Gender comparisons of mechanomyographic amplitude and mean power frequency versus isometric torque relationships.
This study compared the patterns of mechanomyographic (MMG) amplitude and mean power frequency vs. torque relationships in men and women during isometric muscle actions of the biceps brachii. Seven men (mean age 23.9 +/- 3.5 yrs) and 8 women (mean 21.0 +/- 1.3 yrs) performed submaximal to maximal isometric muscle actions of the dominant forearm flexors. Following determination of the isometric maximum voluntary contraction (MVC), they randomly performed submaximal step muscle actions in 10% increments from 10% to 90% MVC. Polynomial regression analyses indicated that the MMG amplitude vs. isometric torque relationship for the men was best fit with a cubic model (R(2) = 0.983),,where MMG amplitude increased slightly from 10% to 20% MVC, increased rapidly from 20% to 80% MVC, and plateaued from 80% to 100% MVC. For the women, MMG amplitude increased linearly (r(2) = 0.949) from 10% to 100% MVC. Linear models also provided the best fit for the MMG mean power frequency vs. isometric torque relationship in both the men (r(2) = 0.813) and women (r(2) = 0.578). The results demonstrated gender differences in the MMG amplitude vs. isometric torque relationship, but similar torque-related patterns for MMG mean power frequency. These findings suggested that the plateau in MMG amplitude at high levels of isometric torque production for the biceps brachii in the men, but not the women, may have been due to greater isometric torque, muscle stiffness, and/or intramuscular fluid pressure in the men, rather than to differences in motor unit activation strategies for modulating isometric torque production.


Impact of Solution Chemistry on Growth and Structural Features of Mo-Substituted Spinel Iron Oxides.
The effect of crystallizing solution chemistry on the chemistry of subsequently as-grown materials was investigated for Mo-substituted iron oxides prepared by thermally activated co-precipitation. In the presence of Mo ions, we find that varying the oxidation state of the iron precursor from Fe(II) to Fe(III) causes a progressive loss of atomic long-range order with the stabilization of 2-4 nm particles for the sample prepared with Fe(III). The oxidation state of the Fe precursor also affects the distribution of Fe and Mo cations within the spinel structure. Increasing the Fe precursor oxidation state gives decreased Fe-ion occupation and increased Mo-ion occupation of tetrahedral sites, as revealed by the extended X-ray absorption fine structure. The stabilization of Mo within tetrahedral sites appears to be unexpected, considering the octahedral preferred coordination number of Mo(VI). The analysis of the atomic structure of the sample prepared with Fe(III) indicates a local ordering of vacancies and that the occupation of tetrahedral sites by Mo induces a contraction of the interatomic distances within the polyhedra as compared to Fe atoms. Moreover, the occupancy of Mo into the thermodynamic site preference of a Mo dopant in Fe2O3 assessed by density functional theory calculations points to a stronger preference for Mo substitution at octahedral sites. Hence, we suggest that the synthetized compound is thermodynamically metastable, that is, kinetically trapped. Such a state is suggested to be a consequence of the tetrahedral site occupation by Mo ions. The population of these sites, known to be reactive sites enabling particle growth, is concomitant with the stabilization of very small particles. We confirmed our hypothesis by using a blank experiment without Mo ions, further supporting the impact of tetrahedral Mo ions on the growth of iron oxide nanoparticles. Our findings provide new insights into the relationships between the Fe-chemistry of the crystallizing solution and the structural features of the as-grown Mo-substituted Fe-oxide materials.


Genome-wide analysis of the sox family in the calcareous sponge Sycon ciliatum: multiple genes with unique expression patterns.
Sox genes are HMG-domain containing transcription factors with important roles in developmental processes in animals; many of them appear to have conserved functions among eumetazoans. Demosponges have fewer Sox genes than eumetazoans, but their roles remain unclear. The aim of this study is to gain insight into the early evolutionary history of the Sox gene family by identification and expression analysis of Sox genes in the calcareous sponge Sycon ciliatum. Calcaronean Sox related sequences were retrieved by searching recently generated genomic and transcriptome sequence resources and analyzed using variety of phylogenetic methods and identification of conserved motifs. Expression was studied by whole mount in situ hybridization. We have identified seven Sox genes and four Sox-related genes in the complete genome of Sycon ciliatum. Phylogenetic and conserved motif analyses showed that five of Sycon Sox genes represent groups B, C, E, and F present in cnidarians and bilaterians. Two additional genes are classified as Sox genes but cannot be assigned to specific subfamilies, and four genes are more similar to Sox genes than to other HMG-containing genes. Thus, the repertoire of Sox genes is larger in this representative of calcareous sponges than in the demosponge Amphimedon queenslandica. It remains unclear whether this is due to the expansion of the gene family in Sycon or a secondary reduction in the Amphimedon genome. In situ hybridization of Sycon Sox genes revealed a variety of expression patterns during embryogenesis and in specific cell types of adult sponges. In this study, we describe a large family of Sox genes in Sycon ciliatum with dynamic expression patterns, indicating that Sox genes are regulators in development and cell type determination in sponges, as observed in higher animals. The revealed differences between demosponge and calcisponge Sox genes repertoire highlight the need to utilize models representing different sponge lineages to describe sponge development, a prerequisite for deciphering evolution of metazoan developmental mechanisms.


Radiosensitization of Salmonella spp. and Listeria spp. in ready-to-eat baby spinach leaves.
The FDA recently approved irradiation treatment of leafy greens such as spinach up to 1 kGy; however, it is important to reduce the dose required to decontaminate the produce while maintaining its quality. Thus, the objectives of this study were: (1) to assess the radiation sensitivities of Salmonella spp. and Listeria spp. inoculated in ready-to-eat baby spinach leaves under modified atmosphere packaging (MAP) and irradiated using a 1.35-MeV Van de Graff accelerator (the leaves were irradiated both at room temperature and at -5 °C); and (2) to understand and optimize the synergistic effect of MAP and irradiation by studying the radiolysis of ozone formation under different temperatures, the effect of dose rate on its formation, and its decomposition. Results showed that increased concentrations of oxygen in the packaging significantly increased the radiation sensitivity of the test organisms, ranging from 7% up to 25% reduction in D(10)-values. In particular, radiosensitization could be effected (P < 0.05) by production of ozone, which increases with increasing dose-rate and oxygen concentration, and reducing temperatures. Radiosensitization was demonstrated for both microorganisms with irradiation of either fresh or frozen (-5 °C) baby spinach. These results suggest that low-dose (below 1 kGy) e-beam radiation under modified atmosphere packaging (100% O(2) and N(2):O(2)[1:1]) may be a viable tool for reducing microbial populations or eliminating Salmonella spp. and Listeria spp. from baby spinach. A suggested treatment to achieve a 5-log reduction of the test organisms would be irradiation at room temperature under 100% O(2) atmosphere at a dose level of 0.7 kGy. Practical Application: Decontamination of minimally processed fruits and vegetables from food-borne pathogens presents technical and economical challenges to the produce industry. Internalized microorganisms cannot be eliminated by the current procedure (water-washed or treated with 200-ppm chlorine). The only technology available commercially is ionizing radiation; however, the actual radiation dose required to inactivate pathogens is too high to be tolerated by the product without unwanted changes. This study shows a new approach in using MAP with 100% O(2), which is converted to ozone to radiosensitize pathogens while improving the shelf life of minimally processed fruits and vegetables. The process results in a high level of microorganism inactivation using lower doses than the conventional irradiation treatments.


Fiducial-aided calibration of a displacement laser probing system for in-situ measurement of optical freeform surfaces on an ultra-precision fly-cutting machine.
Surface metrology is an essential operation to determine whether the quality of manufactured surfaces meets the design requirements. In order to improve the surface accuracy and machining efficiency in the manufacturing of optical freeform surfaces, in-situ surface measurement without re-positioning the workpiece is considered as a promising technique in advanced manufacturing. In this study, a displacement laser scanner is integrated into an ultra-precision fly-cutting machine in order to perform as a coordinate measuring machine. However, some inevitable errors such as motion errors of the machine tool, thermal drift, vibrations, and errors of the laser sensor are introduced due to the manufacturing environment. To improve the performance of the measurement system, calibration of the main error sources is investigated with consideration of the characteristics of the built laser scanner system. Hence, the relationship between the moving speed of the laser scanner and the vibration of the tested signals is studied. Following that, the errors of the z-axis scale could be corrected by measuring a four-step heights artefact. Furthermore, volumetric positioning errors are identified by the proposed modified chi-square method and Gaussian processing prediction method. Simulation and measurement experiments are conducted, and the results indicate that the calibrated measuring system can measure ultra-precision freeform surfaces with micrometre form accuracy.


Chemotherapy-induced cellular senescence suppresses progression of Notch-driven T-ALL.
T cell acute lymphoblastic leukemia (T-ALL) is a serious hematologic malignancy that occurs in children and young adults. Current therapies include intensive chemotherapy and ionizing radiation that preferentially kill malignant cells. Unfortunately, they are frequently accompanied by unintended negative impacts, including the induction of cellular senescence and long-term toxicities in normal host tissues. Whether these senescent cells resulting from therapy increase the susceptibility to relapse or secondary cancers is unknown. Using transgenic and pharmacological approaches to eliminate doxorubicin-induced senescent cells in a Notch-driven T-ALL relapse mouse model, we find that these cells inhibit tumor recurrence, suggesting that senescence in response to treatment suppresses tumorigenesis. This finding, together with extensive evidence from others demonstrating that age-associated health problems develop dramatically earlier among childhood cancer survivors compared to age-matched counterparts, suggests a relationship between therapy-induced senescence and tumorigenesis. Although cancer risk is increased through accelerated premature-aging in the long run, therapy-induced senescence appears to protect survivors from recurrence, at least in the short run.


Mountain desert silicosis.
To evaluate the radiological miliary shadows in the high landers of a Himalayan desert for the presence of silicosis. Seventeen high landers attending the Medicine OPD of a multi-speciality medical camp at Kaza in Himalayas (height 12,500 feet above sea level) having radiological miliary shadows were included in the study. A detailed life time work place history was taken. In the laboratory workup their hemogram, repeat chest skiagram, peak expiratory flow rate, urinalysis and electrocardiogram were undertaken. Their localities were looked for the presence of industries particularly for silicosis prone work place. The silica contents of upper strata of soil were estimated. Silicosis--chronic simple variety was present in nine males and eight females. The youngest person was 43 years of age and oldest person was of 65 years of age. There was no silicosis prone industry in their locality. Four had taken repeated courses of anti-tubercular treatment in adequate doses for these miliary shadows without any change in the radiological shadows. Three had silicosis prone work history. Seven were smokers and two had hypertension. The miliary shadows were 2-4 mm in size and hilar lymphadenopathy was in 11 and fine calcification of lymph nodes in two. Peak expiratory flow rate was reduced. Upper strata of soil had a silica content of 36.8 percent. Silicosis developed in high landers of the Himalayas even without working in silicosis prone work place. They were exposed to silica from the non-work place silica rich environment.


[The Effects of Auditory Hallucination Simulation on Empathy, Knowledge, Social Distance, and Attitudes Toward Patients With Mental Illness Among Undergraduate Students: A Systemic Review and Meta-Analysis].
The negative attitudes of the general public toward mental illness frequently influence the integration of mental illness patients into the community. Auditory hallucination simulation may be considered as a creative teaching strategy to improve the attitudes of learners toward mental illness. However, the empirical effects of auditory hallucination simulation to change the negative attitudes toward mental illness remains uncertain. To compare and analyze, using a systematic review and meta-analysis, the effectiveness of auditory hallucination simulation in improving empathy, knowledge, social distance, and attitudes toward mental illness in undergraduates. A search using the keywords "auditory hallucination" and "simulation" and the 4 outcome indicators of empathy, knowledge, social distance, and attitudes toward mental illness was conducted to identify related articles published between 2008 and 2016 in 6 Chinese and English electronic databases, including Cochrane Library, EBSCO-CINAHL, MEDLINE, PsycINFO, PubMed, and Airiti Library. Research quality was appraised using the Modified Jadad Scale (MJS), the Oxford Centre for Evidence-Based Medicine Level of Evidence (OCEBM LoE), and the Cochrane Risk of Bias tool. Eleven studies were recruited, and 7 studies with sufficient data were included in the meta-analysis. The meta-analysis showed that hallucination simulation significantly improved the empathy and knowledge of participants, with respective effect sizes of 0.63 (95% CI [0.21, 1.05]) and 0.69 (95% CI [0.43-0.94]). However, this intervention also increased social distance, with an effect size of 0.60 (95% CI [0.01, 1.19]), and did not change attitudes toward mental illness significantly, with an effect size of 0.33 (95% CI [-0.11, 0.77]). Auditory hallucination simulation is an effective teaching strategy for improving the empathy and knowledge of undergraduates. However, related evidence for the effects of social distance and attitudes toward mental illness need to be further strengthened. Most of the extant research on this subject was conducted in the United States and Australia and was of moderate quality. Future studies should use sufficiently rigorous research designs to explore the safety issues and the effectiveness of the auditory hallucination simulation intervention in different countries and ethnic populations.


Expression of Plasmodium falciparum blood-stage antigens in Escherichia coli: detection with antibodies from immune humans.
Many proteins produced by blood stages of the malaria parasite Plasmodium falciparum are natural immunogens in man. As an approach to determining which of these are relevant to protective immunity we have constructed an expression library of P. falciparum cDNA sequences, cloned in Escherichia coli. The cDNA sequences were inserted into the beta-galactosidase gene of an ampicillin-resistant derivative of the temperature-sensitive lysogenic bacteriophage lambda gt11. About 5% of the resulting clones expressed P. falciparum sequences as polypeptides fused to beta-galactosidase. We have identified many clones that express P. falciparum antigens by immunological screening in situ with antibodies from immune human sera that inhibit P. falciparum growth in vitro. The antigen-positive clones contain P. falciparum cDNA sequences, as determined by hybridization. Some express polypeptides that are larger than beta-galactosidase and react both with antibodies to beta-galactosidase and with antibodies from humans immune to P. falciparum. The cloned P. falciparum antigens should facilitate new approaches to the identification of potential vaccine molecules.


Zebrafish (Danio rerio) as a model for studying the genetic basis of copper toxicity, deficiency, and metabolism.
Unicellular eukaryotes and cultured cells from several animal species were invaluable in discovering the mechanisms that govern incorporation, handling, and excretion of copper at the cellular level. However, understanding the systemic regulation of copper availability and distribution among the different tissues of an intact multicellular organism has proven to be more challenging. This analysis is made even more difficult if the genetic variability among organisms is taken into account. The zebrafish has long been considered a powerful animal model because of its tractable genetics and embryology, but it has more recently become a player in environmental studies, pharmaceutical screening, and physiologic analysis. In particular, the use of the larvae, small enough to fit into a microtiter well, but developed enough to have full organ functionality, represents a convenient alternative for studies that aim to establish effects of environmental agents on the intact, living organism. Studies by our group and others have characterized absorption and tissue distribution of copper and have described the acute effects of the metal on larvae in terms of survival, organ stress, and functionality of sensory organs. A large body of work has shown that there is strong conservation in mechanisms and genes between fish and mammals, opening the possibility for genetic or small molecule screens or for generating fish models of human diseases related to copper metabolism. The variability within humans in terms of tolerance to copper excess or deficiency requires a genetic approach to be taken to understand the behavior of populations, because markers and vulnerabilities need to be identified. The zebrafish could represent a unique tool to move in this direction.


Pediatric spinal injury patterns and management in all-terrain vehicle and dirt bike crashes, 2010-2019.
Pediatric spinal injuries in all-terrain vehicle (ATV) and dirt bike crashes are relatively uncommon but may be associated with significant morbidity. There are no recent studies examining these injuries, their management, and outcomes. Therefore, a retrospective study was performed to characterize pediatric spinal injuries related to ATV and dirt bike crashes over the last decade. Data on all patients involved in ATV or dirt bike crashes evaluated at a regional level 1 pediatric trauma center over a 10-year period (2010-2019) were analyzed. Descriptive statistics were analyzed and chi-square, Fisher exact, and Mann-Whitney U-tests were performed comparing the demographics, injury characteristics, and clinical outcomes in patients with versus those without spinal injuries. Of 680 patients evaluated, 35 (5.1%) were diagnosed with spinal injuries. Over the study period, both spinal injuries and emergency department visits related to ATV or dirt bike crashes increased in frequency. All spinal injuries were initially diagnosed on CT scans, and 57.9% underwent spinal MRI. Injuries were most commonly thoracic (50%), followed by cervical (36.8%). The injuries of most patients were classified as American Spinal Injury Association (ASIA) grade E on presentation (86.8%), while 2 (5.3%) had complete spinal cord injuries (ASIA grade A) and 3 patients (8.6%) were ASIA grade B-D. Operative management was required for 13 patients (28.9%). Nonoperative management was used in 71.1% of injuries, including bracing in 33% of all injuries. Patients with spinal injuries were older than those without (13.4 ± 3.35 vs 11.5 ± 3.79 years, p = 0.003). Spinal injuries occurred via similar crash mechanisms (p = 0.48) and in similar locations (p = 0.29) to nonspinal injuries. Patients with spinal injuries more frequently required admission to the intensive care unit (ICU; 34.2% vs 14.6%, p = 0.011) and had longer hospital stays (mean 4.7 ± 5.5 vs 2.7 ± 4.0 days, p = 0.0025). Although infrequent among young ATV and dirt bike riders, spinal injuries are associated with longer hospital stays, increased ICU use, and required operative intervention in 29%. Increasing awareness among ATV and dirt bike riders about the severity of riding-related injuries may encourage safer riding behaviors.


Baseline and stress-induced levels of corticosterone in male and female Afrotropical and European temperate stonechats during breeding.
Latitudinal variation in avian life histories falls along a slow-fast pace of life continuum: tropical species produce small clutches, but have a high survival probability, while in temperate species the opposite pattern is found. This study investigated whether differential investment into reproduction and survival of tropical and temperate species is paralleled by differences in the secretion of the vertebrate hormone corticosterone (CORT). Depending on circulating concentrations, CORT can both act as a metabolic (low to medium levels) and a stress hormone (high levels) and, thereby, influence reproductive decisions. Baseline and stress-induced CORT was measured across sequential stages of the breeding season in males and females of closely related taxa of stonechats (Saxicola spp) from a wide distribution area. We compared stonechats from 13 sites, representing Canary Islands, European temperate and East African tropical areas. Stonechats are highly seasonal breeders at all these sites, but vary between tropical and temperate regions with regard to reproductive investment and presumably also survival. In accordance with life-history theory, during parental stages, post-capture (baseline) CORT was overall lower in tropical than in temperate stonechats. However, during mating stages, tropical males had elevated post-capture (baseline) CORT concentrations, which did not differ from those of temperate males. Female and male mates of a pair showed correlated levels of post-capture CORT when sampled after simulated territorial intrusions. In contrast to the hypothesis that species with low reproduction and high annual survival should be more risk-sensitive, tropical stonechats had lower stress-induced CORT concentrations than temperate stonechats. We also found relatively high post-capture (baseline) and stress-induced CORT concentrations, in slow-paced Canary Islands stonechats. Our data support and refine the view that baseline CORT facilitates energetically demanding activities in males and females and reflects investment into reproduction. Low parental workload was associated with lower post-capture (baseline) CORT as expected for a slow pace of life in tropical species. On a finer resolution, however, this tropical-temperate contrast did not generally hold. Post-capture (baseline) CORT was higher during mating stages in particular in tropical males, possibly to support the energetic needs of mate-guarding. Counter to predictions based on life history theory, our data do not confirm the hypothesis that long-lived tropical populations have higher stress-induced CORT concentrations than short-lived temperate populations. Instead, in the predator-rich tropical environments of African stonechats, a dampened stress response during parental stages may increase survival probabilities of young. Overall our data further support an association between life history and baseline CORT, but challenge the role of stress-induced CORT as a mediator of tropical-temperate variation in life history.


Bacillus taxi sp. nov., a novel endophytic bacterium isolated from root of Taxus chinensis (Pilger) Rehd.
A Gram-staining-positive, aerobic, rod-shaped, endospore-forming bacterium, designated strain M5HDSG1-1T, was originally isolated from a surface-sterilized root of Taxus chinensis (Pilger) Rehd. in Guizhou, PR China. This bacterium was tested by a polyphasic approach to determine its taxonomic position. A 16S rRNA gene-based phylogenetic analysis revealed that M5HDSG1-1T had the greatest similarity to the type strain of Bacillus nealsonii DSM 15077T (99.1 %). The average nucleotide identity values between M5HDSG1-1T and Bacillus nealsonii DSM 15077T and Bacillus circulans NBRC 13626T were 73.3 and 72.8 %, respectively. The digital DNA-DNA hybridization values between M5HDSG1-1T and Bacillus nealsonii DSM 15077T and Bacillus circulans NBRC 13626T were 20.1 and 20.6 %, respectively, which were below the recommended thresholds. M5HDSG1-1T grew at a pH range of 6.0-12.0 (optimum, 7.0-8.0), at temperatures between 10 and 45 °C (optimum, 30 °C) and at 0-2 % (w/v) NaCl (optimum, 1 %). Neither substrate nor aerial mycelia was formed, and no diffusible pigments were observed on the media tested. The predominant isoprenoid quinone was menaquinone-7 (MK-7). The major fatty acids were anteiso-C15 : 0, and iso-C15 : 0. The polar lipids comprised diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, an unidentified aminophospholipid and an unidentified phospholipid. The DNA G+C content was 37.5 mol%. According to the phylogeneic, phenotypic and chemotaxonomic data, M5HDSG1-1T was clearly distinguishable from other species with validly published names in the genus Bacillus and should therefore be classified as representing a novel species, and we suggest the name Bacillus taxi sp. nov. The type strain is M5HDSG1-1T (=JCM 33117T=CGMCC 1.13668T).


Review: Livestock production increasingly influences wildlife across the globe.
With the growing human population, and their improving wealth, it is predicted that there will be significant increases in demand for livestock products (mainly meat and milk). Recent years have demonstrated that the growth in livestock production has generally had significant impacts on wildlife worldwide; and these are, usually, negative. Here I review the interactions between livestock and wildlife and assess the mechanisms through which these interactions occur. The review is framed within the context of the socio-ecological system whereby people are as much a part of the interaction between livestock and wildlife as the animal species themselves. I highlight areas of interaction that are mediated through effects on the forage supply (vegetation) - neutral, positive and negative - however, the review broadly analyses the impacts of livestock production activities. The evidence suggests that it is not the interaction between the species themselves but the ancillary activities associated with livestock production (e.g. land use change, removal of predators, provision of water points) that are the major factors affecting the outcome for wildlife. So in future, there are two key issues that need to be addressed - first, we need to intensify livestock production in areas of 'intensive' livestock production in order to reduce the pressure for land use change to meet the demand for meat (land sparing). And second, if wildlife is to survive in areas where livestock production dominates, it will have to be the people part of the socio-ecological system that sees the benefits of having wildlife co-exist with livestock on farming lands (land sharing and win-win).


The effect of bran on bowel function in constipation.
Bran is widely used to treat a variety of gastrointestinal disorders. With few exceptions, investigative work has focused on the effect of bran in normal subjects. Despite its widespread use, there have been no systematic studies of bran in constipation. Wheat bran is the "gold standard" because no other natural fiber has yet been shown to be as effective in increasing fecal bulk. We evaluated the effect of two brans, wheat and corn (20 g/day), on fecal weight, fecal moisture content, bowel movement frequency, intestinal transit time, and symptoms in 10 constipated (less than or equal to 3 movements/wk) but otherwise healthy women. The administration of bran was associated with a significant increase in fecal weight (157%), bowel movement frequency (55%), and decrease in intestinal transmit time (50%). Percentage fecal moisture increased only with wheat bran (67.4-72.1%), whereas corn bran was significantly better than wheat bran in relieving symptoms of constipation.


Assessing calibration uncertainty in molecular dating: the assignment of fossils to alternative calibration points.
Although recent methodological advances have allowed the incorporation of rate variation in molecular dating analyses, the calibration procedure, performed mainly through fossils, remains resistant to improvements. One source of uncertainty pertains to the assignment of fossils to specific nodes in a phylogeny, especially when alternative possibilities exist that can be equally justified on morphological grounds. Here we expand on a recently developed fossil cross-validation method to evaluate whether alternative nodal assignments of multiple fossils produce calibration sets that differ in their internal consistency. We use an enlarged Crypteroniaceae-centered phylogeny of Myrtales, six fossils, and 72 combinations of calibration points, termed calibration sets, to identify (i) the fossil assignments that produce the most internally consistent calibration sets and (ii) the mean ages, derived from these calibration sets, for the split of the Southeast Asian Crypteroniaceae from their West Gondwanan sister clade (node X). We found that a correlation exists between s values, devised to measure the consistency among the calibration points of a calibration set (Near and Sanderson, 2004), and nodal distances among calibration points. By ranking all sets according to the percent deviation of s from the regression line with nodal distance, we identified the sets with the highest level of corrected calibration-set consistency. These sets generated lower standard deviations associated with the ages of node X than sets characterized by lower corrected consistency. The three calibration sets with the highest corrected consistencies produced mean age estimates for node X of 79.70, 79.14, and 78.15 My. These timeframes are most compatible with the hypothesis that the Crypteroniaceae stem lineage dispersed from Africa to the Deccan plate as it drifted northward during the Late Cretaceous.


Gene transfer into hepatoma cell lines via the serpin enzyme complex receptor.
The serpin enzyme complex receptor (SECR) expressed on hepatocytes binds to a conserved sequence in alpha 1-antitrypain (alpha 1-AT) and other serpins. A molecular conjugate consisting of a synthetic peptide (C1315) based on the SECR binding motif of human alpha 1-AT covalently coupled to poly-L-lysine was used to introduce reporter genes into hepatoma cell lines in culture. This conjugate condensed DNA into spheroidal particles 18-25 nm in diameter. When transfected with the SECR-directed complex containing pGL3, Hep G2 cells that express the receptor, but not Hep G2 cells that do not, expressed a peak luciferase activity of 538,731 +/- 144,346 integrated light units/mg protein 4 days after transfection. Free peptide inhibited uptake and expression in a dose-dependent manner. Complexes of DNA condensed with polylysine or LC-sulfo-N-succinimidyl-3-(2-pyridyldithio)propionate-substituted polylysine were ineffective. Transfection with a plasmid encoding human factor IX produced expression in Hep G2 (high) and HuH7 cells that express SECR but not Hep G2 (low) cells that lack the receptor. Fluorescein-labeled C1315 peptide labeled 9-31% of Hep G2 (high), 10-14% of HuH7, and 0.6-3.4% of Hep G2 (low) cells, and when the lac Z gene was transfected, only these cells expressed beta-galactosidase. SECR-mediated gene transfer gives efficient, specific uptake and high-level expression of three reporter genes, and the system merits further study for gene therapy.


Separate prediction of intracerebral hemorrhage and ischemic stroke.
To develop and validate 10-year cumulative incidence functions of intracerebral hemorrhage (ICH) and ischemic stroke (IS). We used data on 27,493 participants from 3 population-based cohort studies: the Atherosclerosis Risk in Communities Study, median age 54 years, 45% male, median follow-up 20.7 years; the Rotterdam Study, median age 68 years, 38% male, median follow-up 14.3 years; and the Cardiovascular Health Study, median age 71 years, 41% male, median follow-up 12.8 years. Among these participants, 325 ICH events, 2,559 IS events, and 9,909 nonstroke deaths occurred. We developed 10-year cumulative incidence functions for ICH and IS using stratified Cox regression and competing risks analysis. Basic models including only established nonlaboratory risk factors were extended with diastolic blood pressure, total cholesterol/high-density lipoprotein cholesterol ratio, body mass index, waist-to-hip ratio, and glomerular filtration rate. The cumulative incidence functions' performances were cross-validated in each cohort separately by Harrell C-statistic and calibration plots. High total cholesterol/high-density lipoprotein cholesterol ratio decreased the ICH rates but increased IS rates (p for difference across stroke types <0.001). For both the ICH and IS models, C statistics increased more by model extension in the Atherosclerosis Risk in Communities and Cardiovascular Health Study cohorts. Improvements in C statistics were reproduced by cross-validation. Models were well calibrated in all cohorts. Correlations between 10-year ICH and IS risks were moderate in each cohort. We developed and cross-validated cumulative incidence functions for separate prediction of 10-year ICH and IS risk. These functions can be useful to further specify an individual's stroke risk.


The anti-infective activity of punicalagin against Salmonella enterica subsp. enterica serovar typhimurium in mice.
Punicalagin, a major bioactive component of pomegranate peel, has been proven to have antioxidant, antiviral, anti-apoptosis, and hepatoprotective properties. The aim of this study was to investigate the anti-infective activity of punicalagin in a mouse model. C57BL/6 mice were initially challenged with Salmonella enterica subsp. enterica serovar typhimurium (S. typhimurium) and then treated with punicalagin. Food and water consumption and body weight were recorded daily. On day 8 post infection, the mice were sacrificed to examine pathogen counts in tissues, hematological parameters, cytokine levels, and histological changes. Compared to mice only infected with S. typhimurium, punicalagin-treated mice had more food consumption and less weight loss. A higher survival rate and lower counts of viable S. typhimurium in feces, liver, spleen, and kidney were found in the punicalagin-treated mice. The enzyme linked immunosorbent assay showed that the levels of IL-6, IL-10, and IFN-γ in serum and the spleen and TNF-α in serum, the spleen and the liver were reduced by punicalagin. Moreover, more neutrophils and higher neutrophil-to-mononuclear cell ratios in the punicalagin-treated mice were observed. Histological examination showed that punicalagin protected cells in the liver and spleen from hemorrhagic necrosis. It is concluded that punicalagin has a beneficial effect against S. typhimurium infection in mice. The anti-infective properties, together with other nutritionally beneficial effects, make punicalagin a promising supplement in human food or animal feeds to prevent disease associated with S. typhimurium.


Activated Microglia Targeting Dendrimer-Minocycline Conjugate as Therapeutics for Neuroinflammation.
Brain-related disorders have outmatched cancer and cardiovascular diseases worldwide as the leading cause of morbidity and mortality. The lack of effective therapies and the relatively dry central nervous system (CNS) drug pipeline pose formidable challenge. Superior, targeted delivery of current clinically approved drugs may offer significant potential. Minocycline has shown promise for the treatment of neurological diseases owing to its ability to penetrate the blood-brain barrier (BBB) and potency. Despite its potential in the clinic and in preclinical models, the high doses needed to affect a positive therapeutic response have led to side effects. Targeted delivery of minocycline to the injured site and injured cells in the brain can be highly beneficial. Systemically administered hydroxyl poly(amidoamine) (PAMAM) generation-6 (G6) dendrimers have a longer blood circulation time and have been shown to cross the impaired BBB. We have successfully prepared and characterized the in vitro efficacy and in vivo targeting ability of hydroxyl-G6 PAMAM dendrimer-9-amino-minocycline conjugate (D-mino). Minocycline is a challenging drug to carry out chemical transformations due to its inherent instability. We used a combination of a highly efficient and mild copper catalyzed azide-alkyne click reaction (CuAAC) along with microwave energy to conjugate 9-amino-minocycline (mino) to the dendrimer surface via enzyme responsive linkages. D-mino was further evaluated for anti-inflammatory and antioxidant activity in lipopolysaccharides-activated murine microglial cells. D-mino conjugates enhanced the intracellular availability of the drug due to their rapid uptake, suppressed inflammatory cytokine tumor necrosis factor α (TNF-α) production, and reduced oxidative stress by suppressing nitric oxide production, all significantly better than the free drug. Fluorescently labeled dendrimer conjugate (Cy5-D-mino) was systematically administered (intravenous, 55 mg/kg) on postnatal day 1 to rabbit kits with a clinically relevant phenotype of cerebral palsy. The in vivo imaging study indicates that Cy5-D-mino crossed the impaired blood-brain barrier and co-localized with activated microglia at the periventricular white matter areas, including the corpus callosum and the angle of the lateral ventricle, with significant implications for positive therapeutic outcomes. The enhanced efficacy of D-mino, when combined with the inherent neuroinflammation-targeting capability of the PAMAM dendrimers, may provide new opportunities for targeted drug delivery to treat neurological disorders.


[Severe bilateral pseudomonas sclerokeratitis in comatose patient (clinical case)].
The article presents a clinical case of severe bilateral pseudomonas sclerokeratitis in a patient with occlusion hydrocephalus and intracranial hypertension, who was in a coma and on a ventilator for 20 days. At first examination (7 days after the onset of purulent keratitis, during which the process had been rapidly progressing), the clinical picture included lagophthalmos, severe purulent corneal ulcer, bilateral purulent scleromalacia, perforated cornea in the left eye. On the same day, in order to maintain eye integrity, urgent reconstructive penetrating sclerokeratoplasty with subsequent sclerocorneal coating was performed in both eyes right in the intensive care unit. Parts of the melted iris and ciliary body pars plana that were left in place were abundantly washed with BSS and moxifloxacin solutions - 150 µg/ml. Postoperative care included forced instillations of antibiotics and antiseptics. Two years after the first surgery, 2 more full-thickness corneal transplantations were performed in the patient's right eye aiming at restoration of its optical system. Thus, immediate sclerokeratoplasty with anterior segment irrigation and intraocular administration of highly diluted antibiotics appeared to be the only chance to save the vision in one eye. The fellow eye, where perforation occurred as a result of severe purulent sclerokeratitis and purulent iridocyclitis, despite all measures taken, lost its sight. After three surgeries (penetrating sclerokeratoplasty and two re-PK), visual acuity in the only seeing (right) eye was 0.1, which can be considered a satisfactory result.


Proteomics, ultrastructure, and physiology of hippocampal synapses in a fragile X syndrome mouse model reveal presynaptic phenotype.
Fragile X syndrome (FXS), the most common form of hereditary mental retardation, is caused by a loss-of-function mutation of the Fmr1 gene, which encodes fragile X mental retardation protein (FMRP). FMRP affects dendritic protein synthesis, thereby causing synaptic abnormalities. Here, we used a quantitative proteomics approach in an FXS mouse model to reveal changes in levels of hippocampal synapse proteins. Sixteen independent pools of Fmr1 knock-out mice and wild type mice were analyzed using two sets of 8-plex iTRAQ experiments. Of 205 proteins quantified with at least three distinct peptides in both iTRAQ series, the abundance of 23 proteins differed between Fmr1 knock-out and wild type synapses with a false discovery rate (q-value) <5%. Significant differences were confirmed by quantitative immunoblotting. A group of proteins that are known to be involved in cell differentiation and neurite outgrowth was regulated; they included Basp1 and Gap43, known PKC substrates, and Cend1. Basp1 and Gap43 are predominantly expressed in growth cones and presynaptic terminals. In line with this, ultrastructural analysis in developing hippocampal FXS synapses revealed smaller active zones with corresponding postsynaptic densities and smaller pools of clustered vesicles, indicative of immature presynaptic maturation. A second group of proteins involved in synaptic vesicle release was up-regulated in the FXS mouse model. In accordance, paired-pulse and short-term facilitation were significantly affected in these hippocampal synapses. Together, the altered regulation of presynaptically expressed proteins, immature synaptic ultrastructure, and compromised short-term plasticity points to presynaptic changes underlying glutamatergic transmission in FXS at this stage of development.


The histologic effects of pulsed and continuous radiofrequency lesions at 42 degrees C to rat dorsal root ganglion and sciatic nerve.
Experimental histologic study of the effects of radiofrequency (RF) or convective heating of the rat dorsal root ganglion or sciatic nerve to 42 degrees C. To determine whether treatment causes neuropathologic changes in an effort to explore the mechanisms and safety of pulsed RF pain therapy. Clinical data suggest that low temperature pulsed RF energy delivered to the DRG is a safe and effective form of therapy for low back pain. However, the mechanism by which this treatment modifies pain is unclear. A total of 118 Sprague-Dawley rats were divided into five groups for different RF and thermal treatments. All treatments increased tissue temperature to 42 degrees C. Treatments of the DRG included pulsed RF, continuous RF, and conductive heat. The generator output was increased until 42 degrees C was obtained in the tissue and was then maintained for 120 seconds. As a positive control, some rat sciatic nerves were treated with continuous RF lesions at 80 degrees C. Animals were killed for histologic study at 2, 7, or 21 days after treatment. Tissue was fixed in gluteraldehyde and embedded in plastic resin for detailed light microscopic neuropathologic evaluation. The methods used to heat the tissue to 42 degrees C caused no significant difference in pathology. However, subclinical changes included endoneurial edema caused by alterations in the function of the blood-nerve barrier, fibroblast activation, and collagen deposition. Tissue returned to normal conditions by 7 days in nerve and 21 days in the DRG. These minor structural changes observed at the light microscopic level in normal animals do not exclude the possibility that there would be nonstructural changes in gene expression or cytokine upregulation in injured tissue. Lesions at 80 degrees C caused consistent thermal injury characterized by Wallerian degeneration of nerve fibers. The data support the hypothesis that pulsed RF treatment does not rely on thermal injury of neurologic tissue to achieve its effect.


Managing the threat of methicillin-resistant Staphylococcus aureus in home care.
Severe fatal complications such as pneumonia, cutaneous abscesses, empyema, necrotizing fasciitis, and sepsis can result from methicillin-resistant Staphylococcus aureus (MRSA) even for healthy people. An overview of managing the threat of MRSA in home care is presented. The home healthcare nurse can be proactive with MRSA infections by following and teaching strict medical asepsis while assessing the slightest signs and symptoms indicating that a patient is deteriorating or not responding to the prescribed antibiotic or wound care. This article aims to increase awareness of community-acquired (CA)-MRSA, to offer suggestions for preventing CA-MRSA, and to describe methods for managing this infectious process in the home.


Defining macropinocytosis.
Macropinocytosis represents a distinct pathway of endocytosis in mammalian cells. This actin-driven endocytic process is not directly co-ordinated by the presence of cargo but can be induced upon activation of growth factor signalling pathways. The capacity to dissect the contribution of macropinocytosis to cellular processes has been hampered by a lack of unique molecular markers and defining features. While aspects of macropinosome formation and maturation are common to those shared by the other endocytic pathways, a number of key differences have recently begun to emerge and will be discussed in this study. It is now well established that macropinocytosis significantly contributes to antigen presentation by the immune system and is exploited by a range of pathogens for cellular invasion and avoidance of immune surveillance.


A multimodal platform for nonlinear optical microscopy and microspectroscopy.
Multimodal nonlinear optical microscopy is a valuable tool to study complex biological samples. We present an easy-to-operate approach to perform coherent anti-Stokes Raman scattering (CARS), two-photon fluorescence (TPF), second harmonic generation (SHG), and third-harmonic generation (THG) imaging using a single laser source composed of an 80 MHz femtosecond (fs) laser, an optical parametric oscillator (OPO), and a PPLN crystal for frequency doubling. The platform allows vibrationally resonant CARS imaging of CH-rich myelin sheath in fresh spinal tissues and lipid bodies in live cells. Multimodal nonlinear optical imaging and microspectroscopy analysis of fresh liver tissues are demonstrated.


Quantification of 18F-Fluoride Kinetics: Evaluation of Simplified Methods.
(18)F-fluoride PET is a promising noninvasive method for measuring bone metabolism and bone blood flow. The purpose of this study was to assess the performance of various clinically useful simplified methods by comparing them with full kinetic analysis. In addition, the validity of deriving bone blood flow from K1 of (18)F-fluoride was investigated using (15)O-H2O as a reference. Twenty-two adults (mean age ± SD, 44.8 ± 25.2 y), including 16 patients scheduled for bone surgery and 6 healthy volunteers, were studied. All patients underwent dynamic (15)O-H2O and (18)F-fluoride scans before surgery. Ten of these patients had serial PET measurements before and at 2 time points after local bone surgery. During all PET scans, arterial blood was monitored continuously. (18)F-fluoride data were analyzed using nonlinear regression (NLR) and several simplified methods (Patlak and standardized uptake value [SUV]). SUV was evaluated for different time intervals after injection and after normalizing to body weight, lean body mass, and body surface area, and simplified measurements were compared with NLR results. In addition, changes in SUV and Patlak-derived fluoride influx rate (Ki) after surgery were compared with corresponding changes in NLR-derived Ki. Finally, (18)F-fluoride K1 was compared with bone blood flow derived from (15)O-H2O data, using the standard single-tissue-compartment model. K1 of (18)F-fluoride correlated with measured blood flow, but the correlation coefficient was relatively low (r = 0.35, P < 0.001). NLR resulted in a mean Ki of 0.0160 ± 0.0122, whereas Patlak analysis, for the interval 10-60 min after injection, resulted in an almost-identical mean Ki of 0.0161 ± 0.0117. The Patlak-derived Ki, for 10-60 min after injection, showed a high correlation with the NLR-derived Ki (r = 0.976). The highest correlation between Ki and lean body mass-normalized SUV was found for the interval 50-60 min (r = 0.958). Finally, changes in SUV correlated significantly with those in Ki (r = 0.97). The present data support the use of both Patlak and SUV for assessing fluoride kinetics in humans. However, (18)F-fluoride PET has only limited accuracy in monitoring bone blood flow.


Pleomorphic lobular carcinoma of the breast: is it a prognostically significant pathological subtype independent of histological grade?
Pleomorphic lobular carcinoma is regarded as a biologically aggressive variant of invasive lobular carcinoma of the breast. However, there is no consensus on the definition and whether this subtype adds useful information to histological grade. Two-hundred and two grade 2 or grade 3 invasive lobular carcinomas were studied. Tumours were categorised according to the components of histological grade: tubules, pleomorphism and mitoses. Pleomorphic lobular carcinoma was defined as a carcinoma with a lobular growth pattern and marked nuclear pleomorphism (pleomorphism 3). Breast cancer-specific survival was used in analysis of prognosis. Grade 3 pleomorphic lobular carcinomas (tubules 3, pleomorphism 3, mitoses 2 and tubules 3, pleomorphism 3, mitoses 3) had a worse prognosis than grade 2 (tubules 3, pleomorphism 2, mitoses 1) carcinomas. Grade 2 lobular carcinomas with marked nuclear pleomorphism (tubules 3, pleomorphism 3, mitoses 1) had a similar prognosis to grade 2 carcinomas with moderate pleomorphism (tubules 3, pleomorphism 2, mitoses 1). Survival was associated with mitotic score, but not with nuclear pleomorphism on both univariate and multivariate analysis. A non-classical growth pattern was seen more frequently in all subgroups with marked nuclear pleomorphism and was associated with worse survival. Histological grade and nodal status were independent of prognostic factors. This study shows that histological grade (in particular the mitotic component) in invasive lobular carcinomas is of prognostic importance, but pleomorphic type does not provide useful additional prognostic information.


A novel mutation in the CD40 ligand gene in a Chinese boy with X-linked hyper-IgM syndrome.
X-linked hyper-IgM Syndrome (XHIGM) is caused by a mutation of CD40 ligand (CD40L), which is normally expressed on activated CD4+ T cells and is responsible for immunoglobulin class switching. A 7-year-old boy with recurrent sino-pulmonary infections since the age of 3 months had normal CD3+, CD4+, CD8+T lymphocytes, and CD19+B lymphocytes and NK cells, but significantly elevated IgM and extremely decreased IgG and IgA. Sequencing of genomic DNA revealed that the patient had a 34 base deletion in intron 3 and exon 4 of CD40L(g.8172_8205del34bp), which lead to the entire deletion of exon 4 in cDNA (c.347_409del63bp, i.e.,exon 4 skipping) and an in-frame deletion of 21 amino acids in CD40L protein. Moreover, the patient had negligible CD40L expression on activated CD3+CD8-T lymphocytes. His mother and sister were carriers of the CD40L mutation. Our studies demonstrated a novel mutation in CD40L, which, to our knowledge, has not been reported previously.


Compartmentalization of telomeres through DNA-scaffolded phase separation.
Telomeres form unique nuclear compartments that prevent degradation and fusion of chromosome ends by recruiting shelterin proteins and regulating access of DNA damage repair factors. To understand how these dynamic components protect chromosome ends, we combine in vivo biophysical interrogation and in vitro reconstitution of human shelterin. We show that shelterin components form multicomponent liquid condensates with selective biomolecular partitioning on telomeric DNA. Tethering and anomalous diffusion prevent multiple telomeres from coalescing into a single condensate in mammalian cells. However, telomeres coalesce when brought into contact via an optogenetic approach. TRF1 and TRF2 subunits of shelterin drive phase separation, and their N-terminal domains specify interactions with telomeric DNA in vitro. Telomeric condensates selectively recruit telomere-associated factors and regulate access of DNA damage repair factors. We propose that shelterin mediates phase separation of telomeric chromatin, which underlies the dynamic yet persistent nature of the end-protection mechanism.


Helmholtz resonator with extended neck.
Acoustic performance of a concentric circular Helmholtz resonator with an extended neck is investigated theoretically, numerically, and experimentally. The effect of length and shape of, and the perforations on the neck extension is examined on the resonance frequency and the transmission loss. A two-dimensional analytical method is developed for an extended neck with constant cross-sectional area, while a three-dimensional boundary element method is applied for the variable area and perforated extension. Lumped and one-dimensional approaches are also included to illustrate the effect of the higher order modes. For a piston-driven model, predicted resonance frequencies using lumped, one-dimensional, and two-dimensional analytical methods are compared with those from multidimensional boundary element method. Analytical and computational transmission loss predictions for pipe-mounted model are compared to the experimental data obtained from an impedance tube setup. It is shown that the resonance frequency may be controlled by the length, shape, and perforation porosity of the extended neck without changing the cavity volume.


Influence of mercurial sulfhydryl agents on aqueous outflow pathways in enucleated eyes.
Enucleated calf and primate eyes were perfused with either p-chloromercuribenzene sulfonate (PCMBS) or p-chloromercuribenzene (PCMB). Both caused a decrease in aqueous outflow that did not result from inhibition of glycolysis in trabecular cells. Morphologic studies of PCMBS-treated primate eyes suggested that the reduction in aqueous outflow occurred as the result of cellular swelling in the trabecular meshwork and, particularly, in the juxtacanalicular connective tissue. These findings suggest that changes in cell volume may influence trabecular outflow resistance and that mercurial-sensitive membrane sulfhydryl groups could participate in the regulation of aqueous outflow.


Mk3.5: a modular, multi-frequency successor to the Mk3a EIS/EIT system.
This paper describes the Sheffield Mk3.5 EIT/EIS system which measures both the real and imaginary part of impedance at 30 frequencies between 2 kHz and 1.6 MHz. The system uses eight electrodes with an adjacent drive/receive electrode data acquisition protocol. The system is modular, containing eight identical data acquisition boards, which contain DSPs to generate the drive frequencies and to perform the FFT used for demodulation. The current drive is in three sequentially applied packets, where each packet contains ten summed sine waves. The data acquisition system is interfaced to a host PC through an optically isolated high speed serial link (RS485) running at 2 Mbaud (2 Mbits s(-1)). Measurements on a saline filled tank show that the average signal to noise performance of the system is 40 dB measured across all frequencies and that this figure is independent of frequency of measurement. These results suggest that the current system is 10 dB better in absolute terms than the previous Sheffield (Mk3a) system.


Acid and non-acid reflux in patients with persistent symptoms despite acid suppressive therapy: a multicentre study using combined ambulatory impedance-pH monitoring.
Empiric proton pump inhibitor (PPI) trials have become increasingly popular leading to gastroenterologists frequently evaluating gastro-oesophageal reflux disease (GORD) patients only after they have "failed" PPI therapy. Combined multichannel intraluminal impedance and pH (MII-pH) monitoring has the ability to detect gastro-oesophageal reflux (GOR) episodes independent of their pH and evaluate the relationship between symptoms and all types of GOR. Using this technique, we aimed to characterise the frequency of acid and non-acid reflux (NAR) and their relationship to typical and atypical GOR symptoms in patients on PPI therapy. Patients with persistent GORD symptoms referred to three centres underwent 24 hour combined MII-pH monitoring while taking PPIs at least twice daily. Reflux episodes were detected by impedance channels located 3, 5, 7, 9, 15, and 17 cm above the lower oesophageal sphincter (LOS) and classified into acid or non-acid based on pH data from 5 cm above the LOS. A positive symptom index (SI) was declared if at least half of each specific symptom events were preceded by reflux episodes within five minutes. A total of 168 patients (103 (61%) females and 65 (39%) males; mean age 53 (range 18-85) years) underwent combined MII-pH monitoring while taking PPIs at least twice daily. One hundred and forty four (86%) patients recorded symptoms during the study day and 24 (15%) patients had no symptoms during testing. Sixty nine (48%) symptomatic patients had a positive SI for at least one symptom (16 (11%) with acid reflux and 53 (37%) with NAR) and 75 (52%) had a negative SI. A total of 171 (57%) typical GORD symptoms were recorded, 19 (11%) had a positive SI for acid reflux, 52 (31%) for NAR, and 100 (58%) had a negative SI. One hundred and thirty one (43%) atypical symptoms were recorded, four (3%) had a positive SI for acid reflux, 25 (19%) had a positive SI for NAR, and 102 (78%) had a negative SI. Combined MII-pH identifies the relation of reflux of all types to persistent symptoms and the importance of NAR in patients taking PPIs.


Inhibition of TRPM7 Attenuates Rat Aortic Smooth Muscle Cell Proliferation Induced by Angiotensin II: Role of Genistein.
Transient receptor potential melastatin 7 (TRPM7) is a Ca, Mg permeable nonselective cation channel of the TRP channel superfamily and plays an important role in cell growth and proliferation. Compounds that alter the activity and expression of the channel protein might be of therapeutic interest. In this study, we investigated the effects of genistein on TRPM7 channels and the proliferation of rat aortic smooth muscle cells (RAoSMCs). In primary cultured RAoSMCs, acute genistein (50 μM) exposure inhibited native TRPM7 currents, whereas chronic expose to genistein (50 μM) downregulated TRPM7 protein expression. The downregulation of TRPM7 protein expression induced by genistein was mimicked by c-Src inhibitor (PP2), but not by epidermal growth factor receptor tyrosine kinase inhibitor (lavendustin A), or daidzein. Additionally, genistein (50 μM) attenuated angiotensin II-induced cell proliferation. This study is the first to demonstrate inhibition of TRPM7 by isoflavone genistein through c-Src tyrosine kinase inhibition in RAoSMCs. Our results not only provide a new modulation mechanism of TRPM7 but also suggest that TRPM7 may serve as a new therapeutic target of genistein in the treatment of vascular diseases.


[Cytogenetic research on micropopulations of the root vole (Microtus oeconomus Pall.) living under various radioecological habitat conditions].
Percentage of aneuploid cells and that of cell with chromosome aberrations in Microtus oeconomus Pall. living in areas with the enhanced radiation background (the dose of external radiation is 50 times and that of internal irradiation of bone marrow by incorporated 226Ra 10 times higher than the controls) exceeds significantly (p less than 0.05) the control values. It is concluded that mutations occur in the experimental animals more frequently than in the controls.


Multiunit bursts in rat pallidum during grooming and stereotyped jaw movements.
Extracellular recordings from the globus pallidus of awake, unrestrained rats showed a distinctive bursting activity during grooming behaviour and in periods of stereotyped jaw movements induced by amphetamine (3 or 5 mg/kg IP) or apomorphine (2 mg/kg SC). During stereotyped licking, there was one burst for each outward movement of the tongue. The bursts were shown to consist of several separate unit spikes firing so as to produce a fusiform envelope of amplitudes, suggesting an ordered recruitment of pallidal neurons related to licking.


Therapeutic effect of Echinococcus granulosus cyst fluid on bacterial sepsis in mice.
The primary pathophysiological process of sepsis is to stimulate a massive release of inflammatory mediators to trigger systemic inflammatory response syndrome (SIRS), the major cause of multi-organ dysfunction and death. Like other helminths, Echinococcus granulosus induces host immunomodulation. We sought to determine whether E. granulosus cyst fluid (EgCF) displays a therapeutic effect on sepsis-induced inflammation and tissue damage in a mouse model. The anti-inflammatory effects of EgCF were determined by in vitro culture with bone marrow-derived macrophages (BMDMs) and in vivo treatment of BALB/C mice with cecal ligation and puncture (CLP)-induced sepsis. The macrophage phenotypes were determined by flow cytometry, and the levels of cytokines in cell supernatants or in sera of mice were measured (ELISA). The therapeutic effect of EgCF on sepsis was evaluated by observing the survival rates of mice for 72 h after CLP, and the pathological injury to the liver, kidney, and lung was measured under a microscope. The expression of TLR-2/MyD88 in tissues was measured by western blot to determine whether TLR-2/MyD88 is involved in the sepsis-induced inflammatory signaling pathway. In vitro culture with BMDMs showed that EgCF promoted macrophage polarization to M2 type and inhibited lipopolysaccharide (LPS)-induced M1 macrophages. EgCF treatment provided significant therapeutic effects on CLP-induced sepsis in mice, with increased survival rates and alleviation of tissue injury. The EgCF conferred therapeutic efficacy was associated with upregulated anti-inflammatory cytokines (IL-10 and TGF-β) and reduced pro-inflammatory cytokines (TNF-α and INF-γ). Treatment with EgCF induced Arg-1-expressed M2, and inhibited iNOS-expressed M1 macrophages. The expression of TLR-2 and MyD88 in EgCF-treated mice was reduced. The results demonstrated that EgCF confers a therapeutic effect on sepsis by inhibiting the production of pro-inflammatory cytokines and inducing regulatory cytokines. The anti-inflammatory effect of EgCF is carried out possibly through inducing macrophage polarization from pro-inflammatory M1 to regulatory M2 phenotype to reduce excessive inflammation of sepsis and subsequent multi-organ damage. The role of EgCF in regulating macrophage polarization may be achieved by inhibiting the TLR2/MyD88 signaling pathway.


A novel non-destructive technique for qualitative and quantitative measurement of dental erosion in its entirety by porosity and bulk tissue-loss.
To explore the potential of combining non-contact profilometry (NCP) and confocal laser scanning microscopy (CLSM) data to measure the entire erosive process non-destructively and to validate findings using inductively coupled plasma-atomic emission spectroscopy (ICP-AES), scanning electron microscopy (SEM) and surface microhardness (SMH) using the same samples throughout. Polished bovine enamel samples (n = 35) were divided into groups (7/group) with similar SMH values. Samples underwent individual erosive challenges (1 % citric acid, pH3.8) for 1, 5, 10, 15 or 30 min under stirring and aliquot extracts were analysed for Ca and P by ICP-AES. SMH was used to measure erosive softening. Profilometry was used to assess bulk volume loss (BVL). Images were captured by SEM. Samples were stained with rhodamine-B (0.1 mM, 24 h) and images captured by CLSM. Image processing was used to determine changes in fluorescent volume for the first 10 μm (ΔFV10) for each enamel sample which were combined with BVL to calculate total lesion volume (TLV). ANOVA, linear regression and Pearson correlation analysis were used where applicable. Surface softening, [Ca], [P], BVL and ΔFV10μm increased with acid erosion duration which were significant by 10 min (P < .01). The Ca:P ratio increased to 1.57 then decreased after 5 min erosion suggesting a sub/surface phase change, which was observed by SEM and CLSM showing significant changes to the enamel surface and subsurface morphology with time. Combination of BVL and ΔFV10 as TLV strengthened the significant correlations with [Ca], [P], and SMH (P < .01). This novel combination of CLSM and NCP allows for concurrent non-destructive quantification of the entire erosive process by mineral loss, and qualitatively characterise microstructural changes during dental erosion.


Spineless-aristapedia: a homeotic gene that does not control the development of specific compartments in Drosophila.
A two-step screen for isolating null mutations of the spineless-aristapedia locus has been performed, and several amorphic mutations, as well as a small deficiency, have been obtained. With the exception of the deficiency, which deletes genes required for viability on either side of the spineless-aristapedia locus, these mutations result in a transformation of only the distal antenna into distal leg, thereby indicating that the spineless-aristapedia gene is required for specifying antennal as opposed to leg development in only the distal portion of the antenna. Because this distal region does not appear to be a developmental compartment, it is probable that the spineless-aristapedia gene, unlike several other homeotic genes, is required for maintaining the correct determined state in a population of cells defined by their relative position, not by their ancestry.


Population pharmacokinetic and pharmacodynamic modelling of the effects of nicorandil in the treatment of acute heart failure.
The aims of the study were 1) to evaluate the pharmacokinetics of nicorandil in healthy subjects and acute heart failure (AHF) patients and 2) to evaluate the exposure-response relationship with pulmonary arterial wedge pressure (PAWP) in AHF patients and to predict an appropriate dosing regimen for nicorandil. Based on the data from two healthy volunteer and three AHF patient studies, models were developed to characterize the pharmacokinetics and pharmacodynamics of nicorandil. PAWP was used as the pharmacodynamic variable. An asymptotic exponential disease progression model was used to account for time dependent changes in PAWP that were not explained by nicorandil exposure. The modelling was performed using NONMEM version V. The pharmacokinetics of nicorandil were characterized by a two-compartment model with linear elimination. CL, V1 and V2 in AHF patients were 1.96, 1.39 and 4.06 times greater than in healthy subjects. Predicted plasma concentrations were assumed to have an immediate concentration effect relationship on PAWP. An inhibitory E(max) model with E(max) of -11.7 mmHg and EC(50) of 423 microg l(-1) was considered the best relationship between nicorandil concentrations and PAWP. PAWP decreased independently of nicorandil exposure. This drug independent decline was described by an asymptotic decrease of 6.1 mmHg with a half-life of 5.3 h. AHF patients have higher clearance and initial distribution volume of nicorandil compared with healthy subjects. The median target nicorandil concentration to decrease PAWP by 30% is predicted to be 748 microg l(-1), indicating that a loading dose of 200 microg kg(-1) and a maintenance dose of 400 microg kg(-1) h(-1) would be appropriate for the initial treatment of AHF.


Exercise training in paediatric congenital heart disease: fit for purpose?
Exercise and physical activity (PA) have been shown to be effective, safe and feasible in both healthy children and children with congenital heart disease (CHD). However, implementing exercise training as an intervention is still not routine in children with CHD despite considerable evidence of health benefits and well-being. Understanding how children with CHD can safely participate in exercise can boost participation in PA and subsequently reduce inactivity-related diseases. Home-based exercise intervention, with the use of personal wearable activity trackers, and high-intensity interval training have been beneficial in adults' cardiac rehabilitation programmes. However, these remain underutilised in paediatric care. Therefore, the aims of this narrative review were to synthesise prescribed exercise interventions in children with CHD, identify possible limitation to exercise training prescription and provide an overview on how to best integrate exercise intervention effectively for this population into daily practice.


Upregulation of reactive oxygen and nitrogen intermediates in Plasmodium berghei infected mice after rescue therapy with chloroquine or artemether.
Plasmodium berghei ANKA infected C57B1/6 mice develop cerebral malaria at a parasitaemia of 15-25%. When parasitaemia reached 10%, P. berghei infected mice were treated with artemether, chloroquine or clindamycin in order to prevent the occurrence of cerebral malaria. Artemether and chloroquine were highly efficient. Functional tests revealed that zymosan stimulated spleen cells from untreated mice with cerebral malaria showed a slight decrease in their capacity to produce reactive oxygen intermediates (ROI) when compared with naive mice. After artemether or chloroquine treatment, the ROI production was significantly enhanced. The interferon-gamma induced production of reactive nitrogen intermediates (RNI) was slightly elevated in mice with cerebral malaria, but markedly elevated in artemether or chloroquine treated mice when compared with naive mice. Moreover, high levels of inducible nitric oxide synthase gene expression could be detected by in-situ hybridization in spleen sections of mice which had been treated with artemether or chloroquine. These findings suggest that increased production of ROI and RNI after chemotherapy may play a protective role for the host during malaria.


[Structure and function changes of oxidized human serum albumin: physiological significance of the biomarker and importance of sampling conditions for accurate measurement].
Human serum albumin (HSA) exists in both reduced and oxidized forms, and the percentage of oxidized albumin increases in several diseases; however, little is known regarding the pathological and physiological significance of oxidation due to poor characterization of the precise structural and functional properties of oxidized HSA. Here, we characterize both structural and functional differences between reduced and oxidized HSA. Using LC-ESITOFMS and FTMS analysis, we determined that the major structural change in oxidized HSA in healthy human plasma is a disulfide-bonded cysteine at the thiol of Cys34 of reduced HSA. Based on this structural information, we prepared standard samples of purified HSA, e.g. nonoxidized (intact purified HSA which mainly exists in reduced form), mildly oxidized and highly oxidized HSA. Using these standards, we demonstrated several differences in functional properties of HSA, including protease susceptibility, ligand-binding affinity and antioxidant activity. From these observations, we conclude that an increased level of oxidized HSA may impair HSA function in a number of pathological conditions. In addition, we determined blood and plasma sampling conditions for accurate measurement of the oxidized albumin ratio in plasma using EST-TOFMS screening.


Psychedelic N,N-dimethyltryptamine and 5-methoxy-N,N-dimethyltryptamine modulate innate and adaptive inflammatory responses through the sigma-1 receptor of human monocyte-derived dendritic cells.
The orphan receptor sigma-1 (sigmar-1) is a transmembrane chaperone protein expressed in both the central nervous system and in immune cells. It has been shown to regulate neuronal differentiation and cell survival, and mediates anti-inflammatory responses and immunosuppression in murine in vivo models. Since the details of these findings have not been elucidated so far, we studied the effects of the endogenous sigmar-1 ligands N,N-dimethyltryptamine (NN-DMT), its derivative 5-methoxy-N,N-dimethyltryptamine (5-MeO-DMT) and the synthetic high affinity sigmar-1 agonist PRE-084 hydrochloride on human primary monocyte-derived dendritic cell (moDCs) activation provoked by LPS, polyI:C or pathogen-derived stimuli to induce inflammatory responses. Co-treatment of moDC with these activators and sigma-1 receptor ligands inhibited the production of pro-inflammatory cytokines IL-1β, IL-6, TNFα and the chemokine IL-8, while increased the secretion of the anti-inflammatory cytokine IL-10. The T-cell activating capacity of moDCs was also inhibited, and dimethyltryptamines used in combination with E. coli or influenza virus as stimulators decreased the differentiation of moDC-induced Th1 and Th17 inflammatory effector T-cells in a sigmar-1 specific manner as confirmed by gene silencing. Here we demonstrate for the first time the immunomodulatory potential of NN-DMT and 5-MeO-DMT on human moDC functions via sigmar-1 that could be harnessed for the pharmacological treatment of autoimmune diseases and chronic inflammatory conditions of the CNS or peripheral tissues. Our findings also point out a new biological role for dimethyltryptamines, which may act as systemic endogenous regulators of inflammation and immune homeostasis through the sigma-1 receptor.


Optimal ilio-sacral screw trajectory in paediatric patients : a computed tomography study.
Pelvic fixation during procedures performed to treat spinal deformities in paediatric patients remains challenging. No computed tomography studies in paediatric have assessed the optimal trajectory of ilio- sacral screws to prevent screw malposition. We used pelvic computed tomography from 80 children divided into four groups : females <10 and ≥10 years and males <10 and ≥10 years. A secure triangular corridor parallel to the upper S1 endplate was delineated based on three fixed landmarks. The optimal screw insertion angle was subtended by the horizontal and the line bisecting the secure corridor. Student's t test was applied to determine whether the optimal screw insertion angle and/or anatomical parameters were associated with age and/or sex. Mean optimal angle was 32.3°±3.6°, 33.8°±4.7°, 30.2°±5.0°, and 30.4°±4.7° in the younger females, younger males, older females, and older males, respectively. The mean optimal angle differed between the two age groups (p=0.004) but not between females and males (p=0.55). Optimal mean screw length was 73.4±9.9 mm. Anatomical spinal canal parameters in the transverse plane varied with age (p=0.02) and with sex in the older children (p=0.008), and those in the sagittal plane varied with sex (p=0.04). Age affected ilio-sacral screw positioning, whereas sex did not. Several anatomical spinal canal parameters varied with age and sex. These results should help to ensure safe and easy ilio-sacral screw placement within a secure corridor.


[Pseudoaneurysm with Infective Endocarditis Ten Years After the Resection of Pseudoaneurysm Complicated with Myocardial Infarction;Report of a Case].
A 76-year-old female was referred to our hospital for fever and chillness. She was diagnosed with pneumonia and sepsis, and methicillin-resistant Staphylococcus aureus (MRSA)was detected by the sputum culture. Echocardiography showed mobile vegetation and left ventricular pseudoaneurysm at the apex. As she had a history of ventricular pseudoaneurysmectomy 10 years before, we considered the patient had developed infective endocarditis at the surgical site. Surgical treatment was needed because of the rapidly growing pseudoaneurysm despite the use of antibacterial agents. She successfully underwent resection of the pseudoaneurysm and MRSA was also detected in the culture of the previous vascular graft patch. Antibacterial agents were administrated for 6 weeks, and she was discharged at 53 days after operation.


Cigarette smoking and Down syndrome.
A matched case-control study of 100 mothers of Down syndrome children, 100 mothers of children with other defects (defect controls), and 100 mothers of children with no defects (normal controls) was carried out. All infants were born in upstate New York in 1980 and 1981. Matching was very close on maternal age for the normal controls but not for the defect controls. The risk ratios for the association of cigarette smoking around the time of conception with Down syndrome was 0.58 (90% confidence interval of 0.34-0.98) in the case-defect control comparison and 0.56 (90% confidence interval of 0.33-0.95) in the case-normal control comparison. Stratification by alcohol ingestion and maternal age did not abolish the negative trend to association. The results are contrary to that of an earlier study of others that found a positive association of older age and trisomy in spontaneous abortions. In fact, among mothers of Down syndrome cases over age 30 in this analysis, the risk ratio was lower than for younger mothers. (For case-normal control comparisons, the value was 0.39 [90% confidence interval of 0.17-0.87]). If not due to chance or confounding, the negative association in our data may be attributable to, among other factors, a selective effect of smoking upon survival or fertilizability of +21 gametes prior to conception or upon survival of +21 conceptuses after fertilization.


The 2023 ACR/EULAR Antiphospholipid Syndrome Classification Criteria.
To develop new antiphospholipid syndrome (APS) classification criteria with high specificity for use in observational studies and trials, jointly supported by the American College of Rheumatology (ACR) and EULAR. This international multidisciplinary initiative included 4 phases: 1) Phase I, criteria generation by surveys and literature review; 2) Phase II, criteria reduction by modified Delphi and nominal group technique exercises; 3) Phase III, criteria definition, further reduction with the guidance of real-world patient scenarios, and weighting via consensus-based multicriteria decision analysis, and threshold identification; and 4) Phase IV, validation using independent adjudicators' consensus as the gold standard. The 2023 ACR/EULAR APS classification criteria include an entry criterion of at least one positive antiphospholipid antibody (aPL) test within 3 years of identification of an aPL-associated clinical criterion, followed by additive weighted criteria (score range 1-7 points each) clustered into 6 clinical domains (macrovascular venous thromboembolism, macrovascular arterial thrombosis, microvascular, obstetric, cardiac valve, and hematologic) and 2 laboratory domains (lupus anticoagulant functional coagulation assays, and solid-phase enzyme-linked immunosorbent assays for IgG/IgM anticardiolipin and/or IgG/IgM anti-β2 -glycoprotein I antibodies). Patients accumulating at least 3 points each from the clinical and laboratory domains are classified as having APS. In the validation cohort, the new APS criteria versus the 2006 revised Sapporo classification criteria had a specificity of 99% versus 86%, and a sensitivity of 84% versus 99%. These new ACR/EULAR APS classification criteria were developed using rigorous methodology with multidisciplinary international input. Hierarchically clustered, weighted, and risk-stratified criteria reflect the current thinking about APS, providing high specificity and a strong foundation for future APS research.


Age-related resistance of Nicotiana benthamiana against hemibiotrophic pathogen Phytophthora infestans requires both ethylene- and salicylic acid-mediated signaling pathways.
Phytophthora infestans, the agent of late blight disease of potato, is a hemibiotrophic pathogen with biotrophic action during early infection and necrotrophic in the later stage of colonization. Mature Nicotiana benthamiana was resistant to P. infestans, whereas relatively young plants were susceptible to this pathogen. Young plants became resistant following a pretreatment with acibenzolar-S-methyl, a functional analog of salicylic acid (SA), indicating that susceptibility of young plants is due to a lack of induction of SA signaling. Further analysis with virus-induced gene silencing indicated that NbICS1 and NbEIN2, the genes for SA biosynthesis and ethylene (ET) signaling, respectively, are required for the resistance of mature N. benthamiana against P. infestans. Furthermore, these genes are required for the production of reactive oxygen species (ROS) induced by treatment of the INF1 elicitor. In NbICS1-silenced plants, cell death induced by either INF1 or necrosis-inducing protein NPP1.1 was significantly accelerated. Expression of genes for phytoalexin (capsidiol) biosynthesis, NbEAS and NbEAH, were regulated by ET, and gene silencing of either of them compromised resistance of N. benthamiana to P. infestans. Together, these results suggest that resistance of N. benthamiana against hemibiotrophic P. infestans requires both SA-regulated appropriate induction of cell death and ET-induced production of phytoalexin.


B cell- and monocyte-activating chemokine (BMAC), a novel non-ELR alpha-chemokine.
A novel alpha-chemokine, designated KS1, was identified from an EST database of a murine immature keratinocyte cDNA library. The EST has 94% similarity to a recently cloned human gene, BRAK, that has no demonstrated function. Northern analysis of mouse and human genes showed detectable mRNA in brain, intestine, muscle and kidney. Tumour panel blots showed that BRAK was down-regulated in cervical adenocarcinoma and uterine leiomyoma, but was up-regulated in breast invasive ductal carcinoma. KS1 bound specifically to B cells and macrophages, as well as two B cell lines, CESS and A20, and a monocyte line, THP-1. KS1 showed no binding to naive or activated T cells. In addition, KS1 stimulated the chemotaxis of CESS and THP-1 cells but not T cells. The s.c. injection of KS1 creates a mixed inflammatory response in Nude and C3H/HeJ mice. The above data indicates that KS1 and its human homologue represents a novel non-ELR alpha-chemokine that may have important roles in trafficking of B cells and monocytes. We propose the name B cell- and monocyte-activating chemokine (BMAC) for this molecule to reflect the described biological functions.


Effect of sodium intake on angiotensin-converting enzyme activity of aorta in rats.
1. The effect of sodium intake on angiotensin-converting enzyme activity in subcellular fractions of aorta (homogenate, mitochondria, microsomes and supernatant) was studied in normotensive and spontaneously hypertensive rats. 2. Angiotensin-converting enzyme activity was extremely high in the supernatant fraction in normotensive and spontaneously hypertensive rats. 3. Total converting enzyme activity in spontaneously hypertensive rats was 34% higher than that in normotensive rats. 4. Increased sodium intake resulted in a marked reduction, almost to zero, of converting enzyme activity in each fraction of aorta in normotensive and spontaneously hypertensive rats. 5. Converting enzyme, widely distributed in subcellular fractions of the aorta, may play a possible rôle in the local control of vascular tone. It is likely that sodium intake inhibits production of the enzyme in vascular tissue.


[Characteristics of medical-demographic processes in rural regions of Russia].
The serious medico-demographic problems of the last decade had a most negative impact on the rural population. A negative natural increment and a high level of aging of the rural population were evident as long ago as in the seventies. The number of children born to unmarried mothers is traditionally higher in villages, and the expected life span is lower there than in towns. Mortality caused by injuries, respiratory diseases, and infections, including the mortality of infants aged under one, is higher in the country than in towns. Urgent reasonable sociodemographic policy is needed to preserve the population of Russia, including its rural population.


[Results of chemotherapy in patients with disseminated testicular tumors].
Results of treatment of 44 patients with disseminated non-seminomatous testicular tumors versus advancement are discussed in the paper. In cases of minimal and moderate advancement (group 1), induction chemotherapy included three cycles of VAB-6 regimen (vinblastine, actinomycin D, bleomycetin, cyclophosphamide plus 120 mg/m2 platidiam) whereas cases of advanced tumor (group 2) received six such cycles; two of them used 150 mg/m2 platidiam dropwise in a 3% sodium chloride solution. Complete regression was observed in 22 of 24 (96.1%) patients of group 1 and in seven out of 20 (35%) cases of group 2; it was registered in 100, 81.8 and 38% of patients with minimal, moderately- and far-advanced disease, respectively. Patients were followed for 4-23 months (average 14.5 months). Eight cases relapsed. At the time of this writing, 38 out of 44 (86.3%) patients are alive, and 23 out of 29 (79.3%) continue in complete remission. Toxicity was moderate. Application of 3% NaCl prevented nephrotoxicity of high-dose (150 mg/m2) platidiam.


Rapid and efficient detection of extra-pulmonary Mycobacterium tuberculosis by PCR analysis.
The diagnosis of extra-pulmonary tuberculosis (EPTB) remains an important clinical problem, primarily because of the inadequate sensitivity of conventional bacteriologic methods for detecting Mycobacterium tuberculosis in extra-pulmonary specimens. To evaluate whether a IS6110-based polymerase chain reaction (PCR) method can be utilized to detect M. tuberculosis in non-pulmonary specimens. Specimens from 286 Mexican patients with a presumptive clinical diagnosis of EPTB were prospectively examined by Ziehl-Neelsen staining, mycobacterial culture on Löwenstein-Jensen slants, and by PCR. The DNA for PCR was extracted by the buffer lysis method and phenol-guanidine thiocyanate-chloroform. Primers that amplify a 200 bp fragment from the insertion-like M. tuberculosis sequence element IS6110 were utilized. Our results demonstrate that this PCR method is highly specific (100%) for identifying M. tuberculosis from a variety of specimens including cerebrospinal fluid (CSF), pleural fluid, ascitic fluid, pericardial fluid, urine, and lymph node exudate. Moreover, the sensitivity of PCR for detecting M. tuberculosis in CSF (94%), pleural fluid (94%), ascitic fluid and other extrapulmonary specimens (93%) greatly exceeds the sensitivity of conventional smear and culture methods. These results demonstrate that PCR can be a highly specific and sensitive aid in the detection of M. tuberculosis from extra-pulmonary specimens.


A properdin system intermediate formed by zymosan and serum at 0 degrees C.
The interaction of guinea pig or rat serum with Z at 0 degrees C leads to the formation of properdin pathway intermediate, ZPI, whose activity is assessed by its capacity to deplete the titer of cobra venom inducible lysis in diluted rat serum. The formation of ZPI does not require C1 or C2 and is not diminished by prior absorption of the serum with Z. In contrast, removal of P suppresses ZPI formation. Both Ca++ and Mg++ are essential cofactors for the formation of this intermediate whose function is abrogated in the presence of anti-C3, anti-P, but not by anti-Factor B. Since C4-deficient guinea pig serum is also effective in ZPI formation, the data suggest that ZPI is an alternative pathway intermediate containing both P and C3.


Patterns of Response to Different Treatment Strategies in Seropositive Rheumatoid Arthritis Patients in a Tertiary Hospital in South-Western Saudi Arabia: A Retrospective Study.
To study the pattern of response to different treatment strategies in seropositive rheumatoid arthritis (RA) patients and to describe our clinical practice in RA management. Over a period of two years from April 2018 to April 2020, we conducted a retrospective analysis of data for 288 consecutive seropositive RA patients attending rheumatology clinics and the daycare unit at Aseer Central Hospital. Data were collected on patient demographics, disease duration, extraarticular manifestations, comorbidities and treatment. Disease activity was assessed using the clinical disease activity index (CDAI). Out of the total 288 patients, 42% (120) are on csDMRADs, while 54% (162) are on bDMRADs and 4% (6) are on tsDMARDs. Of the patients on csDMARDS, 51%, 43% and 7% of them were on remission, low and moderate disease activity, respectively. However, of the patients on non-csDMARDS, 36.3%, 49.4% and 14.3% of them were on remission, low and moderate disease activity, respectively. Failure of csDMARDs was affected by the presence of high disease activity at baseline, extraarticular lung manifestations and coexistent fibromyalgia, with a significant effect of the latter on remission rate. Among patients on non-csDMARDs, 42 (25%) showed one or more therapy changes. Tumor necrosis factor inhibitors were the predominant first-line agents in biologically naive patients (65%) followed by abatacept (18%). Abatacept was the most frequently prescribed second biologic in 52% of cases followed by tocilizumab in 19%. The current clinical practice in our hospital is consistent with the latest American College of Rheumatology (ACR)/The European League Against Rheumatism (EULAR) guidelines. Treat-to-target strategy was achieved in the vast majority of our patients, while remission was observed in almost half of the patients.


Impact of stress on cancer metastasis.
The influence of psychosocial factors on the development and progression of cancer has been a longstanding hypothesis since ancient times. In fact, epidemiological and clinical studies over the past 30 years have provided strong evidence for links between chronic stress, depression and social isolation and cancer progression. By contrast, there is only limited evidence for the role of these behavioral factors in cancer initiation. Recent cellular and molecular studies have identified specific signaling pathways that impact cancer growth and metastasis. This article provides an overview of the relationship between psychosocial factors, specifically chronic stress, and cancer progression.


The metalloregulatory zinc site in Streptococcus pneumoniae AdcR, a zinc-activated MarR family repressor.
Streptococcus pneumoniae D39 AdcR (adhesin competence repressor) is the first metal-sensing member of the MarR (multiple antibiotic resistance repressor) family to be characterized. Expression profiling with a ΔadcR strain grown in liquid culture (brain-heart infusion) under microaerobic conditions revealed upregulation of 13 genes, including adcR and adcCBA, encoding a high-affinity ABC uptake system for zinc, and genes encoding cell-surface zinc-binding pneumococcal histidine triad (Pht) proteins and AdcAII (Lmb, laminin binding). The ΔadcR, H108Q and H112Q adcR mutant allelic strains grown in 0.2 mM Zn(II) exhibit a slow-growth phenotype and an approximately twofold increase in cell-associated Zn(II). Apo- and Zn(II)-bound AdcR are homodimers in solution and binding to a 28-mer DNA containing an adc operator is strongly stimulated by Zn(II) with K(DNA-Zn)=2.4 × 10(8) M(-1) (pH 6.0, 0.2 M NaCl, 25 °C). AdcR binds two Zn(II) per dimer, with stepwise Zn(II) affinities K(Zn1) and K(Zn2) of ≥10(9) M(-1) at pH 6.0 and ≥10(12) M(-1) at pH 8.0, and one to three lower affinity Zn(II) depending on the pH. X-ray absorption spectroscopy of the high-affinity site reveals a pentacoordinate N/O complex and no cysteine coordination, the latter finding corroborated by wild type-like functional properties of C30A AdcR. Alanine substitution of conserved residues His42 in the DNA-binding domain, and His108 and His112 in the C-terminal regulatory domain, abolish high-affinity Zn(II) binding and greatly reduce Zn(II)-activated binding to DNA. NMR studies reveal that these mutants adopt the same folded conformation as dimeric wild type apo-AdcR, but fail to conformationally switch upon Zn(II) binding. These studies implicate His42, His108 and H112 as metalloregulatory zinc ligands in S. pneumoniae AdcR.


MiR-144 functions as tumor suppressor by targeting PIM1 in gastric cancer.
Gastric cancer (GC) is one of the most prevalent types of malignant disease Worldwide. Mounting evidence has demonstrated the involvement of miRNAs in the development of GC. One of these miRNAs, miR-144 has been found aberrantly expressed in a variety of human malignancies. GC tissues were collected from patients, and the level of miR-144 was determined by qRT-PCR. GC cell lines SGC7901 and AGS were used as model cell lines and the anti-tumor effect of miR-144 in both cells were examined. The level of miR-144 was restored in GC cells using miR-144 mimic. Moreover, the target gene of miR-144 wad identified. In this study, our results showed that low miR-144 level significantly correlated with lymph node metastasis stage, TNM stage and differentiation degree. In addition, we found that miR-144 acted as a tumor suppressor in GC. Moreover, our findings showed that miR-144 exerted an anti-tumor effect by directly targeting RLIP76. miR-144 acts as a tumor suppressor in GC and it is a potential therapeutic target for GC treatment.


Grey literature in meta-analyses.
In meta-analysis, researchers combine the results of individual studies to arrive at cumulative conclusions. Meta-analysts sometimes include "grey literature" in their evidential base, which includes unpublished studies and studies published outside widely available journals. Because grey literature is a source of data that might not employ peer review, critics have questioned the validity of its data and the results of meta-analyses that include it. To examine evidence regarding whether grey literature should be included in meta-analyses and strategies to manage grey literature in quantitative synthesis. This article reviews evidence on whether the results of studies published in peer-reviewed journals are representative of results from broader samplings of research on a topic as a rationale for inclusion of grey literature. Strategies to enhance access to grey literature are addressed. The most consistent and robust difference between published and grey literature is that published research is more likely to contain results that are statistically significant. Effect size estimates of published research are about one-third larger than those of unpublished studies. Unfunded and small sample studies are less likely to be published. Yet, importantly, methodological rigor does not differ between published and grey literature. Meta-analyses that exclude grey literature likely (a) over-represent studies with statistically significant findings, (b) inflate effect size estimates, and (c) provide less precise effect size estimates than meta-analyses including grey literature. Meta-analyses should include grey literature to fully reflect the existing evidential base and should assess the impact of methodological variations through moderator analysis.


Chemotherapy is of Value in Second Line and Beyond, Relapsed High-grade, Serous Epithelial Ovarian Cancer: An Analysis of Outcomes Obtained With Oral Etoposide.
Epithelial ovarian cancer is chemotherapy responsive, and multiple lines of chemotherapy are often given. However, there are few data with regard to its effectiveness in later lines. Our aim was to assess its benefit in the high-grade, serous subtype relative to the line of therapy, using etoposide as the example. Women treated with oral etoposide at the British Columbia Cancer Agency upon recurrence/progression in the years 2000 to 2010 were reviewed. Kaplan-Meier and Cox regression methods were used to correlate line of therapy with overall survival, progression-free survival, and interval between etoposide initiation and next progression or death (EPFS). A total of 219 women, median age 61, received etoposide as second (17%), third (30%), fourth (26%), fifth (17%), and sixth to eighth (11%) lines of therapy. The median number of cycles was 2 to 4. Patients who received etoposide as fourth-line to eighth-line treatment had a significantly longer median overall survival and initial progression-free survival (from diagnosis to first relapse) when compared with those who received it as second-line to third-line treatment (47.8 vs. 25.8 mo, P<0.0001; and 16.1 vs. 12.1 mo, P<0.0001, respectively); that is, a selected population of survivors received it later in the course of their disease. On univariate analysis, there was no significant difference in median EPFS (range, 2 to 2.9 mo) on the basis of line of therapy. On multivariate analysis, the hazard ratios improved through the third, fourth, and fifth lines (hazard ratios: 0.82, 0.77, and 0.34, respectively), and was statistically significant in the fifth line. The a priori-defined endpoint of clinical benefit was the "percentage not progressing at 3 months," and this was achieved in 32% to 48%. In this retrospective study, a similar degree of benefit from etoposide, as defined by the percentage remaining progression free at 3 months, was seen in all lines of therapy.


Environmental health in the Baltic region--toxic metals.
Recent reports on concentrations of lead, cadmium, methylmercury, arsenic and nickel in some biological media in populations in the Baltic region are reviewed. In particular, children in parts of Poland, the Czech Republic, and Germany have uptakes of lead sufficient to cause adverse effects on the central nervous system and kidneys. Cadmium exposure is also high in Poland. Slight cadmium-induced effects on the kidneys have been reported from Germany and Sweden. Methylmercury uptake is dependent upon the intake of fish, in particular from contaminated lakes and rivers in Sweden and Finland, as well as the eastern coast of the Baltic Sea. There are some indications of immunotoxic effects associated with the intake of such fish. However, fish also contain other immunomodulating agents. Exposure to arsenic seems to be low everywhere in the Baltic region. There is high nickel exposure in northern Russia.