), epidermal growth factor receptor gene mutation, stage IIIB, adenocarcinoma and chemotherapy were independent predictors of overall survival (hazard ratio (95% CI) 0.36 (0.29-0.44), p<0.001; 0.49 (0.38-0.63), p<0.001; 0.64 (0.51-0.80), p<0.001; 0.81 (0.67-0.99), p=0.044; and 0.40 (0.31-0.52), p<0.001, respectively). Good PS just prior to opting for chemotherapy, but not at emergency admission, was a good independent predictor of overall survival in DFEA patients (hazard ratio (95% CI) 0.26 (0.12-0.55); p<0.001). DFEA is relatively common. DFEA and PS at emergency admission were not independent predictors of overall survival, but good PS just prior to opting for chemotherapy was an independent predictor of longer overall survival. Efforts to improve patient PS after admission should be considered vital in such circumstances.


Transcranial Magnetic Stimulation for the Assessment of Neurodegenerative Disease.
Transcranial magnetic stimulation (TMS) is a noninvasive technique that has provided important information about cortical function across an array of neurodegenerative disorders, including Alzheimer's disease, frontotemporal dementia, Parkinson's disease, and related extrapyramidal disorders. Application of TMS techniques in neurodegenerative diseases has provided important pathophysiological insights, leading to the development of pathogenic and diagnostic biomarkers that could be used in the clinical setting and therapeutic trials. Abnormalities of TMS outcome measures heralding cortical hyperexcitability, as evidenced by a reduction of short-interval intracortical inhibition and increased in motor-evoked potential amplitude, have been consistently identified as early and intrinsic features of amyotrophic lateral sclerosis (ALS), preceding and correlating with the ensuing neurodegeneration. Cortical hyperexcitability appears to form the pathogenic basis of ALS, mediated by trans-synaptic glutamate-mediated excitotoxic mechanisms. As a consequence of these research findings, TMS has been developed as a potential diagnostic biomarker, capable of identifying upper motor neuronal pathology, at earlier stages of the disease process, and thereby aiding in ALS diagnosis. Of further relevance, marked TMS abnormalities have been reported in other neurodegenerative diseases, which have varied from findings in ALS. With time and greater utilization by clinicians, TMS outcome measures may prove to be of utility in future therapeutic trial settings across the neurodegenerative disease spectrum, including the monitoring of neuroprotective, stem-cell, and genetic-based strategies, thereby enabling assessment of biological effectiveness at early stages of drug development.


Effects of melatonin on plasma oxidative stress in rats with streptozotocin induced diabetes.
The aim of this study was to evaluate the effect of single melatonin injection on plasma oxidative stress in rats with streptozotocin induced diabetes. Diabetes was induced after a single intraperitoneal dose of streptozotocin (60 mg/kg), while hyperglycemia was determined 10 days upon injection. Diabetic rats were divided into two groups. In the first group the injection of melatonin was applied intraperitoneally (20 mg/kg), while the second group received physiological solution. Twenty-four hours later the rats were killed and their blood was centrifuged. In the rat plasma the following parameters were evaluated: the glucose level, superoxide radical, lipid peroxidation, reduced glutathione, total antioxidant capacity, antioxidant enzymes and the aldose reductase activity. The injected melatonin decreased the superoxide radical in the rat plasma. Moreover, melatonin increased the total antioxidative capacity and the activity of antioxidative enzymes superoxide dismutase and glutathione peroxidase. These results indicate that melatonin is a strong scavenger, which may diminish negative effects of oxidative stress in diabetic rats 24 hours after its application The findings suggest that melatonin is also a strong antioxidant. It increases the antioxidant enzymes activity, inhibiting the release of superoxide radicals. A high total antioxidative capacity and the lower activity of aldose reductase enlarge melatonin scavenger capacity against reactive oxygen species in diabetic rats.


Modes of exocytosis and electrogenesis underlying canine biphasic insulin secretion.
Biphasic insulin secretion in response to glucose consists of a transient first phase followed by a progressive second phase. It is a well described feature of whole perfused pancreases as well as isolated pancreatic islets of Langerhans. Applying to single cell assays of exocytosis (capacitance monitoring and amperometry) to single canine Beta-cells we have examined the time courses of granule exocytosis in response to voltage-clamp depolarizations that mimic two modes of glucose-induced electrical activity, and then compared these to biphasic insulin secretion. Action potentials evoked in short trains at frequencies similar those recorded during first phase insulin secretion trigger phasic exocytosis from a small pool of insulin granules that are likely docked near voltage-activated Ca²⁺ channels. In contrast, prolonged voltage-clamp pulses mimicking plateau depolarizations occur during second phase insulin secretion and trigger tonic or continuous exocytosis. Comparing the latter results with ones obtained using photorelease of caged Ca²⁺ in other insulin-secreting cells, we suggest that tonic exocytosis likely results from granule release from a highly Ca²⁺-sensitive pool of insulin granules, likely located further from Ca²⁺ channels. Both phasic and tonic modes of exocytosis are enhanced by glucose, via its metabolism. Hence, in canine Beta-cells we propose that two distinct modes of exocytosis, tuned to two types of electrical activity, may underlay biphasic insulin secretion.


Acute alcohol and cognition: Remembering what it causes us to forget.
Addiction has been conceptualized as a specific form of memory that appropriates typically adaptive neural mechanisms of learning to produce the progressive spiral of drug-seeking and drug-taking behavior, perpetuating the path to addiction through aberrant processes of drug-related learning and memory. From that perspective, to understand the development of alcohol use disorders, it is critical to identify how a single exposure to alcohol enters into or alters the processes of learning and memory, so that involvement of and changes in neuroplasticity processes responsible for learning and memory can be identified early. This review characterizes the effects produced by acute alcohol intoxication as a function of brain region and memory neurocircuitry. In general, exposure to ethanol doses that produce intoxicating effects causes consistent impairments in learning and memory processes mediated by specific brain circuitry, whereas lower doses either have no effect or produce a facilitation of memory under certain task conditions. Therefore, acute ethanol does not produce a global impairment of learning and memory, and can actually facilitate particular types of memory, perhaps particular types of memory that facilitate the development of excessive alcohol use. In addition, the effects on cognition are dependent on brain region, task demands, dose received, pharmacokinetics, and tolerance. Additionally, we explore the underlying alterations in neurophysiology produced by acute alcohol exposure that help to explain these changes in cognition and highlight future directions for research. Through understanding the impact that acute alcohol intoxication has on cognition, the preliminary changes potentially causing a problematic addiction memory can better be identified.


Positive selection, relaxation, and acceleration in the evolution of the human and chimp genome.
For years evolutionary biologists have been interested in searching for the genetic bases underlying humanness. Recent efforts at a large or a complete genomic scale have been conducted to search for positively selected genes in human and in chimp. However, recently developed methods allowing for a more sensitive and controlled approach in the detection of positive selection can be employed. Here, using 13,198 genes, we have deduced the sets of genes involved in rate acceleration, positive selection, and relaxation of selective constraints in human, in chimp, and in their ancestral lineage since the divergence from murids. Significant deviations from the strict molecular clock were observed in 469 human and in 651 chimp genes. The more stringent branch-site test of positive selection detected 108 human and 577 chimp positively selected genes. An important proportion of the positively selected genes did not show a significant acceleration in rates, and similarly, many of the accelerated genes did not show significant signals of positive selection. Functional differentiation of genes under rate acceleration, positive selection, and relaxation was not statistically significant between human and chimp with the exception of terms related to G-protein coupled receptors and sensory perception. Both of these were over-represented under relaxation in human in relation to chimp. Comparing differences between derived and ancestral lineages, a more conspicuous change in trends seems to have favored positive selection in the human lineage. Since most of the positively selected genes are different under the same functional categories between these species, we suggest that the individual roles of the alternative positively selected genes may be an important factor underlying biological differences between these species.


Development of Layered Multiscale Porous Thin Films by Tuning Deposition Time and Molecular Weight of Polyelectrolytes.
This work focuses on the design of porous polymeric films with nano- and micro-sized pores existing in distinct zones. The porous thin films are fabricated by the post-treatment of layer-by-layer assembled poly(allylamine hydrochloride) (PAH)/poly(acrylic acid) (PAA) multilayers. In order to improve the processing efficiency, the deposition time is shortened to ≈ 10 s. It is found that fine porous structures can be created even by significantly reducing the processing time. The effect of using polyelectrolytes with widely different molecular weights is also studied. The pore size is increased by using high molecular weight PAH, while high molecular weight PAA minimizes the pore size to nanometer scale. Having gained a precise control over the pore size, layered multiscale porous thin films are further built up with either a microsized porous zone on top of a nanosized porous zone or vice versa.


Microvascular Reactivity Measured by Dynamic Near-infrared Spectroscopy Following Induction of General Anesthesia in Healthy Patients: Observation of Age-related Change.
Background: The purpose of this study was to investigate the effect of general anesthesia on microvascular reactivity and tissue oxygen saturation (StO2) using near-infrared spectroscopy in conjunction with vascular occlusion tests (VOT). Age-related changes of microvascular reactivity, that is, the capacity of capillary recruitment, were examined. Methods: This prospective observational study was performed on 60 patients without comorbidities who underwent elective surgery under general anesthesia. Baseline StO2 on thenar eminence, hemodynamics, and laboratory profile were monitored before (T0) and 30 min after general anesthesia (T1). During VOT, occlusion slope representing oxygen consumption of muscle and recovery slope representing microvascular reactivity were also collected at T0 and T1. Results: Baseline StO2 and minimum / maximum StO2 during VOT increased under general anesthesia. Occlusion slope decreased while the recovery slope increased under general anesthesia. To observe aging effect, Receiver operating characteristic analysis was performed and age less than 65 years old showed a fair performance in predicting the increase of microvascular reactivity after the induction of anesthesia (AUC 0.733, 95% CI 0.594-0.845, P= 0.003). For age-related analyses, 27 patients of younger group (< 65 years) and 26 patients of older group (≥ 65 years) were divided. Recovery slope significantly increased under general anesthesia in younger group (2.44 [1.91-2.81] % ∙ sec-1 at T0 and 3.59 [2.58-3.51] % ∙ sec-1 at T1, P <0.001), but not in older group (2.61 [2.21-3.20] % ∙ sec-1 at T0, 2.63 [1.90-3.60] % ∙ sec-1 at T1, P = 0.949). Conclusions: General anesthesia could improve StO2 through increase of microvascular reactivity and decrease of tissue metabolism. However, microvascular reactivity to capillary recruitment under general anesthesia significantly improves in younger patients, not in older patients.


The regulation of retinoic acid formation.
Rat liver microsomal as well as cytosolic retinol dehydrogenases from cellular retinal-binding protein (CRBP)-retinal from CRBP-retinol. A cytosolic dehydrogenase transforms retinal into retinoic acid. Apo-CRBP and ethanol inhibit the cytosolic but not the microsomal retinol dehydrogenase, the latter being the primary enzyme in the rate-limiting step in retinoic acid synthesis.


Alveolar cell senescence in patients with pulmonary emphysema.
The prevalence of chronic obstructive pulmonary disease (COPD) is age-dependent, suggesting an intimate relationship between the pathogenesis of COPD and aging. In this study we investigated whether the senescence of alveolar epithelial and endothelial cells is accelerated in emphysematous lungs. Samples of lung tissue were obtained from patients with emphysema, asymptomatic smokers, and asymptomatic nonsmokers. Paraffin-embedded lung tissue sections were evaluated for cellular senescence by quantitative fluorescence in situ hybridization to assess telomere shortening, and by immunohistochemistry to assess the expression of senescence-associated cyclin-dependent kinase inhibitors. Tissue sections were also immunostained for proliferating cell nuclear antigen (PCNA), surfactant protein A, and CD31. The patients with emphysema had significantly higher percentages of type II cells positive for p16INK4a and p21CIP1/WAF1/Sdi1 than the asymptomatic smokers and nonsmokers. They had also significantly higher percentages of endothelial cells positive for p16INK4a than the asymptomatic smokers and nonsmokers, and higher percentages of endothelial cells positive for p21CIP1/WAF1/Sdi1 than the asymptomatic nonsmokers. Telomere length in alveolar type II cells and endothelial cells was significantly shorter in the patients with emphysema than in the asymptomatic nonsmokers. The level of p16INK4a expression was negatively correlated with the level of PCNA expression. The level of alveolar cell senescence was positively correlated with airflow limitation. These results suggest that the senescence of alveolar epithelial and endothelial cells is accelerated in patients with emphysema. Cellular senescence may explain the abnormal cell turnover that promotes the loss of alveolar cells in emphysematous lungs.


Male infertility: the focus shifts to sperm manipulation.
Few systemic approaches for treatment of the infertile male are available. Therefore, the focus has shifted to studies of sperm maturation in the female tract, tests of sperm function, and assisted reproduction. New methods of sperm evaluation allow a better determination of those samples that will and will not fertilize ova. These methods include the strict morphology examination, biochemical approaches such as evaluation of creatine kinase levels, and the hemizona assay. Computer assisted semen analysis has not yet proved important for diagnosis but has provided important research information. For example, the drug pentoxifylline significantly increases velocity and has also been shown to enhance in vitro fertilization. The term subfertile is extensively used in the literature but its clear definition and the prognosis for subfertile men have not been established. Approaches that may increase fertilization rates include reducing the gamete culture volume and removal of cumulus oophorus. Assisted reproductive technologies for patients with ejaculatory dysfunction have also been expanded. The use of improved semen processing techniques, advanced oocyte retrieval, and well-timed intrauterine insemination are enabling physicians to use decreasing numbers of viable sperm to achieve pregnancies.


Deep learning method for localization and segmentation of abdominal CT.
Computed Tomography (CT) imaging is widely used for studying body composition, i.e., the proportion of muscle and fat tissues with applications in areas such as nutrition or chemotherapy dose design. In particular, axial CT slices from the 3rd lumbar (L3) vertebral location are commonly used for body composition analysis. However, selection of the third lumbar vertebral slice and the segmentation of muscle/fat in the slice is a tedious operation if performed manually. The objective of this study is to automatically find the middle axial slice at L3 level from a full or partial body CT scan volume and segment the skeletal muscle (SM), subcutaneous adipose tissue (SAT), visceral adipose tissue (VAT) and intermuscular adipose tissue (IMAT) on that slice. The proposed algorithm includes an L3 axial slice localization network followed by a muscle-fat segmentation network. The localization network is a fully convolutional classifier trained on more than 12,000 images. The segmentation network is a convolutional neural network with an encoder-decoder architecture. Three datasets with CT images taken for patients with different types of cancers are used for training and validation of the networks. The mean slice error of 0.87±2.54 was achieved for L3 slice localization on 1748 CT scan volumes. The performance of five class tissue segmentation network evaluated on two datasets with 1327 and 1202 test samples. The mean Jaccard score of 97% was achieved for SM and VAT tissue segmentation on 1327 images. The mean Jaccard scores of 98% and 83% are corresponding to SAT and IMAT tissue segmentation on the same dataset. The localization and segmentation network performance indicates the potential for fully automated body composition analysis with high accuracy.


Epidermal barrier in atopic dermatitis.
Atopic dermatitis (AD) is a complex disease that affects up to 20% of children and impacts the quality of patients and families in a significant manner. New insights into the pathophysiology of AD point to an important role of structural abnormalities in the epidermis combined with immune dysregulation. Filaggrin (FLG) is synthesized as a large precursor, profilaggrin, and is expressed in the upper layers of the epidermis. FLG plays a critical role in the epidermal barrier, and FLG mutations cause abnormal epidermal function. FLG mutations are strongly associated with early-onset, and persistent severe AD. In addition, FLG deficiency in the epidermis is related to allergic sensitization and asthma. The basic skin care including repair and protection of the skin barrier with proper hydration and topical anti-inflammatory therapy is important to control the severity of skin disease in patients with AD.


Efficacy and safety of hypertonic saline solutions fluid resuscitation on hypovolemic shock: A systematic review and meta-analysis of randomized controlled trials.
Fluid resuscitation is a fundamental intervention in patients with hypovolemic shock resulting from trauma. Appropriate fluid resuscitation in trauma patients could reduce organ failure, until blood components are available, and hemorrhage is controlled. We conducted a systematic review and meta-analysis assessing the effect of hypertonic saline/dextran or hypertonic saline for fluid resuscitation on patient outcomes restricted to adults with hypovolemic shock. We conducted a search of electronic information sources, including PubMed, Embase, Web of Science, Cochrane library and bibliographic reference lists to identify all randomized controlled trials (RCTs) investigating outcomes of crystalloids versus colloids in patients with hypovolemic shock. We calculated the risk ratio (RR) or mean difference (MD) of groups using fixed or random-effect models. Fifteen studies including 3264 patients met our inclusion criteria. Survival to hospital discharge rate between research groups varied and amounted to 71.2% in hypertonic saline/dextran group vs. 68.4% for isotonic/normotonic fluid (normal saline) solutions (odds ratio [OR] = 1.19; 95% confidence interval [CI] 0.97-1.45; I2 = 48%; p = 0.09). 28- to 30-days survival rate for hypertonic fluid solutions was 72.8% survivable, while in the case of isotonic fluid (normal saline) - 71.4% (OR = 1.13; 95% CI 0.75-1.70; I2 = 43%; p = 0.56). This systematic review and meta-analysis, which included only evidence from RCTs hypertonic saline/dextran or hypertonic saline compared with isotonic fluid did not result in superior 28- to 30-day survival as well as in survival to hospital discharge. However, patients with hypotension who received resuscitation with hypertonic saline/dextran had less overall mortality as patients who received conventional fluid.


Why do influenza virus subtypes die out? A hypothesis.
Novel pandemic influenza viruses enter the human population with some regularity and can cause disease that is severe and widespread. The emergence of novel viruses, historically, has often been coupled with the disappearance of existing seasonal virus strains. Here, we propose that the elimination of seasonal strains during virus pandemics is a process mediated, at the population level, by humoral immunity. Specifically, we suggest that infection with a novel virus strain, in people previously exposed to influenza viruses, can elicit a memory B cell response against conserved hemagglutinin stalk epitopes and/or neuraminidase epitopes. The anti-stalk and/or anti-neuraminidase antibodies then act to diminish the clinical severity of disease caused by novel influenza viruses and to eliminate seasonal virus strains.


Re-examination of the effects of raphe lesions on the sleep/wakefulness cycle states in cats.
To study the specific effects of central superior raphe nucleus (CeSR) lesions on the different sleep/wakefulness cycle states of the cat, nine animals with implanted electrodes for EOG, EMG and EEG recordings were used. Seven cats received diathermocoagulation lesions that destroyed between 13 and 100 percent of the CeSR; the remaining two cats, which suffered lesions in the paramedial region of the oral pontine reticular nucleus (RPO), were used to determine the effects on sleep/wakefulness states caused by damage to adjacent CeSR structures and/or passage fibres. Three prelesion and five postlesion weekly 24 h recordings were obtained from each cat. Recordings were scored according to the polygraphic criteria for wakefulness (W), drowsiness (D), slow wave sleep (SWS) and paradoxical sleep (PS). Results indicated that insomnia is not produced exclusively by CeSR lesions, since adjacent paramedial RPO lesions also decrease both SWS and PS; however, increased W occurred after the former while increased D occurred after the latter. Correlation coefficient analyses showed that W is the only state that correlates significantly with the volume of CeSR destroyed. The following correlations between different states of the sleep/wakefulness cycle were, however, significant: W-D, W-SWS and SWS-PS. Disinhibition of W, therefore, and not sleep loss seems to be the primary effect of CeSR lesions. Thus, the CeSR nucleus appears to be involved in arousal mechanisms rather than in direct sleep promotion.


[Complete genome sequence analysis of Japanese encephalitis virus newly isolated in China].
To study the complete genome sequence of Japanese encephalitis virus (JEV) strain XJ69 isolated in ZheJiang province and explore its evolution. Overlapping primers were designed according to the full-length genomes from GenBank. RT-PCR was used to amplify the fragments and RT-PCR products were cloned T vector, sequenced and analyzed. The genome of strain XJ69 and XJP613 were 10 964 nucleotides in length with a single open reading frame encoding 3432 amino acids. Comparison of the complete genome sequences of different JEV isolates showed XJ69 and XJP613 were 83.5%-99.2% and 83.4%-99.4% nucleotide sequence homology among them respectively, which resulted in 94.8%-99.7% amino acid sequence homology. Phylogenetic analysis through PrM/C,E and full-length genome showed that the XJ69 and XJP613 strain belonged to genotype I. The nucleotitede sequence and deduced amino acid sequence of XJ69 and XJP613 strain were similar to that of those of genotype I of Japanese encephalitis virus. It belonged to genotype I and were close to the isolates SH17M-07.


Association of city-level walkability, accessibility to biking and public transportation and socio-economic features with COVID-19 infection in Massachusetts, USA: An ecological study.
With people restricted to their residences, neighbourhood characteristics may affect behaviour and risk of coronavirus disease 2019 (COVID-19) infection. We aimed to analyse whether neighbourhoods with higher walkability, public transit, biking services and higher socio-economic status were associated with lower COVID-19 infection during the peak of the COVID-19 pandemic in Massachusetts. We used Walk Score®, Bike Score®, and Transit Score® indices to assess the walkability and transportation of 72 cities in Massachusetts, USA based on availability of data and collected the total COVID-19 case numbers of each city up to 10 April 2021. We used univariate and multivariate linear models to analyse the effects of these scores on COVID-19 cases per 100,000 in each city, adjusting for demographic covariates and all covariates, respectively. In the 72 cities studied, the average Walk Score, Transit Score and Bike Score was 48.7, 36.5 and 44.1, respectively, with a total of 426,182 COVID-19 cases. Higher Walk Score, Transit Score, and Bike Score rankings were negatively associated with COVID-19 cases per 100,000 persons (<0.05). Cities with a higher proportion of Hispanic population and a lower median household income were associated with more COVID-19 cases per 100,000 (P<0.05). Higher Walk Score, Transit Score and Bike Score were shown to be protective against COVID-19 transmission, while socio-demographic factors were associated with COVID-19 infection. Understanding the complex relationship of how the structure of the urban environment may constrain commuting patterns for residents and essential workers during COVID-19 would offer potential insights on future pandemic preparedness and response.


Drivers of gut microbiome variation within and between groups of a wild Malagasy primate.
Various aspects of sociality can benefit individuals' health. The host social environment and its relative contributions to the host-microbiome relationship have emerged as key topics in microbial research. Yet, understanding the mechanisms that lead to structural variation in the social microbiome, the collective microbial metacommunity of an animal's social network, remains difficult since multiple processes operate simultaneously within and among animal social networks. Here, we examined the potential drivers of the convergence of the gut microbiome on multiple scales among and within seven neighbouring groups of wild Verreaux's sifakas (Propithecus verreauxi) - a folivorous primate of Madagascar. Over four field seasons, we collected 519 faecal samples of 41 animals and determined gut communities via 16S and 18S rRNA gene amplicon analyses. First, we examined whether group members share more similar gut microbiota and if diet, home range overlap, or habitat similarity drive between-group variation in gut communities, accounting for seasonality. Next, we examined within-group variation in gut microbiota by examining the potential effects of social contact rates, male rank, and maternal relatedness. To explore the host intrinsic effects on the gut community structure, we investigated age, sex, faecal glucocorticoid metabolites, and female reproductive state. We found that group members share more similar gut microbiota and differ in alpha diversity, while none of the environmental predictors explained the patterns of between-group variation. Maternal relatedness played an important role in within-group microbial homogeneity and may also explain why adult group members shared the least similar gut microbiota. Also, dominant males differed in their bacterial composition from their group mates, which might be driven by rank-related differences in physiology and scent-marking behaviours. Links to sex, female reproductive state, or faecal glucocorticoid metabolites were not detected. Environmental factors define the general set-up of population-specific gut microbiota, but intrinsic and social factors have a stronger impact on gut microbiome variation in this primate species. Video abstract.


Liquid chromatography mass spectrometry based characterization of epitope configurations.
Here we evaluate a quick and easy tool for determination of epitope configuration using immunocapture and liquid chromatography mass spectrometry (LC-MS) subsequent to pre-treatment of the target protein to disrupt its three-dimensional structure. The approach can be a valuable screening tool to identify antibodies that can be used in peptide capture by anti-protein antibodies. The experimental set-up was established using seven monoclonal antibodies (mAbs) with known linear or conformational epitope recognition. The mAbs were developed to target either of the two biomarkers, progastrin releasing peptide (ProGRP) or human chorionic gonadotropin (hCG). Best coherence with established epitope configuration was seen when using both denaturation, reduction and alkylation as pre-treatment method of the proteins (≥70% reduction in MS signal intensity compared to control) prior to immunocapture and LC-MS determination. The final method was used to determine the epitope configuration of four anti-thyroglobulin mAbs with unknown epitope configuration; all four mAbs showed configurational epitope recognition. These results were also supported by western blots of native, and reduced and alkylated protein using three of the evaluated mAbs, and by analysis native, and reduced and alkylated protein in a routine immunofluorometric assay employing the four evaluated antibodies.


Text Message Intervention for Latino Adults to Improve Diabetes Outcomes.
Introduction: Determine the effectiveness of a diabetes text message intervention to improve diabetes outcomes in Latino adults at a free clinic in the southeastern United States. Methods: A pre-post, one-group design was used with a convenience sample (n = 24) of Latino adults with type 2 diabetes and A1C ≥ 7 who consented to participate. Over 8 weeks, participants were sent 23 educational and motivational text messages that were based on the American Association of Diabetes Educators (AADE) 7 self-care tasks and the American Diabetes Association (ADA) Standards of Medical Care in Diabetes Mellitus (DM). Diabetes knowledge, self-efficacy, self-care, and A1C were measured. Results: Diabetes knowledge, self-efficacy, self-care, and A1C improved after 8 weeks. Statistically significant results (p < .05) were seen in two of the four evaluation instruments administered (Spoken Knowledge in Low Literacy in Diabetes Scale [SKILLD], p = .001, Diabetes Self-Efficacy Scale [DSES], p = .000). A1C improvements were significant (0.86 or 10 mmol/mol) from the pre-intervention (M = 9.06 [76 mmol/mol], SD = 1.49) to the post-intervention (M = 8.20 [66 mmol/mol], SD = 1.27, t [23] = 3.02, p = .006). Conclusion: Personalized communication, education, and follow-up via text messages improved diabetes knowledge, self-efficacy, self-care, and A1C among pilot project participants. Text messages are a useful tool for diabetes management.


Inorganic Selenium Induces Nonapoptotic Programmed Cell Death in PC-3 Prostate Cancer Cells Associated with Inhibition of Glycolysis.
Previous studies have shown that selenite, a representative of inorganic form selenium, exerts its anticancer effect by inducing apoptosis in androgen-dependent LNCaP prostate cancer cells, but few studies have determined the nature of cell death induced by selenite in metastatic androgen-refractory PC-3 cells. Our study showed that necrosis-like cell death rather than apoptosis, pyroptosis, or autophagic cell death was caused by selenite in PC-3 cells. Mechanistically, this type of cell death was caused by ATP depletion (26.28 ± 3.39 nmol/mg of control versus 9.12 ± 2.44 nmol/mg of 10 μM selenite treatment) that resulted from phosphofructokinase activity reduction (100.17 ± 0.17% of control versus 21.74 ± 6.65% of 10 μM selenite treatment). Our study also showed that ROS production is necessary for the decrease in cellular ATP levels and in phosphofructokinase activity. To our knowledge, this is the first study showing that selenite can induce necrosis-like cell death in PC-3 cells. Our findings support selenite as an effective compound for the therapy of apoptosis-resistant prostate cancer.


Unexpected Favorable Role of Ca2+ in Phosphate Removal by Using Nanosized Ferric Oxides Confined in Porous Polystyrene Beads.
Polystyrene-based nanoferric oxide composite is a representative nanomaterial successfully applied in scale-up water decontamination for arsenic and phosphorus. However, little is available on the effect of solution chemistry (for instance, the coexisting Ca2+) on the long-term performance of the nanocomposite. In this study, we carried out 20 cyclic runs of phosphate adsorption-desorption on a polymer-supported ferric nanocomposite HFO@201. Unexpectedly, an enhanced phosphate removal was observed in the presence of Ca2+, which is quite different from its adverse effect on phosphate capture by granular ferric oxide. Further mechanistic studies revealed that enhanced phosphate removal was mainly realized via the Ca-P coprecipitation inside the networking pores of HFO@201 as well as the possible formation of the multiple Fe-P-Ca-P complex. The complex formation led to a distinct increase in P adsorption, and the coprecipitation, driven by the accumulated OH- in confined pores during phosphate adsorption and alkaline regeneration, favored P removal via the formation of amorphous calcium phosphate (ACP) and hydroxyapatite inside. TEM-EDS spectra indicated that coprecipitation did not occur on the surface of loaded nano-HFO, greatly mitigating its adverse effect on P adsorption on the surface of nano-HFO. Fixed-bed column study showed that the presence of Ca2+ increased the effective treatable volume of HFO@201 toward P-containing influents by ∼70%. This study is believed to shed new insights into the effect of solution chemistry on similar nanocomposites for advanced water treatment.


Application Analysis of 124I-PPMN for Enhanced Retention in Tumors of Prostate Cancer Xenograft Mice.
In recent years, nuclear medicine imaging and therapy for prostate cancer have radically changed through the introduction of radiolabeled prostate-specific membrane antigen (PSMA)-binding peptides. However, these small molecular probes have some inherent limitations, including high nephrotoxicity and short circulation time, which limits their utility in biological systems. In this study, organic melanin nanoparticles were used to directly label the long half-life radionuclide 124I (t1/2=100.8 h), and PSMA small molecular groups were efficiently bonded on the surface of nanoparticles to construct the PSMA-targeted long-retention nanoprobe 124I-PPMN, which has the potential to increase tumor uptake and prolong residence time. The results showed that the nanoprobe could substantially aggregate in the tumors of prostate cancer xenograft mice and was visible for more than 72 h. Positron Emission Computed Tomography (PET) imaging showed that the nanoprobe could be used for precise imaging of prostate cancer with high expression of PSMA. In addition, organic melanin nanoparticles labeled with an elemental radionuclide achieved a stable, metal-free structure. Cell experiments and mouse toxicity experiments indicated that the nanoprobe has high safety. The new nanoprobe constructed in this study has high specificity and biocompatibility. In the future, combined with the multifunctional potential of melanin nanoparticles, this nanoprobe is expected to be used in the integrated theranostics of prostate cancer.


A Stimuli-Responsive Biosensor of Glucose on Layer-by-Layer Films Assembled through Specific Lectin-Glycoenzyme Recognition.
The research on intelligent bioelectrocatalysis based on stimuli-responsive materials or interfaces is of great significance for biosensors and other bioelectronic devices. In the present work, lectin protein concanavalin A (Con A) and glycoenzyme glucose oxidase (GOD) were assembled into {Con A/GOD}n layer-by-layer (LbL) films by taking advantage of the biospecific lectin-glycoenzyme affinity between them. These film electrodes possess stimuli-responsive properties toward electroactive probes such as ferrocenedicarboxylic acid (Fc(COOH)₂) by modulating the surrounding pH. The CV peak currents of Fc(COOH)₂ were quite large at pH 4.0 but significantly suppressed at pH 8.0, demonstrating reversible stimuli-responsive on-off behavior. The mechanism of stimuli-responsive property of the films was explored by comparative experiments and attributed to the different electrostatic interaction between the films and the probes at different pH. This stimuli-responsive films could be used to realize active/inactive electrocatalytic oxidation of glucose by GOD in the films and mediated by Fc(COOH)₂ in solution, which may establish a foundation for fabricating novel stimuli-responsive electrochemical biosensors based on bioelectrocatalysis with immobilized enzymes.


The Trimeric Major Capsid Protein of Mavirus is stabilized by its Interlocked N-termini Enabling Core Flexibility for Capsid Assembly.
Icosahedral viral capsids assemble with high fidelity from a large number of identical buildings blocks. The mechanisms that enable individual capsid proteins to form stable oligomeric units (capsomers) while affording structural adaptability required for further assembly into capsids are mostly unknown. Understanding these mechanisms requires knowledge of the capsomers' dynamics, especially for viruses where no additional helper proteins are needed during capsid assembly like for the Mavirus virophage that despite its complexity (triangulation number T = 27) can assemble from its major capsid protein (MCP) alone. This protein forms the basic building block of the capsid namely a trimer (MCP3) of double-jelly roll protomers with highly intertwined N-terminal arms of each protomer wrapping around the other two at the base of the capsomer, secured by a clasp that is formed by part of the C-terminus. Probing the dynamics of the capsomer with HDX mass spectrometry we observed differences in conformational flexibility between functional elements of the MCP trimer. While the N-terminal arm and clasp regions show above average deuterium incorporation, the two jelly-roll units in each protomer also differ in their structural plasticity, which might be needed for efficient assembly. Assessing the role of the N-terminal arm in maintaining capsomer stability showed that its detachment is required for capsomer dissociation, constituting a barrier towards capsomer monomerisation. Surprisingly, capsomer dissociation was irreversible since it was followed by a global structural rearrangement of the protomers as indicated by computational studies showing a rearrangement of the N-terminus blocking part of the capsomer forming interface.


Risks Associated with Fish and Seafood.
Fresh fish and seafood are highly perishable, and microbiological spoilage is one of the important factors that limit their shelf life and safety. Fresh seafood can be contaminated at any point from rearing or harvesting to processing to transport or due to cross-contamination by consumer mishandling at home. With the increase in the demand for fish and seafood, aquaculture production is increasing, which could lead to new risks that will need to be addressed in the future to control foodborne pathogens.


Transactivation by Rtg1p, a basic helix-loop-helix protein that functions in communication between mitochondria and the nucleus in yeast.
Rtg1p is a basic helix-loop-helix transcription factor in the yeast Saccharomyces cerevisiae that is required for basal and regulated expression of CIT2, the gene encoding a peroxisomal isoform of citrate synthase. In respiratory incompetent rho degree petite cells, CIT2 transcription is elevated as much as 30-fold compared with respiratory competent rho + cells. Here we provide evidence that Rtg1p interacts directly with a CIT2 upstream activation site (UASr) and that the rho degree/rho + regulation is not due to a change in the levels of Rtg1p. A fusion protein consisting of the DNA binding domain of Gal4p fused to the NH2 terminus of the full-length wild-type Rtg1p was able to transactivate an integrated LacZ reporter under control of the Gal4p-responsive GAL1 UASG in a rho degree/rho(+)-dependent manner. Other Gal4p fusions to deletions or mutations of Rtg1p indicate that the helix-loop-helix domain is essential for transactivation. Regulated expression of CIT2 also requires the RTG2 gene product. The Gal4-Rtg1p fusion was unable to transactivate the LacZ reporter gene in a strain deleted for RTG2, suggesting that the RTG2 product does not act independently of Rtg1p in the rho degree/rho + transcriptional response.


[Epidemiological and microbiological studies on salmonellosis in Russia].
The occurrence of the most important Salmonella serovars in Russia from 1980 to 1993 is reported. Selected Salmonella strains isolated in Russia from 1951 to 1993 were characterized by serotype, phage type, resistance pattern, plasmid profile and detection of aerobactin production. The incidence of salmonellosis was different in the various administrative districts and varied between 0-235 cases/100,000 inhabitants. Since the middle of the eighties both the overall increase of salmonellosis and the replacement of S. typhimurium by S. enteritidis was also demonstrable in Russia. Nosocomial infections due to multiply resistant S. infantis strains and S. typhimurium strains which caused severe septicaemic courses in children were a special epidemic problem. These infections mainly occurred via faecal-oral transmission. In contrast ot Western Europe most of the S. enteritidis strains isolated in Russia belonged to phage type 1/1. Eight phage types were determined additionally. The S. typhimurium strains isolated in nosocomial infections belonged to a definite phage type and carried a 95 MDa plasmid. This plasmid was also detected in the nosocomial S. infantis strains, S. haifa strains, and S. panama strains. All these nosocomial strains formed aerobactin.


Detection of endothelin receptors in human coronary artery vascular smooth muscle cells but not endothelial cells by using electron microscope autoradiography.
Endothelin A (ET(A)) and ET(B) receptors located on vascular smooth-muscle cells mediate potent vasoconstriction in animal and human blood vessels. Although vascular endothelial ET(B) receptors mediate vasodilatation through release of nitric oxide, prostacyclin, or an endothelium-derived hyperpolarizing factor in animal vessels, there is less evidence to support ET-mediated endothelium-dependent relaxation in human arteries. We investigated the regional and subcellular localization of ET-receptor subtypes in human epicardial coronary arteries obtained from patients with congestive heart failure undergoing cardiac transplantation. Radioligand-binding studies revealed a predominance of the ET(A)-receptor subtype in the coronary artery. ET(A) and ET(B) receptors were located in the media and in perivascular structures by using macroautoradiography. Specific binding in the intimal layer was low. Autoradiography at the ultrastructural level revealed the presence of ET(A) and ET(B) receptors on medial vascular smooth-muscle cell plasmalemma and plasmalemmal vesicles with a predominance of the ET(A) subtype. ET receptors were not detected in either luminal endothelial cells of the coronary arteries or endothelial cells lining small adventitial blood vessels. This study is the first to identify the subcellular localization of ET receptors in human coronary artery vascular smooth-muscle cells. These receptors probably mediate the well-established vasoconstrictor response to ET-1. The inability to detect ET receptors on endothelial cells suggests a minor role for endothelial cells in ET-mediated vasodilatation of the coronary vasculature.


The unstable detrusor.
The unstable detrusor (detrusor instability), a common condition in men and women, causes significant morbidity in sufferers and a great financial expense to health care providers. The condition is associated with symptoms of urinary frequency, urgency and urge incontinence. The embarrassing nature of these symptoms means patients are often reluctant to seek medical help and as a consequence there is under-reporting. Epidemiological studies are difficult to perform but appear to show that incidence increases with age and that the unstable detrusor is the commonest cause of male urinary incontinence. The aetiology is still much debated with reasoned arguments for both neurogenic and myogenic causes, though in truth, the likely explanation is multifactorial rather than a single underlying pathological process. The diagnosis can only be made on urodynamic testing. Before deciding on suitable treatment, the clinician must assess the disease severity and also its impact on patient quality of life. This is because perceived improvement in patient quality of life must outweigh any potential morbidity associated with treatment. Pharmacotherapy is based on blocking bladder muscarinic receptors. Unfortunately, no treatment is bladder specific and therefore most drug therapies have unwanted systemic anticholinergic side effects. Surgical treatment often involves a degree of bladder denervation to reduce bladder activity. This can consequently produce voiding difficulties which necessitate patients to self-catheterise on a longterm basis.


GDF-5 deficiency in mice leads to disruption of tail tendon form and function.
Although the biological factors which regulate tendon homeostasis are poorly understood, recent evidence suggests that Growth and Differentiation Factor-5 (GDF-5) may play a role in this important process. The purpose of this study was to investigate the effect of GDF-5 deficiency on mouse tail tendon using the brachypodism mouse model. We hypothesized that GDF-5 deficient tail tendon would exhibit altered composition, ultrastructure, and biomechanical behavior when compared to heterozygous control littermates. Mutant tail tendons did not display any compositional differences in sulfated glycosaminoglycans (GAG/DNA), collagen (hydroxyproline/DNA), or levels of fibromodulin, decorin, or lumican. However, GDF-5 deficiency did result in a 17% increase in the proportion of medium diameter (100-225 nm) collagen fibrils in tail tendon (at the expense of larger fibrils) when compared to controls (p < 0.05). Also, mutants exhibited a trend toward an increase in irregularly-shaped polymorphic fibrils (33% more, p > 0.05). While GDF-5 deficient tendon fascicles did not demonstrate any significant differences in quasistatic biomechanical properties, mutant fascicles relaxed 11% more slowly than control tendons during time-dependent stress-relaxation tests (p < 0.05). We hypothesize that this subtle alteration in time-dependent mechanical behavior is most-likely due to the increased prevalence of irregularly shaped type I collagen fibrils in the mutant tail tendons. These findings provide additional evidence to support the conclusion that GDF-5 may play a role in tendon homeostasis in mice.


Effectively engaging physicians in system change.
What started as a prospective study to support clinical leaders and inform strategies to engage their peers in system change was impacted due to a rapidly evolving political agenda amid a pandemic, affecting both organizations and outcomes. Participants in this mixed methods study in one Local Health Integrated Network (LHIN) in Ontario included clinical leaders and community physicians over a period of 14 months. As the provincial government shifted regional healthcare governance from LHINs to Ontario Health Teams, there was an increase in the engagement of community physicians and leaders identified a noticeable culture shift with the potential to drive change. High-performing healthcare systems are dependent not only on physicians who can lead and engage others but a government that can acknowledge this.


Does tibolone affect serum leptin levels and body weight in postmenopausal women?
Leptin has a significant role in body weight regulation and energy balance. We examined the effect of tibolone on the body weight and serum leptin levels in postmenopausal women. Twenty women (aged 43-60 years) participated in this prospective study. All women in this study protocol received 2.5 mg/day of tibolone. Absolute and body mass index (BMI)-corrected serum leptin concentrations and BMI values were measured at baseline, after 3 months, and after 6 months of the tibolone therapy. Tibolone did not affect absolute and BMI-corrected serum leptin levels, and BMI values during the treatment. A significant linear correlation between BMI values and serum leptin levels was observed (p<0.05, r=0.67). Tibolone seems not to affect serum leptin levels, body weight and BMI values of postmenopausal women. There is a significant correlation between serum leptin levels and BMI values.


An excess of dietary iodine accelerates the development of a thyroid-associated lymphoid tissue in autoimmune prone BB rats.
Previous studies have shown that dietary iodine enhances the severity and incidence of focal thyroiditis in autoimmune BB rats and OS chickens. However, which lymphoid cells are involved in the development of the iodine-induced focal thyroiditis and what the consequences are for the anticolloid antibody production have not been studied in detail. We therefore performed a study in which 3-week-old female BB rats were kept on either an enriched iodine diet (EID; iodine intake, 100 micrograms iodine/day) or a normal iodine diet (NID; iodine intake, 7 micrograms iodine/day) for a period of 18 weeks. The development of the focal thyroiditis was immunohistologically studied. Immunohistological data were compared to the thyroid hormone status and anti-colloid antibody production. Our data confirm that a high dietary iodine intake results in an accelerated development of the focal lymphoid cell infiltrates in the thyroid of the BB rat. After 12-18 weeks of an EID 50% of the BB rats developed these infiltrates. Our data additionally show that: (a) the process starts with increases in the number of infiltrating MHC class II-positive dendritic cells and a clustering of these cells with T cells, B cells, and some macrophages and (b) the focal infiltrates are highly organized and consist of central B cell follicle-like structures surrounded by rims and areas of T cells. The architecture of the focal thyroiditis is hence very similar to mucosa-associated lymphoid tissue and secondary lymphoid organs (spleen and lymph node). Only minor signs of thyrocyte destruction were observed. We therefore consider the term "thyroiditis" as inappropriate and prefer the term "thyroid-associated lymphoid tissue." Since the thyroiditis component was small, it is also not surprising that the BB rats on the EID remained euthyroid. The presence of the thyroid-associated lymphoid tissue in the BB rats was positively correlated to the presence of anti-colloid antibody in the serum of the BB rats. We speculate that the dietary iodine might have direct effects on cells of the immune system or on cells forming the microenvironment of lymphoid tissue (reticulum cells). A role for highly iodinated thyroglobulin in the accelerated development of thyroid-associated lymphoid tissue is also possible.


Suppression by Trypanosoma brucei rhodesiense of the capacities of human T lymphocytes to express interleukin-2 receptors and proliferate after mitogenic stimulation.
We studied the suppressive effects induced in phytohemagglutinin (PHA)-stimulated human peripheral blood mononuclear cells (PBMC) by purified blood forms of Trypanosoma brucei rhodesiense. The parasite was found to markedly impair lymphocyte proliferation (measured in terms of [3H]thymidine incorporation). The extent of this effect increased with parasite concentration and was not due to mitogen absorption, depletion of medium nutrients, or PBMC killing by the parasite. Significant reductions in interleukin-2 receptor (IL-2R) expression, determined by flow cytometric analysis, were also observed in PHA-stimulated PBMC cultured in the presence of T. b. rhodesiense as evidenced by marked decreases in the surface density of the receptor. Concomitant decreases in the percentage of IL-2R+ cells were recorded in approximately half of the experiments. A discrete, dimly stained subpopulation of IL-2R+ cells were consistently demonstrable whether or not a reduction in the percentage of IL-2R+ cells occurred. Living, but not glutaraldehyde-fixed, parasites suppressed IL-2R expression. In kinetic studies, a low but reproducible level of suppression of IL-2R was demonstrable as early as 6 h after PHA stimulation; the extent of this effect became considerably more pronounced as additional culture time elapsed. Levels of IL-2 biological activity in cocultures of T. b. rhodesiense with PHA-stimulated PBMC were comparable with or higher than those present in control cultures lacking the parasite. Therefore, insufficient levels of this cytokine would be an unlikely explanation for the noted suppression of IL-2R expression and lymphoproliferation. These effects of T. b. rhodesiense could represent an important component of the mechanism by which immunosuppression develops in African sleeping sickness.


Recent advances in edge illumination x-ray phase-contrast tomography.
Edge illumination (EI) is an x-ray phase-contrast imaging technique, exploiting sensitivity to x-ray refraction to visualize features, which are often not detected by conventional absorption-based radiography. The method does not require a high degree of spatial coherence and is achromatic and, therefore, can be implemented with both synchrotron radiation and commercial x-ray tubes. Using different retrieval algorithms, information about an object's attenuation, refraction, and scattering properties can be obtained. In recent years, a theoretical framework has been developed that enables EI computed tomography (CT) and, hence, three-dimensional imaging. This review provides a summary of these advances, covering the development of different image acquisition schemes, retrieval approaches, and applications. These developments constitute an integral part in the transformation of EI CT into a widely spread imaging tool for use in a range of fields.


[Protective effects of beta-carotene liposome against rat neutrophile membrane damage caused by intra- or extra-cellular reactive oxygen species].
In this experiment the protective effects of beta-carotene (beta-C) liposome against rat neutrophile membrane damage caused by extracellular or intracellular reactive oxygen were species (ROS) investigated by means of fluorescence labels 1, 6-diphenyl-1,3,5 hexatriane (DPH) and N-(3-pyrene) maleimide (N-(3p)M). The extracellular ROS include the singlet oxygen (1O2) generated by photoreaction of sensitizer rosebengal and the superoxide anion (O2-.) produced by xanthine/xanthine oxidase reactive system. The intracellular ROS was sensitizer rosebengal and superoxide anion (O2-.) produced by xanthine/xanthine oxidase reactive system while the intracellular oxygen was generated by stimulating neutrophile respiratory burst with opsonized zymosan. No change in the peak or shape of fluorescent excitation and emission spectra was found for either of the two labels. The results of fluorescent polarization value in the membrane showed that beta-C liposome could significantly inhibit the damage of rat neutrophile membrane protein reacted either by intracellular generated reactive oxygen species or by extracellular generated O2-. or 1O2. It could also inhibit the injury of the rat neutrophile membrane lipid reacted by extracellular reactive oxygen species. However, the protective effects of beta-C liposome against membrane lipid damage reacted by intracellular reactive oxygen species by at the same concentration as those on membrane protein were not significant, which could be partly explained by relatively higher damage level of rat neutrophile membrane lipid domain reacted by intracellular reactive oxygen species than that of the membrane protein domain.


Two cases of lysergamide intoxication by ingestion of seeds from Hawaiian Baby Woodrose.
We describe two cases of human consumption of seeds from Argyreia nervosa (Hawaiian Baby Woodrose), which resulted in one fatality due to falling from a building and one surviving witness. The principal psychoactive constituent of the seeds, lysergamide (LSA), was recovered from blood and urine samples by mixed-mode cation exchange solid-phase extraction and quantified by ultra performance liquid chromatography-time of flight mass spectrometry (UPLC-ToF/MS). The LSA concentrations were determined by UPLC-ToF/MS to be 4.9 microg/L in blood and 1.0mg/L in urine in the dead person and 1.8 microg/L in blood and 0.50mg/L in urine in the living person. These analytical findings were found to be in accordance with the case story, which indicated that seeds had been ingested and also noted psychological reactions, i.e. the will to jump out of the window. Other findings in the dead person were 22 microg/L THC in blood, 0.71 g/L ethanol in blood and 1.0 g/L ethanol in vitreous humor. Constituents originating from the seeds of A. nervosa, i.e. LSA, ergonovine, lysergic acid alpha-hydroxyethylamide were also identified in the biological samples. The 2-hydroxy-3-oxo metabolites of LSA and ergonovine were identified in the urine sample of the deceased.


[Second phase 99mTc-HMDP accumulation using three phase bone scintigraphy in a case of primary breast cancer].
Accumulation of 99mTc-HMDP was recognized by three-phase bone scintigraphy in a case of primary breast cancer. A 67-year-old female was found to have a 14 x 13 mm mass in the upper lateral area of the left breast. At operation the histopathological finding was a proven solid-tubular carcinoma. On the first phase images, there was no abnormal accumulation in the tumor. On the second phase images, there was abnormal accumulation in the tumor, however, abnormal accumulation was observed followed by a decrease in accumulation. The third phase image showed no abnormal extra-skeletal accumulation. Color Doppler ultrasound sonography showed an abundance of color signals within the breast tumor. Increased 99mTc-HMDP accumulation in the second phase image might reflect tumor hypervascularization. It is concluded that three-phase bone scintigraphy using 99mTc-HMDP has the ability of positive description in primary breast cancer.


Gallotannin-Enriched Extract Isolated from Galla Rhois May Be a Functional Candidate with Laxative Effects for Treatment of Loperamide-Induced Constipation of SD Rats.
Several natural products containing tannins are used as traditional medicines for treatment of constipation; however, their pharmacological mechanism is not well understood. The laxative effects of gallotannin-enriched extract isolated from Galla Rhois (GEGR) were investigated using a constipation model induced by loperamide (Lop) injection. After analysis for antioxidant activity of GEGR, alterations in the excretion parameters, histological structure, mucin secretion, and related protein levels were measured in the transverse colon of Sprague Dawley (SD) rats with Lop-induced constipation following treatment with 250, 500 and 1,000 mg/ml of GEGR. The number and weight of feces increased significantly by 48-79% and 128-159%, respectively, in the Lop+GEGR treated group relative to the Lop+vehicle treated group, while food intake and water consumption were maintained at a constant level. The thickness of mucosa, muscle and flat luminal surface, as well as the number of goblet cells and crypt of lieberkuhn were enhanced in the Lop+GEGR treated group. Moreover, mucin secretion increased significantly in a dose dependent manner in the Lop+GEGR treated group. Furthermore, the downstream signaling pathway of the muscarinic acetylcholine receptors (mAChR) M2 and M3 was recovered by GEGR treatment, although the expression level varied. The levels of Gα expression and inositol triphosphate (IP3) concentration were also recovered in the Lop+GEGR treated group relative to the Lop+vehicle treated group. The results of the present study provide strong evidence that tannins distributed in various medicinal plants are important candidates for improving chronic constipation induced by Lop treatment in animal models.


Flagellin glycosylation with pseudaminic acid in Campylobacter and Helicobacter: prospects for development of novel therapeutics.
Many pathogenic bacteria require flagella-mediated motility to colonise and persist in their hosts. Helicobacter pylori and Campylobacter jejuni are flagellated epsilonproteobacteria associated with several human pathologies, including gastritis, acute diarrhea, gastric carcinoma and neurological disorders. In both species, glycosylation of flagellin with an unusual sugar pseudaminic acid (Pse) plays a crucial role in the biosynthesis of functional flagella, and thereby in bacterial motility and pathogenesis. Pse is found only in pathogenic bacteria. Its biosynthesis via six consecutive enzymatic steps has been extensively studied in H. pylori and C. jejuni. This review highlights the importance of flagella glycosylation and details structural insights into the enzymes in the Pse pathway obtained via a combination of biochemical, crystallographic, and mutagenesis studies of the enzyme-substrate and -inhibitor complexes. It is anticipated that understanding the underlying structural and molecular basis of the catalytic mechanisms of the Pse-synthesising enzymes will pave the way for the development of novel antimicrobials.