Geographical spread of gastrointestinal tract cancer incidence in the Caspian Sea region of Iran: spatial analysis of cancer registry data.
High incidence rates of gastrointestinal tract cancers have been reported in the Caspian region of Iran. This study aimed to: 1) describe the geographical spatial patterns of gastrointestinal tract cancer incidence based on cancer registry data and, 2) determine whether geographical clusters of statistical significance exist. The Babol Cancer Registry, which covers the two major northern Iranian provinces of Mazandaran and Golestan (total population = 4,484,622) was used to identify new gastrointestinal tract cancer cases during 2001 to 2005. Age-specific cancer incidence rates were calculated for 7 gastrointestinal tract cancer sites in 26 wards of the Mazandaran and Golestan provinces. Spatial autocorrelation indices, hierarchical Bayesian Poisson models, and spatial scan statistics were used in measuring the geographic pattern and clusters. There were non-random spatial patterns in esophageal and stomach cancers that were similar for both sexes. Clusters of high incidence were identified in esophageal, stomach, colorectal and liver cancer for both sexes, as well as a possible cluster of pancreas cancer in males. Gastrointestinal tract cancers exhibit significant spatial clustering of risk in northern Iran. Further work is needed to relate these geographical patterns to information on potential life-style and environmental factors.


Clinical assessment of VSR site and size and its relation to the severity of heart failure in post-myocardial infarction ventricular septal rupture patients.
Ventricular septal rupture (VSR) is a rare but well-known mechanical consequence of an acute myocardial infarction (AMI). Even in the later stages of re-perfusion therapy, the result of VSR remains poor. Our aim is to assess the site and size of VSR in relation to the severity of cardiac failure. From January 2016 to December 2022, a total of 71 patients with a diagnosis of post-myocardial infarction VSR were admitted to the First Affiliated Hospital of Zhengzhou University, Zhengzhou, China. Data records were retrospectively included in this registry. In all patients, clinical and echocardiographic data were gathered, and statistical analyses were performed. A total of 71 consecutive patients (mean age: 66.27 ± 8.88 years); 50.7% male, 49.3% female, with (M:F) ratio of almost (1:1). Left ventricular ejection fraction (LVEF) was (48.55 ± 10.44%) on echocardiography, and apical VSR was the most common site (69.0%). Overall, the VSD site was strongly related to the VSD size (p = .016), LVEF (p = .012), AMI site (p = .001), and affected coronary vessel (p = .004). Prodromal angina (p = .041), intra-aortic balloon pump (p = .002), affected coronary vessels (p = .020), pro-BNP (p = .000), and LVEF (p = .017) were predictors of the severity of heart failure. Diabetes mellitus is a common risk factor for post-myocardial infarction VSR. VSR site and size had no relation to the severity of heart failure. A presentation with prodromal angina predicted severe heart failure and a worse prognosis.


Quantification of human platelet antigen-1a antibodies with the monoclonal antibody immobilization of platelet antigens procedure.
Fetal-neonatal alloimmune thrombocytopenia results from a maternal alloimmunization against fetal platelet (PLT) antigens. In Caucasian persons, human PLT antigen-1a (HPA-1a) is the most frequently implicated antigen. Our aim was to develop a method supported by a mathematical approach to quantify HPA-1a antibodies. The monoclonal antibody immobilization of PLT antigens (MAIPA) protocol was applied on serial dilutions of a serum with high concentration of HPA-1a antibodies used as reference. Two mathematical procedures were used: the standard curve was constructed with a linear and a logistic regression. These regressions were used to calculate the amount of antibody in 14 test serum samples from mothers with detectable anti-HPA-1a. Similar quantifications were obtained with both mathematical procedures. In 11 serum samples, the results, expressed in arbitrary units (AU), ranged from 62 +/- 4 to 1096 +/- 19 with the linear regression and from 62 +/- 4 to 1117 +/- 38 with the logistic regression. In addition, the linear regression allowed us to measure 3 test serum samples with low antibody concentrations from 4 +/- 2 to 44 +/- 3 AU, whereas the logistic regression was not suitable for the quantification of antibodies in these serum samples. A simple quantification method was developed for anti-HPA-1a, based on the MAIPA procedure, allowing further studies concerning correlations between anti-HPA-1a quantification and clinical relevance.


[Methanogenic destruction of (amino)aromatic compounds by anaerobic microbial communities].
Destruction of a number of aromatic substrates by anaerobic microbial communities was studied. Active methanogenic microbial communities decomposing aminoaromatic acids and azo dyes into CH4 and CO2 were isolated. Products of primary conversion were found to be 2-hydroxybenzyl and benzyl alcohols gradually transforming into benzoate. It was shown that isolated microbial communities are capable of converting the initial substrates--benzyl alcohol, benzoate, salicylic acid, and golden yellow azo dye--into biogas without a lag-phase but with different velocities. Aromatic and linear intermediates of biodestruction of aromatic amines by obtained enrichment cultures were determined for the first time. Selective effect of aromatic substrates on a microbial community that was expressed in decrease in diversity and gradual change of dominant morphotypes was revealed.


SMR proteins SugE and EmrE bind ligand with similar affinity and stoichiometry.
Suppressor of a groEL mutation protein E (SugE) is a small multidrug resistance (SMR) homologue. In comparison with other SMR proteins, SugE promotes bacterial resistance to a narrow range of quaternary ammonium compounds (QACs). Isothermal titration calorimetry was used to study the binding of QACs to Escherichia coli SugE in different membrane mimetic environments. In this study, the binding stoichiometry of SugE to drug was found to be 1:1, and the binding of SugE to drug was observed with the dissociation constant (K(D)) in the micromolar range for each of the drugs in the membrane mimetic environments explored. This interaction appears to be enthalpy-driven with enthalpies of 8-12 kcal/mol for each of the drugs. These results are similar to those found with drug binding to the SMR protein EmrE in an earlier study.


Association of adiponectin gene polymorphisms with the risk of ischemic stroke in a Chinese Han population.
Adiponectin is inversely associated with the risk of ischemic stroke through its anti-inflammatory and anti-atherogenic effects. Genetic variations in the adiponectin gene (ADIPOQ) have been shown to be associated with the risk of ischemic stroke in Caucasians and Japanese populations. However, it was unknown whether variations in the ADIPOQ gene were associated with the risk of ischemic stroke in Chinese population. A case-control study was performed among 302 patients with ischemic stroke and 338 unrelated controls in a Chinese Han population. The single-nucleotide polymorphisms (SNPs) rs266729 (-11377C/G), rs2241766 (+45T/G), rs1501299 (+276G/T) in the ADIPOQ gene were genotyped by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The frequencies of GG genotype and G allele of rs266729 in the patients with ischemic stroke were significantly higher than those in the controls (P = 0.034, P = 0.010, respectively). In univariate logistic analysis, compared with CC genotype, GG genotype of rs266729 increased the risk of ischemic stroke (odds ratio (OR) = 2.062, 95% confidence interval (CI) = 1.145-3.715, P = 0.016). After adjustment for potential risk factors by the multivariate logistic analysis, rs266729 remained positive correlation with ischemic stroke (OR = 2.165; 95% CI = 1.116-4.197, P = 0.022). However, no significant association was observed among rs2241766, rs1501299 and ischemic stroke. In addition, no significant difference was found in haplotype frequencies between the patients with ischemic stroke and control subjects. The present study demonstrated that the promoter polymorphism rs266729 of the ADIPOQ gene was associated with an increased risk of ischemic stroke in the Chinese Han population.


Anti-Ferroptotic Effects of bone Marrow Mesenchymal Stem Cell-Derived Extracellular Vesicles Loaded with Ferrostatin-1 in Cerebral ischemia-reperfusion Injury Associate with the GPX4/COX-2 Axis.
Accumulating evidence of the critical role of Ferrostatin-1 (Fer-1, ferroptosis inhibitor) in cerebral ischemia has intrigued us to explore the molecular mechanistic actions of Fer-1 delivery by bone marrow mesenchymal stem cells-derived extracellular vesicles (MSCs-EVs) in cerebral ischemia-reperfusion (I/R) injury. In vivo middle cerebral artery occlusion (MCAO) in mice and in vitro oxygen-glucose deprivation/reperfusion (OGD/R) in hippocampal neurons were developed to simulate cerebral I/R injury. After Fer-1 was confirmed to be successfully delivered by MSCs-EVs to neurons, we found that MSCs-EVs loaded with Fer-1 (MSCs-EVs/Fer-1) reduced neuron apoptosis and enhanced viability, along with curtailed inflammation and ferroptosis. The regulation of Fer-1 on GPX4/COX2 axis was predicted by bioinformatics study and validated by functional experiments. The in vivo experiments further confirmed that MSCs-EVs/Fer-1 ameliorated cerebral I/R injury in mice. Furthermore, poor expression of GPX4 and high expression of COX-2 were witnessed in cerebral I/R injury models. MSCs-EVs/Fer-1 exerted its protective effects against cerebral I/R injury by upregulating GPX4 expression and inhibiting COX-2 expression. Taken together, our study indicates that MSCs-EVs/Fer-1 may be an attractive therapeutic target for the treatment of cerebral I/R injury due to its anti-ferroptotic properties.


Structure of the capsular polysaccharide of Klebsiella serotype K40.
The capsular polysaccharide from Klebsiella Serotype K40 contains D-galactose, D-mannose, L-rhamnose, and D-glucuronic acid in the ratios of 4:1:1:1. Methylation analysis of the native and carboxyl-reduced polysaccharide provided information about the glycosidic linkages in the repeating unit. Degradation of the permethylated polymer with base established the identity of the sugar unit preceding the glycosyluronic acid residue. The modes of linkages of different sugar residues were further confirmed by Smith degradation and partial hydrolysis of the K40 polysaccharide. The anomeric configurations of the different sugar residues were determined by oxidation of the peracetylated native and carboxyl-reduced polysaccharide with chromium trioxide. Based on all of these results, the heptasaccharide structure 1 was assigned to the repeating unit of the K40 polysaccharide. (Formula: see text)


Effects of ACTH4-10 on synaptic transmission in frog sympathetic ganglion.
The influence of ACTH4-10, a behaviourally active fragment of adrenocorticotropic hormone (ACTH) devoid of endocrine activity, on synaptic transmission in the paravertebral sympathetic ganglion of the frog was investigated. Postsynaptic potentials evoked by electrical stimulation of pregnanglionic nerves were recorded using a sucrose gap method. Fast excitatory postsynaptic potentials (EPSPs), which are mediated via nicotinic cholinergic synapses, were not affected by 10(-6) M ACTH4-10. Application of ACTH4-10 in a concentration as low as 10(-8) M for 60 min caused a marked augmentation of the amplitude of slow inhibitory postsynaptic potentials (IPSPs) which are mediated via dopaminergic synapses. The increase in amplitude developed gradually after a latency of 60--90 min and outlasted the application of the peptide. In addition, ACTH4-10 at 10(-6) M increased the hyperpolarising response of the ganglion to exogenous dopamine, as studied by a micro-application method. There was no significant effect of ACTH4-10 on the muscarinic cholinergic depolarising response of the ganglion towards exogenous acetylcholine. The behaviourally active vasopressin fragment DG-LVP (10(-6) M) had no effect on slow IPSPs. The results demonstrate that ACTH4-10 specifically affects slow synaptic inhibition in frog sympathetic ganglion, probably by acting upon the postsynaptic membrane. The possibility is discussed that ACTH4-10 affects one of the intermediate steps between dopaminergic receptor interaction and generation of the slow IPSP.


Peri-infarction block, a marker for VT risk?
We present a monomorphic ventricular tachycardia (VT) in a patient with a history of myocardial infarction and syncope. The ECG shows an acceleration-dependent peri-infarction block inferiorly. The authors suggest that this old (but forgotten) ECG-entity may serve as a marker for assessing the risk of post-infarction VT.


Cerebral and Muscle Oxygenation During Intermittent Hypoxia Exposure in Healthy Humans.
To evaluate changes in muscle and cerebral oxygenation during intermittent hypoxia (IH). Fifteen healthy subjects were exposed to 45-min IH (2-min cycles). Arterial blood oxygen saturation (SpO2), prefrontal cortex and brachial biceps muscle oxygenation (assessed by near-infrared spectroscopy), heart rate, and ventilation were continuously recorded. During 2-min IH cycles, changes in SpO2 (9.2% ± 3.3%) were associated with significant changes in cortex oxygenation (3.2% ± 1.8%), minute ventilation, and heart rate, but no change in muscle oxygenation (0.2% ± 1.0%). Fluctuations of blood oxygen levels comparable to severe obstructive sleep apnea translate into distinct pattern of oxygenation changes in the muscle and cortex.


Smelling the roses and seeing the light: gene therapy for ciliopathies.
Alterations in cilia formation or function underlie a growing class of pleiotropic disorders termed ciliopathies. The genetic basis of ciliopathies is remarkably complex, with an incomplete but expanding list of more than 89 loci implicated in various disorders. Current treatment of ciliopathies is limited to symptomatic therapy. However, our growing understanding of ciliopathy genetics, coupled with recent advances in gene delivery and endogenous gene and transcript repair demonstrated thus far in tissues of the eye, nose, and airway, offers hope for curative measures in the near future. This review highlights these advances, as well as the challenges that remain with the development of personalized medicine for treating a very complex spectrum of disease, penetrant in a variety of organ systems.


Pulsed electric field exposure of insulin induces anti-proliferative effects on human hepatocytes.
The purpose of this study was to examine the effects and the mechanism of a pulsed electric field (PEF) on insulin and its subsequent mediation of proliferative changes in human hepatocytes in vitro. The PEF, the electric field intensity coupled into the culture medium, was about 0.7 V/m with a repeating frequency of 50 Hz. Insulin solution was exposed to PEF for 20 min and added to the culture medium of human hepatocytes. Combining fluorescence spectroscopy, immunocytochemistry, microarrays, RT-PCR and MTT, several important events of the insulin signaling pathways were investigated, including ligand-receptor binding capacity, intracellular tyrosine phosphorylation level, gene transcription, and cell proliferation. PEF produced a conformational change of insulin molecule. The binding capacity of insulin to its receptors was reduced to 87% of the control level after PEF treatment, and the average intracellular tyrosine phosphorylation level decreased by 11%. The expression of 55 of 12,000 genes examined was modified, including an increase in the expression of human tyrosine phosphatase and the small GTP-binding protein. Based on these results, a mechanism is proposed to explain the bio-effects of PEF on hepatic cell proliferation through the insulin signaling pathway.


Cardiovascular mortality in bipolar disorder: a population-based cohort study in Sweden.
To estimate the cardiovascular mortality among persons with bipolar disorder in Sweden compared to the general population. Population register-based cohort study with a 20-year follow-up. Sweden. The entire population of Sweden (n=10.6 million) of whom 17 101 persons were diagnosed with bipolar disorder between 1987 and 2006. Mortality rate ratios (MRR), excess mortality (excess deaths), cardiovascular disorder (CVD) and specifically cerebrovascular disease, coronary heart disease, acute myocardial infarction, sudden cardiac deaths and hospital admission rate ratio (ARR). Persons with bipolar disorder died of CVD approximately 10 years earlier than the general population. One third (38%) of all deaths in persons with bipolar disorder were caused by CVD and almost half (44%) by other somatic diseases, whereas suicide and other external causes accounted for less than a fifth of all deaths (18%). Excess mortality of both CVD (n=824) and other somatic diseases (n=988) was higher than that of suicide and other external causes (n=675 deaths). MRRs for cerebrovascular disease, coronary heart disease and acute myocardial infarction were twice as high in persons with bipolar disorder compared to the general population. Despite the increased mortality of CVD, hospital admissions (ARR) for CVD treatment were only slightly increased in persons with bipolar disorder when compared to the general population. The increased cardiovascular mortality in persons with bipolar disorder calls for renewed efforts to prevent and treat somatic diseases in this group. Specifically, our findings further imply that it would be critical to ensure that persons with bipolar disorder receive the same quality care for CVD as persons without bipolar disorder.


Impacts of oxygen deficiency on embryo life-history traits of migratory locust Locusta migratoria from low and high altitudes.
Hypoxia challenges aerobic organisms in numerous environments, and hypoxic conditions may become more severe under future climate-change scenarios. The impact of hypoxia on the development of terrestrial insect embryos is not well understood. Here, to address this gap, embryonic life-history traits of migratory locust Locusta migratoria from low-altitude and high-altitude regions were compared under 2 oxygen levels: normoxia (i.e., 21 kPa oxygen partial pressure and mild hypoxia (i.e., 10 kPa oxygen partial pressure). Our results demonstrated that, whether reared under normoxia or mild hypoxia, L. migratoria from high-altitude populations had longer developmental times, reduced weight, and lower mean relative growth rate as compared with those from low-altitude populations. When transferred from normoxia to mild hypoxia, nearly all the tested life-history traits presented significant negative changes in the low-altitude populations, but not in the high-altitude populations. The factor 'strain' alone explained 18.26%-54.59% of the total variation for traits, suggesting that the phenotypic differences between L. migratoria populations from the 2 altitudes could be driven by genetic variation. Significant genetic correlations were found between life-history traits, and most of these showed differentiation between the 2 altitudinal gradients. G-matrix comparisons showed significant structural differences between L. migratoria from the 2 regions, as well as several negative covariances (i.e., trade-offs) between traits in the low-altitude populations. Overall, our study provides clear evidence that evolutionary divergence of embryonic traits between L. migratoria populations from different altitudes has occurred.


Lexical and gestural development in 5p deletion syndrome-A case report.
Individuals with 5p deletion syndrome (also known as cri du chat syndrome) have various speech and language problems. The aim of this work was to examine early gestural and lexical development in a boy with this syndrome and to see to what extent his skills in these areas were delayed and/or deviant when compared to typically developing children. The participant's parents completed the Norwegian adaptation of the MacArthur-Bates Communicative Development inventories (CDI) ten times over a period of five years. His scores were compared to those of typically developing infants aged eight to 20 months. It was found that the subject followed a considerably delayed, but not deviant, developmental trajectory in three areas, receptive vocabulary, productive vocabulary and communicative gestures, compared to typically developing infants and toddlers. The speech and language problems of individuals with 5p deletion syndrome, which have been documented in the domains of phonetics and phonology and grammar, also extend to gestural and lexical development. The findings of this study will have clinical implications for assessment, in that a broad assessment of gestural and lexical skills should be carried out as early as possible as a basis for interventions to improve communicative skills.


Rose spring dwarf-associated virus has RNA structural and gene-expression features like those of Barley yellow dwarf virus.
We determined the complete nucleotide sequence of the Rose spring dwarf-associated virus (RSDaV) genomic RNA (GenBank accession no. EU024678) and compared its predicted RNA structural characteristics affecting gene expression. A cDNA library was derived from RSDaV double-stranded RNAs (dsRNAs) purified from infected tissue. Nucleotide sequence analysis of the cloned cDNAs, plus for clones generated by 5'- and 3'-RACE showed the RSDaV genomic RNA to be 5808 nucleotides. The genomic RNA contains five major open reading frames (ORFs), and three small ORFs in the 3'-terminal 800 nucleotides, typical for viruses of genus Luteovirus in the family Luteoviridae. Northern blot hybridization analysis revealed the genomic RNA and two prominent subgenomic RNAs of approximately 3 kb and 1 kb. Putative 5' ends of the sgRNAs were predicted by identification of conserved sequences and secondary structures which resembled the Barley yellow dwarf virus (BYDV) genomic RNA 5' end and subgenomic RNA promoter sequences. Secondary structures of the BYDV-like ribosomal frameshift elements and cap-independent translation elements, including long-distance base pairing spanning four kb were identified. These contain similarities but also informative differences with the BYDV structures, including a strikingly different structure predicted for the 3' cap-independent translation element. These analyses of the RSDaV genomic RNA show more complexity for the RNA structural elements for members of the Luteoviridae.


Silver ions trigger Ca2+ release by acting at the apparent physiological release site in sarcoplasmic reticulum.
Rapid Ca2+ release from Ca2+ -loaded sarcoplasmic reticulum vesicles (SR) was previously shown to occur upon the addition of micromolar concentrations of heavy metals, and the extent of Ca2+ release was dependent on the binding affinity of the metal to sulfhydryl group(s) on an SR protein (Abramson, J.J., Weden, L., Trimm, J.L., and Salama, G. (1982) Biophys. J. 37, 134a; Abramson, J.J., Trimm, J.L., Weden, L., and Salama, G. (1983) Proc. Natl. Acad. Sci. U.S.A. 80, 1526). The nature of this Ca2+ release site was examined further and found to be predominantly distributed in heavy SR (HSR) rather than light SR fractions. Ag+ -induced Ca2+ release from heavy SR was blocked by local anesthetics and ruthenium red which are known to inhibit Ca2+ release in skeletal fibers and in heavy SR, respectively. The rate of Ca2+ efflux from SR triggered by Ag+ was dependent on pH, Mg2+, and ionic strength of the medium. Efflux rates increased by a factor of 4 from pH 6.0 to 7.0 and then decreased in more alkaline reaction mixtures. Efflux rates from actively or passively loaded SR increased by a factor of 2.5 with increasing Mg2+ from 0 to 1 mM and then decreased in the range of 1 to 10 mM Mg2+. ATP-dependent Ca2+ uptake by SR was similar in 100 mM KCl and in 200 mM sucrose solutions, but the extent and rate of Ca2+ efflux induced by Ag+ were dramatically reduced with decreasing ionic strength of the medium. In solutions containing 5 mM Mg2+, the rate of Ca2+ efflux from heavy SR averaged over the first 1.5 s after the addition of Ag+ was 58 nmol of Ca2+/mg of SR/s, a value comparable to the fast initial rate of ATP-dependent Ca2+ uptake. The maximum initial rate of Ag+ -induced Ca2+ efflux from heavy SR in 1 mM Mg2+ may be comparable to the rate of Ca2+ release and tension development in muscle fibers. Our data indicate that Ag+ reacts with a protein or proteins in the SR, probably not the (Ca2+, Mg2+)-ATPase, to induce a rapid release of Ca2+, possibly from the physiological Ca2+ release site.


Neurophysiological basis of contrast dependent BOLD orientation tuning.
Recent work in early visual cortex of humans has shown that the BOLD signal exhibits contrast dependent orientation tuning, with an inverse oblique effect (oblique > cardinal) at high contrast and a horizontal effect (vertical > horizontal) at low contrast. This finding is at odds with decades of neurophysiological research demonstrating contrast invariant orientation tuning in primate visual cortex, yet the source of this discrepancy is unclear. We hypothesized that contrast dependent BOLD orientation tuning may arise due to contrast dependent influences of feedforward (FF) and feedback (FB) synaptic activity, indexed through gamma and alpha rhythms, respectively. To quantify this, we acquired EEG and BOLD in healthy humans to generate and compare orientation tuning curves across all neural frequency bands with BOLD. As expected, BOLD orientation selectivity in V1 was contrast dependent, preferring oblique orientations at high contrast and vertical at low contrast. On the other hand, EEG orientation tuning was contrast invariant, though frequency-specific, with an inverse-oblique effect in the gamma band (FF) and a horizontal effect in the alpha band (FB). Therefore, high-contrast BOLD orientation tuning closely matched FF activity, while at low contrast, BOLD best resembled FB orientation tuning. These results suggest that contrast dependent BOLD orientation tuning arises due to the reduced contribution of FF input to overall neurophysiological activity at low contrast, shifting BOLD orientation tuning towards the orientation preferences of FB at low contrast.


Effect of Low-Load Blood Flow Restriction Training on Patients With Functional Ankle Instability: A Randomized Controlled Trial.
Decreased muscle strength and balance in patients with functional ankle instability (FAI) can be effectively improved by ankle strength training. Low-load blood flow restriction (LL-BFR) training increases muscle size and strength, but there is limited evidence from studies on muscle strength and balance in FAI patients. To study the effects of LL-BFR training versus high-load training (HLT) on muscle strength and balance in FAI patients. Randomized controlled trial. Forty-six young adults with a history of FAI. Participants in the LL-BFR and HLT groups performed 4 sets (30 × 15 × 15 × 15) of ankle training at 20% to 40% of the one-repetition maximum and 70% to 85% one-repetition maximum, respectively, twice a week for 6 weeks. Plantar flexion, dorsiflexion, inversion, and eversion muscle strength, and the Y-balance test scores were assessed at baseline and after 3 and 6 weeks; the thickness of the tibialis anterior, triceps surae, and peroneus longus muscles were assessed at baseline and after 6 weeks. Inversion, eversion, dorsiflexion, and plantar flexion muscle strength; tibialis anterior, triceps surae, and peroneus longus thickness; and Y-balance test scores were significantly increased in the LL-BFR group after 3 and 6 weeks compared with baseline (P < .05), with no significant difference between the LL-BFR and HLT groups after 6 weeks (P > .05). However, at the end of 3 weeks, eversion muscle strength and Y-balance test scores were significantly higher in the LL-BFR group than in the HLT group (P < .05). Over 6 weeks, LL-BFR training was as effective as HLT in improving ankle muscle strength, muscle thickness, and balance in FAI patients, but LL-BFR training improved the ankle eversion muscle strength and dynamic balance more than HLT did in the early stages of the intervention. This finding will provide a new intervention strategy for the clinical rehabilitation of FAI patients.


Development of Algorithm for Discriminating Hydrometeor Particle Types with a Synergistic Use of CloudSat and CALIPSO.
We developed a method for classifying hydrometeor particle types, including cloud and precipitation phase and ice crystal habit, by a synergistic use of CloudSat/Cloud Profiling Radar (CPR) and Cloud-Aerosol Lidar and Infrared Pathfinder Satellite Observations (CALIPSO)/Cloud-Aerosol LIdar with Orthogonal Polarization (CALIOP). We investigated how the cloud phase and ice crystal habit characterized by CALIOP globally relate with radar reflectivity and temperature. The global relationship thus identified was employed to develop an algorithm for hydrometeor type classification with CPR alone. The CPR-based type classification was then combined with CALIPSO-based type characterization to give CPR-CALIOP synergy classification. A unique aspect of this algorithm is to exploit and combine the lidar's sensitivity to thin ice clouds and the radar's ability to penetrate light precipitation to offer more complete picture of vertically resolved hydrometeor type classification than has been provided by previous studies. Given the complementary nature of radar and lidar detections of hydrometeors, our algorithm delivers thirteen hydrometeor types: warm water, supercooled water, randomly-oriented ice crystal (3D-ice), horizontally-oriented plate (2D-plate), 3D-ice+2D-plate, liquid drizzle, mixed-phase drizzle, rain, snow, mixed-phase cloud, water+liquid drizzle, water+rain and unknown. The global statistics of three-dimensional occurrence frequency of each hydrometeor type revealed that 3D-ice contributes the most to the total cloud occurrence frequency (53.8%), followed by supercooled water (14.3%), 2D-plate (9.2%), rain (5.9%), warm water (5.7%), snow (4.8%), mixed-phase drizzle (2.3%), and the remaining types (4.0%). This hydrometeor type classification provides useful observation-based information for climate model diagnostics in representation of cloud phase and their microphysical characteristics.


Regenerative Endodontic Treatment as a Biologically Based Approach for Non-Surgical Retreatment of Immature Teeth.
The goal of regenerative endodontic treatment (RET) is to regenerate the pulp-dentin complex in immature teeth with pulp necrosis. There is no consensus in using RET in previously treated teeth. This article reports on long-term outcomes of RET in previously treated immature teeth. Previously treated immature teeth with signs/symptoms of failure were included. After local anesthesia and rubber dam isolation, the root canal filling material was removed without the use of chemical solvents. The root canals were irrigated with 6% sodium hypochlorite using an EndoVac (Kerr Endodontics, Culver City, CA), and a creamy mix of metronidazole and ciprofloxacin was placed in the canals for 4 weeks. At the second visit, the antibiotic dressing was removed using 17% EDTA. Bleeding was induced into the root canal space, and SynOss Putty (Collagen Matrix Inc, Oakland, NJ) was placed. Bioceramic putty was placed over the SynOss Putty, and the teeth were restored at a subsequent visit. Five teeth in 4 patients were included. The patients' ages ranged from 14-46 years. Recall examinations were performed from 20 to 72 months with an average of 54.4 months. The apical openings of the teeth ranged from 1-3.7 mm. All teeth were functional and asymptomatic at the recall visits. All periapical lesions resolved with partial or complete mineralization of the root canals, and all teeth showed partial to complete apical closure at the follow-up visits. RET can be a viable option for nonsurgical retreatment of immature teeth. This article adds a new perspective to the field of regenerative endodontics.


Evaluation of nursing students' health fatalism, knowledge, and attitudes toward patients with epilepsy.
This study was conducted to determine the relationship between nursing students' health fatalism, epilepsy knowledge, attitudes, and some sociodemographic variables. The descriptive cross-sectional study was conducted with 423 nursing department students who were studying at a state university and who agreed to participate in the study in the Elazığ province in eastern Turkey during the 2020-2021 academic year. The data were collected using a Sociodemographic Information Form, the Health Fatalism Scale (HFS), the Epilepsy Knowledge Scale (EKS), and the Epilepsy Attitude Scale (EAS). The nursing students had total mean scores of 46.95 ± 12.98, 6.94 ± 3.81, and 50.19 ± 8.47 for the HFS, EKS, and EAS, respectively. A positive and significant correlation was found between the EKS and EAS scores. First-year students had lower HFS scores than students in their second and fourth years. Third- and fourth-year students, those living in rural areas, and those who had patients with epilepsy in their families or immediate environments had higher EKS scores. Female students and those who had patients with epilepsy in their families or immediate environments had higher EKS and EAS scores. A significant positive correlation was found between age and the EKS score, whereas a significant negative correlation was found between age and the EAS score. While nursing students generally had low levels of epilepsy knowledge, they had positive attitudes toward epilepsy and moderate levels of health fatalism. The study found that students' positive attitudes toward epilepsy increased as their levels of knowledge about the disorder increased. Age, gender, educational status, place of residence, the presence of a chronic disease, and knowing someone with epilepsy all affected HFS, EKS, and EAS scores.


A high-throughput dual system to screen polyphosphate kinase mutants for efficient ATP regeneration in L-theanine biocatalysis.
ATP, an important cofactor, is involved in many biocatalytic reactions that require energy. Polyphosphate kinases (PPK) can provide energy for ATP-consuming reactions due to their cheap and readily available substrate polyphosphate. We determined the catalytic properties of PPK from different sources and found that PPK from Cytophaga hutchinsonii (ChPPK) had the best catalytic activity for the substrates ADP and polyP6. An extracellular-intracellular dual system was constructed to high-throughput screen for better catalytic activity of ChPPK mutants. Finally, the specific activity of ChPPKD82N-K103E mutant was increased by 4.3 times. Therefore, we focused on the production of L-theanine catalyzed by GMAS as a model of ATP regeneration. Supplying 150 mM ATP, GMAS enzyme could produce 16.8 ± 1.3 g/L L-theanine from 100 mM glutamate. When 5 mM ATP and 5 U/mL ChPPKD82N-K103E were added, the yield of L-theanine was 16.6 ± 0.79 g/L with the conversion rate of 95.6 ± 4.5% at 4 h. Subsequently, this system was scaled up to 200 mM and 400 mM glutamate, resulting in the yields of L-theanine for 32.3 ± 1.6 g/L and 62.7 ± 1.1 g/L, with the conversion rate of 92.8 ± 4.6% and 90.1 ± 1.6%, respectively. In addition, we also constructed an efficient ATP regeneration system from glutamate to glutamine, and 13.8 ± 0.2 g/L glutamine was obtained with the conversion rate of 94.4 ± 1.4% in 4 h after adding 6 U/ mL GS enzyme and 5 U/ mL ChPPKD82N-K103E, which further laid the foundation from glutamine to L-theanine catalyzed by GGT enzyme. This proved that giving the reaction an efficient ATP supply driven by the mutant enzyme enhanced the conversion rate of substrate to product and maximized the substrate value. This is a positively combination of high yield, high conversion rate and high economic value of enzyme catalysis. The mutant enzyme will further power the ATP-consuming biocatalytic reaction platform sustainably.


The correlation between bismuth and uranyl staining and phosphorus content of intracellular structures as determined by electron spectroscopic imaging.
Four groups of intracellular structures can be recognized according to bismuth and uranyl staining and phosphorus content. (1) Those which contain phosphorus and stain strongly with uranyl acetate but not with bismuth (ribosomes, heterochromatin and mature ribosomal precursor granules), presumably because of their nucleic acid content. (2) Those which contain phosphorus and stain with uranyl acetate and bismuth (interchromatin granules, immature ribosomal precursor granules and mitochondrial granules), presumably because at least some of their phosphate is available to react with bismuth. (3) Those which contain little phosphorus but which stain strongly with bismuth and weakly with uranyl acetate (Golgi complex beads), perhaps because some ligand in addition to phosphate reacts with bismuth, and (4) those which do not contain phosphorus and stain with neither uranyl acetate nor bismuth (portasomes). Uranyl staining correlates strongly with the phosphorus content of nucleic acids, proteins and inorganic deposits. Bismuth will stain some phosphorylated molecules but not all. Thus only some phosphates stain with bismuth.


Identification of adaptor proteins using the ANOVA feature selection technique.
The adaptor proteins play a crucially important role in regulating lymphocyte activation. Rapid and efficient identification of adaptor proteins is essential for understanding their functions. However, biochemical methods require not only expensive experimental costs, but also long experiment cycles and more personnel. Therefore, a computational method that could accurately identify adaptor proteins is urgently needed. To solve this issue, we developed a classifier that combined the support vector machine (SVM) with the composition of k-Spaced Amino Acid Pairs (CKSAAP) and the amino acid composition (AAC) to identify adaptor proteins. Analysis of variance (ANOVA) was used to select the optimized features which could generate the maximum prediction performance. By examining the proposed model on independent data, we found that the 447 optimized features could achieve an accuracy of 92.39% with an AUC of 0.9766, demonstrating the powerful capabilities of our model. We hope that the proposed model could provide more clues for studying adaptor proteins.


Small Molecule Inhibition of Ligand-Stimulated RAGE-DIAPH1 Signal Transduction.
The receptor for advanced glycation endproducts (RAGE) binds diverse ligands linked to chronic inflammation and disease. NMR spectroscopy and x-ray crystallization studies of the extracellular domains of RAGE indicate that RAGE ligands bind by distinct charge- and hydrophobicity-dependent mechanisms. The cytoplasmic tail (ct) of RAGE is essential for RAGE ligand-mediated signal transduction and consequent modulation of gene expression and cellular properties. RAGE signaling requires interaction of ctRAGE with the intracellular effector, mammalian diaphanous 1 or DIAPH1. We screened a library of 58,000 small molecules and identified 13 small molecule competitive inhibitors of ctRAGE interaction with DIAPH1. These compounds, which exhibit in vitro and in vivo inhibition of RAGE-dependent molecular processes, present attractive molecular scaffolds for the development of therapeutics against RAGE-mediated diseases, such as those linked to diabetic complications, Alzheimer's disease, and chronic inflammation, and provide support for the feasibility of inhibition of protein-protein interaction (PPI).


Transcriptional analysis of dynamic heat-shock response by the hyperthermophilic bacterium Thermotoga maritima.
The thermal stress response of the hyperthermophilic bacterium Thermotoga maritima was characterized using a 407-open reading frame-targeted cDNA microarray. Transient gene expression was followed for 90 min, following a shift from 80 degrees C to 90 degrees C. While some aspects of mesophilic heat-shock response were conserved in T. maritima, genome content suggested differentiating features that were borne out by transcriptional analysis. Early induction of predicted heat-shock operons hrcA-grpE-dnaJ (TM0851-TM0850-TM0849), groES-groEL (TM0505-TM0506), and dnaK-sHSP (TM0373-TM0374) was consistent with conserved CIRCE elements upstream of hrcA and groES. Induction of the T. maritima rpoE/ sigW and rpoD/ sigA homologs suggests a mechanism for global heat-shock response in the absence of an identifiable ortholog to a major heat-shock sigma factor. In contrast to heat-shock response in Escherichia coli, the majority of genes encoding ATP-dependent proteases were downregulated, including clpP (TM0695), clpQ (TM0521), clpY (TM0522), lonA (TM1633), and lonB (TM1869). Notably, T. maritima showed indications of a late heat-shock response with the induction of a marR homolog (TM0816), several other putative transcriptional regulators (TM1023, TM1069), and two alpha-glucosidases (TM0434 and TM1068). Taken together, the results reported here indicate that, while T. maritima shares core elements of the bacterial heat-shock response with mesophiles, the thermal stress regulatory strategies of this organism differ significantly. However, it remains to be elucidated whether these differences are related to thermophilicity or phylogenetic placement.


Knowledge and Beliefs of Breast Self-Examination and Breast Cancer among Market Women in Ibadan, South West, Nigeria.
In most resource constrained settings like Nigeria, breast self-examination self-breast examination (BSE) is culturally acceptable, religious friendly and attracts no cost. Women's knowledge and beliefs about breast cancer and its management may contribute significantly to medical help-seeking behaviours. This study aimed to assess knowledge and beliefs of BSE among market women. A descriptive cross-sectional study was conducted among 603 market women in Ibadan, Nigeria. Data was collected using semi-structured interviews and analyzed using descriptive and analytic statistical methods. The mean age of the respondents was 34.6±9.3 years with 40% of the women aged between 30-39years. The proportion of married women was 339 (68.5%) with 425 (70.8%) respondents reporting that they do not know how to perform BSE. However, 372 (61.7%) women strongly agreed that BSE is a method of screening for breast cancer. Highest proportion 219 (36.3%) reported that the best time for a woman to perform BSE was 'anytime'. Most of the respondents believed breast cancer is a dangerous disease that kills fast and requires a lot of money for treatment. More efforts are needed in creating awareness and advocacy campaigns in the grassroots in order to detect early breast cancer and enhance prevention strategies that would reduce the burden of breast cancer in Nigeria.


Characterization of muscarinic receptors in the rat nucleus tractus solitarius.
Crude membrane preparations from the dorsomedial medulla and whole heart of adult rats were used to characterize muscarinic receptors (mChRs) in the nucleus tractus solitarius (NTS). Isotherm binding assays with [3H]quinuclidinyl benzilate ([3H]QNB) indicated a uniform population of muscarinic cholinergic receptors in both tissues with a Kd of 60.3 pM and a Bmax of 401 fmol/mg protein for NTS membranes while heart membranes had a Kd of 56.6 pM and a Bmax of 193 fmol/mg protein. Competitive binding assays with selective muscarinic antagonists on NTS membranes using 80 pM [3H]QNB showed the following relative Ki values consistent with M2 mChRs: atropine, 1.9 nM; methoctramine, 7.7 nM; 4-diphenylacetoxy-N-methylpiperidine methobromide, 9.4 nM; AF-DX 116, 87.6 nM; and pirenzepine, 657 nM. Similarly, two agonists, carbachol and oxotremorine, showed Ki values of 1.82 microM and 0.379 microM, respectively. All of the competitive binding assays exhibited Hill coefficients close to 1.0 and InPlot analysis showed a single binding site for each agonist and antagonist examined.


MTF of columnar phosphors with a homogenous part: an analytical approach.
A method for the theoretical estimation of the MTF of columnar phosphors with a homogeneous part at the end used in X-ray imaging has been developed. This method considers the light transport inside the scintillator through an analytical modelling, the optical photon beams distribution on the scintillator-optical sensor interface, and uses the definition of the PSF and a Gauss fitted LSF to estimate the MTF of an indirect detector. This method was applied to a columnar CsI:Tl scintillator and validated against experimental results found in literature, and a good agreement was observed. It was found that, by increasing the pixel size of the optical detector and the thickness of the scintillator, the MTF decreased as expected. This method may be used in evaluating the performance of the columnar phosphors used in medical imaging, given their physical and geometrical characteristics.Graphical abstract (a) Side view of a part of the scintillator where five crystal columns with homogeneous ends attached to an optical sensor is shown. (b) Propagation of two random optical photon beams emitted from point K with different angles of emission is shown. All the symbols are explained analytically in the text.


The Effect of Supportive Implementation of Healthier Canteen Guidelines on Changes in Dutch School Canteens and Student Purchase Behaviour.
We developed an implementation plan including several components to support implementation of the "Guidelines for Healthier Canteens" in Dutch secondary schools. This study evaluated the effect of this plan on changes in the school canteen and on food and drink purchases of students. In a 6 month quasi-experimental study, ten intervention schools (IS) received support implementing the guidelines, and ten control schools (CS) received only the guidelines. Changes in the health level of the cafeteria and vending machines were assessed and described. Effects on self-reported purchase behaviour of students were analysed using mixed logistic regression analyses. IS scored higher on healthier availability in the cafeteria (77.2%) and accessibility (59.0%) compared to CS (60.1%, resp. 50.0%) after the intervention. IS also showed more changes in healthier offers in the cafeteria (range -3 to 57%, mean change 31.4%) and accessibility (range 0 to 50%, mean change 15%) compared to CS (range -9 to 46%, mean change 9.7%; range -30 to 20% mean change 7% resp.). Multi-level logistic regression analyses on the intervention/control and health level of the canteen in relation to purchase behaviour showed no relevant relations. In conclusion, the offered support resulted in healthier canteens. However, there was no direct effect on students' purchase behaviour during the intervention.


Coronary anomaly with a left ventricular accessory chamber: Is it left ventricular aneurysm or diverticulum?
A 60-year-old male hypertensive was suspected to have post myocardial infarction left ventricular aneurysm. His transthoracic echocardiogram performed elsewhere reported an inferior wall left ventricular aneurysm. He was referred for coronary angiography. His coronary angiogram and repeat transthoracic echocardiogram demonstrated an accessory chamber arising from mid-inferior wall synchronously contracting with the left ventricle suggesting a diverticulum. In addition, he had anomalous coronary arteries with coronary artery disease. His electrocardiogram did not reveal any features suggestive of a post infarct aneurysm. This case illustrates the importance of a comprehensive clinical and echocardiographic examination with a broad differential thought process.


Revealing cellular and molecular complexity of the central nervous system using single cell sequencing.
The mammalian central nervous system (CNS) is one of the most complex systems, with thousands of cell types and subtypes with distinct and unique morphology and gene expression profiles. Based on classic histological methods and conventional cellular and molecular approaches, single cell sequencing is becoming a powerful tool to uncover the complexity of the CNS. In this review, we summarize the principle of single cell sequencing and highlight its use for studying the development of neural stem cells, neural progenitors, and distinct neurons. By revealing transcriptomes in each individual cell using single cell sequencing, we are now able to dissect the cellular heterogeneity of a hundred billion cells in the CNS and comprehensively investigate mechanisms of brain development and function at the cellular and molecular levels.


Heterophyes species in Greece: record of H. heterophyes, H. aequalis and H. dispar from the first intermediate host, Pirenella conica.
Heterophyes heterophyes is reported for the first time from the snail host Pirenella conica in Greece. H. aequalis and H. dispar are reported for the first time from Greece and from Europe. The results presented, together with the available data on necropsies of dogs and cats, human infections and the biotope preference of the snail, suggest that Greece is an important focus of infection in Europe.


Recombinant AAV-mediated gene delivery to the central nervous system.
Various regions of the brain have been successfully transduced by recombinant adeno-associated virus (rAAV) vectors with no detected toxicity. When using the cytomegalovirus immediate early (CMV) promoter, a gradual decline in the number of transduced cells has been described. In contrast, the use of cellular promoters such as the neuron-specific enolase promoter or hybrid promoters such as the chicken beta-actin/CMV promoter resulted in sustained transgene expression. The cellular tropism of rAAV-mediated gene transfer in the central nervous system (CNS) varies depending on the serotype used. Serotype 2 vectors preferentially transduce neurons whereas rAAV5 and rAAV1 transduce both neurons and glial cells. Recombinant AAV4-mediated gene transfer was inefficient in neurons and glial cells of the striatum (the only structure tested so far) but efficient in ependymal cells. No inflammatory response has been described following rAAV2 administration to the brain. In contrast, antibodies to AAV2 capsid and transgene product were elicited but no reduction of transgene expression was observed and readministration of vector without loss of efficiency was possible from 3 months after the first injection. Based on the success of pioneer work performed with marker genes, various strategies for therapeutic gene delivery were designed. These include enzyme replacement in lysosomal storage diseases, Canavan disease and Parkinson's disease; delivery of neuroprotective factors in Parkinson's disease, Huntington disease, Alzheimer's disease, amyotrophic lateral sclerosis, ischemia and spinal cord injury; as well as modulation of neurotransmission in epilepsy and Parkinson's disease. Several of these strategies have demonstrated promising results in relevant animal models. However, their implementation in the clinics will probably require a tight regulation and a specific targeting of therapeutic gene expression which still demands further developments of the vectors.


Anti-CD40 Ab- or 8-oxo-dG-enhanced Treg cells reduce development of experimental autoimmune encephalomyelitis via down-regulating migration and activation of mast cells.
This study investigated whether anti-CD40 Ab and 8-oxo-dG attenuate mast cell migration and EAE development. Anti-CD40 Ab and 8-oxo-dG reduced EAE scores, mast cell numbers, expression of adhesion molecules, OX40L and Act1, levels of TNF-α, LTs, expression of cytokines, and co-localization of Treg cells and mast cells, all of which are increased in EAE-brain tissues. Each treatment enhanced Treg cells, expression of OX40, and cytokines related to suppressive function of Treg cells in EAE brain tissues. Act-BMMCs with Treg cells reduced expression of OX40L and CCL2/CCR2, VCAM-1, PECAM-1, [Ca²⁺]i levels, release of mediators, various signaling molecules, Act1 related to IL-17a signals versus those in act-BMMCs without Treg cells. The data suggest that IL-10- and IL-35-producing Foxp3⁺-Treg cells, enhanced by anti-CD40 Ab or 8-oxo-dG, suppress migration of mast cells through down-regulating the expression of adhesion molecules, and suppress mast cell activation through cell-to-cell cross-talk via OX40/OX40L in EAE development.


Screening of Siderophore-Producing Bacteria and Their Effects on Promoting the Growth of Plants.
Iron deficiency is a major global agricultural problem. Siderophores can help organisms to uptake iron in form of siderophore-Fe3+ complexes and then in the cell cytosol, iron is reducted and released in ferrous form. This research aimed to obtain some efficient siderophore-producing bacterial strains and evaluate their plant growth-promoting effects in the iron-deficit environment. Two strains, Brucella sp. E7 and Pseudomonas brassicae W7, were isolated from rhizosphere soil. Both strains could produce maximum siderophores under the optimal conditions. Plant promoting experiment showed that many indicators of Vigna radiata seedling were all increased significantly by strain E7/W7 or the consortium of E7 + W7. Under no-iron and high iron stress, the inoculation treatment also showed growth promotion effects on both Vigna radiata and Lolium multiflorum. These results indicated that the potential ability of strain E7 and W7 in increasing agricultural production as a growth-promoting agent in iron-deficit soil.


Abnormal inhibin A and inhibin B secretion in obese women with and without insulin resistance.
The present study aimed to establish inhibin A and B serum levels during the menstrual cycle of obese women, and its usefulness as an index of luteal-phase follicular development. Twenty-one obese patients (mean body mass index: 34.9+/-3.7 kg/m2; range: 30.0-45.0 kg/m2) were submitted to basal hormonal measurements and an oral glucose tolerance test after challenge with 75 g glucose. Progesterone and inhibin A and B levels were determined 5-7 days after the menstrual cycle and 7 days prior to expected menses. As expected, an increase in inhibin A and a decrease in inhibin B were observed when first-phase samples were compared with samples obtained after 15-20 days. On the other hand, the percentage variation of both inhibin A and B was at least four times smaller than the values for normal women described previously by other authors employing the same enzyme-linked immunosorbent assays. A small number of obese women presented ovulatory cycles characterized by progesterone concentration higher than 5.8 ng/ml. The percentage elevation (>190%) of inhibin A in the second samples was in agreement with the progesterone levels, but it seemed to be more sensitive for the detection of follicle luteinization. We conclude that obese women present less percentage variation of both inhibin A and B during the menstrual cycle, associated with a low frequency of ovulatory cycles. In obese women, the percentage increase of inhibin A can represent an additional marker to recognize follicle luteinization.


Combining human STR and microbial population profiling: Two case reports.
In forensic case investigations involving human traces, cell type identification has become increasingly important. No longer only the donor of a trace (sub-source level), but also the actions which could have led to the deposition of the trace ('beyond-the-source'/activity level) need to be evaluated by forensic experts. For this evaluation determining the cellular source of a DNA profile can be beneficial. In this report two criminal cases are described where both human STR profiling and microbial population profiling were applied to the same trace sample. Human STR profiling was used to evaluate the sub-source question and microbial population profiling was used to evaluate the source question. The Bayesian framework was used to evaluate the evidence.


Diagnosis of unknown nonhematological tumors by bone marrow biopsy: a retrospective analysis of 10,112 samples.
To investigate how unselected bone marrow (BM) biopsy examinations could help in the diagnosis of clinically unknown nonhematological tumors. 10,112 plastic-embedded BM biopsy sections were retrospectively analyzed. In the analysis we focused on the following aspects: (1) the frequency of BM involvement arising from clinically unknown nonhematological malignancies, (2) the clinical indication for BM biopsy examination in cases with tumor BM metastasis, (3) the primary sites of the metastatic tumors, and (4) the advantage of plastic-embedded biopsy sections over paraffin-embedded samples and the complementarity of biopsy with aspiration smears. Of the 10,112 BM samples analyzed, 101 (1.0%) were interpreted as being nonhematological tumor metastases. In cases with metastatic tumors, the nonhemocyte-related changes, such as skeletal pain (25%) and bone destruction (5%), were considerably higher than in those without metastasis (3.7 and 0.32%, respectively; P < 0.001). Primary lesions were documented antemortem in 50 of the 101 biopsy-positive cases (49.5%); the most frequent being in the lung, gastric tract, and breast. Using this assay, a higher incidence of metastatic tumors was detected when compared with previously reported paraffin-embedded samples. The frequency of metastatic tumors based on aspiration smears when the positive results for biopsy sections were used as a reference was 74.3%. All the 101 sections with metastatic tumors showed various degrees of myelofibrosis. We concluded that the routine BM biopsy examination is helpful in detecting insidious metastatic nonhematological malignancies. Skeletal pain and bone destruction are critical indications for susceptible patients to undergo BM examination. Plastic embedding of biopsy sections appears to be more sensitive than the paraffin embedding of samples and is an excellent complementation to isolated aspiration smears.


Isocitrate lyase plays important roles in plant salt tolerance.
Isocitrate lyase (ICL) is a key enzyme in the glyoxylate cycle. In a previous study in rice, the expression of the ICL-encoding gene (OsICL) was highly induced by salt stress and its expression was enhanced in transgenic rice lines overexpressing OsCam1-1, a calmodulin (CaM)-encoding gene. CaM has been implicated in salt tolerance mechanisms in plants; however, the cellular mechanisms mediated by CaM are not clearly understood. In this study, the role of OsICL in plant salt tolerance mechanisms and the possible involvement of CaM were investigated using transgenic plants expressing OsICL or OsCam1-1. OsICL was highly expressed in senesced leaf and significantly induced by salt stress in three OsCam1-1 overexpressing transgenic rice lines as well as in wild type (WT). In WT young leaf, although OsICL expression was not affected by salt stress, all three transgenic lines exhibited highly induced expression levels. In Arabidopsis, salt stress had negative effects on germination and seedling growth of the AtICL knockout mutant (Aticl mutant). To examine the roles of OsICL we generated the following transgenic Arabidopsis lines: the Aticl mutant expressing OsICL driven by the native AtICL promoter, the Aticl mutant overexpressing OsICL driven by the 35SCaMV promoter, and WT overexpressing OsICL driven by the 35SCaMV promoter. Under salt stress, the germination rate and seedling fresh and dry weights of the OsICL-expressing lines were higher than those of the Aticl mutant, and the two lines with the icl mutant background were similar to the WT. The Fv/Fm and temperature of rosette leaves in the OsICL-expressing lines were less affected by salt stress than they were in the Aticl mutant. Finally, glucose and fructose contents of the Aticl mutant under salt stress were highest, whereas those of OsICL-expressing lines were similar to or lower than those of the WT. OsICL, a salt-responsive gene, was characterized in the transgenic Arabidopsis lines, revealing that OsICL expression could revert the salt sensitivity phenotypes of the Aticl knockout mutant. This work provides novel evidence that supports the role of ICL in plant salt tolerance through the glyoxylate cycle and the possible involvement of OsCam1-1 in regulating its transcription.


Dopaminergic Dependency of Cholinergic Pallidal Neurons.
We employed the whole-cell patch-clamp method and ChAT-Cre mice to study the electrophysiological attributes of cholinergic neurons in the external globus pallidus. Most neurons were inactive, although approximately 20% displayed spontaneous firing, including burst firing. The resting membrane potential, the whole neuron input resistance, the membrane time constant and the total neuron membrane capacitance were also characterized. The current-voltage relationship showed time-independent inward rectification without a "sag". Firing induced by current injections had a brief initial fast adaptation followed by tonic firing with minimal accommodation. Intensity-frequency plots exhibited maximal average firing rates of about 10 Hz. These traits are similar to those of some cholinergic neurons in the basal forebrain. Also, we examined their dopamine sensitivity by acutely blocking dopamine receptors. This action demonstrated that the membrane potential, excitability, and firing pattern of pallidal cholinergic neurons rely on the constitutive activity of dopamine receptors, primarily D2-class receptors. The blockade of these receptors induced a resting membrane potential hyperpolarization, a decrease in firing for the same stimulus, the disappearance of fast adaptation, and the emergence of a depolarization block. This shift in physiological characteristics was evident even when the hyperpolarization was corrected with D.C. current. Neither the currents that generate the action potentials nor those from synaptic inputs were responsible. Instead, our findings suggest, that subthreshold slow ion currents, that require further investigation, are the target of this novel dopaminergic signaling.


Environmental and meteorological factors linked to malaria transmission around large dams at three ecological settings in Ethiopia.
A growing body of evidence suggests that dams intensify malaria transmission in sub-Saharan Africa. However, the environmental characteristics underpinning patterns in malaria transmission around dams are poorly understood. This study investigated local-scale environmental and meteorological variables linked to malaria transmission around three large dams in Ethiopia. Monthly malaria incidence data (2010-2014) were collected from health centres around three dams located at lowland, midland and highland elevations in Ethiopia. Environmental (elevation, distance from the reservoir shoreline, Normalized Difference Vegetation Index (NDVI), monthly reservoir water level, monthly changes in water level) and meteorological (precipitation, and minimum and maximum air temperature) data were analysed to determine their relationship with monthly malaria transmission at each dam using correlation and stepwise multiple regression analysis. Village distance to reservoir shoreline (lagged by 1 and 2 months) and monthly change in water level (lagged by 1 month) were significantly correlated with malaria incidence at all three dams, while NDVI (lagged by 1 and 2 months) and monthly reservoir water level (lagged by 2 months) were found to have a significant influence at only the lowland and midland dams. Precipitation (lagged by 1 and 2 months) was also significantly associated with malaria incidence, but only at the lowland dam, while minimum and maximum air temperatures (lagged by 1 and 2 months) were important factors at only the highland dam. This study confirmed that reservoir-associated factors (distance from reservoir shoreline, monthly average reservoir water level, monthly water level change) were important predictors of increased malaria incidence in villages around Ethiopian dams in all elevation settings. Reservoir water level management should be considered as an additional malaria vector control tool to help manage malaria transmission around dams.


Measuring government commitment to vaccination.
Vaccination is among the most cost-effective health interventions and has attracted ever greater levels of funding from public and private donors. However, some countries, mainly populous lower-middle income countries, are lagging behind on vaccination financing and performance. In this paper, we discuss the rationale for investing in vaccination and construct a metric to measure government commitment to vaccination that could promote accountability and better tracking of performance. While noting the limitations of available data, we find that populous middle-income countries, which stand to gain tremendously from increased vaccination uptake, perform poorly in terms of their vaccination outcomes.


Cyclophosphamide as an alternative to azathioprine in cardiac transplant recipients with suspected azathioprine-induced hepatotoxicity.
AZA has been reported to cause liver dysfunction in some recipients of solid organ transplants. To assess the safety and efficacy of cyclophosphamide in maintenance immunosuppression in the setting of AZA-induced liver dysfunction, we retrospectively reviewed the records of 320 surviving cardiac transplant recipients in Utah. Cyclophosphamide was substituted for AZA in 29 patients due to elevated liver enzymes. Patients were switched to cyclophosphamide 689 +/- 104 days after transplantation; total follow-up after initiation of cyclophosphamide was 540 +/- 56 days. The dose of cyclophosphamide after 2 and 6 months of cyclophosphamide therapy was 62 +/- 6 mg/day (0.8 +/- 0.1 mg/kg/day) and 48 +/- 5 mg/day (0.6 +/- 0.1 mg/kg/day), respectively, compared with 233 +/- 20 mg/day (2.9 +/- 0.2 mg/kg/day) of AZA. The substitution of cyclophosphamide for AZA was associated with a significant improvement in liver function tests. Liver enzymes decreased by up to 49% (P = 0.027), while serum bilirubin decreased by 58% (P < 0.001). Rejection frequency did not increase; neither corticosteroid nor CsA dosage was altered significantly after the substitution of cyclophosphamide. Significant bone marrow suppression was not observed; specifically, no significant change in white blood cell count or hematocrit occurred. Complications of treatment with cyclophosphamide were few; only 1 patient discontinued cyclophosphamide because of alopecia. We conclude that cyclophosphamide appears to be safe in maintenance immunosuppression, permitting the discontinuation of AZA in patients with AZA-induced hepatic dysfunction without necessitating the augmentation of either corticosteroids or CsA.


How many lives do COVID vaccines save? Evidence from Israel.
Israel began a mass vaccination program with the rapid rollout of the Pfizer-BioNTech COVID-19 BNT162b2 vaccine for adults. At the same time, government stringency measures in terms of closing public life were decreased. Our objective was to estimate the total number of Covid-19 deaths avoided due to the massive vaccination campaign in the elderly Israeli population. We examined the effect of vaccination on mortality when at least 90% of the population over age 70 were vaccinated for less than seven months. Projected deaths as expected from vaccine efficacy and actual mortality data were compared for the study population to account for potential confounding effects of government stringency. The average Oxford Stringency Index was calculated in the study period and the preceding period of the pandemic. Potential confounding effects of an age shift in the distribution of deaths were examined by analyzing the distributions before and after the study period. Confirmed deaths from COVID-19 in the population over 70 after mass vaccination were recorded as 370, versus 5,120 estimated without vaccinations. Vaccines against COVID-19 saved more lives than expected by simply applying individual vaccine efficacy to the vaccinated population in Israel, despite a loosening of government stringency. Our findings support the worldwide efforts of governments to improve vaccination rates, especially in the elderly population.


Genomic and fossil windows into the secret lives of the most ancient fungi.
Fungi have crucial roles in modern ecosystems as decomposers and pathogens, and they engage in various mutualistic associations with other organisms, especially plants. They have a lengthy geological history, and there is an emerging understanding of their impact on the evolution of Earth systems on a large scale. In this Review, we focus on the roles of fungi in the establishment and early evolution of land and freshwater ecosystems. Today, questions of evolution over deep time are informed by discoveries of new fossils and evolutionary analysis of new genomes. Inferences can be drawn from evolutionary analysis by comparing the genes and genomes of fungi with the biochemistry and development of their plant and algal hosts. We then contrast this emerging picture against evidence from the fossil record to develop a new, integrated perspective on the origin and early evolution of fungi.


Regulation of the human prostacyclin receptor gene in megakaryocytes: major roles for C/EBPδ and PU.1.
The prostanoid prostacyclin plays a central role in haemostasis and vascular repair. Recent studies investigating the regulation of the human prostacyclin receptor (hIP) gene identified an upstream repressor region (URR) within its regulatory promoter, herein termed the PrmIP. This study aimed to identify the main trans-acting factors that bind within the URR to transcriptionally repress PrmIP-directed gene expression in the megakaryoblastic human erythroleukemia (HEL) 92.1.7 cell line. Of the putative cis-acting elements examined, disruption of C/EBP and PU.1 elements within the URR substantially increased PrmIP-directed gene expression. Chromatin immunoprecipitation (ChIP) confirmed that C/EBPδ and PU.1, but not C/EBPβ, bind to the URR in vivo, while ectopic expression of C/EBPδ substantially reduced hIP mRNA levels and PrmIP-directed gene expression. Phorbol 12-myristate 13-acetate (PMA)-induced megakaryocytic differentiation increased hIP mRNA and PrmIP-directed reporter gene expression and hIP-mediated cAMP generation in HEL cells. Two PMA-responsive regions, termed PRR1 and PRR2, were identified within PrmIP. Disruption of C/EBPδ and PU.1 cis-elements within the overlapping PRR1/URR and of Sp1, PU.1 and Oct-1 cis-elements within the overlapping PRR2/core PrmIP, revealed that both PRR1 and PRR2 contribute to the PMA- induction of hIP mRNA and gene expression in HEL cells. Furthermore, ChIP analysis established that induction of PrmIP-directed gene expression during megakaryocytic differentiation is largely regulated by PMA-induced dissociation of C/EBPδ and enhanced binding of PU.1 to PRR1 in addition to increased binding of Sp1, PU.1 and Oct-1 to elements within the core promoter/PRR2 in vivo. Taken together, these data provide critical insights into the transcriptional regulation of the hIP gene within the vasculature, including during megakaryocytic differentiation.