Strain differences in Echinococcus granulosus, with special reference to the status of equine hydatidosis in the United Kingdom.
The genetics of speciation in the genus Echinococcus are briefly discussed. As the organism is a hermaphrodite and multiplies asexually in the larval stage, the genetic mechanism for the ready production of new strains is inherent in the life-cycle. Some biological, biochemical and nutritional differences between the horse and sheep strains are examined. The sheep strain may be grown to sexual maturity, in vitro, in a diphasic system; the horse strain fails to grow in such a system. Differences have also been demonstrated between the soluble proteins of the two strains. The sheep strain is infective to man but, probably, non-infective to horses. The horse strain (which is now the major strain in the U.K and Ireland) appears to be poorly infective to sheep and may prove to be non-infective to man. The importance of determining the basic characteristics of other strains of E. granulosus are also stressed.


The 'tilt deformity' of the femoral head in osteoarthritis of the hip: a poor indicator of previous epiphysiolysis.
Apparent superomedial migration of the femoral head producing the 'tilt deformity' is the most frequent pattern of femoral head deformity in osteoarthritis of the hip. Previous reports have suggested a relationship to minor degrees of epiphysiolysis in the adolescent. Radiological and pathological examination of femoral heads obtained at the time of total hip replacement for osteoarthritis demonstrate characteristic abnormalities accompanying the 'tilt deformity'. It would appear that, in most patients, this pattern of femoral head migration results not from epiphysiolysis but from a remodelling process in the osteoarthritic joint.


One-pot synthesis of ternary-emission molecularly imprinted fluorescence sensor based on metal-organic framework for visual detection of chloramphenicol.
In this paper, a ternary-emission fluorescence imprinted polymer was one-pot synthesized by sol-gel method after mixing luminescence metal organic framework, green CdTe and near infrared red CdTe for visual detection of chloramphenicol in food. The ternary-emission fluorescence imprinted sensor showed wider linear range within concentration of 10 pM-0.5 nM and 0.5 nM-4.5 nM, with rapid response time of 3 min and the lower detection limit of 3.8 pM toward chloramphenicol. Meanwhile, NH2-UiO-66 improved the fluorescence sensitivity and response speed, the near-infrared CdTe enhanced the anti-interference ability of the imprinted sensor. Compared with the traditional single-emission and dual-emission fluorescence imprinted sensors, the ternary-emission imprinted sensor provided richer color changes from yellowish green to apricot to orange-salmon to amaranth to purple to final blue. It was applied to detect trace chloramphenicol in food with the recoveries of 98.2-101.2 %, which provided a new way for rapid visual detection of chloramphenicol.


The limit dextrinase from malted sorghum (Sorghum vulgare).
A limit dextrinase, free from contaminating carbohydrases, has been purified from malted sorghum flour. The enzyme readily hydrolysed alpha-limit dextrins having maltosyl or maltotriosyl side-chains, pullulan, and amylopectin beta-limit dextrin. Glycogen beta-limit dextrin and amylopectin were more slowly hydrolysed, the detection of the hydrolysis of amylopectin being dependent on enzyme concentration. No significant debranching of glycogen could be detected.


1.5-T MR relaxometry in quantifying splenic and pancreatic iron: retrospective comparison of a commercial 3D-Dixon sequence and an established 2D multi-gradient echo sequence.
To compare the quantitative measurement of splenic and pancreatic iron content using a commercial 3D-Dixon sequence (qDixon) versus an established fat-saturated R2* relaxometry method (ME-GRE). We analyzed splenic and pancreatic iron levels in 143 MR examinations (1.5 T) using the qDixon and a ME-GRE sequence (108 patients: 65 males, 43 females, mean age 61.31 years). Splenic and pancreatic R2* values were compared between both methods using Bland-Altman plots, concordance correlation coefficients (CCC), and linear regression analyses. Iron overload (R2* > 50 1/s) was defined for both organs and compared using contingency tables, overall agreement, and Gwet's AC1 coefficient. Of all analyzable examinations, the median splenic R2* using the qDixon sequence was 25.75 1/s (range: 5.6-433) and for the ME-GRE sequence 35.35 1/s (range: 10.9-400.8) respectively. Concerning the pancreas, a median R2* of 29.93 1/s (range: 14-111.45) for the qDixon and 31.25 1/s (range: 14-97) for the ME-GRE sequence was found. Bland-Altman analysis showed a mean R2* difference of 2.12 1/s with a CCC of 0.934 for the spleen and of 0.29 1/s with a CCC of 0.714 for the pancreas. Linear regression for the spleen/pancreas resulted in a correlation coefficient of 0.94 (p < 0.001)/0.725 (p < 0.001). Concerning iron overload, the proportion of overall agreement between the two methods was 91.43% for the spleen and 93.18% for the pancreas. Our data show good concordance between R2* values obtained with a commercial qDixon sequence and a validated ME-GRE relaxometry method. The 3D-qDixon sequence, originally intended for liver assessment, seems to be a reliable tool for non-invasive evaluation of iron content also in the spleen and the pancreas. • A 3D chemical shift imaging sequence and 2D multi-gradient echo sequence show good conformity quantifying splenic and pancreatic R2* values. • The 3D chemical shift imaging sequence allows a reliable analysis also of splenic and pancreatic iron status. • In addition to the liver, the analysis of the spleen and pancreas is often helpful for further differential diagnostic clarification and patient guidance regarding the iron status.


Determination of thiamine (vitamin B1) in maternal blood during normal pregnancies and pregnancies with intrauterine growth retardation.
The concentration of thiamine (vitamin B1) was measured in blood cells and plasma from mothers with normal pregnancy and from mothers whose pregnancy was complicated by intrauterine growth retardation (IUGR). Thiamine concentrations were estimated by the thiochrome method using HPLC separation and fluorimetric detection according to Weber et al. (2). During normal pregnancies the thiamine values in blood cells fall in the 28th to the 39th week of gestation from 230 nmol/l to 170 nmol/l. In cases with severe IUGR there is only a slight decrease in the thiamine levels from 140 nmol/l in the 30th week of gestation to a level of 130 nmol/l in the 39th week of gestation. During this period the thiamine values in plasma remain constant. To compare the thiamine concentrations in normal pregnancies and in those with IUGR we separated the thiamine values in a group from 28/0 to 34/6 and one from 35/0 to 39/6 weeks of gestation. In both groups mothers with normal pregnancy had significantly higher thiamine levels in the blood cells than mothers whose pregnancy was complicated by IUGR (p = 0.0001 and p = 0.0005). However, the thiamine values in plasma were not significantly different in normal pregnancies and pregnancies with IUGR. The results indicate that maternal thiamine deficiency might be one cause of IUGR.


Comparison of patterns of pulmonary venous blood flow in the functional single ventricle heart after operative aortopulmonary shunt versus superior cavopulmonary shunt.
In this study we investigated the patterns of pulmonary venous flow in children with functional single ventricles to obtain a better understanding of the determinants of transpulmonary blood flow. Sixty-eight patients with functional single ventricles and aortopulmonary shunt (n = 34, group I), or superior cavopulmonary connection (n = 34, group II) underwent transesophageal Doppler echocardiographic assessment of flow in the left upper pulmonary vein before undergoing the next stage of surgery. Twelve patients from group II also underwent simultaneous evaluation of superior vena caval flow. Biphasic forward pulmonary venous flow was noted in 62 patients in sinus rhythm (S wave in systole, D wave in diastole); in 6 patients with junctional rhythm, significant early systolic reversal of flow was present. Both the S- and D-wave velocity-time integrals (VTI) were greater in group I than in group II (S(VTI) 9.9 +/- 4.2 vs 8.0 +/- 2.6, p = 0.02; D(VTI) 8.0 +/- 3.5 vs 4.2 +/- 2.6, p <0.001). In both groups, pulmonary venous flow was predominantly systolic; however, the proportion of flow during ventricular systole was significantly greater in group II than in group I (S(VTI)/D(VTI) group II: 2.4 +/- 1.5; group I 1.4 +/- 0.5, p = 0.001; percent systolic fraction of pulmonary venous flow group II = 67%, group I = 56%, p <0.001). Analysis of superior vena caval flow in group II revealed a single predominant wave with onset at early systole and peak in late systole at a mean of 150 ms after the pulmonary venous S-wave peak. Our data suggest that ventricular systole (i.e., atrial relaxation, atrioventricular valve descent) asserts great influence on transpulmonary blood flow in the functional single ventricle.


Substantial decline in hospital admissions for heart failure accompanied by increased community mortality during COVID-19 pandemic.
We hypothesized that a decline in admissions with heart failure during COVID-19 pandemic would lead to a reciprocal rise in mortality for patients with heart failure in the community. We used National Heart Failure Audit data to identify 36 974 adults who had a hospital admission with a primary diagnosis of heart failure between February and May in either 2018, 2019, or 2020. Hospital admissions for heart failure in 2018/19 averaged 160/day but were much lower in 2020, reaching a nadir of 64/day on 27 March 2020 [incidence rate ratio (IRR): 0.40, 95% confidence interval (CI): 0.38-0.42]. The proportion discharged on guideline-recommended pharmacotherapies was similar in 2018/19 compared to the same period in 2020. Between 1 February-2020 and 31 May 2020, there was a 29% decrease in hospital deaths related to heart failure (IRR: 0.71, 95% CI: 0.67-0.75; estimated decline of 448 deaths), a 31% increase in heart failure deaths at home (IRR: 1.31, 95% CI: 1.24-1.39; estimated excess 539), and a 28% increase in heart failure deaths in care homes and hospices (IRR: 1.28, 95% CI: 1.18-1.40; estimated excess 189). All-cause, inpatient death was similar in the COVID-19 and pre-COVID-19 periods [odds ratio (OR): 1.02, 95% CI: 0.94-1.10]. After hospital discharge, 30-day mortality was higher in 2020 compared to 2018/19 (OR: 1.57, 95% CI: 1.38-1.78). Compared with the rolling daily average in 2018/19, there was a substantial decline in admissions for heart failure but an increase in deaths from heart failure in the community. Despite similar rates of prescription of guideline-recommended therapy, mortality 30 days from discharge was higher during the COVID-19 pandemic period.


Rapid liquid chromatography-ultraviolet determination of organic acids and phenolic compounds in red wine and must.
A reversed-phase liquid chromatography-ultraviolet (LC-UV) method is proposed for the rapid simultaneous analysis of the main carboxylic acids and polyphenols in must and wine. Good resolution was obtained for citric, tartaric, malic, lactic, acetic, caffeic, ellagic and gallic acids, (-)-epicatechin, quercetin and resveratrol. A novel silica-based column containing ether-linked phenyl groups, with polar end-capping and suitable for low-pH aqueous mobile phases was used and found to be superior to others tested. The method employed a mixture of 0.2% TFA in water and acetonitrile as eluents, showed linearity and precision, and was applied to samples of must and wine.


Effect of risk-reducing salpingo-oophorectomy on sex steroid hormone serum levels among postmenopausal women: an NRG Oncology/Gynecologic Oncology Group study.
Risk-reducing salpingo-oophorectomy is an effective ovarian cancer risk reduction strategy. However, bilateral oophorectomy has also been associated with increased long-term nonneoplastic sequelae, effects suggested to be mediated through reductions in systemic sex steroid hormone levels. Currently, it is unclear whether the postmenopausal ovary contributes to the systemic hormonal milieu or whether postmenopausal ovarian volume or other factors, such as body mass index and age, affect systemic hormone levels. We examined the impact of oophorectomy on sex steroid hormone levels in postmenopausal women. Furthermore, we explored how well ovarian volume measured by transvaginal ultrasound correlated with direct ovarian measures obtained during surgical pathology evaluation and investigated the association between hormone levels and ovarian volumes. Postmenopausal women who underwent risk-reducing salpingo-oophorectomy (180 cases) or ovarian cancer screening (38 controls) enrolled in an international, prospective study of risk-reducing salpingo-oophorectomy and risk of ovarian cancer algorithm-based screening among women at increased risk of ovarian cancer (Gynecologic Oncology Group-0199) were included in this analysis. Controls were frequency matched to the cases on age at menopause, age at study entry, and time interval between blood draws. Ovarian volume was calculated using measurements obtained from transvaginal ultrasound in both cases and controls and measurements recorded in surgical pathology reports from cases. Serum hormone levels of testosterone, androstenedione, androstenediol, dihydrotestosterone, androsterone, dehydroepiandrosterone, estrone, estradiol, and sex hormone-binding globulin were measured at baseline and follow-up. Spearman correlation coefficients were used to compare ovarian volumes as measured on transvaginal ultrasound and pathology examinations. Correlations between ovarian volumes by transvaginal ultrasound and measured hormone levels were examined using linear regression models. All models were adjusted for age. Paired t tests were performed to evaluate individual differences in hormone levels before and after risk-reducing salpingo-oophorectomy. Ovarian volumes measured by transvaginal ultrasound were only moderately correlated with those reported on pathology reports (Spearman rho [ρ]=0.42). The median time interval between risk-reducing salpingo-oophorectomy and follow-up for the cases was 13.3 months (range, 6.0-19.3), and the median time interval between baseline and follow-up for the controls was 12.7 months (range, 8.7-13.4). Sex steroid levels decreased with age but were not correlated with transvaginal ultrasound ovarian volume, body mass index, or time since menopause. Estradiol levels were significantly lower after risk-reducing salpingo-oophorectomy (percentage change, -61.9 post-risk-reducing salpingo-oophorectomy vs +15.2 in controls; P=.02), but no significant differences were seen for the other hormones. Ovarian volumes measured by transvaginal ultrasound were moderately correlated with volumes directly measured on pathology specimens and were not correlated with sex steroid hormone levels in postmenopausal women. Estradiol was the only hormone that declined significantly after risk-reducing salpingo-oophorectomy. Thus, it remains unclear whether the limited post-risk-reducing salpingo-oophorectomy changes in sex steroid hormones among postmenopausal women impact long-term adverse outcomes.


On the susceptibility of human platelets to aggregation by concanavalin A and the effect of this lectin on their response to ADP.
Concanavalin A aggregated gel-filtered platelets in 0.9% NaCl solution signifying cross-bridging by the lectin. Aggregation of these platelets by concanavalin A was temperature dependent; it did not occur at 0-4 degrees C unless the platelets were previously trypsinized. The level of aggregation of trypsinized platelets by concanavalin A at 0-4 degrees C was similar to that of untreated platelets at 37 degrees C. It is suggested that trypsin facilitates platelet aggregation by concanavalin A at 0-4 degrees C by causing a configurational change in membrane glycoproteins which orientates concanavalin A receptor sites into positions that favour lectin cross-bridging. Concanavalin A failed to aggregate platelets in plasma. Radioisotope studies showed that the amount of [3H]concanavalin A which combined with platelets in plasma was extremely low compared with gel-filtered platelets in saline. The aggregation of Ehrlich ascites cells by concanavalin A was considerably reduced when platelet-free plasma was added to the medium suggesting that it was due to the presence of concanavalin A-reactive components in the plasma. Concanavalin A inhibited the ADP-induced aggregation of platelets suspended in plasma or in a salts solution supplemented with calcium and fibrinogen, although the inhibitory effect was more conspicuous in the latter case. The results suggests that concanavalin A produces its inhibitory effect on ADP-induced platelet aggregation by interacting with membrane glycoproteins, and this further suggests their involvement in aggregation.


Multigenerational study of the obesogen effects of bisphenol S after a perinatal exposure in C57BL6/J mice fed a high fat diet.
Bisphenol S is an endocrine disruptor exhibiting metabolic disturbances, especially following perinatal exposures. To date, no data are available on the obesogen effects of BPS in a mutligenerational issue. We investigated obesogen effects of BPS in a multigenerational study by focusing on body weight, adipose tissue and plasma parameters in male and female mice. Pregnant C57BL6/J mice were exposed to BPS (1.5 μg/kg bw/day ie a human equivalent dose of 0.12 μg/kg bw/day) by drinking water from gestational day 0 to post natal day 21. All offsprings were fed with a high fat diet during 15 weeks. Body weight was monitored weekly and fat mass was measured before euthanasia. At euthanasia, blood glucose, insuline, triglyceride, cholesterol and no esterified fatty acid plasma levels were determined and gene expressions in visceral adipose tissue were assessed. F1 males and females were mated to obtain the F2 generation. Likewise, the F2 mice were cross-bred to obtain F3. The same analyses were performed. In F1 BPS induced an overweight in male mice associated to lipolysis gene expressions upregulation. In F1 females, dyslipidemia was observed. In F2, BPS exposure was associated to an increase in body weight, fat and VAT masses in males and females. Several plasma parameters were increased but with a sex related pattern (blood glucose, triglycerides and cholesterol in males and NEFA in females). We observed a down-regulation in mRNA expression of gene involved in lipogenesis and in lipolysis for females but only in the lipogenesis for males. In F3, a decrease in VAT mass and an upregulation of lipogenesis gene expression occurred only in females. BPS perinatal exposure induced sex-dependent obesogen multigenerational effects, the F2 generation being the most impacted. Transgenerational disturbances persisted only in females.


The mechanism of the search for homology promoted by recA protein. Facilitated diffusion within nucleoprotein networks.
recA protein promotes the homologous pairing of single strands with duplex DNA by polymerizing on the single strands to make presynaptic nucleoprotein filaments which are polyvalent with respect to duplex DNA and which consequently form large networks or coaggregates when duplex DNA is added. Previous work has shown that efficient homologous pairing occurs within these networks. In the experiments described here, we observed that the length of the duplex DNA determined the stability of coaggregates, their steady state level, and the yield of joint molecules. Correspondingly, heterologous duplex DNA when preincubated with presynaptic filaments excluded subsequently added homologous duplex DNA from coaggregates and inhibited homologous pairing; the extents of exclusion and inhibition were determined by the length of the heterologous duplex DNA. On the other hand, long heterologous duplex DNA when added together with short homologous duplex DNA was capable of stimulating the absorption of the homologous molecules into coaggregates and increasing the rate of homologous pairing. In reactions involving short duplex molecules, polyamines exerted comparable effects on coaggregation and homologous pairing. We conclude that coaggregates are instrumental in homologous pairing, that they constitute distinct domains that are responsible for the processive or first order character of the pairing reaction, and that they act by concentrating DNA and facilitating diffusion.


Monocyte to macrophage differentiation-associated (MMD) targeted by miR-140-5p regulates tumor growth in non-small cell lung cancer.
Monocyte to macrophage differentiation-associated (MMD) is identified in macrophages as a gene associated with the differentiation from monocytes to macrophages. Recent microarray analysis for non-small cell lung cancer (NSCLC) suggests that MMD is an important signature associated with relapse and survival among patients with NSCLC. Therefore, we speculate that MMD likely plays a role in lung cancer. In this study, we found that the protein level of MMD was increased in lung cancer compared to benign lung tissues, and knockdown of MMD inhibited the growth of A549 and Lewis lung cancer cells (LLC) in vitro and in vivo. Integrated analysis demonstrated that MMD was a direct functional target of miR-140-5p. Furthermore, we found that miR-140-5p/MMD axis could affect the cell proliferation of lung cancer cells by regulating Erk signaling. Together, our results highlight the significance of miR-140-5p/MMD axis in lung cancer, and miR-140-5p/MMD axis could serve as new molecular targets for the therapy against lung cancer.


Effects of asbestos and smoking on gas diffusion in people exposed to crocidolite.
To examine the effects of asbestos exposure and tobacco smoking on the level and rate of change of the diffusing capacity of the lung for carbon monoxide (DLCO). A cohort study of 934 people (including both mine workers and town residents) exposed to crocidolite (blue asbestos) at the asbestos mines and in the town of Wittenoom, Western Australia, between 1943 and 1966. DLCO measurements were taken during a follow-up period from 1992 to 2002. Baseline levels of DLCO and change in levels over time. 2980 DLCO measurements were done on 934 people (of whom 818 were men and 724 were workers) who underwent a median of 2 (range, 1-17) measurements during the follow-up period. Radiographic asbestosis at baseline and asbestos exposure at a younger age were associated with lower DLCO values. The average rate of decline in DLCO was 0.33 (95% CI, 0.31-0.35) units per year, plus an additional decrement of 0.22 (95% CI, 0.12-0.32) units per year if the participant had radiographic asbestosis at the beginning of the follow-up period. Compared with never-smokers, current smokers and ex-smokers had lower DLCO at baseline, but smoking status did not affect the change in DLCO during the follow-up period. Our results confirm a continuous deleterious effect of crocidolite on DLCO, especially on people with asbestosis. Smoking was associated with lower DLCO levels, but was not a significant predictor of rate of change in DLCO. Smoking status did not affect the relationships between crocidolite exposure and the level or rate of change of DLCO in this population.


Quality of life and its predictors in adults with tuberous sclerosis complex (TSC): a multicentre cohort study from Germany.
Tuberous sclerosis complex (TSC) is a monogenetic, multisystemic disease characterised by the formation of benign tumours that can affect almost all organs, caused by pathogenic variations in TSC1 or TSC2. In this multicentre study from Germany, we investigated the influence of sociodemographic, clinical, and therapeutic factors on quality of life (QoL) among individuals with TSC. We assessed sociodemographic and clinical characteristics and QoL among adults with TSC throughout Germany using a validated, three-month, retrospective questionnaire. We examined predictors of health-related QoL (HRQoL) using multiple linear regression analysis and compared the QoL among patients with TSC with QoL among patients with other chronic neurological disorders. We enrolled 121 adults with TSC (mean age: 31.0 ± 10.5 years; range: 18-61 years, 45.5% [n = 55] women). Unemployment, a higher grade of disability, a higher number of organ manifestations, the presence of neuropsychiatric manifestations or active epilepsy, and a higher burden of therapy-related adverse events were associated with worse QoL, as measured by two QoL instruments (EuroQoL-5 dimensions [EQ-5D] and Quality of Life in Epilepsy Patients [QOLIE-31]). Neuropsychiatric and structural nervous system manifestations, the number of affected organs, and therapy-related adverse events were also associated with higher depression, as measured by the Neurological Disorders Depression Inventory for Epilepsy (NDDI-E). In multiple regression analysis, more severe therapy-related adverse events (large effect, p < 0.001), active epilepsy (large effect, p < 0.001), and neuropsychiatric manifestations (medium effect, p = 0.003) were independently associated with worse HRQoL, explaining 65% of the variance (p < 0.001). The HRQoL among patients with active TSC-associated epilepsy was worse than that among patients with drug-refractory mesial temporal lobe epilepsy (p < 0.001), and the generic QoL among patients with more than three TSC organ manifestations was similar to those of patients with severe migraine and uncontrolled asthma. Active epilepsy, neuropsychiatric manifestations (such as anxiety and depression), and therapy-related adverse events are important independent predictors of worse quality of life among adults with TSC. Generic quality of life in TSC with several manifestations is similar to uncontrolled severe chronic diseases and significantly negatively correlates with TSC severity. DRKS, DRKS00016045 . Registered 01 March 2019.


Effect of intermittent treatment with amodiaquine on anaemia and malarial fevers in infants in Tanzania: a randomised placebo-controlled trial.
Malaria is a major cause of infant morbidity and mortality in sub-Saharan Africa, and is often complicated by severe anaemia. Resistance of Plasmodium falciparum to most affordable antimalarial drugs is an impediment to intermittent chemotherapy. We investigated the effect of presumptive intermittent treatment with amodiaquine and daily iron supplementation in infants on malarial fevers and anaemia, in a holoendemic area of Tanzania where malaria is largely resistant to chloroquine and sulfadoxine/ pyrimethamine. 291 infants aged 12-16 weeks who attended three clinics were randomised to receive amodiaquine, iron supplementation, amodiaquine plus iron supplementation, or placebo. Over 6 months, we gave amodiaquine three times with intervals of 60 days; oral iron supplementation was given daily. Malarial fevers and anaemia were monitored at bimonthly treatment visits and by self-reporting to health centres. The protective efficacy of intermittent amodiaquine treatment in prevention of malarial fevers and anaemia was 64.7% (95% CI, 42.4-77.2) and 67.0% (95% CI, 34.5-83.4), respectively. Protective efficacy was similar in the group receiving amodiaquine plus iron supplementation. Infants receiving iron supplementation only were partly protected against anaemia (protective efficacy 59.8%; 95% CI, 23.4-78.9), but not against malarial fevers. 4 months' follow-up did not show rebound morbidity. We noted no haematological or clinical adverse effects. Presumptive intermittent treatment for malaria with amodiaquine reduced malarial fevers and anaemia in infants, in an area with high resistance to other antimalarials. Intermittent treatment strategies for malaria in highly endemic areas could be of great benefit to public health.


Impact of Different Saccharides on the In-Process Stability of a Protein Drug During Evaporative Drying: From Sessile Droplet Drying to Lab-Scale Spray Drying.
Solid biopharmaceutical products can circumvent lower temperature storage and transport and increase remote access with lower carbon emissions and energy consumption. Saccharides are known stabilizers in a solid protein produced via lyophilization and spray drying (SD). Thus, it is essential to understand the interactions between saccharides and proteins and the stabilization mechanism. A miniaturized single droplet drying (MD) method was developed to understand how different saccharides stabilize proteins during drying. We applied our MD to different aqueous saccharide-protein systems and transferred our findings to SD. The poly- and oligosaccharides tend to destabilize the protein during drying. The oligosaccharide, Hydroxypropyl β-cyclodextrin (HPβCD) shows high aggregation at a high saccharide-to-protein molar ratio (S/P ratio) during MD, and the finding is supported by nanoDSF results. The polysaccharide, Dextran (DEX) leads to larger particles, whereas HPBCD leads to smaller particles. Furthermore, DEX is not able to stabilize the protein at higher S/P ratios either. In contrast, the disaccharide Trehalose Dihydrate (TD) does not increase or induce protein aggregation during the drying of the formulation. It can preserve the protein's secondary structure during drying, already at low concentrations. During the drying of S/P formulations containing the saccharides TD and DEX, the MD approach could anticipate the in-process (in) stability of protein X at laboratory-scale SD. In contrast, for the systems with HPβCD, the results obtained by SD were contradictory to MD. This underlines that depending on the drying operation, careful consideration needs to be applied to the selection of saccharides and their ratios.


The Design and Optimization of Plasmonic Crystals for Surface Enhanced Raman Spectroscopy Using the Finite Difference Time Domain Method.
We present computational studies of quasi three-dimensional nanowell (NW) and nanopost (NP) plasmonic crystals for applications in surface enhanced Raman spectroscopy (SERS). The NW and NP plasmonic crystals are metal coated arrays of cylindrical voids or posts, respectively, in a dielectric substrate characterized by a well/post diameter (D), relief depth (R D), periodicity (P), and metal thickness (M T). Each plasmonic crystal is modeled using the three-dimensional finite-difference time-domain (FDTD) method with periodic boundary conditions in the x- and y-directions applied to a computational unit cell to simulate the effect of a periodic array. Relative SERS responses are calculated from time-averaged electric field intensity enhancements at λ exc and λ scat or at λ mid via G SERS 4 = g 2 ( λ exc ) × g 2 ( λ scat ) or G mid 4 = g 4 ( λ mid ) , respectively, where g 2 = | E | 2 / | E 0 | 2 . Comparisons of G SERS 4 and G mid 4 are made to previously reported experimental SERS measurements for NW and NP geometries. Optimized NW and NP configurations based on variations of D, P, R D, and M T using G SERS 4 are presented, with 6× and 2× predicted increases in SERS, respectively. A novel plasmonic crystal based on square NP geometries are considered with an additional 3× increase over the optimized cylindrical NP geometry. NW geometries with imbedded spherical gold nanoparticles are considered, with 10× to 10 3 × increases in SERS responses over the NW geometry alone. The results promote the use of FDTD as a viable in silico route to the design and optimization of SERS active devices.


Pomegranate (Punica granatum) peel fractions obtained by supercritical CO2 increase oxidative and colour stability of bluefish (Pomatomus saltatrix) patties treated by UV-C irradiation.
The sequential fractionation by supercritical-CO2 (SC-CO2) was applied to obtain fractions enriched in bioactive compounds of pomegranate peel, and we investigated if pomegranate peel extract and fractions would be effective to inhibit lipid and protein oxidation, and discolouration of bluefish patties stored at 4 °C for 9 days, after UV-C irradiation. The non-fractionated SC-CO2 extract from pomegranate peel was rich in phenolic compounds, mainly ellagitannins, besides, it possessed lipophilic compounds such as tocopherols and β-carotene. These compounds were successfully separated by the fractionation protocols, in a lipid fraction concentrated in lipophilic compounds, and one or two fractions enriched with phenolic compounds, especially ellagitannins. The lipid fraction and the high phenolics fraction from pomegranate peel were then as effective as the synthetic antioxidant BHT in avoiding bluefish patties oxidation during refrigerated storage. Our data indicates that pomegranate peel fractions could be used to replace a synthetic antioxidant in fish meat.


Imaging of gallbladder and biliary tract before laparoscopic cholecystectomy: comparison of intravenous cholangiography and the combined use of HASTE and single-shot RARE MR imaging.
To compare intravenous cholangiography (i.v.c.) and magnetic resonance imaging (MRI) as preoperative imaging techniques in patients scheduled for laparoscopic cholecystectomy. Twenty patients underwent i.v.c. and MRI, 40 axial 'localizer' images were first obtained with a half-Fourier single-shot turbo spin echo (HASTE) sequence. Next, an extremely high T2-weighted rapid acquisition relaxation enhancement (RARE) acquisition (TE = 1100 msec) was used for MR cholangiography. All images obtained with i.v.c. and MRI were independently analyzed by two observers. The relative visibility of the (normal or abnormal) gallbladder (GB), cystic duct (CD), and bile ducts (BD) on both types of images was scored as follows: 1 = MRI better than i.v.c.; 2 = no difference; 3 = i.v.c. better than MRI. We observed 3 anatomic variants: 1 of the cystic duct and 2 of the intrahepatic bile ducts; 15 patients had gallstones; one had cholecystitis. Magnetic resonance images were considered more informative than i.v.c.-images for visualization of GB in 11 patients (55%), for CD in 9 patients (45%), and in 8 patients (40%) for visualization of the CBD. Intravenous cholangiography outperformed MRI in the evaluation of the CBD and CD in one patient (5%). The combined use of half-Fourier single-shot RARE and high T2-weighted RARE MR imaging is a valuable alternative to i.v.c. in the preoperative evaluation of gallbladder and biliary tract.


[Clinical effects of pre-expanded anterior perforator flap of transverse cervical artery and pre-expanded thoracic random flap in reconstructing extensive facial and cervical scar in relay].
Objective: To explore the clinical effects of pre-expanded anterior perforator flap of transverse cervical artery in extensive facial and cervical scar reconstruction and contralateral pre-expanded thoracic random flap in relay in donor site repair. Methods: A retrospective cohort study was conducted. From May 2008 to December 2018, 10 patients with extensive facial and cervical scar after burns were treated in the Fourth Medical Center of PLA General Hospital, including 8 males and 2 females, aged 10-55 years. In the first stage of operation, two skin and soft tissue expanders of the same volume (with rated capacity of 250-600 mL) were respectively placed in the right side and left side of the chest according to the size of scar, and then the skin was expanded. The total amount of normal saline injected was 2 to 4 times of the rated capacity of the expander. In the second stage, the defect with area of 12 cm×8 cm-23 cm×15 cm caused by scar resection and release was repaired with unilateral pre-expanded anterior perforator flap of transverse cervical artery with area of 12 cm×9 cm-24 cm×16 cm. The contralateral pre-expanded thoracic random flap with the same area as that of the above-mentioned perforator flap was extended to repair the secondary defect with area of 8 cm×6 cm-17 cm×14 cm formed after transfer of the above-mentioned perforator flap. The exploration of perforating branch of transverse cervical artery, flap transfer and survival, injury repair, and complications were observed. The appearance and related function of donor and recipient sites and satisfaction of patients were followed up. Results: The perforating branches of transverse cervical artery appeared stably in the 10 patients. All the flaps were transferred to the recipient area without tension and survived. Both facial and cervical injuries were repaired successfully with no common complications. During the follow-up of 6 months-8 years, the color and texture of the pre-expanded anterior perforator flap of transverse cervical artery matched with the surrounding tissue, the functions of head raising and neck rotation of patients were significantly improved compared with those before operation, the color and texture of the flap transplanted in the first donor site matched with the original skin, linear scar left at the surgical incision, and 9 patients were satisfied with the restoration of the appearance and function of donor and recipient sites. Conclusions: The color and texture of the pre-expanded anterior perforator flap of transverse cervical artery match well with the face and neck, and the repairable area is large. After the perforator flap is removed, the secondary wound can be repaired with the pre-expanded thoracic random flap at the same time, and the injury of the chest donor site is alleviated. This relay repair method is a good choice for reconstructing extensive facial and cervical scar.


Medium-chain triglyceride supplementation exacerbates peritonitis-induced septic shock in rats: role on cell membrane remodeling.
Lipid emulsions for parenteral nutrition interfere with immunity and may alter the cell plasma membrane and microparticle release, thus modulating their biological effects. Our aim was to evaluate the effect of two lipid emulsions for parenteral nutrition containing either a mixture of long- and medium-chain triglycerides (LCTs and MCTs) or LCTs only, to assess their role on microparticle release and acute inflammation during septic shock in rats. Septic rats (cecal ligation and puncture) and sham rats were infused with 5% dextrose or a lipid emulsion during 22 h. After 18 h, rats were resuscitated during 4 h and hemodynamic parameters monitored. Circulating microparticles and their phenotype were measured by prothrombinase assay; heart and aorta were collected for Western blotting and electron paramagnetic resonance measurements. No significant effect of lipid emulsions was observed in sham rats. In septic rats, norepinephrine requirements were increased in MCT/LCT-infused rats compared with 5% dextrose- or LCT-infused rats (2.7 ± 0.2 vs. 1.9 ± 0.8 and 1.2 ± 0.3 μg/kg per minute, respectively; P < 0.05) with increased procoagulant microparticle generation (38.6 ± 5.8 vs. 18.8 ± 3.1 and 19.2 ± 3.0 nM equivalent phosphatidylserine [Eq PhtdSer]; P < 0.05), leukocyte- (17.4 ± 3.5 vs. 7.7 ± 1.8 and 6.0 ± 1.1 nM Eq PhtdSer; P < 0.05), platelet- (13.9 ± 2.5 vs. 4.4 ± 0.7 and 5.4 ± 1.3 nM Eq PhtdSer; P < 0.05), and endothelial-derived microparticles (16.9 ± 3.6 vs. 6.4 ± 1.4 and 5.6 ± 0.8 nM Eq PhtdSer; P < 0.05). The mixture of MCTs/LCTs significantly increased cardiac and vascular nitric oxide and superoxide anion production, phosphorylated IκB, and cyclooxygenase 2 expression compared with the lipid emulsion containing only LCTs. Compared with 5% dextrose, MCT/LCT supplementation during septic shock in rats induced deleterious effects with increased inflammation and cell activation, associated to vascular hyporeactivity. During septic shock, LCT supplementation seemed to be neutral compared with 5% dextrose infusion.


Discriminative stimulus effects of flumazenil in rhesus monkeys treated chronically with chlordiazepoxide.
Discriminative stimulus effects of the benzodiazepine antagonist flumazenil were studied in two rhesus monkeys receiving 3.2 mg/kg/12 h of chlordiazepoxide while discriminating between vehicle and 0.056 mg/kg of flumazenil. In a drug discrimination component responding was maintained under a FR 10 schedule of stimulus-shock termination; in a non-discrimination component responding was maintained under a FR 10 schedule of food presentation. Flumazenil and Ro 15-4513 occasioned >80% flumazenil-lever responding at doses larger than 0.032 and 0.056 mg/kg, respectively. Pentylenetetrazole, ethyl-beta-carboline-3-carboxylate (betaCCE), ketamine and spiradoline failed to substitute for flumazenil although >80% drug-lever responding was observed for two of the compounds in one monkey. Flumazenil, Ro 15-4513, pentylenetetrazole, betaCCE but not ketamine or spiradoline decreased rates of responding in the food component at doses that had little effect on rates in the stimulus-shock termination component. When chlordiazepoxide injections were discontinued and saline was administered before the session, monkeys did not respond on the flumazenil lever; when flumazenil was administered under the same conditions, monkeys responded on the flumazenil lever despite not having received chlordiazepoxide for nine days. Drug stimulus control was established with flumazenil in monkeys receiving chlordiazepoxide and substitution studies suggest that this effect of flumazenil might result from antagonist actions at benzodiazepine receptors: however, lack of withdrawal-related effects after termination of chlordiazepoxide treatment precludes validation of this procedure for studying benzodiazepine dependence.


Healthy nutritional behavior during COVID-19 lockdown: A cross-sectional study.
Ongoing outbreak of the novel coronavirus infectious disease 2019 (COVID- 19) constitutes a major global pandemic health care system challenge. Shortly following the disease outbreak, like SARS-CoV virus, all affected countries are implementing various preventive and control measures to mitigate the spread of the disease. Optimizing public health system during COVID-19 pandemic requires not only advanced medical and biological sciences knowledge, but also all human sciences related to social, as well as nutritional behavior, and lifestyle practices. To investigate the effect of COVID-19- quarantine on healthy nutritional behavior and lifestyle practices among Jordanian population. A cross-sectional study among Jordanian population was conducted using an online questionnaire between March and April 2020. Participants were kindly requested to answer a standardized and validated structured questionnaire. Demographic information (age, gender, place of residence, and occupation), anthropometric data (reported weight and height); nutritional behavior information (number of meals per day, snakes, water intake), and physical activity behavior were requested. A total of 4473 respondents were included in the study; obese (n = 1135), normal body weight (n = 1561), and underweight (n = 116). During COVID-19 quarantine, a significant increase in body weight (12.9% underweight, 28.5% normal body weight, 36.4% overweight' and 41.1% of obese (p < 0.001) was reported. Moreover, the number of snacks between meals (p < 0.001), number of main meals (p < 0.001), and the smoking rate was also increased significantly; however, there was no significant difference regarding physical activity among various weight status groups (p < 0.05). In this study, we have provided evidence, for the first time, that there were significant negative changes in healthy nutritional behavior among Jordanians during COVID-19 quarantine who encountered significantly increased body weight, appetite, and smoking. Hence, future larger cross-sectional studies are warranted.


[Fast fluorimetric assay of chlortetracycline in meat and bone (author's transl)].
Antibiotics in biological materials, including tetracyclines, are determined usually be means of microbiological methods. Because of the low specificity and the time consuming procedure of the microbiological assays, we developed a fluorimetric method, which enables us to determine specifically the tetracyclines and to reduce the time for the assay down to 4 hours. Our method is based on the determination of tetracyclines published by Kohn (1961), which we modified for the determination of chlortetracycline in meat and bone. Tetracyclines are fluorescent compounds, whose quantum yields are very much enhanced in most cases by forming a complex with Ca2+ ions and by extracting it into an organic solvent. This transfer into acetic acid ethylester separates the chlortetracycline from interfering substances. The yield of chlortetracycline with the described method was about 80%, the sensitivity of the detection was about 0.5 ppm.


Stimulation of murine bone-marrow colony formation by conditioned medium from human fetal liver cells.
A substance that stimulates growth of colonies of mononuclear granulocytic cells derived from the bone marrow of mice was produced by incubating fetal liver cells (conditioned medium). This substance appears to have the same properties described elsewhere as colony-stimulating factor (CSF). The enhanced stimulatory ability of the conditioned medium from human fetal liver cells compared to medium not conditioned suggests that fetal liver is a potent source of colony-stimulating factor.


Estimating species trees using approximate Bayesian computation.
Development of methods for estimating species trees from multilocus data is a current challenge in evolutionary biology. We propose a method for estimating the species tree topology and branch lengths using approximate Bayesian computation (ABC). The method takes as data a sample of observed rooted gene tree topologies, and then iterates through the following sequence of steps: First, a randomly selected species tree is used to compute the distribution of rooted gene tree topologies. This distribution is then compared to the observed gene topology frequencies, and if the fit between the observed and the predicted distributions is close enough, the proposed species tree is retained. Repeating this many times leads to a collection of retained species trees that are then used to form the estimate of the overall species tree. We test the performance of the method, which we call ST-ABC, using both simulated and empirical data. The simulation study examines both symmetric and asymmetric species trees over a range of branch lengths and sample sizes. The results from the simulation study show that the model performs very well, giving accurate estimates for both the topology and the branch lengths across the conditions studied, and that a sample size of 25 loci appears to be adequate for the method. Further, we apply the method to two empirical cases: a 4-taxon data set for primates and a 7-taxon data set for yeast. In both cases, we find that estimates obtained with ST-ABC agree with previous studies. The method provides efficient estimation of the species tree, and does not require sequence data, but rather the observed distribution of rooted gene topologies without branch lengths. Therefore, this method is a useful alternative to other currently available methods for species tree estimation.


Role of denosumab before resection and reconstruction in giant cell tumors of bone: a single-centered retrospective cohort study.
Denosumab has been approved by Food and Drug Authority in 2013 for use in surgically unresectable Giant cell tumor (GCT) to achieve resectable tumor margins. The aim of this study is to investigate the functional outcome and surgical convenience with the use of neoadjuvant denosumab before resection and reconstruction in Campanacci grade III GCT. We retrospectively reviewed 70 cases of Campanacci grade III GCT receiving resection and reconstruction between January 2014 and December 2019. They were stratified into two groups: one group of 29 patients received once-weekly denosumab 120 mg for 4-weeks before resection and reconstruction, while the other group of 41 patients did not receive denosumab before resection and reconstruction. Quality of life by musculoskeletal tumor society score where 0-7 means poor, 8-14 means fair, 15-22 means good; above 22 means excellent, incidence of tumor recurrence, intraoperative duration in minutes and postoperative positive margins were assessed for each cohort after 12 months follow-up. There was no significant difference in musculoskeletal tumor society score (25.75 vs. 27.41; P = 0.178), incidence of recurrence (3.45% vs. 4.88%; P < 0.001), and postoperative positive margins (10.34% vs. 4.88%; P = 0.38) for both groups. However, the intraoperative duration (133.38 vs. 194.49; P < 0.001) was significantly higher in the non-denosumab group compared with denosumab group. Neoadjuvant denosumab is equally effective considering postoperative functional outcomes and surgical convenience except intraoperative duration where it is highly helpful in saving the operating time duration. Easier identification, resection and lesser reconstruction are the key surgical convenience offered by neoadjuvant denosumab.


Upper extremity tourniquet tolerance.
Twenty unsedated volunteers were tested to compare the relative tolerance of an arm tourniquet on one side and of a forearm tourniquet on the other. The forearm tourniquet was tolerated an average of 13 minutes (45%) longer and was consistently rated as less painful during and immediately after the test. No subject tolerated the arm tourniquet longer than the forearm tourniquet. Two peaks of discomfort were found, one just before deflation beneath the tourniquet and one in the hand 2 minutes later. Ulnar nerve distribution paresthesias were the earliest changes experienced; however, complete numbness occurred first in the median nerve distribution. Complete paralysis occurred 7 minutes later (24%) with the forearm tourniquet. Pulse rate did not correlate with tourniquet pain, but blood pressure did. No measurable tourniquet-induced edema occurred on either side.


Bone mineral measurements and Tc-99m IDP retention in the dog.
The reproducibility of various procedures for the noninvasive assessment of the skeleton in dogs has been measured. In one animal, dual photon measurements of bone mineral mass were made in the whole body, the lumbar spine, and the proximal femur on 10 occasions during a 15-week period. In two other dogs, repeat measurements of Tc-99m imidodiphosphate (IDP) retention were made. For the total body and the proximal femur, the reproducibilities were 2 and 2.5%, respectively. The reproducibility of the lumbar spine was approximately 10% and was influenced by the variable fraction of the posterior spinous processes included in the measurement. The reproducibility of IDP retention measurements was not established. It was about 8% when all 10 measurements were considered but fell to 1.5% when three consecutive measurements were excluded. Dual photon measurements of the whole body and the proximal femur can be used to assess bone mass changes in the canine skeleton.


Radiation Therapy Alone for Human Papillomavirus-Related Squamous Cell Carcinoma of the Oropharynx: A Single-Arm, Phase 2 Study.
Human papillomavirus (HPV)-related squamous cell carcinoma of the oropharynx (OPSCC) is extremely radiosensitive. Radiation therapy plus high-dose cisplatin remains the standard of care but causes long-term toxicity. Treatment deintensification approaches that reduce toxicity while maintaining survival are desirable for HPV-related OPSCC. We conducted a single-arm, multicenter, phase 2 trial. Patients with newly diagnosed, biopsy-proven, American Joint Committee on Cancer (seventh edition) stage III or IV OPSCC positive for both p16 and HPV DNA were eligible. Patients with T4, N3, or T1N1 disease were excluded. Smoking history was not included in eligibility criteria. Patients received intensity modulated radiation therapy (IMRT) of 70 Gy in 35 fractions or 70.4 Gy in 32 fractions without chemotherapy. The primary endpoint was complete response or complete metabolic response 10 weeks after IMRT completion. Between September 13, 2013, and November 15, 2016, 39 patients were enrolled according to a 2-stage Simon design. Twenty-three patients (59%) had smoked for more than10 pack-years. Thirty-six patients (92%) had tumors genotyped as HPV16. Thirty-seven patients (95%) received full-dose radiation therapy and 35 (90%) had complete response or complete metabolic response. Median follow-up was 51 months (interquartile range, 41-63 months). One patient (3%) had regional recurrence and 3 (8%) had distant metastasis. One patient died of disease. The 2-year progression-free survival rate was 94% (95% CI, 81%-99%), and the 2-year overall survival rate was 100%. Common grade 3 adverse events during IMRT included mucositis in 10 patients (26%) and dysphagia in 7 patients (18%). No patients were dependent on a feeding tube at 1 month after IMRT completion. No grade 3 or 4 late adverse events were observed. IMRT alone is associated with excellent response as well as reduced toxicity and could be a treatment option for carefully selected patients with locally advanced "true" HPV-related OPSCC. Further studies are warranted.


Effects of eCG and FSH in timed artificial insemination treatment regimens on estrous expression and pregnancy rates in primiparous and multiparous Bos indicus cows.
Effects were evaluated in Bos indicus cows of eCG and FSH on follicular growth, estrous expression, and pregnancy per artificial insemination (P/AI) as a result of fixed-time artificial insemination (TAI). In Experiment 1, extent of timing-of-ovulation synchronization among cows was evaluated after imposing an estrogen/progesterone-based treatment regimen. At progesterone device removal (D8), cows were administered: eCG, or FSH or served as untreated Controls. In Experiment 2, percentage of cows P/AI was evaluated when the Experiment 1-treatment regimen was imposed. On D10, all cows were artificially inseminated. In Experiment 3, cows were assigned to two treatment groups (Control and eCG) on D8 to evaluate percentage of cows P/AI and estrous expression. In Experiment 1, follicular dynamics were similar among treatment groups. In Experiment 2, follicular growth was greater (P = 0.0001) with the eCG treatment. There was an interaction of treatment × parity (P = 0.007) on percentage of cows P/AI. There was a greater percentage of primiparous cows P/AI in the eCG-treated than Control and FSH-treated cows. There was a greater percentage of eCG-treated multiparous cows pregnant as a result of TAI than Control cows. There was an interaction of treatment × parity (P = 0.005) on P/AI in Experiment 3, in which the eCG effect was more pronounced in primiparous cows. Treatment with FSH, therefore, was not as effective as eCG in stimulation of follicular growth or enhancing percentage of cows pregnant as a result of TAI. Physiological effects of eCG, however, were also more evident in primiparous cows.


Development of a CAFs-related gene signature to predict survival and drug response in bladder cancer.
As one of important components of tumor microenvironment, CAFs (cancer-associated fibroblasts) play a vital role in the development and metastasis of bladder cancer. The present study aimed to develop a CAFs-related gene signature to predict the prognosis of patients and the response to chemotherapy and immunotherapy based on research of multidatabase. Expression data and clinical information were obtained from TCGA and GEO databases. Different bioinformatic and statistical methods were combined to construct the robust CAFs-related gene signature for prognosis. The model was explored from four aspects: single-cell source, immune infiltration, correlation with cancer-related genes and pathways, and prediction of drug response. After screening, five genes (BNC2, LAMA2, MFAP5, NID1, and OLFML1) related to CAFs were used for constructing the signature to divide patients into high- and low-risk groups. Patients in low-risk group had better prognosis and multidatabase analysis confirmed the predictive value. The five genes were mainly expressed by fibroblasts and involved in regulation of pathways related with glycolysis, hypoxia, and epithelial-mesenchymal transition (EMT). BNC2, LAMA2, and NID1 were strongly associated with drug sensitivity. Moreover, the immunological status was different between high- and low-risk groups. High-risk patients had poor response to chemotherapy or immunotherapy. The CAFs-related gene signature might help to optimize risk stratification and provide a new insight in individual treatment for bladder cancer.


Azide-Based High-Energy Metal-Organic Frameworks with Enhanced Thermal Stability.
We describe the structure and properties of [Zn(C6H4N5)N3] n , a new nonporous three-dimensional high-energy metal-organic framework (HE-MOF) with enhanced thermal stability. The compound is synthesized by the hydrothermal method with in situ ligand formation under controlled pH and characterized using single-crystal X-ray diffraction, elemental analysis, and Fourier transform infrared. The measured detonation temperature (T det = 345 °C) and heat of detonation (ΔH det = -0.380 kcal/g) compare well with commercial explosives and other nitrogen-rich HE-MOFs. The velocity and pressure of denotation are 5.96 km/s and 9.56 GPa, respectively. Differential scanning calorimetry analysis shows that the denotation of [Zn(C6H4N5)N3] n occurs via a complex temperature-dependent mechanism.


Tourism as a driver of conflicts and changes in fisheries value chains in Marine Protected Areas.
Although critical tools for protecting ocean habitats, Marine Protected Areas (MPAs) are sometimes challenged for social impacts and conflicts they may generate. Some conflicts have an economic base, which, once understood, can be used to resolve associated socioenvironmental problems. We addressed how the fish trade in an MPA that combines no-take zones and tourist or resident zones creates incentives for increased fisheries. We performed a value chain analysis following the fish supply and trade through interviews that assessed consumer demand and preference. The results showed a simple and closed value chain driven by tourism (70% of the consumption). Both tourists and local consumers preferred high trophic level species (predators), but the former preferred large pelagics (tuna and dolphinfish) and the latter preferred reef species (barracuda and snapper). Pelagic predators are caught with fresh sardines, which are sometimes located only in the no-take zone. Pelagic species are mainly served as fillet, and the leftover fish parts end up as waste, an issue that, if properly addressed, can help reduce fishing pressure. Whereas some of the target species may be sustainable (e.g., dolphinfish), others are more vulnerable (e.g., wahoo) and should not be intensively fished. We advise setting stricter limits to the number of tourists visiting MPAs, according to their own capacity and peculiarities, in order to avoid conflicts with conservations goals through incentives for increased resource use.


Methionine restriction alleviates high-fat diet-induced obesity: Involvement of diurnal metabolism of lipids and bile acids.
Circadian misalignment induced by a high-fat diet (HFD) increases the risk of metabolic diseases. Methionine restriction (MR) is known to have the potential of alleviating obesity by improving insulin sensitivity. However, the role of the circadian clock in mediating the effects of MR on obesity-related metabolic disorders remains unclear. Ten-week-old male C57BL/6 J mice were fed with a low-fat diet (LFD) or a HFD for 4 wk., followed with a full diet (0.86% methionine, w/w) or a methionine-restricted diet (0.17% methionine, w/w) for 8 wk. Our results showed that MR attenuated insulin resistance triggered by HFD, especially at ZT12. Moreover, MR led to a time-specific enhancement of the expression of FGF21 and activated the AMPK/PGC-1α signaling. Notably, MR upregulated the cyclical levels of cholic acid (CA) and chenodeoxycholic acid (CDCA), and downregulated the cyclical level of deoxycholic acid (DCA) in the dark phase. MR restored the HFD-disrupted cyclical fluctuations of lipidolysis genes and BAs synthetic genes and improved the circulating lipid profile. Also, MR improved the expressions of clock-controlled genes (CCGs) in the liver and the brown adipose tissue throughout one day. In conclusion, MR exhibited the lipid-lowering effects on HFD-induced obesity and restored the diurnal metabolism of lipids and BAs, which could be partly explained by improving the expression of CCGs. These findings suggested that MR could be a potential nutritional intervention for attenuating obesity-induced metabolic misalignment.


Solution structure of the inhibitor of cysteine proteases 1 from Entamoeba histolytica reveals a possible auto regulatory mechanism.
The genome of Entamoeba histolytica encodes approximately 50 Cysteine Proteases (CPs) whose activity is regulated by two Inhibitors of Cysteine Proteases (ICPs), EhICP1 and EhICP2. The main difference between both EhICPs is the acquisition of a 17 N-terminal targeting signal in EhICP2 and three exposed cysteine residues in EhICP1. The three exposed cysteines in EhICP1 potentiate the formation of cross-linking species that drive heterogeneity. Here we solved the NMR structure of EhICP1 using a mutant protein without accessible cysteines. Our structural data shows that EhICP1 adopts an immunoglobulin fold composed of seven β-strands, and three solvent exposed loops that resemble the structures of EhICP2 and chagasin. EhICP1 and EhICP2 are able to inhibit the archetypical cysteine protease papain by intercalating their BC loops into the protease active site independently of the character of the residue (serine or threonine) responsible to interact with the active site of papain. EhICP1 and EhICP2 present signals of functional divergence as they clustered in different clades. Two of the three exposed cysteines in EhICP1 are located at the DE loop that intercalates into the CP substrate-binding cleft. We propose that the solvent exposed cysteines of EhICP1 play a role in regulating its inhibitory activity and that in oxidative conditions, the cysteines of EhICP1 react to form intra and intermolecular disulfide bonds that render an inactive inhibitor. EhICP2 is not subject to redox regulation, as this inhibitor does not contain a single cysteine residue. This proposed redox regulation may be related to the differential cellular localization between EhICP1 and EhICP2.


Ocular involvement in facial nerve paralysis: risk factors for severe visual impairment and ocular surface exposure in 1870 patients.
To identify the risk factors associated with the development of ocular surface exposure and severe visual impairment (SVI) in patients with facial nerve palsy (FNP). Ocular data of all consecutive FNP patients (N = 1870) presenting to an eye-care network over the past 10 years were reviewed. Risk factors associated with SVI (best-corrected visual acuity <20/200) and ocular surface exposure at presentation were evaluated using multivariate analysis and odds ratios (OR). The prevalence of SVI was 15%, and 47% had ocular surface exposure at the first presentation. The presence of corneal scar (28% vs. 10.2%, p < .001; OR 3.05), corneal ulcer (12.9% vs. 2.3%, p < .001; OR 4.67), older age at presentation (p < .001; OR 1.02), >10 mm lagophthalmos (p < .001; OR 8.7), male sex, and duration of FNP (p = .021; OR 1) were independent risk factors for developing SVI. Of the 893 eyes with ocular surface exposure, 75 (3.9%) had a corneal ulcer, and 11 (0.6%) were perforated, with the rest having epithelial defects and punctate keratopathy. Patients with neoplastic etiology (15.9% vs. 5.7%; p < .001; OR 2.39), lagophthalmos (49% vs. 29.7%; p < .001; OR 2.25) and poor Bell's phenomenon (9.4% vs. 4.2%; p = .005; OR 1.8) had twofold risk for developing ocular surface exposure. Eyes with lagophthalmos, poor Bell's phenomenon, and FNP of neoplastic etiology have a higher risk of developing ocular surface exposure. Male sex, increasing age, longer duration of FNP, and >10 mm lagophthalmos are associated with SVI in FNP patients; hence, early and timely intervention is necessary.


Delivery of MiR335-5p-Pendant Tetrahedron DNA Nanostructures Using an Injectable Heparin Lithium Hydrogel for Challenging Bone Defects in Steroid-Associated Osteonecrosis.
Corticosteroids-induced Dickkopf-1 (DKK1) upregulation and Wnt signaling inhibition result in bone metabolism disorder and steroid-associated osteonecrosis (SAON). Implanting biomaterials to regulate the Wnt pathway is a promising method to repair challenging bone defects associated with SAON. Here, tetrahedral DNA nanostructures (TDNs) are fabricated as gene carriers to deliver MiR335-5p, which targets DKK1 translation. Heparin lithium hydrogel (Li-hep-gel) is synthesized to act as a lithium and MiR@TDNs delivery agent. Finally, the repair effects on challenging bone defect in SAON using a MiR@TDNs/Li-hep-gel composite are assessed in vivo. The results reveal that MiR@TDNs are absorbed by bone mesenchymal stem cells (BMSCs) and increase cell viability and reduce apoptosis. Moreover, MiR@TDNs promote alkaline phosphatase expression and calcium nodular deposition, decrease lipid droplet expression of BMSCs, and improve vascular endothelial growth factor secretion and vascular-like structure formation in vitro. After MiR@TDNs/Li-hep-gel is implanted into the SAON model, the internal bone defect of osteonecrosis is repaired with a large area of new bone accompanied with neovascularization and reduced empty lacunae. In conclusion, MiR@TDNs/Li-hep-gel can provide dual delivery of lithium and MiR@TDNs, which synergistically upregulate the Wnt signaling pathway, enhancing bone regeneration in challenging bone defects, and can be potentially used in SAON repair.


Mission thrombectomy 2020 (MT2020+) surveys of Caribbean stroke services: A call for action in our region.
Stroke is the leading cause of death and adult-onset disability in the Caribbean region. Despite the tremendous advances in acute stroke care in the past 25 years, treatment in the Caribbean lags stroke standards of care. Mission Thrombectomy 2020+ (MT2020+) is a metrics based global campaign focused on increasing access to organized stroke care and mechanical thrombectomy. We conducted two consecutive online surveys across the MT2020+ Caribbean Region assessing access to thrombolytics, mechanical thrombectomy and stroke centers. The first survey - Mechanical Thrombectomy Access-Caribbean sub-study, allowed a comparison between global services and those in the MT2020+ Caribbean region. This survey provided a snapshot of the current state of stroke centers worldwide and confirmed lower available stroke resources in the MT2020+ Caribbean Region which has one certified stroke center, and 9 mechanical thrombectomy capable centers. The second survey - MT2020+ Caribbean Region infrastructure survey - was designed to determine the availability of stroke resources that are key components of stroke care in participating hospitals. Key infrastructural components such as 24/7 computerized tomographic scanners, access to thrombolytic therapy and access to mechanical thrombectomy capable centers were scarce. There were low volumes of mechanical thrombectomy performed in the Caribbean compared to other countries around the world. Limited resources and inadequate stroke infrastructure remains a major challenge in the Caribbean. We advocate for governmental support and investment, public and private partnerships and legislation to increase access and availability to acute stroke treatments to allow for equal access to care for all Caribbean citizens.


Phenylalanyl-tRNA synthetases as an example for comparative and evolutionary aspects of aminoacyl-tRNA synthetases.
Aminoacyl-tRNA synthetases are indispensable components of protein synthesis in all three lines of evolutionary descent, eubacteria, archaebacteria and eukaryotes. Furthermore they are also present in the translational apparatus of the semi-autonomous organelles, mitochondria and chloroplasts, of the eukaryotic cell. Therefore aminoacyl-tRNA synthetases are appropriate objects for comparative molecular biology in order to obtain a comprehensive picture of the evolution of the translational process. The analysis of the phenylalanyl-tRNA synthetase in a large variety of organisms and organelles in this respect is the most advanced. In addition to comparison of quaternary structure, analysis includes functional aspects of accuracy mechanisms (proofreading) and comparison of structural features by means of substrate analogs. Evolutionary relationships are furthermore elucidated using the immunological approach and heterologous aminoacylation.


Reaching under-screened/never-screened indigenous peoples with human papilloma virus self-testing: A community-based cluster randomised controlled trial.
Indigenous women in the high-income countries of Canada, Australia, New Zealand and USA, have a higher incidence and mortality from cervical cancer than non-Indigenous women. Increasing cervical screening coverage could ultimately decrease cervical cancer disparities. To increase cervical screening for under-screened/never-screened Māori women. This study was a cluster randomised controlled trial. Inclusion criteria were women aged 25-69, last screened ≥4 years ago, in Northland, New Zealand. The intervention arm was the offer of a human papilloma virus (HPV) self-test and the control arm was the usual offer of standard care - a cervical smear. The primary outcome was rate of cervical screening in the intervention group compared to control in Māori, the Indigenous peoples of New Zealand. Six primary care clinics were randomly allocated to intervention or control. Of 500 eligible Māori women in the intervention arm, 295 (59.0%) were screened. Of 431 eligible Māori women in the control arm, 94 (21.8%) were screened. Adjusting for age, time since last screen, deprivation index, Māori women in the intervention arm were 2.8 times more likely to be screened than women in the control arm (95% CI: 2.4-3.1, P-value <0.0001). Offer of HPV self-testing could potentially halve the number of under-screened/never-screened Māori women and decrease cervical morbidity and mortality. These results may be generalisable to benefit Indigenous peoples facing similar barriers in other high-income countries.


Electric-powered indoor/outdoor wheelchairs (EPIOCs): users' views of influence on family, friends and carers.
This study explored the effects of electric-powered indoor/outdoor wheelchair (EPIOC) provision to users on their family and carers. EPIOC users receiving their chairs between February and November 2002 (N = 74) were invited to participate in a telephone questionnaire/interview, and 64 agreed. This study examined the responses to the question 'Has the use of your EPIOC affected your family or friends in any way?' and related comments. Interviews were analysed using a qualitative framework approach to identify emergent themes. In addition, the proportion of individuals raising issues related to each theme was determined. Participants were interviewed 10-19 (mean = 14.3) months after chair delivery. The following themes emerged: reduced physical burden on family/friends and increased independence and freedom. However, an EPIOC does not eliminate other practical problems particularly during transportation and negotiating kerbs and slopes. Users also reported anxiety/worry in relationship to EPIOC use, e.g., weather conditions, personal safety (muggings), use of ramps and kerbs. There are considerable benefits to families and carers associated with powered wheelchair use. A reduction in the physical demand for pushing and increased freedom were identified. These benefits appear to outweigh the residual practical difficulties and worries.


Maturation of thyroidal dendritic cells in Graves' disease.
Because thyroidal dendritic cells (t-DC) may be implicated in the pathogenesis of Graves' disease (GD), we compared t-DC in thyroid sections of patients with GD (n = 15) and control patients with toxic (TG; n = 12) or non-toxic goitre (NG; n = 12). Goitres in GD, but not TG or NG, were populated with three discernible t-DC phenotypes. (i) Immature t-DC (major histocompatibility complex (MHC) II+/CD40-/CD80-) were located perifollicularly (95% of the patients with GD, but only 55% of TG and 51% of NG patients); numbers of such t-DC were significantly elevated in GD (P < 0.001). (ii) Partially matured CD80+ t-DC were present in connective tissue (73% of the patients) and focal interstitial clusters (40% of the patients). In 53% of the patients with GD, single as well as clustered interstitial t-DC expressed CD40. (iii) However, phenotypically mature t-DC (MHC II+/CD40+/CD80+/RFD1+) were only present in clusters and colocalized with activated CD4+/MHC class II+ T-helper (Th) cells. Expression of CD54 and CD83 did not significantly differ among the groups. The phenotype of intrathyroidal DC in GD thus supports their role as potential (co)stimulators of thyroid autoimmunity.


Predictive value of eosinophils and neutrophils on clinical effects of ICS in COPD.
Inflammation is present to a variable degree and composition in patients with COPD. This study investigates associations between both eosinophils and neutrophils in blood, sputum, airway wall biopsies and bronchoalveolar lavage (BAL) and their potential use as biomarkers for clinical response to inhaled corticosteroids (ICS). In total, 114 steroid-naïve COPD patients of the Groningen Leiden Universities Corticosteroids in Obstructive Lung Disease (GLUCOLD) study using ICS or placebo during 30-month follow-up were included. Cell counts in blood, sputum, biopsies and BAL were evaluated at baseline. In addition, at baseline, 6 and 30 months, forced expiratory flow in 1 s (FEV1 ), residual volume/total lung capacity (hyperinflation) and Clinical COPD Questionnaire were evaluated. Cross-sectional analyses at baseline showed that higher blood eosinophils were significantly associated with higher eosinophil counts in sputum, biopsies and BAL. However, blood neutrophils did not significantly correlate with neutrophil counts in the other compartments. Baseline eosinophils and neutrophils, in whichever compartment measured, did not predict longitudinal FEV1 changes. Higher baseline biopsy eosinophils were associated with an increase in symptoms during 6- and 30-month ICS treatment. In addition, higher biopsy neutrophils were associated with a more marked reduction in hyperinflation during 6-month ICS treatment compared with placebo. Our findings indicate that blood eosinophils reflect eosinophils in other compartments, in contrast to neutrophils, in ICS-naïve COPD patients. Both baseline eosinophils and neutrophils do not predict ICS-induced lung function changes over a period of 6-30 months. The associations of biopsy eosinophils with worsening respiratory symptoms and biopsy neutrophils with improvement in hyperinflation during ICS treatment deserve further investigation.


The interaction of respiration and visual feedback on the control of force and neural activation of the agonist muscle.
The purpose of this study was to compare force variability and the neural activation of the agonist muscle during constant isometric contractions at different force levels when the amplitude of respiration and visual feedback were varied. Twenty young adults (20-32 years, 10 men and 10 women) were instructed to accurately match a target force at 15% and 50% of their maximal voluntary contraction (MVC) with abduction of the index finger while controlling their respiration at different amplitudes (85%, 100% and 125% normal) in the presence and absence of visual feedback. Each trial lasted 22s and visual feedback was removed from 8-12 and 16-20s. Each subject performed three trials with each respiratory condition at each force level. Force variability was quantified as the standard deviation of the detrended force data. The neural activation of the first dorsal interosseus (FDI) was measured with bipolar surface electrodes placed distal to the innervation zone. Relative to normal respiration, force variability increased significantly only during high-amplitude respiration (∼63%). The increase in force variability from normal- to high-amplitude respiration was strongly associated with amplified force oscillations from 0 to 3 Hz (R(2) ranged from .68 to .84, p< .001). Furthermore, the increase in force variability was exacerbated in the presence of visual feedback at 50% MVC (vision vs. no-vision: .97 vs. .87N) and was strongly associated with amplified force oscillations from 0 to 1 Hz (R(2)= .82) and weakly associated with greater power from 12 to 30 Hz (R(2)= .24) in the EMG of the agonist muscle. Our findings demonstrate that high-amplitude respiration and visual feedback of force interact and amplify force variability in young adults during moderate levels of effort.


Effects of ClO2 on the absorption and distribution of dietary iodide in the rat.
Aqueous chlorine dioxide (ClO2), an alternative disinfectant for drinking water, was found to decrease gastrointestinal (GI) bioavailability of dietary iodide. It has been previously reported that subchronic exposure to ClO2 decreases thyroxine (T4) levels in nonhuman primates. In this study in vitro experiments with animal feed, isolated rat stomachs, as well as in vivo studies with intact rats, showed that ClO2 in drinking water (at in situ concentrations as low as 2 ppm) oxidizes iodide to its reactive elemental (radical) state, binding it to organic substances present in the GI tract. A single instance of acute exposure to ClO2, however, did not decrease blood iodide levels, or thyroid glandular uptake of iodine.


Profiles of eicosanoid production by superficial and proliferative colonic epithelial cells and sub-epithelial colonic tissue.
The profile of cyclooxygenase and lipoxygenase products in normal rat colonic epithelium and subepithelium was examined. Colons were thoroughly perfused to eliminate contamination with blood. Two preparations of colonic epithelium were employed. The first consisted of intact colonic crypts and epithelial sheets. The second yielded single cell suspensions of superficial versus proliferative epithelial cells. Lipoxygenase product formation by colonic epithelium as measured by hydroxyeicosatetraenoic acid (HETE) and leukotriene B4 (LTB4) production (5-HETE greater than 12-HETE greater than 15-HETE greater than LTB4) accounted for 58% of the total colonic production of these moieties, whereas epithelium accounted for only 20% of total colonic protein. By contrast, prostaglandin (PG) E2 and PGF2 alpha production occurred predominantly (greater than 97%) in the subepithelial layers. The present studies also demonstrate markedly higher levels of accumulation of lipoxygenase products in proliferative versus superficial epithelial cells, whereas prostaglandin accumulation was greater in superficial cells. Previous studies have supported a role for lipoxygenase and cyclooxygenase products in the control of colonic secretion, inflammatory cell infiltration and proliferative activity. The present results raise the possibility that the striking differences in the sites of production of these products within the colon has functional implications.


Tasting and smelling as active, exploratory sensory processes.
Tasting and smelling in terrestrial vertebrates usually involve intermittent, active sampling of the chemosensory environment. This active sampling is produced by ingestive and/or respiratory behavior initiated by the central nervous system. Vascular and secretory factors modulate access to chemosensitive regions. For olfaction, per se, respiratory rhythms provide the basic pattern of intermittent odorant input. In humans and many other species, sniffing alters the respiratory pattern and moves relatively brief, higher-flow-rate samples toward the olfactory epithelium. Sniffing may be triggered by nonchemosensory events or by odorants obtained during normal respiration and can be stereotyped with regard to rate and duration of the sniffs. Within the central nervous system, processing of olfactory input can differ as a function of its temporal relationship to respiratory and sniffing rhythms. For gustation, ingestion of food and liquids provides the major source of stimulation. Since these ingestive acts are intermittent, although often repetitive and stereotyped, natural taste stimulation also is necessarily both patterned and distributed in time. Swallowing and the interingestion interval that follows tend to return the taste receptors toward a salivary baseline. During the drinking of liquid by humans, retention of a bolus in the mouth is much briefer than intersip intervals. Simulated drinking with alternating flows of artificial saliva and a sufficiently different stimulus yields constant taste intensity. Conversely, unchanging gustatory stimulation produces reductions in intensity.


Chronic migraine headache and multiple dental pathologies causing cranial pain for 35 years: the neurodental nexus.
We report a case that illustrates how chronic migraine headaches and multiple dental pathologies caused severe and long-standing cranial pain that affected the quality of life of a man for more than 35 years. His case was investigated at several settings including the neurology outpatient clinic of the hospital without a definitive diagnosis or resolution. After investigations, multiple oral pathologies including two occult dental abscesses were diagnosed. Once both affected teeth and associated abscesses were surgically removed, with subsequent antibiotic therapy the headaches resolved.


Koyanagi urethroplasty for proximal hypospadias: A stage procedure?
To report experience and technical refinements with the Koyanagi urethroplasty for proximal hypospadias. A retrospective study was conducted between 2004 and 2017. Medical records of patients treated by the Koyanagi technique were investigated. The penile ventral skin was closed either with a Byars flap or using an "Ombredanne's chasuble" (OC). The cohort was divided chronologically into two groups of the same number of patients (early -E- and late experience group-LEG-). Demographics, anatomical findings and surgical outcomes were compared between groups using univariate analysis. A logistic regression was performed to assess factors associated with the occurrence of a postoperative urethrocutaneous fistula or urethroplasty dehiscence (UD). The Koyanagi urethroplasty was performed in 67 patients, 33 in the EEG and 34 in the LEG. The overall surgical complication rate was 64.2% (n=43), including 42 fistula or UD, higher in the EEG (81.8%) than in the LEG (44.1%, P<0.01). However, in the LEG, patients underwent the surgery older and the use of OC more frequent. After multivariate analyses, factors associated with a postoperative fistula or UD were the year of surgery (OR=0.71 [0.53-0.96] P=0.02), the age at surgery (OR=1.11 [1.01-1.22], P=0.03); contrary to the skin coverage method or the stenting duration (P>0.05). An urethral stenosis occurred in 1 patient (1.5%). At last follow-up, 64.2% of patients required a further procedure and 80.6% of urethral meatus were glandular. In this study the complication rate, particularly the urethrocutaneous fistula, remained high. Urethral stenosis, were rare but the follow-up was too short to clearly identified them. Considering as a 2 stage procedure koyanagi uretroplasty allows to obtain finally good results. III.


13C-CP/MAS NMR studies of the cyclomalto-oligosaccharide (cyclodextrin) hydrates.
The 13C-CP/MAS NMR spectra of cyclomaltohexaose (alpha-cyclodextrin) hexahydrate, cyclomaltoheptaose (beta-cyclodextrin) "undecahydrate", cyclomalto-octaose (gamma-cyclodextrin) "octadecahydrate", and of the same materials at lower levels of hydration are compared with solution NMR data, structures obtained from single crystal diffraction studies, and with previous reports of the 13C-CP/MAS NMR spectra. The chemical shifts of the C-1 and C-4 resonances can be correlated with the conformation about the (1----4) linkage. The chemical shifts of the C-6 resonances are also sensitive to hydrogen-bonding interactions, as shown by the spectral changes on loss of water from the structures. The results suggest that, for resonances of carbon atoms close to a centre of significant conformational change, chemical shifts may be predicted on the basis of conformation alone, but for the resonances of more distant atoms, changes in chemical shift due to conformational change may be masked by the effects of alterations in the local environment.


Maximizing the outcome of early ADMET models: strategies to win the drug-hunting battles?
Despite substantial changes in the drug discovery paradigm leveraged from the advancement of early ADMET technologies, an open debate remains on how full the return on investment is, along with where to balance risks to costs of lost opportunities in the clinic. Here, the recent advancement of ADMET tools, the areas where they seem to work and where their application and connection with physiology in man remain challenging are briefly reviewed. While the 'more is better' type of 'box-checking' profiling strategy is no longer viable, the key to success lies in an intelligent integration of existing in silico, in vitro and in vivo ADMET data to help generate and test hypotheses that are critical for projecting the benefits and risks of a drug candidate in the clinic. The improvement of in silico, in vitro and in vivo correlations (ISIVIVC) and best utilization of early ADMET data are far more critical and urgent than expanding capacity and portfolio in leveraging ADMET to win the drug-hunting battles in the post-genome era.


Studies on the pseudo-exfoliation material: a review.
Pseudo-exfoliation (PE) material may be found on several surfaces of the anterior segment of the eye. The material contains typical cross-banded PE fibrils distributed at random. These fibrils have been studied with transmission and scanning electron microscopy. Negative staining of dispersed PE fibrils indicated that the fibrils have a core consisting of a small number of filamentous subunits. The core is surrounded by a fuzzy material which protrudes from the fibrils at regular intervals. This is the basis of the cross-bands of the fibrils. Staining with ruthenium red and alcian blue indicates that glycosaminoglycans are present, mainly on the surface of the PE fibrils. Tracer particles have been applied in vitro on lenses with PE. The distribution of the tracers in the PE material indicates that the fibrils are embedded in an interfibrillar matrix consisting of filamentous units. It is concluded that the PE material is made up of filamentous proteo-glycosaminoglycans. The fibrils represent relatively dense aggregates of such filaments, while the interfibrillar matrix is a gel of the same filamentous units in a loose, random arrangement.


Immunogenicities of haemocyanins from the giant African snail (Achatina fulica) and the keyhole limpet (Megathura crenulata).
1. The immunogenicities of the giant African snail (Achatina fulica) haemocyanin (AFH) and the keyhole limpet (Megathura crenulata) haemocyanin (KLH) were compared by determining their capacities to induce humoral and cell-mediated immune response in rats. 2. KLH was found to be more immunogenic than AFH, and this was attributed to the fact that KLH contains more moieties, and probably more antigenic determinants than AFH. 3. Since AFH was found capable of stimulating both humoral and cell-mediated immunity, it was suggested that it could be used as an antigen for the investigation of immune responses.


Aberrantly expressed Wnt5a in nurse-like cells drives resistance to Venetoclax in chronic lymphocytic leukemia.
Chronic lymphocytic leukemia (CLL) is characterized by the accumulation of neoplastic B lymphocytes with high levels of Wnt5a in the plasma. Currently, the cell source of Wnt5a remains controversial. The receptor of Wnt5a is ROR1, whose expression is associated with disease progression and resistance to venetoclax, a BCL-2 inhibitor approved for the treatment of CLL. In this study, we found that the levels of Wnt5a in the plasma of CLL patients were positively correlated with absolute monocyte counts, but not lymphocyte counts. We cultured monocyte-derived nurse-like cells (NLCs) from patients with CLL, and detected Wnt5a expressed in NLCs. Flow cytometry and transwell assays showed that the antibody neutralizing Wnt5a inhibited the enhanced survival and migration in CLL cells co-cultured with NLCs. Furthermore, we performed a drug screening with CLL cells cultured with or without NLCs with a library containing 133 FDA-approved oncology drugs by using high-throughput flow cytometry. We observed a significant resistance to venetoclax in CLL cells co-cultured with NLCs. Immunoblot revealed the activation of NF-κB with enhanced expression of MCL-1 and BCL-XL in CLL cells co-cultured with NLCs. Neutralizing Wnt5a or blocking NF-κB pathway significantly decreased the expression of MCL-1 and BCL-XL, which leads to enhanced sensitivity to venetoclax in CLL cells co-cultured with NLCs. In conclusion, our data showed that NLCs could be one of the sources of Wnt5a detected in patients with CLL, and Wnt5a-induced NF-κB activation in the CLL microenvironment results in resistance to venetoclax in CLL cells.


Capgras syndrome: a novel probe for understanding the neural representation of the identity and familiarity of persons.
Patients with Capgras syndrome regard people whom they know well such as their parents or siblings as imposters. Here we describe a case (DS) of this syndrome who presents several novel features. DS was unusual in that his delusion was modality-specific: he claimed that his parents were imposters when he was looking at them but not when speaking to them on the telephone. Unlike normals, DS's skin conductance responses to photographs of familiar people, including his parents, were not larger in magnitude than his responses to photographs of unfamiliar people. We suggest that in this patient connections from face-processing areas in the temporal lobe to the limbic system have been damaged, a loss which may explain why he calls his parents imposters. In addition, DS was very poor at judging gaze direction. Finally, when presented with a sequence of photographs of the same model's face looking in different directions, DS asserted that they were "different women who looked just like each other'. In the absence of limbic activation, DS creates separate memory "files' of the same person, apparently because he is unable to extract and link the common denominator of successive episodic memories. Thus, far from being a medical curiosity. Capgras syndrome may help us to explore the formation of new memories caught in flagrante delicto.


Enzymatic repair of UV-irradiated DNA in vitro.
Excision repair of UV-damaged Bacillus subtilis transforming DNA has been carried out by a sequential enzyme system in vitro. Incision adjacent to the pyrimidine dimer in the DNA strand by correndonuclease II-initiated excision of the damage by the 5' in equilibrium 3'-directed exonuclease of the Micrococcus luteus DNA polymerase. Reinsertion of nucleotides into the gap in the strand by the DNA polymerase at 10 degrees C terminated in a single-strand break which was sealed by a polynucleotide ligase, thereby repairing the DNA strand. This restored biological activity to damaged DNA up to doses resulting in 60% inactivation of transforming activity. At higher doses, less repair was achieved, due to the development of double-strand breaks during the in vitro incision and excision steps.


Differential effects of potassium on the potency of different opioids in the mouse vas deferens.
The effect of potassium on the potency of different opioids to inhibit electrically evoked twitches of the mouse vas deferens has been studied. In general, opioids lose activity as the concentration of potassium increases. Distinct differences, however, were observed between specific opiate receptor agonists. Least sensitive to the effect of potassium proved mu- and delta-opiate receptor agonists, showing only a 2-fold increase in the concentration required to cause a half-maximal inhibition at 14 mM K+. In contrast, the supposed dynorphin receptor/kappa-receptor agonists exhibited at this concentration up to an 8-fold loss of potency. Dynorphin1-13 was found to be intermediate with a 5-fold loss of inhibitory activity. These data suggest a modulatory action of potassium for dynorphin- and kappa-receptors.


Suppression of MD2 inhibits breast cancer in vitro and in vivo.
To explore the effects of the intervening measure targeting myeloid differentiation 2 (MD2) on breast cancer progression in vitro and in vivo. The expression of MD2 in normal breast cells (Hs 578Bst) and three kinds of breast carcinoma cell lines (MCF-7, MDA-MB-231 s and 4T1) were detected by western blot. MTT assay was used to detect the proliferation of 4T1 cells treated by L6H21, cell migration and invasion was measured by wound healing assay and trans-well matrigel invasion assay, respectively. In addition, to further study the role of MD2 in tumor progression, we assessed the effects of inhibition of MD2 on the progression of xenograft tumors in vivo. The expression of MD2 is much higher in MDA-MB-231 s and 4T1cells than that in normal breast cells (Hs 578Bst) or MCF-7 cells (p < 0.05). In vitro, suppression of MD2 by L6H21 has a significant inhibition of proliferation, migration and invasion in 4T1 cells in dose-dependent manner. In vivo, L6H21 pretreatment significantly improved survival of 4T1-bearing mice (p < 0.05). Additionally, we also observed that none of the mice died from the toxic effect of 10 mg kg-1 L6H21 in 60 days. Overall, this work indicates that suppression of MD2 shows progression inhibition in vitro and significantly prolong survival in vivo. These findings provide the potential experimental evidence for using MD2 as a therapeutic target of breast carcinoma.


Malaria therapeutics: are we close enough?
Malaria is a vector-borne parasitic disease caused by the apicomplexan protozoan parasite Plasmodium. Malaria is a significant health problem and the leading cause of socioeconomic losses in developing countries. WHO approved several antimalarials in the last 2 decades, but the growing resistance against the available drugs has worsened the scenario. Drug resistance and diversity among Plasmodium strains hinder the path of eradicating malaria leading to the use of new technologies and strategies to develop effective vaccines and drugs. A timely and accurate diagnosis is crucial for any disease, including malaria. The available diagnostic methods for malaria include microscopy, RDT, PCR, and non-invasive diagnosis. Recently, there have been several developments in detecting malaria, with improvements leading to achieving an accurate, quick, cost-effective, and non-invasive diagnostic tool for malaria. Several vaccine candidates with new methods and antigens are under investigation and moving forward to be considered for clinical trials. This article concisely reviews basic malaria biology, the parasite's life cycle, approved drugs, vaccine candidates, and available diagnostic approaches. It emphasizes new avenues of therapeutics for malaria.


Expression profiles of a banana fruit linker histone H1 gene MaHIS1 and its interaction with a WRKY transcription factor.
Chromatin remodeling-related proteins, such as linker histone H1, involving in fruit ripening and stress responses are poorly understood. In the present study, a novel cDNA encoding linker histone H1 gene, designated as MaHIS1 was isolated and characterized from banana fruit. The full-length cDNA sequence was 1,253 bp with an open-reading frame (ORF) of 948 bp, encoding 315 amino acids with a molecular weight of 31.98 kDa and a theoretical isoelectric point of 10.67. Subcellular localization analysis showed that MaHIS1 was a nucleus-localized protein. Real-time PCR analysis indicated that expression of MaHIS1 gene is induced by external and internal ethylene during fruit postharvest ripening. Accumulation of MaHIS1 transcript was also obviously enhanced by exogenous hormones, including methyl jasmonate, abscisic acid, and hydrogen peroxide (H₂O₂), as well as stresses, such as chilling and pathogen Colletotrichum musae infection. Moreover, yeast two-hybrid and bimolecular fluorescence complementation assays showed that MaHIS1 could interact with a transcription factor (TF) MaWRKY1. Taken together, our results suggest that MaHIS1 may be related to ripening and stress responses of banana fruit, and be likely functionally coordinating with MaWRKY1 in these physiological processes. MaHIS1 may be related to ripening and stress responses of banana fruit, and it also could interact with WRKY TF, which expands the very limited information regarding the functions of linker histone H1 in fruits.


[Optic neuritis in juvenile idiopathic arthritis patient].
Optic neuritis (ON) was rarely reported in juvenile idiopathic arthritis (JIA) patients, particularly in those under anti-tumor necrosis factor alpha blockage. However, to our knowledge, the prevalence of ON in JIA population has not been studied. Therefore, 5,793 patients were followed up at our University Hospital and 630 (11%) had JIA. One patient (0.15%) had ON and was reported herein. A 6-year-old male was diagnosed with extended oligoarticular JIA, and received naproxen and methotrexate subsequently replaced by leflunomide. At 11 years old, he was diagnosed with aseptic meningitis, followed by a partial motor seizure with secondary generalization. Brain magnetic resonance imaging (MRI) and electroencephalogram showed diffuse disorganization of the brain electric activity and leflunomide was suspended. Seven days later, the patient presented acute ocular pain, loss of acuity for color, blurred vision, photophobia, redness and short progressive visual loss in the right eye. A fundoscopic exam detected unilateral papilledema without retinal exudates. Orbital MRI suggested right ON. The anti-aquaporin 4 (anti-AQP4) antibody was negative. Pulse therapy with methylprednisolone was administered for five days, and subsequently with prednisone, he had clinical and laboratory improvement. In conclusion, a low prevalence of ON was observed in our JIA population. The absence of anti-AQP4 antibody and the normal brain MRI do not exclude the possibility of demyelinating disease associated with chronic arthritis. Therefore, rigorous follow up is required.


Reframing the Wilderness Concept can Bolster Collaborative Conservation.
Indigenous territories represent ~45% of land categorized as wilderness in the Amazon, but account for <15% of all forest loss on this land. At a time when the Amazon faces unprecedented pressures, overcoming polarization and aligning the goals of wilderness defenders and Indigenous peoples is paramount, to avoid environmental degradation.